Clinical Chemistry G. R. Kingsley, Clinical Chemistry Consulfant, 62 I Bonhill Road, 10s Angeles, Calif. 90049
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of significant developments in clinical chemistry ( 1 0 A ) last reported by the author forthe period December 1964 to December 1966 are continued for the period December 1966 t o December 1968. HE REVIEWS
REVIEWS
D'Eustachio (6A) reviewed biochemical analysis under t h e subjects of: dialysis, centrifugation, electrophoresis, column chromatography, ion exchange and gel filtration, paper chromatography, thin layer chromatography. spectrophotometry, ultraviolet and infrared, atomic absorption, fluorescence and lu mini s c e n c e, radiometric analysis , electron microscopy, neutron activation analysis, mass spectrophotometry, lasers, magnetic resonance, electrochemical methods, gasometric analysis, instrumentation, and automation. Anan et a l . reviewed clinical a n a l y t i c a l methods ( I A ) , and Pinkerton ( 1 3 A ) revien-ed criteria for the selection of a colorimetric analytical clinical chemistry method, such as: economy, ease of performance, speed, precision, sample size, general acceptance, and inherent laboratory error. Strickland ( 1 6 A ) reviewed electrophoresis under the subtitles of interest to the clinical chemist such as: biological applications, human serum, specialized blood proteins, hemoglobin, immunoglobulins, lipoproteins, enzymes, hormones, forensic, tosicological, and pharmaceuticalapplications. hlargoshes and Scribner (11-4) reviened emission spectrometry under the titles: instrumentation. standards, calibration, escitation sources, trace analysis, lasers, and microanalysis. Whiteand Keissler ( I 7.4) reviewed fluorometric analysis under the subjects: apparatus, metals, atomic fluorescence, organic and biological, vitamins, catecholamines, amino acids, proteins, enzymes, steroids. hormones, and immunofluorescence. Rubin ( 1 4 A ) presented a resume of fluorescence analysis as applied to clinical chemistry which included instrumental characteristics, variables affecting thc fluorescence of molecules in solution, and a selected survey of procedures. Phillips and Elevitch ( I Z A ) reviewed and discussed fluorimetric techniques in clinical pathology including a great variety of methodologies such as: electrolytes, steroids, lipids, proteins, amino acids, enzymes, drugs, metabolites, etc. Fluorometric methods for calcium, steroids, porphyrins, adrenaline, noradrenaline, and 14 R
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amino acids were reviewed by Hajdu (7A). Infrared spectrometry was reviewed b y Chihara (SA) a n d Crisler ( 4 A ) . Kanabrocki et al. (9A) reviewed neutron activation analysis as a tool in clinical and forensic applications i n medicine. Wnefordner, IlcCarthy, and St. John ( 1 8 A ) reviewed the theoretical considerations and instrumentation of phosphorimetry and its application in biochemical analysis, such as: drugs, anticoagulants, sulfonamides, amino acids, proteins, and enzymes. Ultraviolet spectrometry as applied to organic and inorganic analysis was reviewed by Crummett and Hummel ( 5 A ) . An exhaustive historical, clinical, chemical, and bacteriological essay on the art of urinalysis was presented by Berman ( 2 A ) . Scott and Melville ( 1 5 A ) presented a symposium on the identification, significance, and automated analysis of u r i n a r y c o n s t i t u e n t s of low molecular weight. The future of analytical chemistry as a profession was reviewed by Hallett (8A). APPARATUS AND EQUIPMENT
Several new t y p e s of a u t o m a t e d equipment for application to clinical chemistry were developed since the presentation of the last review of clinical chemistry. Reference to this equipment will be made in the section on automation. The impact of technological development on medical instrumentation was revien-ed by DeBakey ( 1 2 B ) . Lauer, Abel, and Anson (28B) described a system for electrochemical data acquisition and analysis based on a digital computer. Frazer (16B) discussed the use of digital control computers in analytical laboratories. Clark ( I O B ) described a simpledataanalyzerforlaboratory statistics. The technical characteristics of commercial clinical electrophoresis e q u i p m e n t was listed by Martinek ( S I B ) .Greater reproducibility of electrophoretic runs were obtained by Dike and Bew ( 1 4 B ) when a constant potential across the separation field was used. lluller ( S 5 B )discussed Beckman's new continuous particle electrophoresis system and its application in biological and industrial uses. An electronic cyclical scanner for the automatic analysis of stained electrophoretic strips on cellulose acetate membranes was developed by Yanzetti, Palatucci, and Cosi ( 5 5 B ) . The theory and application of membrane electrodes was reviewed b y Pungor (S9B) and Sollner (52B). Sekelj and
Goldbloom ( 4 7 B ) described the clinical applications of glass electrodes for the measurement of pH, pCOl and Na+. Rechnitz (4OB) cited the need of more research on the functioning of specific ion electrodes in complex practical media. A simple portable apparatus was developed for argentometric titration of chlorides using a simple electron tube millivoltmeter by Kic (27'B). Blaedel and Olson (6B)obtained a patent on a n apparatus for the automated amperometric determination of glucose. Jones ( 2 5 B ) patented a gas analyzer for the determination of CO, expired from lungs. X patent was obtained on a n instrument for the determination of O2 saturation of whole blood by Goldberg (2OB).Rispens et al. ( 4 1 B ) designed an apparatus for the direct determination of HCO-3 concentration in blood and plasma by displacing CO, from sample into a n electrode chamber for measurement. A new method was described by Rochkind (42%) for the infrared analysis of multicomponent gas mixtures which involves a condensed phase sampling technique employing commercially available cryogenic equipment. Demiaux and Badinand (1SB)described a n apparatus for the extraction of CO from 1-ml samples of blood. Freeman, Lampo, and Windsor (1'7B) developed a n instrument for the semiautomatic infrared analysis of mixtures of triglycerides and cholesterol esters. -1simple rapid, precise method for the detection, identification, and quantitation of drugs in blood serum by gas chromatography was developed by Thompson and Decker ( 6 4 B ) . Nitruka and Alexander (SZB) described a n ultrasensitive electron capture detector for detection of certain microbial metabolites. An economical electronic integrator for gas chromatography was developed by Bartle and Jleckstroth ( 4 B ) . Morin ( S S B ) described a new tube for concentrating samples for gas chromatography and thin layer chromatography. Schlunegger ( 4 5 B ) described a sample introduction apparatus for direct gas chromatographic determination of volatile constituents of whole blood. Karger (26B) reviewed new developments in chemical selectivity in gas-liquid chromatography. Harkness and Torrance (2SB)described a simple cheap device for the automatic introduction of solid samples into a gas chromatographic column. Schell and Ghetie ( 4 4 B ) described the preparation and use of cross-linked agarose in gel filtration. Scott (46B) described a n automatic high resolution
analytical system for the quantitative determination of the ultraviolet absorbing molecular constituents of urine which included a heated high pressure anion-exchange column for the separation step. Martinek (SOB) reviewed the technical characteristics of commercial flame photometers. An integrating micro flame photometer for simultaneous determination of &‘a and K was described by Carlsson, Giacobini, and Hovmark (8B). Multichannel flame photometers were described by Fuwa and Vallee (18B)and HaagenSmit andRamirez-Munoz ( 2 2 B ) . Bache and Lisk ( S B )usedamicrowave-powered helium plasma to fragment and excite organic bromine, chlorine, iodine, phosphorus, and sulfur compounds eluting from a gas chromatograph to determine nanogram quantities. West (56B) described an ultrasonic sprayer for atomic emission and absorption spectrochemistry. Martinek (29B) listed the tcchnical characteristics of commercial ultramicro analysis systems. Bull (7B) described a microsample dilutor which utilizes a modified microliter syringe for sample and a syringe pump dispenser for diluent. Abelson (1B) developed a double-path quartz micro cell for solutions of unknown absorbance. S a telson (S6B) designed a capillary dispenser for automated microchemical analysis. A micromethod for obtaining small quantities of biological fluids was described by Solomon and Sonnenberg (5SB). Siggaard-Andersen and Bull (49B) described an apparatus for semiautomatic pipetting of ultramicro volumes of sample and reagent. A procedure and apparatus (24B) for examination of biological cells was patented by International Business Machines. A tubular dropper for microtitration mas patented by Duff and Hayward (15B). Groulade and Ollivier (21B) described an apparatus for measuring small biological samples. Damminger and Schuck (1 1B ) patented an apparatus for sampling and preparation of blood. An apparatus for separation of cellular bodies from serum was patented by Bio-Consultants, Inc. (5B). Skeggs (61B) patented an apparatus which provides a method for detection and removal of clots and other solid matter from human blood. Mortensen (34B) described a photoelectric instrument in which clotting end point is assessed by means of the optical changes of the reaction mixture as occurs in prothrombin analysis. Serlupi-Crescenzi and Paolini (48B) developed a double beam spectrophotometer with selective phasesensitive lock-in amplifier. Pro and Hoffman (S8B) reviewed modern instrumentation available for ultrasensitive analytical techniques such as: atomic absorption, spectrometry, neutron activation analysis, x-ray fluorescence, etc. Siggaard-Andersen, and
Komarmy (60B) developed a simple interference filter photometer for measurement of both total and conjugated bilirubin in 20 pl of plasma. Chance and Legallais (9B) patented a n aparat u s t o measure in vivo intracellular oxidation-reduction by irridation and measurement of resultant fluorescence in intact organs. Andersen (BB) reviewed the biochemical applications of the electron probe micro analyzer. Glick (19B) reviewed the construction and application of the laser microprobe in elemental analysis in histrochemistry. Rybak and LeCamus (43B) patented a probe or catheter capable of being ininserted into the blood stream to measure the pressure and the content of 0 a n d CO, dissolved gases. Olehy, Schmitt, and Bethard (S7B) described thermal neutron activation and rapidion exchange radiochemical separation for the quantitation of metals in erythrocytes and plasma. AUTOMATION
The continued increasing interest in automation in clinical chemistry is indicated by the numerous reports in the literature concerning new and improved applications of methodology, especially in fluorometry and enzymology. New systems of automation have been proposed by major instrument manufacturers to challenge the dominant position of Technicon Instruments in the field of automated instrumentation for analytical chemistry. The publication, “Automation in Analytical Chemistry,’’ contains the proceedings of the 1967-68 Technicon International Symposia on automation in analytical chemistry which were held in New York City and Brighton, England. Automated methods in use in clinical biochemistry with the AutoAnalyzer were reviewed by Hankiewicz (37C). A limited number of references will be made t o the large number of papers appearing in “Technicon Symposium on Automation in Analytical Chemistry” as edited by Skeggs (9OC) as examples of progress in various kinds of methodologies. Thiers, Cole, and Kirsch (9SC) studied the kinetic parameters of continuous flow analysis (AutoAnalyzer) and found the transition between steady states obeyed first order kinetics which permits correlation of time required to change from base-line steady state to sample steady state; the characteristics of this change, time interval between samples, proportionality of sampling and washing time, fraction of steady state reached in any given sampling time, and interaction between samples. Kallace (97C) derived equations from graphs for calculation of cross contamination in AutoAnalyzer analysis. Considerable improvement was made by Technicon in the development of the
sequential multiple apparatus, SMA 12-60 over the former SMA 12-30 in that; some blanks could be run, smaller samples of serum required, volume of reagents reduced considerably, continuous monitoring of flow system by oscilloscope, and the number of specimens analyzed increased from 30 t o 60 per hour (each specimen analyzed for 12 different constituents). The older SMA 12-30 was described by Whitehead (99C). Scholtis (847)described a 4-channel analytical system in which signals from 4 colorimeters mere received on a one-pen recorder. An assessment of the SMA 12-30 for mass screening procedures in metabolic profile studies was made by Ratliff, Casey, and Thrasher (77C) and Broughton et al. ( 5 C ) . Gal, Hanok, and Savignano (28C) reported a quality control study of a 15-channel AutoAnalyzer System (SMA-12+3) which showed satisfactory day-to-day standard deviations and coefficients of variation. Kingsley, Coutts, and Betz (51C) studied the precision of a SMA-12 Technicon AutoAnalyzer which was expanded to perform 4 additional analyses, LDH, uric acid phosphorus, and cholesterol, in which all 16 tests were recorded on one chart and data obtained were within approximately two standard deviations. New Automation Analyzers. Rohte and Meyer (81C) described the design of the new Beckman automatic chemical analyzer. This apparatus employs a pneumatic system, uses disposable containers t o p r e p a r e protein free filtrates, a n d performs discrete wet chemistry analysis by conventional colorimetric methods which may be adapted from routine procedures. One hundred and twenty analyses per hour may be performed. Conventional, SGOT, LDH, alkaline phosphorus, glucose, urea N, phosphorus, bilirubin, and creatinine procedures were satisfactorily performed with the automatic chemical analyzer as reported by Gallwas, Gray, and Carne (29C) and Ray and Gray ( 7 8 2 ) . Sterling, Westover, and Nagao (91C) reported the new Beckman automatic chemical analyzer gave satisfactory performance as a microchemical analyzer. Nadeau (69C) described the development of the DuPont automatic clinical analyzer (ACA) system. I n this system the reagents for each separate determination are packaged in a single plastic container as liquids or tablets. The sample is injected directly through a gel, Sephadex, or ion exchange adsorption column as needed and passes into a plastic chamber in which it is machine processed with reagents to form a colored solution in part of the plastic container which acts as a plastic optical cuvette. Reagents for different analytical procedures can be changed to any sequence desired. The system VOL. 41, NO. 5, APRIL 1969
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has specimen identification and printout for 60 determinations per hour. Evenson (22C) made a field evaluation of the DuPont ACA and found excellent precision and recoveries for analytical procedures and compared the DuPont system with the Technicon SMA 12-30. Larsen (58C) and Boutwell, Winsten, and Wilkinson (4C) evaluated the Eskalab clinical chemistry system in which the reagents are combined in a prepackaged, dry, stable, one-assay form. Cotlove, et al. (14C) designed a fully automated discrete sample processing enzyme assay system for elinical chemistry. A patent was obtained for a multiple sample analyzer by Warner-Lambert Pharmaceutical Co. (98C) which is suited for automation. The sample is measured and treated a t one or more stations by carrying the sample on a moving tape. A patent (75C) was obtained for an automatic colorimetric analysis system for small samples of liquid by N.V. Philips’ Gloeilampenfabriken. Dahms (18C) developed a system containing specific electrodes for inorganic constituents, Na+, K+, H f , C1- in blood which is automated for sampling, calibration, and data acquisition and processing. Natelson (7OC) patented an automatic biochemical analyzer for microquantities of blood and urine which consisted of a circular distributor for capillary tubes, the contents of which are delivered onto a ribbon consisting of an upper porous strip pressed onto a test strip proper; the porous strip may act as a filter and the strip proper as a reagent carrier. Vecerek et al. (96C) described a device for serial automatic deproteinization of biological materials on a micro scale. A semiautomatic syringe type pipetting system for measurement of ultra micro volumes of sample and reagent was described by SiggaardAndersen and Bull (86C). An instrument employing a multiple flow-cell system was developed by Tsuji, Griffith, and Sperry (94C) which permits the automatic quantitation of the turbidities of microbiological assay specimens. Brown (6C) investigated the reaction of 2,4,6-trinitrobenzenesulfonic acid with amino acids to develop optimum conditions for application to automated amino acid chromatography. Carles (9C) described the automated determination of organic acids after separation on a Celite 535 anion-exchange column. Dreux and Leymarie (21C) developed a procedure for the estimation of free and total urinary hydroxyproline using the AutoAnalyzer. Automated continuous fluorometric estimation of lactic and pyruvic acid was reported by Forichon, Minaire, and Studievic (26C) and Cramp (15C) and ultraviolet by Freund (27C). Hill, Summer, and Hill (4OC) modified the automated proce16 R
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dure for blood phenylalanine by use of glycyl-1-leucine. Hochella (42C) modified the Waalkes and Udenfriend blood tyrosine method in which tyrosine is coupled to 1-nitroso-2-napthol in the presence of nitrous acid to form a fluorescent compound. Looye, Kwarts, and Groen (65C) devised an automated procedure for xanthurenic acid in urine based on its reaction with 2,6-dichloroquinone chlorimide. Simmons (87C) described an automated method for both direct and indirect serum bilirubin which uses a method based on that of Michaelssonin which ascorbic acid, diazo reagent, and accelerator were included in total and direct bilirubin and blanks. Wilson, Guillan, and Hocker (100C) presented an enzymatic automated method for the determination of ceruloplasmin in serum. An automated alcoholic dehydrogenase method for the determination of ethyl alcohol in blood and urine mas reported by Goldberg and Rydberg (35C). Schersten and Tibbling (8SC) described a semiautomated, enzymic fluorometric method for the determination of low urinary glucose by a procedure based on the use of hexokinase and glucose-6-phosphate dehydrogenase. The use of the .iutodnalyzer in determination of total carbohydrate was described by Jolley and Freeman (47C) and Robyt and Bemis (8OC). Kaldor and Schiavone (48C) automated the manual serum aldolase method of Bruns. h more stable orcinol reagent was substituted by Daly and Levine (2OC) for a-napthol in the automated creatine phosphokinase method of Siege1 and Cohen. An A u t o h a l y z e r method for the colorimetric determination of serum creatine kinase was described by Fleisher ( 2 5 C ) . Hunter, Knight, and Nolan (44C) described an automated system in which serum lactic dehydrogenase and glutamic oxaloacetic transaminase may be analyzed in series. Runstedler (82C) automated a procedure for serum lactic dehydrogenase based on the “Forward” (lactate to pyruvate) reaction coupled with a redox terazolium indicator. Pribor, Kirkham, and Fellows (76C) described how serum protein and LDH isozyme electrophoretic patterns developed on cellulose acetate support medium are scanned in a densitometer and the analog signals transmitted on line to a digital computer which prepares a complete report. Brown and Ebner ( 7 C ) adapted the advantages of continuous flow analysis to the kinetic assay of multiple alkaline phosphatase samples which was used to demonstrate the superiority of such a system over single-point enzyme analysis. Klein and Kaufman (52C) and Hviid (45C) automated the Babson procedure for the determination of serum alkaline phosphatase in which
phenolphthalein monophosphate substrate is used. Hill, Summer, and Waters (41C) used 3-0-methyl fluorescein phosphate in an automated fluorometric assay for alkaline phosphatase, Miyada et al. (67C) found a linear relationship when the manual and automated procedures for determination of serum alkaline phosphatase by the phosphastrate, King-Armstrong and the Bessey-Lowry methods were compared. Loewen et al. (6SC) developed a n automated method for acid phosphatase using disodium phenyl phosphate substrate. Morgenstern, Kaufman, and Klein (68C) described an automated serum glutamic oxaloacetic transaminase method for adaptation to the Robot Chemist in which enzymically generated oxaloacetic acid is coupled with fast ponceau L to form a colored solution. Conaill and Muir (13C) adapted the total urinary estrogen method of Ittrich for the AutoAnalyzer using a continuous digester for the combined operation of phase exchange and color development. Rush (8C)devised a general system of automated biochemical analysis for urinary steroids based on direct photometry or scanning of thin-media chromatogram using a small laboratory digital computer. Van der Honing, Saarloos, and Stip (95C) described a fully automated method for determination of free and esterified cholesterol in which saponification is carried out. Levine (60C) reviewed the developments in automated instrumentation which apply to the determination of blood cholesterol, triglycerides, and phospholipids. Baird, Black, and Faulkner (2C) described a semiautomated method for the determination of free fatty acids in plasma in which the fatty acids are extracted manually from plasma into heptane and then analyzed automatically by using color. the reaction of Mosinger. Dalton and Kowalski (19C) recommended a modification of the automated colorimetric method of Antonis for the routine determination of free fatty acids. Claude, Corre, and Levallois (11C) proposed some modifications of the semiautomated fluorometric serum triglyceride method of Kessler and Lederer. Jacobs (46C) and Siriwardene et al. (89C) reported that automated Kjeldah1 analysis performed on the AutoAnalyzer gave accurate results. Litchfield (62C) reported the automated analysis of nitrite and nitrate in blood. Crouch and Nalmstadt (16C) described an automatic reaction rate method for determination of phosphate in blood serum which utilizes a digital readout of the initial rate of formation of molybdenum blue. Hernandez, Murray, and Doumas (38C) adapted the manual procedure of Rodkey for the direct determination of serum albumin with
bromcresol green to the AutoAnalyzer. Kelson (71C) described an automated cyanmethaemoglobin method for determination of haemoglobin. Strumia and Eusebi (92C) described a n automated haemoglobin method which mas dependent on the measurement of oxyhemoglobin a t 543 mp in a standard AutoAnalyzer. Nyssen and Dorche (74C) applied automation to the determination of urinary iron and Kauppinen and Gref (49C) t o the determination of serum iron. Giovanniello et al. (32C) described a n automated method for the determination of serum iron and total iron-binding capacity in which continuous filter resin loaded paper is employed to remove unbound iron. An evaluation of the automated AutoAnalyzer method for the determination of serum organic iodine was made by Gambino, Schreiber, and Covolo (SOC), Simpson ( S 8 C ) , and Comoy (12C). M a r t i n a n d H a r r i s o n ( 6 6 C ) described a n automated trihydroxyindole method for the determination of noradrenaline and adrenaline. Gaumer, Sprague, and Slavin ( 3 I C ) described a n automatic instrument for the atomic absorption determination of l I g , Ca, Cu, Zn, Fe, and K in serum. Automated techniques for the atomic absorption spectrophotometry of serum calcium were described by Raynaud and Griffiths (79C), Klein, Kaufman, and Morgenstern (55C), Klein, Kaufman, and Oklander (5SC) and Klein and Kaufman (5%). Continuous automated fluorometric methods for determination of serum calcium were described by Classen, Marquardt, and Spaeth (IOC), and Fingerhut and Poock (24C). Automated colorimetric calcium methods employing glyoxal bis (2-hydroxyanil) and cresolphthalein complesone were described by Farese, Schmidt, and Llager (2%) and Gitelman (3%’). Lelir and Broglia (59C) determined ionized calcium in serum by a n Auto.inalyzer dialysis technique. Gray and Pruden (36C) reported that the automated direct fluorometric serum magnesium method appears to be as specific as atomic absorption. Klein and Oklander ( 5 4 C ) presented two automated flow systems for the fluorometric determination of serum magnesium. Glick (34C) studied the variables affecting the determination of sodium and potassium when using the Autohnalyzer. Searcy et al. (85C) applied the Berthelot method for urea S determination to the A u t o h a l y z e r . Annino ( I C ) presented a n improved automated urea S method in which thiosemicarbazide color reagent is used to obtain greater sensitivity and linearity. Hunter (43C) developed a n automated method for the simultaneous determination of blood glucose and urea. Nishi (7SC) adapted the Archibald colorimetric uric acid method to the Auto-
Analyzer. Lofland and Crouse (S4C) reported a n automated procedure for uric acid determination using the cupric-phenanthroline method. Laessig, Underwood, and Basteyns (57C) developed a n automated colorimetric microprocedure suitable for determination of blood uric acid levels. Crowley and Alton (17C) automated the carbonate method for uric acid analysis. Barron and Bouley ( 3 C ) utilized a direct automated method for the determination of uric acid by reading absorbance at 292 mp from a glycine buffered sample and one in which the enzyme uricase is added. On-line computers for the clinical laboratory LTere described by Levy, Kanon, and Lubin ( S I C ) and Hicks et al. (SQC). An electronic device to simplify the calculation of enzyme activity from slopes generated by the continuous technique on recording spectrophotometers was described by Kindig, Nielson, and Inman (50C). Sesset et al. (72C) automated the Mendlin-Butler ascorbic acid method. CONTROL A N D PRECISION OF CLINICAL CHEMISTRY METHODS
Hill and Brown ( 1 7 0 ) reviewed statistical methods in chemistry and attempted t o provide a guide to application of statistics in chemistry. Dybkaer ( 1 3 0 ) reported on the recommendations of 1966 on quantities and units in clinical chemistry which consists of a systematic and thorough discussion of the basic and derived kinds of quantities and their appropriate units of main interest to the clinical chemist. Cooper (100) outlined how evaluation of performance in a clinical laboratory should be carried out. Anastassiadis and Common ( 3 0 ) made a technical differentiation of specificit?; accuracy, precision, sensitivity, their relationships, and assignment of a n appropriate statistical parameter. Young and Nears ( 3 0 0 ) and Nears and Young ( 2 2 0 ) stated that proper daily operation of the laboratory depends upon basic measurement, purity of reagents and standards employed and noted the availability of new standard reference material such as cholesterol, uric acid, urea, and creatinine from the National Bureau of Standards. Radin ( 2 5 0 ) reviewed the definition and the use of standards and the standards available for clinical chemists and discussed the limitations of various so-called standards. Logan and Allen ( 1 9 0 ) examined 17 different commercial control serums prepared by six manufacturers in several lots a t 6month intervals and u p to two years and recommended that these commercial serums should not be used in place of a primary standard when a stable solution of the latter is available.
Copeland et al. ( 1 1 D ) reported results of a national comprehensive laboratory survey 1965, chemistry section, in which over 2300 laboratories participated in performing glucose, urea K, chloride, Na, K, cholesterol, bilirubin, uric acid, calcium, and phosphorus determinations. (A report of the Standards Committee of t h e College of American Pathologists.) Hanok and Kuo (160) investigated the stability of reconstituted serum for 15 constituents when stored a t different temperatures. Baer and Krause (40)found definite changes in serum SGOT, protein electrophoresis, and T-3 uptake when holding time, temperature, and containers were varied to simulate conditions of mailing. Reynolds ( 6 8 0 ) made a survey of the adequacy of the clinical chemistry laboratories of 172 selected hospitals with a bed capacity of less than 70 and found most laboratories adequately equipped to do the common clinical chemical assays. Marsteller, Leifheit, and Kiener ( 2 0 0 ) described a method of preparing artificial blood serum for teaching situations. Roberts ( 2 9 0 ) determined the normal ranges of 17 blood constituents, both male and female, using automated methodology and applying a statistical analysis. Barnett and Pinto ( 5 0 ) evaluated 10 commonly performed clinical chemical analyses by a quality control system, based on mixed patient samples as described by Youden and found that standard deviations, coefficient of variation, and confidence limits were close t o those found by the pooled serum technique. Berry ( 6 D ) reported t h a t standard deviation derived from duplicate sample analysis by several techniques are different from day to day samples. Kilgariff and Owen ( 1 8 0 ) found in a majority of cases the “average of normals” quality control method failed t o detect error. Maynard ( 2 1 0 ) reviewed sources of error and their control in clinical chemistry. Owen and Campbell ( 2 4 0 ) evaluated the statistics derived from the analysis of patient specimens over a three-month period for quality control purposes and examined the sensitivity of the various daily means to systematic error. Seumann ( 2 3 0 ) made a n estimation of normal range on probability paper using a purposely truncated form of the “normal” distribution. Amador ( 1 0 ) made a study of quality control by the reference sample method and concluded that errors had t o be 15 times larger than the coefficient of variation before they became obvious. Amador, Hsi, and Massod ( 2 0 ) designed a study t o test the sensitivity to errors of the “average of normals,” “number plus,” and the “clinical specimens” methods and concluded that the “average of normals” and related methods do not provide the sensitivity to errors which is VOL. 41, NO. 5, APRIL 1969
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necessary. Frankel and Ahlvin ( 1 4 0 ) tested the number plus system of quality control and found that it indicated lack of control a t a time when the procedures were apparantly in control. Griffin (160) described a practical method of usingthe cumulative sumplot for detecting trends, shifts, and out of control situations. Ressler and Whitlock (26D)described applications of computer produced frequency distribution curves in quality control and the evaluation of the diagnostic significance of test results by multidimensional analysis ( 2 7 D ) . Davis, Schonfeld, and Kibbey (12D) presented information on application of digital computers to the problem of urinary analysis to determine the quantity of each UV-absorbing urinary component in a multicomponent solution by linear-least-squares resolution of the UV spectrum of the solution. Conn et al. ( 9 D ) described a system for computer analysis of quality control data which permits extensive analysis of control data with a net saving of technologist's time. Cillo et al. (80) described an operational computer assisted laboratory information system for more effective laboratory control. Blackburn ( 7 D ) developed a method by which a composite spectrum may be transformed and brought into coincidence with standard spectra. AMINO ACIDS
Gruette and Kohnke (IOE)described four chromatographic systems for the quantitative determination of 12 amino acids by one-dimensional paper chromatography. A method for the separation and estimation of dinitrophenyl derivatives of 13 amino acids by a gas chromatographic technique was described by Ikekawa, Hoshino, and Watanuki ( I I E ) . Goodwin (8E, 9 E ) presented a simple procedure for the estimation of plasma and urine amino nitrogen based on the reaction of amino groups with 2, 4-dinitrofluorobenzene and colorimetric measurement of the resultant dinitrophenyl amino derivatives. Mokrasch ( M E ) used 2,4,6-trinitrobenzenesulfonic acid for the coestimation of amines, amino acids, and proteins in mixtures. Krzeczkowska, Czernaik, and Burzynski (15E) determined the free amino acids in human blood by the photometric measurement of the negative chromatographic prints. Bottomley ($E)described a rapid method forurine, plasma, and tissue amino acid analysis employing electrophoresis and chromatography on ion-exchange paper. Young (28E)described a scheme for the systematic screening of amino acids and sugars in urine using high voltage electrophoresis to separate amino acids and thin layer chromatography for sugar separation. Clayton and Steel 18 R
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(6E) modified the copper method of Pope and Stevens for estimation of alpha-amino N by a change in buffer and the use of tetraethylenepentamine as a color intensifier. Messineo (18E) used 2,4-dichloro-lnapthol reagent to determine arginine in unhydrolyzed proteins. Schneider, Bradley, and Seegmiller (2SE),studied the colorimetric assay method of Roston for cystine using noradrenochrome. MacDonald and Fellers (I7E) developed a simple semiquantitative method for the determination of urinary cystine by modification of the Sullivan test. Kivirikko, Laitinen, and Prockop (ICE) made several modifications for a rapid specific chemical assay method for hydroxyproline in urine. Miyahara and Jikoo (2OE) determined the optimum conditions for the colorimetric determination of lysine by the method of Kibrick. Seegmiller (25E) outlined chemical methodologies for the detection of human inborn errors of metabolism by examination of altered composition of urinary metabolites. Searle et al. (24E) described a manual fluorometric paper disc method for detecting phenylketonuria. Ambrose et al. ( 1 E ) modified the fluorometric technique for the determination of phenylalanine in serum to improve reproducibility, stability, and sensitivity. Irwin, Unanue, and Notrica (12E) described a microfluorometric screening technique (modified McCaman & Robins) for phenylketonuria using filter paper sample collection. Jones (1SE) described a n accurate colorimetric assay of serum phenylalanine which was suitable for routine lab use in dietary control of phenyl-ketonurics. Meyer, Droii, and Klingmueller ( I Q E )described a screening test for phenylketonuria and a semiquantitative estimation of phenylalanine in plasma by thin layer chromatography. Uchiyama et al. (27E) described the enzymic determination of L-phenylalanine and L-tyrosine. -4rapid quantitative chromatographic determination of phenylalanine and tyrosine in blood was described by Ballarini, Tosi, and Bandi (,%'E).Berry (SE) found a high degree of correlation between paper chromatography and fluorometric procedures for the measurement of phenylalanine. Spies (26E) compared two methods of hydrolysis of proteins for the determination of tryptophan. Rio and Sanahuja (22%) described a new technique for the determination of tryptophan in plasma based on the dimethylaminobenzaldehyde reaction. The spectrophotometric determination of tryptophan with mercuric chloride was reported by Leslie (16E). Gibbs, Saunders, and Sweeney (YE) described a new method for the quantitative estimation of tryptophan in plasma and urine using thin layer chromatography and induced
fluorescence. A fluorometric method for the measurement of tryptophan in biological materials based on the formation of the fluorophore norharman was developed by Denckla and Dewey
(6E)* BLOOD PRESERVATION, CLOTTING FACTORS, GASOMETRIC ANALYSIS, pH, AND VOLUME
Bradley and Barr ( I F ) compared the blood volume indocyanine green dye method and the standard isotopic techniques and found significant correlation. Graystone et al. ( 8 F ) used four methods: falling drop, freezing point elevation, infrared absorption, and gas chromatography to determine deuterium oxide concentration in plasma water. Miale and Lafond ( 1 6 F ) made a prothrombin time test survey and from the data obtained proposed criteria for acceptable commercial thromboplastin. Seegers, McCoy, and Marciniak (208') reviewed blood-clotting enzymology. il, system of prothrombin assay using a disposable semimicro chamber was described by McCormick and Kopp (15F). A comparative study of the Fibrometer, Seratek, and manual techniques for measuring prothrombin time was made by Gudaitis and Donauer (108'). Gilmour and Notrica ( 7 F ) described a single substrate for prothrombin activity and prothrombin proconvertin analyses. Burrows ( 2 F ) developed a slide rule for simple calculation of percentage of prothrombin activity. Severinghaus (,%'IF)reviewed blood gas analyses and discussed the problems of their measurement, recent developments, and improvements. Siemon (22F) compared various membrane electrodes for the measurement of blood gases. Conway and Masterson ( 4 F ) presented a method for the measurement of combined blood 0 content and capacity with the use of a modified Conway microdiffusion unit. Clauvel and Schwartz ( J F ) made adaptations of the Van Slyke oxygen capacity method to reduce error. Lenfant and Aucutt ( 1 4 F ) measured the blood gases 0, CO,, and ?r' by gas chromatography. Ledsome et al. (IBF) developed an apparatus consisting of 3-electrode systems for the measurement of pH, pCOz and p 0 2 . Sinnema ($3) described a colorimetric method for the determination of bicarbonate in serum using methyl red. Rispens et al. ( f 9 F ) described a direct method for the determination of the HCO-a concentration as total COz in plasma. Reyes and Neville (18F) described a rapid electrochemical technique for measuring COZ content of blood which required a membrane covered pH electrode. Dubowski (6F)presented a procedure for the electrometric determination of pC02 in se-
rum which employs a Severinghaus pCOz electrode. Thews and Eschweiler (25F) patented a n apparatus for the determination of acid-base equilibrium in blood. Szakacs and Cramer (24F) discussed the principles involved in the electrometric method for blood p H determination with glass electrodes. Whitehead (26F) reviewed the terminology, physiology, and clinical applications of blood hydrogen ion. S a u m a n n ( 1 7 F ) reviewed and discussed the chemical methods for the measurement of total base in serum. Le Massena ( 1 S F ) patented a hollow hypodermic needle containing a reference and a measuring electrode to measure pH in vivo. Jaques and Wollin ( 1 1 F ) described a sensitive accurate quantitative colorimetric assay for heparin with Azure A. Grzyb-Skorzynska ( Q F )utilized the reaction of the conjugation of Karfarin and diazotized p-nitroaniline for the photometric determination of Warfarin in serum. Corn and Berberich (68’) described a rapid fluorometric assay method for plasma Karfarin based on the principle of decrease of fluorescence of Warfarin in acetone by acidification. CARBOHYDRATES
Anno and Sen0 ( I G ) made a comprehensive review of several methods for carbohydrate determination such as: reducing sugars, hexoses, pentoses, tetroses, deoxy sugars, uronic acid, hexosamine, etc. Ashwell (2G) reviewed new colorimetric methods of sugar analysis such as: tetroses, aldopentoses, aldohexoses, ketohexoses, deoxy sugars, etc. Reinauer and Hollmann (SSG) reviewed titrimetric, colorimetric, enzymic, chromatographic, and electrophoretic methods for the determination of D-glucose and low molecular weight carbohydrates. Goffrini, Regolisti, and Veggetti (1SG) compared chemical and enzymic blood glucose methods and reported 8% lower glucose values by the enzymic technique. Lorentz and Leudemann (21G) also made a comparative study of blood sugar methods and recommended triphenyltetrazolium chloride and the glucose oxidase-peroxidase methods. Barletta, Scolari, and Testi (4G) found lower blood glucose values with the enzymic Keilin and Hartree than with the Hogedorn-Jensen and Folin-Ku methods. Christensen (1OG) made a report on some methodological studies of the enzymatic determination of blood sugar. Puetter and Strufe (SZG) improved the enzymic determination of glucose by stabilization of the 0-dianisidine color by the addition of poly(viny1pyrrolidione). R a r e , Marbach, and E‘en (40G) made a direct application of a glucose oxidase method in which the HzOz produced reacts with catalyzed iodine to form iodine which is measured
photometrically. Tammes and Nordschow (SQG) measured true glucose by glucose oxidase employing titanium and xylenol orange to form a highly colored metal-chelate complex in the presence of H 2 0 2 . Phillips and Elevitch (S1G) measured plasma glucose enzymically by measuring the peroxide formed with peroxidase and homovanillie acid. Middleton (26G) stabilized a glucose-oxidase-peroxidase reagent by addition of polyethylene glycol (“Carbowax 20 M”).h microspectrophotometric method for the determination of glucose with fluorescein was described by Braun and Wadman (8G). Kamel, Hart, and Anderson (2OG) described an ultraspecific micro method for D-glucose employing a purified stereospecific D-glucokinase coupled with glucose-6phosphate dehydrogenase. Hexokinase coupled reaction methods for the determination of true glucose were reported by Xitchell and Rydalch (27G), Stork and Schmidt (S8G), Peterson (29G), Peterson and Young (SOG) and Schersten and Tibbling (35G). Methods for the determination of glucose with 0toluidine were reported by Yamamoto (42G), Wuest (GIG), Luess (22G), Zender and Xolen (4SG), Horak, Kchouk, and Hawas (ITG) and Braun (9G). Shats, Filov, and SIaksimova (S6G) described a photometric method for the determination of reducing sugars in blood and urine with 3, j-dinitrosalicylic acid. Deckert (12G) determined glucose directly in p l a s m a , s p i n a l fluid and urine with p-brom-aniline. Clark and Timms (11G) substituted reduced 2,6-dichlorophenolindophenolfor 0-tolidine in the enzymic determination of blood glucose. Avigad (SG) increased the sensitivity of the alkaline ferricyanide glucose method by use of 2,4,6tripyridyl-S-triazine as the Fez+ reagent. Patents for glucose oxidase techniques, tests, and procedures for the detection of glucose were obtained by , Gretton Mast (24G), Kallies ( I Q G ) and and Rees (14G). A patent for a glucose method in which glucose is determined enzymically by a transferase and a phosphate donor was obtained by Boehringer (TG). Bittner (6G) obtained a patent for a copper-neocuproine indicator for glucose. Rupe and Hue (S4G) obtained a patent for the qualitative detection and determination of galactose. Aletras (25G) and Becker and May (5G) presented procedures for the identification of sugar in urine by thin layer chromatography. Hjelm (16G) developed a simple reliable procedure for the determination of glucose in serum, similar to the glucose oxidase method. Heinz and Wistuba (16G) developed a n enzymatic spectrophotometric method for the assay of fructose l-phosphate. Passonneau et al. (28G) described an enzymic method for the
measurement of glycogen using the TPN+ system. Kadish, Litle, and Sternberg (18G) determined glucose levels in biological fluids by a glucose oxidase method employing a polarographic oxygen sensor, with a circuit modified to record the rate of oxygen consumption. Makino and Konno (2SG) measured blood glucose by oxygen electrodes. Simon, Christian, and Purdy (STG) combined the coulometric titration method with an enzymatic analytic reagent for determination of glucose in human serum. CATIONS A N D ANIONS
Ingram and Hogben (25H) made electrolyte analyses with electron probe. Teloh ( 6 5 H ) reviewed the history and theory of emission photometry for the serum cations: Na, K, Ca, and Mg. MacFate ( S 4 H ) reviewed the chemical methods for the determination of serum calcium. Sarkar and Chauhan ( 4 S H ) worked out a new method for the micro colorimetric determination of serum calcium employing O-cresolphthalein complexone. The glyoxal bis(2-hydroxyanil) direct spectrophotometric calcium reagent was studied by Lindstrom and Milligan ( S I H ) and Kuczerpa (SOH). Chen and Dotti, ( Q H , 1OH) used resin Chelex 100 for separation of calcium from jaundiced, hemolyzed, and lipemic sera and urine and stools. Borle and Briggs ( 6 H ) described a method for determination of very small amounts of calcium in biological material by automatic fluorimetric titration of a calcium-calcein complex with EDTA. Arnold, Stansell, and Malvin ( 2 H ) evaluated a calcium ion electrode for routine measurement of serum Ca*+. Turpin and Bethune ( 5 8 H ) described a simplified method for determination of radioactive calcium-45 in biological material by gel scintillation counting. Studies of the factors affecting the determination of calcium in serum and urine by atomic absorption spectrophotometry were reported by Bowers, Smith, and Feldman ( 7 H ) , Kocian and Rubeska ( 2 9 H ) and Trudeau and Freier ( 5 7 H ) . Rodgerson and Abraham (& H ) obtained better calcium recoveries by atomic absorption than by fluorimetry. Studies of the determination of calcium and magnesium in serum, urine, etc. by atomic absorption spectrophotometry were reported by Gimblet, Marney, and Bonsnes ( 1 7 H ) . Monder and Sells ( S 6 H ) , Sunderman and Carrol (52H) and Thin and Thompson ( 5 6 H ) . Osmun ( 3 9 H ) and Sunderman ( 6 1 H ) described improved methods for the determination of ultrafiltratable calcium and magnesium in human plasma and serum. Lomax ( S S H ) determined protein-bound magnesium and calcium by gel filtration. Sunderman and SunderVOL. 41,NO. 5.,APRIL 1969 * 19 R
man ( 5 4 H ) reviewed the chemical measurement of magnesium in biological fluids. Ryan and Hingerty ( 4 2 H ) compared flame emission, atomic absorption, fluorimetry, and titan yellow magnesium methods and found magnesium ranged from 1.71-1.76 MEq for normal adults for the first three methods and 1.92 MEq for the latter. Oreopoulos, Soyannwo, and McGeown (38H) studied the influence of gluconate and glycogalacto-gluconate on the fluorometric estimation of magnesium with 0,O-dihydroxyazobenzene. Gusev ( 1 9 H ) used lumomagneson (2-hydroxy-3-sulfo-5-chlorophenylazo) barbituric acid for the fluorometric determination of trace quantities of magnesium in serum and urine. Connerty and Briggs (12H) described a spectrophotometric method for the determination of serum magnesium using Erochrome black T with perchlorate filtrates and the barium salt of ethylene bis(oxyethylenenitri1o) tetraacetic acid to supress calcium. Clark and Hou ( 1 1 H ) reported an improved fluorometric procedure for the determination of magnesium in biological materials in which interfering substances were eliminated. Bauditz ( 5 H ) made titrations of calcium and magnesium subsequently by changing the indicator from murexide to pyrocatechol violet. Studies of the determination of magnesium in biological material by atomic absorption spectrophotometry were made by Iida, Fuwa, and Wacker ( 2 4 H ) and Hansen and Freier (2OH). Sinnema ( 4 7 H ) described a colorimetric method for the determination of bicarbonate in plasma or serum which employed methyl red. Senterre and Sodoyez-Goffaux ( 4 5 H ) compared blood pCO2 measurements by Astrup and Siggaard-Andersen and pCOz electrode methods and found little statistical difference. de la Huerga, Smetters, and Sherrick (1414) reviewed chloride methodology; iodometric, absorption indicator, mercurimetric, microdiff usion, electrometric, colorimetric, and titration methods. Sitzmann (48H)described a micro-chloride method based on the photometric measurement of chloranilic acid which is formed from Hg chloranilare in the presence of C1- in acid media. Siggaard-iindersen ( 4 6 H ) determined chloride in 8 pl of serum by employing a Cotlove chloride titrator with ultra micro-dilutor. Gindler ( 1 8 H ) found that 2-( 8-hydroxyquinolyl-5-azo)benzoic acid was suitable as a mercurimetric indicator for the determination of chloride in human serum. Kist, Lobanov, and Kryzhnenkova (27”) described a simplified activation determination of chloride in biological materials. Zipf and Katchman ( 6 1 H ) reviewed the gravimetric, titrimetric, and photometric methods for the determination 20 R
ANALYTICAL CHEMISTRY
of serum Na. Sunderman (60H)recommended a modification of Barber and Kolthoff’s gravimetric sodium method for reference purposes. Pommer, Simon, and Calcagno (40H) and Annino ( 1 H ) determined urinary sodium with the sodium electrode. Oberg, U1fendahl, and Wallin (S7H) gave a detailed description of a low cost integrating flame photometer for simultaneous determination of Na and K. Knauff (28H) measured the interference in various biological fluids on flame photometric determination of sodium and potassium. Sunderman and Sunderman ( 5 d H ) investigated the platinic chloride and the tetraphenylboron methods for the measurement of serum potassium. Hillmann and Beyer ( 2 I H ) made a rapid determination of serum potassium by turbidity measurement with sodium tetraphenyl borate after precipitation of protein. Cannon and Gadsden (8H)and Gadsden and Cannon ( 1 6 H ) reviewed the evolution and development of the cobaltinitrite method for the measurement of serum potassium. Fried and Hoeflmayr ( 1 5 H ) patented a variation of the cobaltinitrite method for serum potassium measurement. Kaltenbach, Dean, and Hoffman ( 2 6 H ) reviewed the problems associated with the measurement of inorganic phosphorous in serum. Skogerboe, Gravatt, a n d Morrison ( 4 9 H ) described the flame conditions which permit the excitation of the atomic spectrum of phosphorus. Schulz, Passonneau, and Lowry (44H) described an enzymatic method for measuring inorganic phosphorus in which T P N H is finally produced from TPH+ for fluorometric measurement. Hynie, Stepan, and Vecerek (92H) and Hynie, Vecerek, and Stepan, ( 2 d H ) described a new, simple, and sensitive method for detection of P043- on paper based on its reaction with methyl violet and ammonium molybdate in dilute HCl. Bastiaanse and Meijers ( 4 H ) presented a micromethod for the colorimetric determination of inorganic phosphate in which malachite green forms a complex with phosphomolybdate a t a low pH. Crouch and Malmstadt ( I S H ) made a study of the chemistry of the molybdenum blue method for the determination of phosphate. Baginski, Foa, and Zak ( 3 H ) described the microdetermination of inorganic phosphate, phospholipids, and total phosphate in biological materials. Yoshimatsu (60H) presented an indirect, isotopic method for the estimation of serum inorganic iodine. Leonard ( S 1 H ) patented a method for the removal of inorganic iodine from blood by the addition of solid AgC1. Wheeler, Brierre, and Dickson ( 6 9 H ) reviewed titrimetric, gravimetric, turbidimetric, and colorimetric methods for the measurement of inorganic sulfate
in biological fluids. Mathies and Lund ( d 6 H ) described an X-ray emission spectographic procedure for the quantitative analysis of calcium, sulfur, and phosphorus in urinary calculi. LIPIDS
Methods for the microdetermination of total lipids, cholesterol, and lipid phosphorous in spinal fluid were described by Pilz ( 1 9 J ) . Martinek ( 1 5 4 proposed a simple, rapid, and precise turbidimetric procedure for serum total lipids. A scheme was developed by Krehl, Lopez-S, and Good ( 9 J ) for a rapid semiautomatic determination of serum lipids in which a single isopropyl alcohol extract is used. Friedman ( 5 4 determined serum total lipids by a method based on the lipid extraction technique of Van Slyke and Plazin. Markunas ( 1 4 4 presented a method for the rapid extraction of lipids from serum. Spectrophotometric methods for the determination of free fatty acids were described by Porges, Oravcova, and Bozek (2OJ), Laurell and Tibbling ( I I J ) and Massion and Seligson ( 1 7 4 . Titration methods for the determination of free fatty acids in serum were employed by Mueller and Ulrich ( I 8 4 and Goss and Lein (7‘J). Louis-Ferdinand et al. ( 1 3 4 and Free and Free ($4employed thin layer and gas chromatography for the quantitation and fractionation of fatty acids in serum. Williams and Soeling ( 2 5 4 described the pitfalls in the enzymic determinationof serum triglycerides. Wolf, Kochsiek-Schuster, and Loehr ( 2 6 4 made an ultramicro modification of Kreutz’s enzymic method for the determination of free glycerol and triglycerides in serum. Szostak (23sJ) compared the van Handel and Zilversmit and the Carlsen spectral serum triglyceride methods and found good agreement. Spectrophotometric serum triglyceride methods were described by Laurell ( I O J ) , Martinek ( I S J ) , Sardesai and Manning (IbJ), Freeman, Lampo, and Windsor ( 4 4 and Fletcher ( 2 J ) . Kanter ( 8 4 suggested mannitolasaprimarystandardin thedetermination of triglycerides. Ryan and Rasho ( S l J )describedasimplemethodof preparing extracts of plasma or serum for determination of both triglycerides and cholesterol. Fuehr and Schacht ( 6 4 prepared a secondary cholesterol serum standard by adding a known amount of pure cholesterol to a solution of serum containing 3% Thesit prewarmed to 65-70 “C. Leibman and Ortiz ( I N ) reported a procedure for assay of glyceric acids in which acidic glycol is oxidized with periodic acid and the p-nitrophenylhydrazone of the resulting carbonyl acid is prepared for colorimetric measurement. de Freitas (1J) described a paper chromatographic procedure for the quantitative
isolation of glycerol from 100 r l of plasma. Thaxton and Bowie ( 2 4 4 described a method for the determination of lecithin in plasma as cholin using lecithinase D in an in vitro system. ENZYMES
Guilbault (22K) reviewed the factors affecting reaction rates, methods of monitoring these rates, and their application to enzymic and nonenzymic reactions. Henley, Schmidt, and Schmidt ( 2 9 K ) and Latner ( 4 S K ) reviewed isoenzymes, their distribution, technical methods, and clinical applications. S e w substrates for the fluorometric determination of oxidative enzymes was described by Guilbault, Brignac, and Juneau ( 2 4 K ) and Guilbault, Brignac, and Zimmer ( 2 5 K ) . Guilbault ( 2 S K ) reviewed the determination of enzymes; fluorometric methods, oxidative enzymes. dehydrogenases, electrochemical, and radiocheniical methods, determination of suhstrates, coenzymes and inhibitors. Juul ( S 6 K )reported t h a t all plasma enzymes are stable a t 20 "C for 8 days except lactic dehydrogenase. Fletcher and Verdi (17K) developed an analytical method for determining serum acid phosphatase utilizing a p nitrophenyl phosphate substrate. Amador and Price ( 2 K ) reported that alphanapthyl phosphate was not specific for prostatic acid phosphatase. Inglis, Ghosh, and Fishman ( S S K ) developed a method for separation of the variants of human serum alkaline phosphatase by horizontal Sephadex gel electrophoresis. Smith, Lightstone, and Perry ( 7 I K ) described a new procedure for the separation of alkaline phosphatases using a continuous tris-borate buffer on a 5% acrylamidt, disc gel. -1 4% hour method for the quantitative separation of serum alkaline phosphatase isoenzymes using the Beckman microzone system was reported by Fisher and Nixon ( I 5 K ) . Epstein et al. ( I S K ) used a synthetic para-toluidine salt of 5-bromo-4-chloro-3-indolyl phosphate as a substrate for alkaline phosphatase determination after electrophoretic separation. Romel, Lallancusa, and DuF r e n e ( 6 2 K ) used phenolphthalein monophosphate and Fischl, Segal, and Rabiah ( 1 4 K ) , phenolphthalein diphosphate as substrates for phosphatase determination. Seumann and Van Vreedendaal ( 5 S K ) improved Bessey's alkaline phosphatase method by use of a SHaOH-TH4Clbuffer. Johnson ( S 4 K ) used a naphthyl AS-phosphoric acid ester to provide a fluorogenic substrate for a very sensitive alkaline phosphatase method. Coleman ( 9 K ) synthesized a magnesium thymol-phthalein monophosphate substrate for alkaline phosphatase determination. Ban-ers, Kelley, and RlcComb ( 8 K ) studied the activity and sensitivity of nine
self-indicating phenolic substrates for spectrophotometric measurement of alkaline phosphatase. Bowers, Kelley, and McComb ( 7 K ) reported variability of analytic results in a survey related to the use of nonhuman serum alkaline phosphatase. Berk ( 6 K )reviewed advances in methodology of serum amylase and lipase. Searcy, Kilding, and Berk (69K) made an appraisal of the amylase methods and concluded that the saccharogenic methods appear to possess the greatest intrinsic validity. Strumeyer (72K) described a simple reproducible procedure for the preparation of a n amylase substrate which was suitable for use with various reducing sugar methods. A novel substrate, dyed amylopectin was employed for serum amylase determination by Babson, Kleinman, and Alegraiv ( 4 K ) . Jlellerup ( 4 9 K ) combined the enzymic formation of urea with the method of Coulombe for measuring serum arginase. Satoh and Ito (66K) reported a rapid micromethod for arginase which utilizes diacetylmonoxime in an oxidizing medium. K a r d and Srere ( 7 7 K ) developed a n e x spectrophotometric arginase assay method based on the fact t h a t the absorbancy of arginase below 210 mp is larger than the combined absorbancies of ornithine and urea. Krainev ( 4 1 K ) described a method for the simultaneous colorimetric detection of blood catalase and hemoglobin on paper. Juul ( S 7 K ) described a method for the separation and quantitation of cholinesterase isoenzymes using disk electrophoresis. Gerarde ( I 9 K ) patented a n improved test for cholinesterase activity in blood. Johnson, Osaki, and Frieden ( 3 5 K ) reported a sensitive method for the detection of ceruloplasmin (ferroxidase) in human serum using Fe*+ as the substrate. Kilkinson and Steciw (79K) evaluated a new creatine kinase procedure in which the reagents of the method of Oliver rrere prepared in tablet form. Sax and Moore (6827) compared the Sax-Noore and Conn-Anido fluorometric creatine kinase methods and found less interference in t h e Sax-Moore method. Pon and Bondar (58K) nieasured pyruvate kinase by following the enzymic transphosphorylation reaction, catalyzed by pyruvate kinase and recording the disappearance of phosphoenol pyruvate a t 230 mp. Methods for the analysis of creatine phosphokinase isoenzymes after electrolytic separation were described by Trainer and Gruenig (1610 and Sherwin, Siber, and Elhilali (7OK). Rosalki ( 6 S K ) improved a procedure in which adenosine triphosphate liberated by creatine phosphokinase is linked to the reduction of nicotinamide-4 adenine dinucleotide phosphate. similar method was described by AleCrimmon and Lewin ( 4 8 K ) for the de-
termination of serum creatine phosphokinase. Methods for the spectrophotometric determination of creatine phosphokinase were described by Hess, Murdock, and Natho ( S I K ) , Hess et al. (SOK), and Bigazzi and Ciampi ( 6 K ) . Graig, Smith, and Foldes ( 2 1 K ) reported that creatine phosphokinase activity is increased upon dilution of serum which can be prevented by dilution with heat inactivated serum or the incubation time shortened. Fishinan ( 1 6 K ) reviewed the assay methods for 6-glucuronidase. Szasz ( S 6 K ) compared the p-nitrophenyl glucuronide and the phenolphthalein glucuronide substrates for assay of 6-glucuronidase. Menken, Barrett and Berlin (O'OK) assayed hepatic glucuronyl transferase activity using [14c] bilirubin as a substrate. Syssen and Dorche ( 5 6 K ) described a semiautomatic method for the determination of guanine deaminase activity in human serum in which the ammonia formed is estimated by the Berthelot reaction. McComb and Gay ( 4 7 K ) examined the physical and chemical properties of sereral commercial sources of reduced S A D and compared relative rates of oxidation with t n o human lactic dehydrogenase (LDH) isoenzymes. Marymont, Cawley, and Hoffman (45K) compared four L D H methods and found methods employing oxidation of lactate to pyruvate gave best correlation. Krieg, Rosenblum, and Henry ( 4 2 K ) compared pyruvate to lactate and lactate to pyruvate assay methods for serum L D H isoenzymes. Methods for electrophoretic separation and idcntification of L D H isoenzymes were described by Pribor, Iiirkham, and Fellows ( 6 9 K ) , Hemmingsen and Skov ( 2 8 K ) , Holeysouska ( S Z K ) , Papadopoulos and Kintzios ( 5 7 K ) , Gay. N e and Myers Comb, and Bowers (18K), and Van Remortel ( 6 2 K ) . The differential determination of L D H isoenzymes was reported by Lubran and Jensen ( 4 4 K ) , Kelshman and Rixon (78K) and Emery, lloores, and Hodson ( I Z K ) . Kelly, Belorit, and Copeland (S8K) studied factors affecting LDH isozyme patterns of muscle. Szasz (74K) reported a kinetic method for the determination of leucine aminopeptidase (LAIP)activity in serum using L-leucyl-p-nitroanilide in tris buffer as substrate. Knight and Hunter (39K) described a new method for LAP in which LAP reacts with substrate Lleucyl-L-alanine to generntc alanine which forms pyruvate in the presence of excess a l p h a - k e t o g l u t a r i c acid. Haschen, Farr, and Keichelt ( 2 6 K ) described a new photometric method they used for the determination of serum L.4P. S e w rapid methods for the determination of serum lipase were described by Massion and Seligson ( 4 6 K ) and VOL. 41, NO. 5, APRIL 1969
21 R
Dirstine, Sobel, and Henry (11K) in which olive oil substrates were used. Amador, Massod, and Franey ( 1 K ) optimized all reagents and conditions for the spectrophotometric glutamic oxalacetic transaminase (GOT) method and compared it with the colorimetric. Healy (27K) commented on the necessity of serial dilutions of serum of high GOT activity. Sax and Moore ( 6 7 K ) described a method for measuring GOT activity in which substrate concentrations are more nearly optimal than in previous colorimetric methods. Rinderknecht et al. (61K) developed sensitive fluorometric and colorimetric methods for the estimation of pancreatic elastase. Paglia and Valentine ( 5 6 K ) presented an assay in which blood cell glutathione peroxidase may be measured by a direct spectrophotometric procedure. Kg, Bergren, and Donne11 ( 5 4 K ) described an improved procedure for assay of hemolysate galactose-l-phosphate uridyl transferase. Simple methods for the estimation of ornithine carbamoyl-transferase activity were described by Moore ( 5 I K ) and Konttinen (4OK). Rinderknecht et al. (60K) described an ultrasensitive method for the colorimetric determination of proteolytic activity in biological fluids based on an insoluble substrate derived from hide powder labelled covalently with Remazol-Brilliant-Blue. Rutenburg, Smith, and Fischbein ( 6 4 K ) investigated the electrophoretic mobilites of serum alpha-glutamyl transpeptidase in serum. Rybak et al. (65K) presented a sensitive and specific method for the determination of enzymes splitting S-acetyl tyrosine ethyl ester. Dietz et al. (1OK) developed a method for the estimation of serum trypsin inhibitory capacity to permit consistent results with various trypsin sources. Swaim ( 7 S K ) proposed an indirect caseinolytic assay for plasminogen, utilizing acid treated serum and optimal concentration of streptokinase. Arawaka et al. ( S K ) devised an improved zero order reaction method for the determination of plasma renin in which human plasma protein fraction IV-4 was used as substrate. Glen (ZOK) described a method for the measurement of DX.1 and RXA in human peripheral lymphocytes. FUNCTION TESTS
Liver. Roth ( S 5 L ) presented a simple sensitive assay method for total bilirubin in serum in which 0.05 ml of serum is mixed with 0.6 ml of H3P04 (85%) 3 ml of water and fluorescence measured. Kulhanek and Appelt (WSL) developed a simple method for total bilirubin determination in serum based on the coupling of bilirubin with a stable diazonium salt (Fast red RC). Gindler and Ishizaki ( 1 6 L ) determined 22
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ANALYTICAL CHEMISTRY
total serum bilirubin colorimetrically by formation of azo compounds with diazotized 2-amino-5-nitrobenzenesulfonic acid. Methods for the determination of serum bilirubin were patented by Fried and Hoeflmayr ( 1 4 L ) and Ferro and Ham (11L). h’ew methods for the determination of total and direct bilirubin were published by Ichida and Nobuoka (2OL) and Ferro and Ham (1OL). Becker (1L) described the criteria for the preparation of bilirubin standards for microbilirubin determination. Gallo ( 1 5 L ) made spectrophotometric determination of plasma bilirubin fractions by solvent separation. Brodersen ( S L ) described a radioisotope derivative method for quantitative determination of bilirubin diglucuronide in serum. Krylov (22L) suggested a method combining the procedures of Jendrassik and Van den Berg for the determination of 5 serum bilirubin fractions. Fog and Bakken (12L) used the difference between the absorption curve of conjugated and unconjugated bilirubin to determine their ratio in serum. Fog and Bakken ( I S L ) determined the absorptivity of alkaline solutions of highly purified bilirubin. Roovers, Evrard, and Vanderhaeghe ( S 4 L ) developed a new method for blood bile acids employing an enzyme from Clos-
tridium perfringes Welchii. Castrom et al. (5L) described a rapid screening test for urinary porphyrins using anion-exchange resin. Rapid thin layer chromatographic methods for isolation and determination of urine porphyrins u-ere described by Scott, Labbe, and Nutter ( S 6 L ) , Doss and llannheim ( 8 L ) ,and Chu and Chu ( 6 L ) . Chu and Chu (7L) used simple paper chromatography of the methyl ester of the total porphyrin of a urine sample to reveal a chromatographic pattern of the porphyrins characteristic of the type of porphyria. Spandrio and Albertini (S7L) compared the results of oxidation with and without iodine in the spectrophotometric determination of urinary coproporphyrins. Methods for the resin, column, thin layer and electrophoretic separation of urinary 3methoxy-4-hydroxy-mandellic acid (VMA) and its quantitative determination was described by Kybenga and Pileggi (4WL), Weil-Malherbe ( 4 1 L ) , Preston ( S I L ) ,and Butler ( 4 L ) . O’Gorman (28L) estimated vanilmandelic acid in serum and urine by a radioactive 14C labelling technique. Hingerty (18L) described a rapid method for vanilmandelic acid in which interfering chromogens are removed by acidic amyl alcohol extraction and color developed with diazotized p-nitroaniline. Duke and Demopoulos (9L) described a one-dimensional paper chromatographic method for determination of urinary homovanillic acid. Mellinger ( 2 6 L ) determined homovanillic
acid in urine by potassium permanganate oxidation to produce strong fluorescence. Mikulecky (27L) made new experimental findings and theoretical considerations in a detailed study of the kinetics of plasma bromsulphalein. Ott and Pirrwitz (29L) improved the bromsulphalein method by use of NanS 2 0 ain blank to discolor the dye and retain the same pH as sample to minimize interferences. Tovey (S8L) maintained that 5 mg of BSP/Kg of body weight is not sufficient to allow accurate monitoring of the excretory function of the liver and that 7 . 5 mg/Kg should be used. Kidney. Beyermann, Gerok, a n d Groth (WL)reported that direct useof the Jaffe reaction without preliminary separation from noncreatinine chromagens gives variable results. Husdan and Rapoport (19L) investigated the total chromogen, true creatinine, and AutoAnalyzer methods of measuring serum and urine creatinine by the Jaffe reaction. Pokrovskaya, Semenova, and Tereshchenko (SOL) purified urinary extracts for creatinine determination by use of a cationite ( S a + ) column. Watkins (4OL) discussed the precautions required to prevent the interference of ketone bodies on the determination of serum creatinine. Varma, Yadava, and Chand (S9L) used ferricyanide hydrolysis and ferric alum color development for the micro determination of creatinine in urine. Marymont, Smith, and Klotsch ( 2 5 L ) adapted a creatine phosphokinase enzymic method for the determination of creatine in urine. London. Freiberger, and Marymont ( 2 4 L ) used the heat-clot method of preparing filtrates for serum creatinine determination. Grafnetter, Janosova, and Cervinkova ( 1 7 L ) modified Slot’s method for true creatinine determination in which apparent creatinine value is corrected for nonspecific chromagen by photometer measurements in the alkaline and acetified sample. Rockerbie and Rasmussen (3SL) determined serum creatinine by a rapid ion exchange batch technique in which creatinine is absorbed by a strong cationexchange resin and finally eluted with p H 12.4 phosphate buffer. Rink and Krebber (S2L) used thin layer chromatography for separation of urinary creatinine and creatine. Krees, Baukal, and Wolff (21L) described a semiautomated method for the simultaneous colorimetric determination of inulin and p-amino hippuric acid in blood and urine. HEMOGLOBIN
Noeller (6M) patented an apparatus for the determination of hemoglobin in blood in which the electrical resistance of a mixture of acid hematin is sensed. Klein et al. (5M)developed a new pro-
cedure for hemoglobin in which blood is mixed with sodium hypochlorite and benzodiazepin chelating agent in buffer and ascorbic acid and the absorbance of the violet blue solution is measured a t 580 mp. Katchman et al. (41M) determined the total body hemoglobin from the measurement of alveolar carbon monoxide concentration. Eilers ( 3 M ) reported notification of final adoption of a n international method and standard solution for hemoglobinometry and specifications for preparation of standard solution. Pantlitschko a n d Weippl ( 7 M ) described a new method for quantitative haptoglobin determination based on the measurement of hemoglobin in the Soret zone. Chong and Owen (ZM) determined methaemalbumin in plasma by use of dithionite. A method by which a sample of whole blood may be applied to acrylamide gel to obtain electrophoresis of hemoglobins was described by Smith and E v a t t ( 1 0 M ) . Bjerre ( 1 M ) determined simultaneously the concentration of two abnormal hemoglobin pigments by a simple spectrophotometric method. Rosenbaum ( Q M ) reported t h a t electrophoresis on cellulose acetate is a rapid reproducible method for fractionation and quantitation of Hbs -4,S, C, F, and A*. Zettner and Mensch ( 1 4 M ) used atomic absorption spectroscopy for the determination of iron in hemoglobin. Rand, Lacombe and Barker (8M)evaluated the effect of hemoglobin concentration on the percentage of oxygen saturation in blood as determined by the spectrophotometric method of Gordy and Drabkin. Zettner ( 1 3 M ) believed t h a t the internationally recommended value of 11.0 for the 1/4 m M absorptivity constant of cyanmethemoglobin mas too high, but Williams’ (12M) analyses yielded a mean value of 43.96 for m M . Theil and .luer (1IM) studied hemoglobin oxidizing agents. METALS
Bedrosian, Skogerboe, and Morrison developed a rapid comprehensive spectrographic procedure for the direct determination of a large number of trace elements in 25-50 mg of dried blood serum. Berman (5X) gave procedures for the atomic absorption determination of cadmium, thallium, and mercury in biological materials. Howe (33147) reviewed atomic absorption spectrometry, theory, instrumentation, and application. The determination of trace elements by activation analysis was reviewed by Heydorn (ZQAlr), Comar and Desenne (12N) and Kanabrocki et al. (SYN). Diehl (16-V) reviewed the work on EDTA titrations of calcium and magnesium, and the development of metallochromic indicators. Huey and Hargis ( S 4 S ) determined cesium spectrophotometrically (4147)
with 12-molybdophosphoricacid. Chromium was determined in bilological materials by coulometric titration by Feldman, Christian, and Purdy (81N). Parker and Boltz (46N) proposed a new ultraviolet spectrophotometric method for the determination of trace amounts of chromium as the peroxychromic acid-2,2’-bipyridine complex. Binnerts and Boom ( 7 N ) determined cobalt in liver by atomic absorption, and Binnerts (6N)by photometry. Todd, Thorpe, and Rosenoer (55N) described a method for handling of tissue samples for copper determination by neutron activation analysis. Sunderman and Roszel (63A7 described atomic absorption spectrometry for the d e t e r m i n a t i o n of copper in urine, serum, feces, and tissues. Heller, Guyon (88N)developed a sensitive spectrophotometric method for the determination of trace amounts of copper based on the catalytic effect of copper on the ascorbic acid reduction of the isopolymolybdate species a t pH 1.85. Dmitrieva (17A7 proposed new specific organic reagents for photometric determination of copper as a trace element. A new calcichrome method for colorimetric determination of copper in serum was described by Deguchi ( 1 5 N ) . Budziszewski (ION) described the best cation exchangers for the adsorption of copper ions. Van De Bogart and Beinert (56.V) described micromethods for the quantitative determination of iron and copper in biological material. A simple rapid method for the isolation of ceruloplasmin was described by Stokes (50N). Methods for the determination of copper and zinc in biological material by atomic absorption spectrophotometry were described by Parker, Humoller, and Mahler (4Y.V), Girard (ZSN) and Dawson, Ellis, and SemtonJohn (1SA7). Holland and Bozic (32N) found t h a t 2,2‘-dipyridyl ketoxime mas a highly selective colorimetric reagent for gold. Zaino (5YAV) compared a n atomic absorption iron method and a new colorimetric method employing 2,4,6-tripyridyl-S-triazine and found good agreement. The colorimetric method was devised by Ichida, Osaka, and Kojima (35N). Schlit and Hoyle (48147 found a ferroin reagent, 3-(4-phenyl-2 pyridyl)-5,6-diphenyl-l,2,4-triazineto be a sensitive chromogen for iron (Fez+), McDonald and Bedenbaugh (44N) described a spectrophotometric method for the determination of iron based on the blue color formed when ethyl 4,6dihydroxy-5-nitrosonicotinate is added to a slightly acidic solution of iron (Fez+). Johnson and Young ( S S N ) evaluated the interferences in the spectrophotometric determination of iron with ethylenediamine di(0-hydroxyphenylacetic acid). Gandhi and Desai (22W) observed t h a t 2,4-dihydroxypropiophe-
none oxime forms a stable purple colored complex with ferric iron between p H 2.0 and 3.0. Deguchi (14N) used calcichrome as a chromagen for the determination of 0.2 to 2 pg of serum iron. The use of Teepol 710 (Shell) as an anionic surfactant to prevent protein precipitation in direct colorimetric serum iron methods was described by Dronski and Galts (18 N ) . Kitaguchi and Nishimoto (38N),Schmidt (4QLV), and Askevold and Vellar ( 2 s ) . Babson and Kleinman (3N) stated t h a t erroneously high values for serum was given by the AutoAnalyzer because of the Donnan equilibrium effect on the rate of dialysis of ferrous ions. Zettner and Mensch (58iV) described a rapid method for iron analysis of hemoglobin by atomic absorption spectrophotometry. Serum iron binding capacity methods were described by Burrows ( f l S ) , Brownstein ( 9 5 ) , Hillman, Morgan, and Finch (3012’), Hoeflmayr and Fried (3f4V) and Xielsen ( 4 5 5 ) . The determination of lithium in serum and urine by atomic absorption spectroscopy was described by Zettner, Rafferty, and Jarecki (59S), Hansen (87h’), Lehmann (4O.V), and Little, Platman, and Fieve ( 4 1 S ) . Blijenberg and Leijnse (8s)found t h a t atomic absorption spectroscopy is not superior to flailse emission spectroscopy for routine lithium determination. Titov and Kh (54.V) reported that flame photometric determination of lithium in biological material is more accurate than chemical and photometric methods. Amdisen (IX ) studied the chemical and physical variables which may influence the flame photometric lithium determination. Spectrophotometric methods for the determination of manganese in biological samples was reported by Sulochana (51N ) and Hamaguchi, Horiuchi, and Tanaka ( 6 6 s ) . Feldman. Bosshart, and Christian (ZOS) described a study of the atomic absorption spectrometry of manganese in four solvents; acetone, methyl isobutyl ketone, ethanol, and water. Sunderman (5ZN) determined nickel in serum spectrophotometrically by chloroform extraction of nickel from serum digest and colorimetric measurement as the bisdiethyldithiocarbamate. Markham (43N) determined silver in the concentration range of lO-6-lO-4M by using crystal violet to extract the silver cyanide complex into benzene. El-Ghamry and Frei (19,V) proposed two simple rapid sensitive, selective methods for the photometric determination of trace amounts of silver based on the formation of a ternary complex between silver ions, l,10-phenantholine and 2,4,5,7-tetrabromofluorescein. Methods for the determination of zinc in serum by atomic absorption spectrophotometry were described by Hackley, Smith, and Halsted (26N) and Haas, Lehnert, and VOL. 41, NO. 5, APRIL 1969
9
23 R
Schaller ( 2 d N ) . Mahanand and Houck ( 4 S N ) determined zinc in biological fluids fluorometrically with 8-quinolinol. Klaus ( S 9 N ) determined zinc and cadmium in biological material by resonance fluorescence. NITROGEN COMPOUNDS
Walther ( 2 9 P ) reviewed methods of ammonia detection in microbiology. Rubin and Knott (WbP) described a n enzymatic fluorometric method for ammonia determination in which ammonia and alpha-ketoglutaric acid react in the presence of glutamic dehydrogenase to form glutamic acid and NADH which is oxidized to S A D + in the process. Leffler (18P) described a new method for plasma ammonia in which di-K(ethylenedinitri1o) tetraacetic acid (di-K EDTA) was added to the blood as an anticoagulant. Seely, Petitclerc, and Benoiton (Z7P)reported the interference of protein and amines in the determination of ammonia by the isocyanurate method. Horn and Squire (12P) investigated the catalytic action of both sodium nitroprusside and acetone on the indophenol blue reaction for the estimation of ammonia X. Reatherburn (SOP) determined the optimum concentrations of reagents and sequence of addition to use in the Berthelot method for serum ammonia determination. Gangolli and Kicholson (1OP) prepared blood filtrates for ammonia analysis by precipitating blood with a mercuric chloride-lead acetate solution buffered to p H 4.1 which stops new ammonia formation. Lorentz and Ossenberg ( 1 9 P ) investigated different methods of isolation of ammonia from blood. Jenkins, Cheek. and Linnenbom ( 1 4 P ) described a new gas chromatographic ammonia met hod based on the measurement of S 2liberated by the oxidation of ammonia by alkaline hypobromite. The estimation of plasma ammonia was improved by ion exchange by Fenton and T$7illiams ( 8 P ) . Acland and Strong (1P) compared microdiffusion and ion-exchange methods for the determination of plasma ammonia and found more consistent results obtained by ion exchange. Gips and n'ibbens-Alberts ( I I P ) claimed t h a t heparinized blood could be stored a t 20 "C for 2 hours without appreciable rise in ammonia and 14 days at -20 "C which is contrary to most stability reports. Lowe (WOP) reported that ammonia is stabilized in blood frozen a t -20 "C. Enzymic methods for the determination of creatine in urine and serum and spinal fluid were described by Marymont, Smith, and Klotsch (ZIP) and Berlet (ZP). Parker (2SP) des c r i be d s p e et r o pho t o m et r i c methods for the determination of carnosine and anserine in muscle. Dambacher, Gubler, and Haas ( 7 P ) utilized 24 R
ANALYTICAL CHEMISTRY
a n improved ashing procedure and the Berthelot reaction to determine nitrogen in biological material in less than 4 hours. Bohley ( S P ) carried out ultramicro determination of nitrogen by Kjeldahl digestion and phenol-hypochlorite reaction. The hypobromitephenosafranine KPN method of Rappaport was modified by Kameoka and Shibata (15P). Franzen (9P) determined histidine in urine by paper chromatography. Klinenberg et al. (17P) and Chalmers and Watts (5P)described an enzymatic spectrophotometric method for the determination of xanthine and hypoxanthine in urine and blood. Simmonds and Wilson (28P) determined xanthine and hypoxanthine specifically in biological material by use of uricase and xanthine oxidase. Searcy, Reardon, and Foreman (26P) described a new color reaction for serum urea K determination in which salicylate replaces phenol and dichloroisocyanurate, hypochlorite in the Berthelot reaction. Kaftalin, Rhitaker, and Stephens ( 2 2 P ) estimated urea N by use of sodium hypobromite and thymol in a reaction similar to that of Berthelot direct methods. Direct serum urea N methods employing diacetyl monoxine and semicarbazide and diacetylmonoximephenazone were described by Crocker ( 6 P ) and Bohuon, Delarue, and Comoy (4P). Hughes (1SP) determined urinary urea by use of a cation responsive electrode. Khramov and Koltavskova (16P) determined blood urea with p-dimethylaminobenzaldehyde. Roch-Ramel ( 2 4 P ) described an ultramicro enzymic fluorimetric method for urea determination. HORMONES
Bravo and Travis (4Q) described a relatively simple and rapid method for urinary aldosterone utilizing paper chromatography, liquid-liquid partition, and gas chromatography. Bacchus ( d Q ) presented a simple method for urinary total 17-hydroxy-21-deoxycorticosteroids in which these compounds are selectively extracted with carbon tetrachloride from beta-glucuronidase hydrolyzed urine. Jansen, Hvidberg, and Schou (15Q) presented a modified spectrophotofluorometric micromethod by which quantitation of cortisol in 30-70 mg of tissue or 100 pg of plasma is carried out. Nugent and Mayes (19Q) used corticosteroid-binding globulin and dextran coated charcoal to determine plasma corticosteroids. Green, Hallaways, and Blacker ( 1 1 9 ) estimated tetrahydrocortisol and tetrahydrocortisone in urine by onestage paper chromatography and quantitation with blue tetrazolium reaction on paper. Greer, Sprinkle, and Williams (12Q) developed a gas chromatographic meth-
od for the determination of urinary 3-methoxytyramine, metanephrine, and normetanephrine. A new modification of the trihydroxyindole method for the fluorometric estimation of mixtures of epinephrine and norepinephrine was described by Weil-Malherbe and Bigelow ( 2 9 Q ) . Viktora, Baukal, and Wolff (27Q) described two new improved automated fluorometric methods for the estimation of small amounts of adrenaline and noradrenaline in plasma, urine, and tissue extracts. Kahane et a2. (16Q) improved hydrolytic and analytical techniques for the estimation of conjugated epinephrine and norepinephrine in urine. Van Baelen, Hayns, and De Moor (26Q) reported a sensitive specific fluorometric method for the determination of urinary estrogens which utilized gel filtration as a purification step. Schindler, Ratanasopa, and Herrmann (24Q) carried out urinary estriol determination by acid hydrolysis and gas-liquid chromatography. Gas chromatographic urinary pregnanediol methods were described by Wallach and Touchstone (2SQ) and Guarnieri and Barry ( I S Q ) . Colorimetric pregnancy tests were patented by Fossel (7Q) and Eberhard, Friedl, and Stephens ( 6 Q ) . Methods for the spectrofluorimetric estimation of serotonin in urine and brain were described by Dreux and Halter ( 6 Q )and \ h e (SOQ). Ahuja, Kaplan, and Van Dreal ( 1 Q ) described an anion exchange resin column for removal of serum inorganic iodide prior to the determination of protein bound iodine (PBI). Perko and Simonovic (209) described a modified isotope dilution method for the determination of plasma inorganic iodine. Frischauf and Altmann ( S Q ) showed that the determination of iodine in serum proteins by activation analysis was simple and rapid. Yee, Katz, and Jenest ( S I & ) and Isola (14Q) described an improved semimicro method for the determination of PBI by alkaline incineration. Pileggi and Kessler (21Q) and Kessler and Pileggi (17'Q) reported that pretreatment of acetic acid column eluates of serum obtained in the determination of serum thyroxine by column chromatography with Br, or C1, permits the direct assay of thyroxine by the ceric-arsenite system without incineration or wet digestion. This method of determining thyroxine iodine in blood serum was patented by Masen (18Q). Rosenbaum et al. ( S S Q ) reported that measurement of free thyroxine concentration is indicated when PBI and T 3 measurements may be misleading when there is an increase or decrease of thyroxine binding globulin. Gimlette (IO@ compared three simple methods for the assessment of "free" thyroid hormone. Roitt and Torrigiani (2WQ) described a radioimmunoassay for the detection of thyroglobulin in
human serum. Gimlette (9Q) described a method for assessing thyroid status using Sephadex column chromatography of serum samples with added 1251 triiodothyronine. dsimple specific method for the determination of iodoamino acids and hormonal iodine in serum by means of cation exchange resin and chloric acid digestion was presented by Backer, Postmes, and Kiener (SQ). Tang and Tomlinson (25Q) determined serum P B I by activation analysis. ORGANIC ACIDS AND COMPOUNDS
Free and Free (8R)reviewed the measurement of organic acids in serum. Legault-Demare and Bisagni (188) micro measured alpha-ketocarbosylic acids as their thiosemicarbnzides a t 290 mp. Stoner and Evans (SUR) determined alpha-ketoglutaric acid enzymatically. Micromethods for the enzymic determination of acetoacetic acid and Dbeta-hydroxybutyrate were described by Stein and Baessler (89R)and Gibbard and Katkins (9R).Routh rt al. (24R) described a LX7 spectral absorption method for the, determination of acetyl salicylic acid in blood. Chisoim (4R) reported a method for determination of n-amino levulinic acid in plasma ivhich n a s adapted from the ion-exchange resin chromatographic urinary procedure of Manzmall and Granick. Roovers, Evrard, and Tanderhaeghe (2SR)developed a new enzymatic method for human blood bile acids. Jones ( I S R ) described a rnicromethod for the detection of 0.02 ~g of citrate in biological fluids by a modification of the method of Fujin-ara. Camp and Farmer (SR)described a swum citric acid method based on the formation of pentabromoacetone and color dcwlopment n ith thiourea. Hla-Pe. and l u n g Than-Batu (12R) determined formiminoglutamic acid in urine by using a n aluminum oxidc adsorption column and the alkaline ferricyanide nitroprusside color reaction. Gompertz (IOR) measured urinary methylmalonic acid by a combination of thin layer and gas chromatography and Dreyfus and Dube (6R) by thin layer chromatography. Strassman, Ceci, and Tucci ( S I R ) described a fluorometric assay of malic acid. Leonard (19R) described a chromatographic method for indole acids in urine and spinal fluid. Anton and Sayre ( I R ) improved the specificity of the nitrosonapthol procedure for urinary 5-hydrouyindoleacetic acid. -1method for the estimation of S-hydroxy-anthranilic acid based upon the degradation of the compound by 3-hydroxyanthranilic acid oxidase was described by Schievelbein and Buchfink (25R). Enzymatic methods for the determination of lactate and pyruvate were described by Thompson and Richardson (32R),Hadjivassiliou and Rieder ( I I R )
and Fornari, Marchesi, and Sorbini (7R). Marbach and Weil ( 2 I R ) described the use and significance of metaphosphoric acid as a common precipitant i n enzymatic measurement of blood lactate and pyruvate. Kehl(I6R) determined urinary hippuric acid by converting to N-bromobenzamide and measuring a t 300 mp. Sinha and Gabrieli (27R) described a simple and sensitive gel filtration-spectrophotometric technique for simultaneous determination of benzoic and hippuric acids in urine. Bjornesjo and Jarnulf (2R) described a method for the deterinination of isonicotinic acid hydrazide in blood serum by use of sodium nitritopentacyanoferroate reagent. Lee, Lee, and Hivang (IYR) used 5-nitrosalicylic acid to determine a number of organic acids such as malic, tartaric, malomic, citric, oxalic, tartronic, and ascorbic. Stajner, Suva, and Musil (28R) described a spectrophotometric method for serum orotic acid. Kawasaki. Ito, and Tanino ( I 5 R ) described a new colorimetric method for mtimation of retinoic acid .rvith 747, sulfuric acid. Wachsmuth and Denissen ( S S R ) found 2,6-dichloroquinone chlorimide to be a highly sensitive reagent for quantitative determination of uanthurenic acid in urine. Simmonds (26R) determined urinary uric acid by using anion exchange resin and measuring total uric acid before and after incubation with uricase. Enzymatic methods for uric acid based on quantitative development of H 2 0 2by uricase followed by oxidation of a chromogenic acceptor by peroxidast m r e described by Lorentz and Berndt (2UR) and Domagk and Schlicke (5R). Kells (SbR) improved the determination of uric acid in blood and urine by reduction of phosphotungstate in the presence of sodium glycinate. Pate1 (d2R) described a uric acid method in which uric acid reduces phosphotungstic acid in a n alkaline medium of E D T B tungstate and hydrazine sulfate. Kanter ( I @ ) studied the optimum conditions for the reaction between uric and phosphotungstic acids in the presence of g1y c e r 01-si 1ic a t e re agent . K a ch t e r (34R) patented a uric acid method based on a n enzyme reaction and a peroxide detecting system. PROTEINS
Soejima and Sugawara (37s)reviewed the quantitative methods for the determination of proteins. Watson (38s) stated t h a t the factor for calculating serum albumin and total protein from nitrogen content should be 6.4 instead of 6.25. Peters ( 3 2 s ) reported that the Standards Committee of the A h C C presented proposals for the standardization of total protein assays, suggesting bovine serum albumin produced to
rigid specifications and stabilized at 7%. A new dye binding procedure for the determination of albumin in serum using bromcresol purple mas described by Louderback, Mealey, and Taylor ( 2 7 s ) . Keyser ( 1 9 s ) found good agreement between electrophoretic and immunoprecipitation methods for serum albumin but the methyl orange dye binding method gave slightly higher results. A rapid method for quantitative isolation of human serum albumin by means of trichloroacetic acid and ethanol was described by Iwata, Iwata, and Holland (17s). Martinek ( 2 9 s ) investigated the interchangeability of refractometric and biuret methods for the determination of serum total proteins. Burgi, Richterich, and Briner (6s) increased the sensitivity of the biuret method by UV measurement at 334 mp. Kingsley (21s) reported that bases such as sodium carbonate, sodium orthophosphate, sodium metasilicate, and lithium hydroxide could be substituted for sodium hydroxide. Kingsley (2US) obtained a patent on a stable dry biuret reagent. Cronkleton, Appelman, and Batsakis (IUS)investigated the effect of bilirubin on three methods of serum albumin determination. Diamant, Von Redlich, and Glick (11s) found no significant difference in sensitivity or reproducibility of the phenol reagent and broinsulfalein binding methods for the determination of protein. llrskos, Tovarek, and Kahovcc (318) patented dye reagents for the determination of proteins in biological fluids. colorimetric micromethod for the determination of total proteins was described by Kaltwasser, Kolters, and Pieper ( I S S ) . Goodwin (1%”) modified the sulfite precipitation technique for fibrinogen to assay microvolumes of plasma and eliminate interference of bilirubin. Goodn-in (148) evaluated turbidimetric techniques for estimation of plasma fibrinogen. Pryce (34s) simplified the microestimation of fibrinogen and seromucoid in plasma. Lezy (25s) reviewed the electrophoresis of serum proteins. Leaback and Rutter (24s)described a new technique for electrophoresis of proteins in which a 3% polyacrylamide gel is used. Busse (?S)made a paper chromatographic study of seven dyes used in coloring electrophoresis strips. Pristoupil and Fricova (33s)studied microelectrophoresis of serum proteins on nitrocellulose membranes. Kremers, Briere, and Batsakis ( 2 2 s ) studied the precision of reflectance densitometry of cellulose acetate protein electrophoresis. Igou (168) evaluated a gel filtration-spectrophotometric method for spinal fluid protein determination. Methods for the acrylamide gel electrophoresis of serum proteins were described by M a n and Whitehead ( 2 8 S ) , Groulade and Pichot (15S), and Gofman (12s). MiVOL. 41, NO. 5, APRIL 1969 * 25 R
lesterol is reacted with a reagent comgita (SOS) fractionated serum proteins posed of ethyl acetate, HISO,, and ferby thin layer chromatography on Seric perchlorate. Eskelson, Dunn, and phadex G-200 superfine with p H 7.0 Cazee ( 8 T ) reported that tomatine inphosphate-NaC1 buffer. Blatt et al. terferes in the colorimetric determina(58)evaluated five membranes for fraction of cholesterol by the Zak procetionation of protein solutions by memdure. Methods to eliminate the effect brane partition chromatography. Memof bilirubin on the determination of sebrane filtration for determination of rum cholesterol were described by proteins was described by Kuno and Ichida, Osaka and Nishikage (122') and Kihara (238)and Bennett (88).Searcy Jordan (13T). Simple rapid methods (36s) developed a technique for assessfor the determination of total estrogens ing the cholesterol-bearing lipoproin pregnancy urine were described by teins separated electrophoretically in Brombacher, Gijzen, and Verheesen serum. (4T), and Nordstrom (182'). BrombaCohenandDjordjevich (98)improved cher, Gijzen, and Verheesen ( S T ) delipoprotein staining and starch gel septermined pregnandiol in urine by aluaration methods to reveal different semina-column chromatography followed rum alpha-lipoprotein patterns. Chin by H2S04reaction and gas chromatogand Blankenhorn ( 8 s ) described a meraphy. Roberts, Olson, and Herrmann thod for lipoprotein electrophoresis on (292') described a new relatively simcellulose acetate which gives complete ple method for simultaneous determinaresolution of chylomicrons, beta-lipotion of urinary testosterone and epiproteins, pre-beta-lipoporteins and altestosterone utilizing periodic oxidapha-lipoproteins. ai new method for tion, chromatographic purification, and the electrophoretic separation of sethin layer chromatography. Whigham rum lipoproteins which permits a sta(232') described a simple method for ble resolution of lipoproteins into 4 bands 11as described by Bellone (2s). fractionating 17-ketosteroids by solvent partition. Bacchus ( I T ) described a Lou and Shanbrom (268) reviewed the procedure which permits the measurelaboratory techniques involving immument of Corti-costeroid metabolites in noelectrophoresis. Ritchie (858)dethree solvent extracts of hydrolyzed scribed a simple, direct, sensitive techurine. l l a s a d a et al. (162') developed nique for measurement of specific proa better separation method for three tein in dilute solution. Agostoni, Verisomers of urinary 11-deoxy-17-ketogani, and Lomanto ( I S ) described a steroids by gas chromatography. Soloneiv method which combines in a single mon, Strummer, and S a i r ( 2 I T ) and sensitive preparation; fractionation of Cawley, Musser, and Tretbar (52') deserum proteins by thin layer gel filtrascribed analytical procedures for the tion and immunological characterizaanalysis of ll-deoxy-17-ketosteroids, tion. Bernier (4s) estimated serum pregnanediol and pregnanetriol by gashaptoglobin levels by a method using liquid chromatography. Bjerre and high-voltage electrophoresis in agar Kita (25") replaced the unstable ethagel and benzidine staining. nolic potassium hydroxide in the Zimmerman reaction with the stable orSTEROLS ganic base, hydroxide of Hyamine 10X. Georges and Politzer (92') comMethods for the estimation of cholespared four different procedures for terol in serum were reviewed by Tonks measuring the Zimmermann color in (%?I"). A new technique for detection the estimation of total neutral 17-ketoof cholesterol based on the property of steroids. Kimura, Kishina, and SakaLE-3 hydroxysteroids yielding after acid mot0 (142') developed a new microoxidation products which are deeply method forthe determination of ketostestained by aldehydefuchsin, toluidine roids using p-nitrophenylhydrazine reblue or .Ucian blue, was proposed by agent. Russell, Raymer, and Lobstein, Hadler et al. (112'). Zlatkis and Zak (202'),described a method for concur(252') reported that 1,2 benzene dicarrent determination of urinary 17-ketobonal or naphthalene-2,3, dialdehyde genic and 17-ketosteroids. A study of reacts rapidly with cholesterol to yield acid hydrolysis of free and conjugated a colored substance which permits exurinary 17-ketosteroids by Goldzieher tremely sensitive measurement of choand de la Pena (102') indicated that a lesterol. Clark, Rubin, and Arthur compromise time of 30 minutes of hy(62') described a new colorimetric sedrolysis appeared to be optimal. Luporum cholesterol reagent composed of vitch ( 1 5 T ) studied the interference of FeC13,ethanol, KOH,andH2SOa.Eavenphenothiazine on the Porter-Silber reson et al. (72') developed a Liebermann action. cholesterol color reagent which was stable for 8 hours. Moriguchi and Oba (172') compared the Sperry-Webb seTOXICOLOGY rum cholesterol method with 4 other Analytical methods in forensic chemmethods. Wybenga et al. (242') deistry were reviewed by Ohkuma (40U). scribed a simple, specific, direct methBryan, Guinn, and Settle ( 8 U ) discusod for total cholesterol in which cho26 R
ANALYTICAL CHEMISTRY
sed the activation analysis of biological samples of forensic interest. Samsahl (48 U ) described radiochemical methods for the determination of arsenic, bromine, mercury, antimony, and selenium in neutron irradiated biological material. Thompson and Decker (59U) developed a simple rapid and precise method for identification and quantitation of drugs present in human blood serum by gas chromatography. The significance of thin layer chromatography in chemical toxicological analysis was reviewed by Raudonat ( 4 5 U ) . Machata (35U) stated that thin layer chromatography was the simplest and most efficient method for the routine determination of organic poisons in the fieldof toxicology. Kirk ( S o t ' ) reviewed the method of pyrolysis combined with gas chromatography for the detection and identification of poisons and drugs. Weber (64 L') described the application of chemiluminescence of luminol in judicial medicine and toxicology. Gas chromatographic methods for the determination of meprobamate in biological fluid were described by Douglas et al. ( I r L ' ) ,Maddock and Bloomer ( S 6 U ) and Skinner (54c'). Gas chromatogrnphic methods for the determination of doriden were described by Sunshine, Maes, and Faracci (67c') and Winsten and Brody (66c'). Knowlton and Goldbaum ( S I C ) described a n improved method for the determination of doriden in blood. Curry ( I S C ) determined nanogram quantities of chlorpromazine in plasma by gas-liquid chromatography. Tewari (58t') described separation and identification of alkaloids by paper electrophoresis and its application in medico-legal cases. Yoshimura, Oguri, and Tsukamoto (67C) determined morphine in urine by a n improved t h i n layer chromatographic method utilizing potassium platinum iodide as reagent for both coloration and fluorescence. Oka (41 I;) described a simple method for the isolation of morphine glucuronide from urine. Three substances, nicotinamide, uracil, and thymine were found by Kaempe ( 2 9 U ) to interfere in the determination of poisons in autopsy material. Bors et al. (6C) reported that strychnine can be detected in tissues and separated by paper electrophoresis more easily than by chemical methods. Xoirfalise et al. (39 C ) proposed a n analytical procedure for the psycho-stimulating amines. Comstock, Comstock, a n d Ellison ( 1 2 U ) presented a turbidimetric method for t h e determination of pentachlorophenol in urine. Guidotti, Borghetti, and Loreti (26U) presented a simple colorimetric method for the determination of thiazolidine-4-carboxylic acids, Bruce, Pitts, and Pinchbeck (YC) described a gas-liquid chromatographic method for the determination of brompheniramine in blood and urine.
Routh et al. (47C') compared t h r e e methods for the determination N-acetylp-aminophenol in plasma; Brodie, differential absorbance, and diphenylpicrylhydrazoyl dye methods, and recommended the dye method as the most simple, sensitive, and rapid. Popa and Popa (44 C) determined dinitro-o-cresol by a method based on the violet color given by the reduction product of I with ICl. Funk (23U) reviewed the most useful photometric procedures for the determination of sulfonamides in biological material. Methods for the determination of arsenic in animal tissues and urine were described by Stone (56C.) and Simon, Christian, and Purdy (52C). Dunlop (18C) described a simple colorimetric method for the determination of bromide in urine. Eisenberg and Tillson (29C) identified barbiturates and amphetamines by microscopic chemical and instrumental techniques. Sprung (55C) and Olesen (42C') used thin layer chromatography to identify barbiturates. Bogan and Smith (5C) reviewed analytical methods for investigation of barbiturate poisoning. Agazzi (1 C) extracted boron into methyl isobutyl ketone for emission spectrophotometry. De Leeuw et al. (16C) reviewed the methods for carbon monoxide determination in blood. Wieczorek (65V) determined small amounts of CO in blood by reaction with 120ja t 100 "C to liberate I2 which is spertrophotometrically measured with starch. Nakaaki (38r)compared the accuracy of the detector tube, gas chromatograph, and the spectrophotometric methods and found the latter preferable for carboxyhemoglobin in blood. A method for the determination of carboxyhemoglobin in blood in which CO is liberated by H2SOa a n d pumped i n t o indicator tubes containing PdSO, and the color change recorded, was described by Gassman and Wranne (Z4U). Ciuhandu and Rusu (9C) determined CO in air and blood by reaction with .Ig+p-sulfamoylbenzoate in alkaline solution. Collison, Rodkey, and O'Neal (11 L-) determined CO content of blood by gas chromatography. A specific spectrophotometric method for the determination of small amounts of CNin biological material was described by Wawschinek, Paletta, and Beyer (63U). A colorimetric micro determination of cyanide in food m s made by Kondo, Kawashiro, and Furuhashi (32C') using p-nitrobenzaldehyde and o-dinitrobenzene. Methods for the gas chromatographic determination of blood ethanol were described by Savory et al. ( 4 9 C ) Bassette and Glendening (SG), Curry, Walker, and Simpson ( 1 4 C ) , Le Blanc (34U) and Roach and Creaven (46U). Colehour (10 C ) investigated the use of water as an internal standard in the determination of blood ethanol by gas
chromatography. Jones, Gerber, and Drell (28U) determined ethanol in body fluids by a method based on alcohol dehydrogenase catalyzing the oxidation of ethanol to acetaldehyde with the concomitant reduction of NAD which results in an increase of light absorbance a t 340 mp. Vidic ( 6 1 U ) described a rapid blood alcohol method based on the reduction of vanadic acid to form a blue colored product for spectrophotometric measurement. Singer and Armstrong (53 C ) developed electrodes made from single crystal sections of rare earth fluoride to measure fluoride ion activity in solutions. McCann (37 C) determined fluorideinmineralized tissues with a fluoride ion electrode. Sereda (50U) separated fluorine in body fluids by column anion exchange resin. Auentin ( 2 U ) determined fluoride colorimetrically by the formation of a red chelate by alizarin complexon and fluorides. Davis and Andelman ( 1 5 U ) designed ion-exchange chromatographic columns for the rapid isolation of delta-aminolevulinic acid for the early detection of lead poisoning. Truff ert and GirardWallon ( 6 0 U ) described a dry mineralization method for detecting lead in urine by polarography. Kopito and Shwachman (33U) determined lead in urine by coprecipitation with bismuth nitrate prior to atomic absorption spectroscopy. Hessel (27 U) presented a simplified procedure for blood lead determination by hemolyzing the blood with Triton X-100 solution, complexing with ammonium pyrrolidine dithiocarbamate and extracting the lead complex with methyl isobutyl ketone for atomic absorption. Girard et al. (25C) applied polarography by anodic redissolution as a new procedure for determination of lead in urine. Berman, Valavanis, and Dubin (4 C) determined lead in a 260 p1 blood sample by extraction with 1% sodium diethyldithiocarbamate in methyl isobutyl ketone and atomic absorption a t 2830 resonance line. Fischer (21 C ) described a method for the rapid detection of acute iron toxicity. Fischl, Pinto, and Gordon (22U) detected organic phosphorus poisons with a filter paper strip impregnated rrith a buffered acetylcholine substrate solution containing phenol red as an indicator and 1,2-propylene glycol as a wetting agent. Serra (51 C ) assayed small amounts of mercury in urine by a method based on the use of dithizone. Watkinson (62U) reviewed analytical methods for selenium in biological material and reported that micromethods of choice were fluorescence and radioactivation. Ewan, Baumann, and Pope (20C) described a fluorometric micromethod for selenium in biologicaI materials based on wet digestion, coprecipitation of Se with As and measurement of fluorescence with 2,3-
diaminonaphthalene. Petrow and Strehlow (@U) determined thorine in bone ash using arsenazo 111. Zsoldos et al. (68U)studied the optimum conditions of wet acid digestion for determination of natural uranium in urine. VITAMINS
Drujan, Castillon and Guerrero ( 2 V ) described a micromethod for the determination of blood and tissue Vitamin A based on the application of fluorometric and t h i n layer chromatographic techniques. Other methods for the fluorimetric determination of Vitamin A in small samples of blood and urine were described by Kahan ( 6 V ) and Hansen and Warmick (5V). McLaren et al. ( 7 V ) revielved the extraction and separation procedures and critically evaluated and recommended micromethods for the determination of Vitamin A and carotenoids in blood, body tissue, and milk. Ostlund and Bjorlin ( 9 V ) described a semimicro method for the determination of Vitamin h and carotene in blood serum. Sherman (11V) describcd the use of ultraviolet irradiation in the estimation of retinol (Vitamin A alcohol) and its derivatives. Contractor and Shane (1 V ) described a sensitive and specific method for the estimation of Bg compounds in blood and urine in which the fluorescence of their cyanohydrin and lactone derivatives are utilizcd. Yamane, l l o t o miya, and Yamada (221') determined thiamine by a sensitive method based on the red-fluorescent product formed by the reaction of thiolized thiamine with Cu2+ which was carried out as a spot test on filter paper. Frodynia and Lieu ( 4 V ) developed a proccdurr in which five B-group vitamins could be resolved on thin layer plates and analyzed by means of ultraviolet reflectance spectrometry. llichaelsson and Michaelsson (8V) described a new blood plasma ascorbic acid method based on a color reaction between the ZnC12 double salt of diazotized 4-nitroaniline2,5 dimethoxyaniline and ascorbic acid. Pelletier (10V) determined ascorbic acid and dehydroascorbic acid in biological materials by an accurate and rapid method based on the 2,4-dinitrophenyl-hydrazine reaction Ivithout interference from diketoglulonic acid. Fabianek et al. (3V) improved a microprocedure for blood serum tocopherol based on the FeCL oxidation of tocopherol and the spectrophotometric determination of the bathophenanthroline complex. LITERATURE CITED
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(6J) Fuehr, J., Schacht, U., Aerztl. Lab., 13, 298 (1967). Chem. Abstr., 68, 736 (19681.
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(1968) ~ - -
(32K) Holeysouska, H., Clin. Chim. Acta, 15, 353 (1967). (33K) Inglis, N. R., Ghosh, S. K., Fishmdn. W. H.. Anal. Biochem.., 22,. 382 (1068). (34K) Johnson, R. B., Jr., Clin. Chem., 14, 801 (1968). (35K) Johnson, D. A,, Osaki, S., Frieden, E., ibid., 13, 142 (1967). (36K) J u u ~P., , ibid., p 416. (37K) Juul. P.. Clin. Chim. Acta, 19. 205 (1968). (38K) Kelley, S., Belorit, A., Copeland, W.,ibid., 18, 483 (1967). (39K) Knight, J. A., Hunter, D. T., Clin. Chem., 14, 555 (1968). (40K) Korittinen. A,. Clin. Chim. ilcta, 21. 29 (1968). (41K) Krainev, S. I., Lab. Delo, 8, 471 (1967). Chem. Abstr., 67, 97562 (1967). (42K) Krieg, A. F., Rosenblum, L. J., Henry, J. B., Clin. Chem., 13,196 (1967). (43K) Latner, A. L., Adv. Clin. Chem., 9, 69 (1967). (44IC) Lubran, M.,Jensen, 1%'. E., Clin. Chim. Acta, 22, 125 (1968). (45K) Marymont, J. H., Jr., Cawley, L. P., Hoffman, R. G., Am. J . Clin. Pathol., 49, 431 (1968). (46K) Illassion, C. G., Seligson, D., ibid., 48, 307 (1967). 147K) lIcComb, R. B., Gay, R. J., Clin. Chem.. 14. 754 11968). (48K) $lcC&mmon, A:, Lewin, E., Am. J . Med. TechnoZ., 33, 269 (1967). (49K) llellerup, B., Clin. Chem., 13, 900 11967). (50K) hlenken, ll., Barrett, P. V. D., Berlin, N. I., Clin. Chim. Acta, 14, 777 (1966). (51K) Moore, F. A l . L., ibid., 15, 103 (1967). (52K) Myers, R. C., Van Remortel, H., Clin. Chem., 14, 1131 (1968). (5310 Neumann, H., Van Vreedendaal, 11..Clin. Chim. Acta. 17. 183 11967). (54Kj Ng., W. G., Bergren', W.K., Donnell, G. N.,ibid., 15, 489 (1967). (55K) Nyssen, bl., Dorche, J., ibid., 22, 363 (1968). (56K) Paglia, D. E., Valentine, W. N., J . Lab. Clin. Med., 70, 158 (1967). ,
Enzymes
(1K) Amador, E., Massod, M. F., Franey, R. J., A m . J . Clin. Pathol., 47, 419 (1967). (2K) Amador, E., Price, J. W., Clin Chem., 14, 185 (1968). (3K) Arawaka, K., Minohara, A., Yamada, J., Uemura, N., Nakamura, V., Clin. Chim. Acta, 22, 309 (1968). (4K) Babson, A. L., Kleinman, N. AI., Meeraw, R. E., Clin. Chem., 14. 802 (1988). ' (5K) Berk, J. E., J . A m . Med. Assoc., 199, 98 11967). (Sic) Bigazzi, P. L., Ciampi, G. P., Clin. Lab. (Italy), 2, 507 (1966). Chem. Abstr., 68, 783 (1968). 17K) Bowers. G. N., Jr., Kellev, M.L., McComb, R . B., CUn: Chem:,' 13, 595 11967) . (8K) Bowers, G. N., Jr., Kelley, hl. L., McComb, R. B., Ibid., p, 608. (9K) Coleman, C. hl., Clzn. Chim. Acta, 13, 401 (1966). (10K) Dietz, A. A., Hodges, L. K., Rubinstein. H. M.. Brinev. R. R.. Clin. Chem., 13, 242 (1967). (11K) Dirstine, P. H., Sobel, C., Henry, R. J., Ibid., 14, 1097 (1968). (12K) Emery, A. E. H., Moores, G. E., Hodson, V., Clin. Chim. Acta, 19, 159 (1968). (13K) Epstein, E., Wolf, P. L., Horwitz, J. P., Zak, B., A m . J . Clin. Pathol., 48, 530 (1967). (14K) Fischl, J., Segal, S., Rabiah, S., Clin. Chem., 13, 941 (1967). 115K) Fisher. C. L.. Nixon. J. C.. ibid.. 14,'820 (1968). ' I
\ - - -
I
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Lipids
,
(16K) Fishman, W. H., Methods of Biochem. Anal., XV, 77 (1967). (17K) Fletcher, D. C., Verdi, J. L., Clin. Chem., 13, 695 (1967). (18K) Gay, R. J., McComb, R. B., Bowers, G. N., Jr., ibid., 14, 740 (1968 (19K) Gerarde, H. W., Fr. 1,462,521 G Oln), (Dec. 16, 1966,) (U.S. Jan. 28,1965:, 8- rDn.1 r (20K) Glen, A . C. A., Clin. Chem., 13,299 110fi7). j - _ _ . ,. (21K) Graig, F. A., Smith, J. C., Foldes, F. F., Clin. Chim., Acta, 15, 103 (1967). (22K) Guilbault, G. G., Fluorescence, 297 (1967). Chem. Abstr., 68, 36531 (1968). (23K) Guilbault, G. G., ANAL.CHEM.,40, No. 5, 459R (1968). (24K) Guilbault, G. E., Brignaca, P. J., Jr., Juneau, hl., ibid., p 1256. (25K) Guilbault, G. G., Brignac, P. J., Jr., Zimmer, M., ibid., p 190. (26K) Haschen, R. J., Farr, W., Reichelt, D., Z. Klin. Chem. Klin. Biochem., 6, 11 (1968). Chem. Abstr., 68,66286 (1968). (27K) Healy, P. J., Clin. Chim. Acta, 20, 165 (1968). (28K) Hemmingsen, L., Skov, F., ibid., 19, 81 (1968). 129K) Henlev. K. S.. Schmidt. E.. Schmidt, F. W:, A m . J. Clin. Pathol., 48, 364 (1967). (30K) Hess, J. W.,MacDonald, R. P., Natho, G. J. W.,Murdock, K. J., Clin. Chem., 13, 994 (1967). 131K) Hess, J. W., Murdock, K. J., Natho, G. J. IT.,A m . J. Clin. Pathol.. 50. 89
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VOL. 41, NO. 5, APRIL 1969
31 R
(57K) Papadopoulos, N. M., Kintzios, J. A., Am. J. Clin. Pathol., 47, 96 (1967). (58K) Pon. N. G., Bondar, R. J. L.. -4nal. Biochem.; 19, 272 (1967): (59K) Pribor, H. C., Kirkham, W. R., Fellows, G. E., Am. J . Clin. Pathol., 50, 67 (1968'1. (60K) -Rinderknecht, H., Geokas, M. C., Silverman, P., Haverbach, B. J., Clin. Chim. Acta, 21, 197 (1968). (61K) Rinderknecht, H., Geokas, L1. C., Silverman, P., Lillard, Y., Haverback, B. J., ibid., 19, 327 (1968). (62K) Romel, W. C., LaMancusa, S. J., DuFrene, J. K., Clin. Chem., 14, 47 (1968). (63K) Rosalki, S. B., J . Lab. Clin. Med., 69, 696 (1967). (64K) Rutenburg, A., >I., Smith, E. E., Fischbein, J. W.,zbad., p 504. (65K) Rybak, M.,Mansfeld, V., Petakova, hl., Simonianov, A., Clin. Chim. Acta, 19, 405 (1968). (66K) Satoh, P. S., Ito, Y., Anal. Biochem., 23, 219 (1968). (67K) Sas, S. M., Moore, J. J., Clin. Chem., 13, 175 (1967). (68K) Sax, S. >I., Moore, J . J., ibid., 14, 660 (1968). (69K) Searcy, R. L., Wilding, P., Berk, J. E., Clin. Chim. Acta, 15, 189 (1967). (70K) Sherwin, A. L., Siber, G. R., Elhilali. 11. hI.. ibid.. 17. 245 11967). (71Kj Smith; I., Lightstone; P. J., Perry, J. D., ibid., 19, 499 (1968). (72K) Strumeyer, D. H., Anal. Biochem., 19, 61 (1967). (73K) Swaim, R.R., Clin. Chem., 14,262 (1968). (74K) Szasz, G., Am. J . Clin. Pathol., 47, 607 (1967). (75K) Szasz, G., Clin. Chem., 13, 752 (1967). (76K) Trainer, T. D., Gruenig, D., Clin. Chim. Acta, 21, 151 (1968). (77K) Ward, R. L., Srere, P. A., Anal. Biochem., 18, 102 (1967). (78K) Welshman, S . G., Rixon, E. C., Clin. Cham. Acta, 19, 121 (1968). (79K) Wilkinson, J. H., Steciw, B., Clin. Chem., 14, 797 (1968).
(16L) Gindler, E. M., Ishizaki, R. I., Clin. Chem., 13,711 (1967). (17L) Grafnetter, D., Janosova, Z., Cervinkova. I.. Clin. Chim. Acta, 17. 493 (1967). ' ' (18L) Hingerty, D., Irish J. Med. Sn'., 491, 525 (1966). Chem. Abstr., 66, 3352 I
.
(19671.
(lgLj-Husdan, H., Rapoport, A,, Clin. Chem., 14, 222 (1968). (20L) Ichida, T., Nobuoka, M., Clin. Chim. Acta, 19, 249 (1968). (21L) Krees, S. V., Baukal, A. J., Wolff, F. W.,Am. J . Clin. Pathol., 48, 95 (1 \ -967). --.,-
(22L) Krylov, A. A., Lab. Delo, 9, 565 (1967). Chem. Abstr., 68, 46899 (1968). (23L) Kulhanek, V., Atmelt, J., Clin. Chim. Acta, 20, 29 (1968). (24L) London, >I., Freiberger, I. A,, Marymont, J. H., Jr., Clin. Chem., 13, 970 (1967). (25L) Marymont, J. H., Jr., Smith, J. N., Klotsch, S., Am. J . Clin. Pathol., 49, 289 (1968). (26L) Mellinger, T. J., ibid., p 200. (27L) Mikulecky, M,,Clin. Chim. Acta, 21, 43 (1968). (28L) O'Gorman, L. P., ibid., 19, 485 (1968). (29L) Ott, H., Pirrwitz, D., ibid., 22, 439 (1968). (30L) Pokrovskaya, E. I., Semenova, T. D., Tereshchenko, A. N.,Lab. Delo, 3, 154 (1968). Chem. Abstr., 69,623 (1968). (31L) Preston, J. A., Clin. Chem., 13, 19 (1967). (32L) Rink, hl., Krebber, D., J. Chromatogr., 25, 80 (1966). (33L) Rockerbie, R. A., Rasmussen, K. L., Clin. Chim. Acta, 15, 475 (1967). (34L) Roovers. J.. Evrard. E.. Vander' haeghe, H., &id.; 19, 449'(1968). (35L) Roth, V., ibid., 17, 487 (1967). (36L) Scott, C. R., Labbe, R. F., Nutter, J., Clin. Chem., 13, 493 (1967). (37L) Spandrio, L., Albertini, A., Biochim. A d . . 14. 68 (19671. Chem. Abstr..
Function Tests
(1L) Becker, S., Clin. Chem., 12, 637 (1966). (2L) Beyermann, K., Gerok, W.,Groth, U., Klin. Wochenschr., 45, 950 (1967). Chem. Abstr., 68, 19392 (1968). (3L) Brodersen, R., Scand., J . Clin. Lab. Invest., 18, 361 (1966). (4L) Butler, T. J., Clin. Chem., 13, 488 (1967). (5L) Castrow, F. F., Mullins, J. F., Flowers, R. H., Mills, G. C., Arch. Dermatol., 96. 204 (1967). Chem. Abstr., 67. 88212
32
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ANALYTICAL CHEMISTRY
(1968): 14OLl Ratkins. P. J.. Clin. Chim. Acta. 18; 191 (1967). (41L) Weil-Malherbe, H., J . Lab. Clin. Med., 69, 1025 (1967). (42L) Wybenga, D., Pileggi, V. J., Clin. Chim. Acta, 16, 147 (1967). \ -
~
Hemoglobin (111) Bjerre, S., Clin. Biochem., 1, 299
(1968). (211) Chong, G. C., Owen, J. A., J. Clin. Pathol., 20, 211 (1967). (3M) Eilers, R. J., A m . J . Clin. Pathol., 47, 212 (1967). (411) Katchman, B. J., Murphy, J. P. F., Offner, K. M., Fox, J. C., Clin. Chem., 13, 877 (1967). (531) Klein, B., Kelly, B., Lucas, L. B., Foreman, J. A., Searcy, R. L., ibid., 14, 827 (1968). (611) Noeller, H. G., U.S. 3,374,063 (Cl. 23-230), (March 19, 1968,) (Ger. Appl. May 29, 1963; 3 pp.) (711.1) Pantlitschko, M., W-eippl, G., Clin. Chim. Acta, 19 439 (1968). (8x1) Rand, P. &., Lacombe, E., Barker, N., J . Lab. Clin. Med., 69, 862 (1967). (931) Rosenbaum, D. L., Am. J . Med. Sci., 252, 726 (1966). (lOM) Smith, E. W., Evatt, B. L., J.Lab. Clin. .We+, 69,1018 (1967). (11R.I) Theil, G. B., Auer, J. E., Clin. Chem., 13, 1010 (1967).
(12M) Williams, G. Z., Am. J . Clin. Pathol., 48, 333 (1967). (13M) Zett'ner, A,, ibid., p 332. (14M) Zettner, A., Mensch, A. H., ibid., p 225. Metals
(IN) Amdisen, A., Scand. J . Clin. Lab. Invest., 20, 104 (1967). (2N) Askevold, R., Vellar, 0. D., ibid., (3R)?abson, A. L., Kleinman, N. M., Clin. Chem., 13, 163 (1967). (4N) Bedrosian, A. J., Skogerboe, R. K., Morrison, G. H., ANAL.CHEM.,40, 855 (1968). (5N) Berman, E., At. Absorption Newsletter, 6 , 57 (1967). Chem. Abstr., 67, 61502 (1967). (6N) Binnerts, W. T., Tijdschr. Diergeneesk, 93, 558 (1968). (7Xj Binnerts, W. T., Boom, W. G., ibid., p 573. (8N) Blijenberg, B. G., Leijnse, B., Clin. Chim. Acta, 19, 97 (1968). (9K) Brownstein, H., Am. J . Clin. Pathol., 47, 714 (1967). (10N) Budziszewski, A,, Arch. Zmmunol. Ther. Exp., 15,936 (1967). Chem. Abstr., 68, 46872 (1968). (11X) Burrows, S., Am. J . Clin. Pathol., 47, 326 (1967). (12N) Comar, D., Desenne, J. J., Acta Cient. Venez, 18, 152 (1967). (13N) Dawson, J. B., Ellis, D. J., NewtonJohn, H., Clin. Chim. Acta, 21, 33 (1968). (14s) Deguchi, M., Hiroshima Daigaku Kogakubu Kenkyu Hokoku, 16, 287 (1968). (15N) Deguchi, &I.,ibid., p 135. (16N) Diehl, H., ANAL.CHEM.,39, No. 3, 30A (1967). (17N) Dmitrieva, T. G., Tr. Khar'kov Med. Inst., 67, 30 (1966). (Zh. Khim. Pt. 1, Abstr., No. 14G50 (1967)). (18N) Dronski, H., Galts, G., Deut. Gesundheitsw., 21, 2457 (1966). Chem. Abstr., 66 92287 (1967). (19N) El-Ghamry, M. T., Frei, R. W., AKAL.CHEM., 40, 1986 (1968). (20N) Feldman, F. J., Bosshart, R. E., Christian, G. D., ibid., 39, 1175 (1967). (21N) Feldman, F. J., Christian, G. D., Purdv. W. C.. Am. J . Clin. Pathol.. 49. 826 (1968). ' (22rlT) Gandhi, h'f. H., Desai, M. N., ANAL.CHEM.,39, 1643 (1967). (23N) Girard, &L., 'I. Clin. Chim. Acta, 20. 243 (1968). (24s) Haas, T., Lehnc?rt,, G., Schaller, K. H.. 2. Klin. Chem. Klin.'Biocher n.. 5. 218 (1967). Chem. Abstr., 68, 75647 (1968). (25N) Hackley, B. &I., Smith, J. C., Halsted, J. A., Clin. Chem., 14, 1 (1968). (26N) Hamaguchi, T., Horiuchi, K., Tanaka, N., Bunseki Kagaku, 15, 1264 (1966). Chem. Abstr., 66, 102386 (1967). (27N) Hansen, J. C., Am. J . Med. Tech., 34, 625 (1968). (28N) Heller, R. L., Jr., Guyon, J. C., ANAL.CHEM.,40,773 (1968). (29N) Heydorn, K., Arch. Pharm. Chemi, 74, 4 (1967). Chem. Abstr., 66, 73078 (1967). (30K) Hillman, R. S., Morgan, E. H., Finch, C. A., J . Lab. Clin. Med., 69,874 (1967). (31X) Hoeflmayr, J., Fried, R., Med. Klin. (Munich),61,1820 (1966). Chem. Abstr., 66, 838 (1967). (32N) Holland, W. J., Bozic, J., ANAL. CHEM.,39, 109 (1967). I.) J ., Med. (33N) Howe, (Sister M. &Am. Technol., 33, 105 (1967). (34N) Huey, F., Hargis, L. G., ANAL. CHEM.,39, 125 (1967). I
,
(35N) Ichida, T., Osaka, T., Kojima, K., Clin. Chim. Chzm. Acta, Acta. 22. 22, 271 11968). (1968). (36N) Johnson, G. V., Young, R. A., ANAL.CHEW.,40, 354 (1968). (37N) Kanabrocki, E. L., Scheving, L. E., Kaplan, E., Pauly, J. E., Chicago M e d . Xch. Quart., 27, 151 (1968). (38N) Kitaguchi, IC., Nishimoto, S., Rinsho Byori, 16, 233 (1968). (39N) Klaus, R., 2. Klin. Chem., 4, 299 (1966). Chem. Abstr., 67, 79562 (1967). (40N) Lehmann, V., Clin. Chim. Acta, 20, 523 (1968). (41K) Little, B. R., Platman, S.R., Fieve, R. R., Clin. Chem. 14, 1211 (1968). (42N) lfahanand, D., Houck, J. C., ibid., nA
(45@ Markham, J. J., ANAL.CHEM.,39, 241 (1967). (44N) hIcDonald, C. IT., Bedenbaugh, J. H., ibid., p 1476. 145N) Nielsen. I.. 2. Klin. Chem. Klin. ‘ Biochem.. 6.’10$ (1968’1. (46N) Parker: G. A,, Boltz, D, F., ASAL. CHEM.,40, 420 (1968). (47N) Parker, RI. 11, Humoller, F. I,., Mahler. D. J.. Clin. Chem., 13, 40 (1967). (48N) Schlit, A. A., Hoyle, IV. C., ANAL. CHEM.,39, 114 (1967). (49N) Schmidt, G., Deut. Gesundheitsw., 22, 1686 (1967). Chem. Abstr., 68, 46924 (1968). (50N) Stokes, R. P., Clzn. Chim. Acta, 15, 517 (1967). i5lN) Sulochnna, C. B., Curr. Sci., 36, 508 (1967). Chenz. Abstr., 68, 10107 (1968). (52N) Sunderman, F. IT., Jr. Clin. Chem., 13. -115 - - 11067’1. , (5iN) Sunderman, F. W.,Jr., Roszel, N. O., Am. J . Clin. Pathol., 48, 286 (1967). (54N) Titov, 11.B., Kh., S., Gig. Tr. Prof. Zabol, 11. 55 (1967). Chern. Abstr.., 67., 61502 (1967). (55N) Todd, A. P., Thorpe, 11. E. C., Rosenoer, V. XI.,J . Clin. Pathol., 20, 276 (1967). (56N) Van De Bogart, hl., Beinert, H., Anal. Biochem., 20, 325 (1967). (57N) Zaino, E. C., At. Absorption Nezcsletter, 6, 93 11967). (58Ni Zettner. A.. Mensch. A. H..‘ Am. J . Clin. Pathol.,‘48, 225 (1967). (59N) Zettner, A . , Rafferty, K., Jarecki, H. J., At. Absorption Newsletter, 7, 32 ( 1968). I
\ - - -
Nitrogen Compounds (1P) Acland, J. D., Strong, R., J . Clin. Pathol., 21, 12 (1966). (2P) Berlet, H. H., Clin. Chini. Acta, 20, 149 (1968). (3P) Bohley, P., Hoppe Seylers Z . Physiol. Chem.. 348. 100 (1967). (Ger.\. Chem. Abstr.: 66. 52867 ‘1196?!,.
UP)’Dambacher, M.,Gubler, A., Haas, H. G.. Clin. Chem.. 14. 61.5 (1968’1.
Chim. Ada, 14, 585 (1966).’ (11P) Gips, C. I%., Wibbens-Alberts, XI.,
zbtd., 22, 183 (1968). (12P) Horn, D. B., Squire, C. R., ibid., 17, 99 (1967). (13P) Hughes, AI. G., J. Sci. Technol., 13, 131 (1967). Chem. Abstr., 68, 102419 (1968).
(14P) Jenkins, R. R., Jr., Cheek, C. H., Linnenbom, V. J., ANAL. CHEM., 38,
1257 (1966). (15P) Kameoka, M., Shibata, S., Rinsho Byori, 14,239 (1966). Chem. Abstr., 68, 27439, (1968). (16P) Khramov, V. A., Koltavskova, L. S., Vop. Med. Khim., 13, 208 (1967). Chem. Abstr., 67, 8575 (1967). ( l i p ) Klinenberg, J. R., Goldfinger, S., Bradley, K. H., Seegmiller, J. E., Clin. Chem., 13, 834 (1967). (18P) Leffler, H. H., Am. J . Clin. Pathol., 48,233 (1967). (19P) Lorentz, K., Ossenberg, W.,Med. Lab., 20, 77 (1967). Chem. Abstr., 67, 70977 (1967): (20P) Lowe, 11. C., Clin. Chem., 14, 1074 (1968). (21P) Marymont, J. H., Jr., Smith, J. N., Klotsch, S.,Am. J . Clin. Pathol., 49, 289 (1968). (22P) Naftalin, L., Whitaker, J. F., Stephens, A., Clin. Chim. Acta, 14, 771 (1966). (23P) Paiker, C. J., Jr., ASAL. CHEM.,38, 1359 (1966). (24P) Roch-Ramel, F., Anal. Biochem., 21, 372 11967). (25P) Rubin, l f . , Knott, L., Clin. Chim. Acta, 18, 409 (1967). (26P) Searcy, R. L., Reardon, J. E., Foreman, J. A., Am. J . Med. Technol., 33, 15 (1967). (27P) Seely, J. H., Petitclerc, J. C., Benoiton, L , Clin. Chim. Acta, 18, 85 (1967). (28P) Simmondi, H. A , , Wilson, J. B., ibid., 16, 155 (1967). (29P) Kalther, H J., Arch. Hyg. Bakteriol, 152, 202 (1968). (30P) Reatherburn, 11.W., ANAL.CHEM., 39, 971 flY67). Hormones (1Q) Ahuja, J. K.,Kaplan, A., Van Dreal, P., Clin. Chem., 14, 664 (1968). (2Q) Bacchus. H.. Am. J . Clin. Pathol..
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(19Q) Nugent, C. A., Mayes, D. M., J . Clzn. Endocr., 26, 1116 (1966). (20Q) Perko, Z., Simonovic, I., Lijemicki Viesn., 89. 937 (1967). (Yugoslavia). Chem. ’Ahsir.. 69. 16756’(1968’1: (2lQ) Pileggi, ’ V. ‘ J., Kessler, ’ G., Clin. Chem., 14, 339 (1968). (22Q) Roitt, I. LI., Torrigiani, G., Endocrinology, 81, 421 (19167). (23Q) Rosenbaum, J. I f . , Brie-!a, A. F., Henry, J. B., Yiozley, J. M.,l\lcAfee, J. G.. -4m. J . Clin. Pctthol.’., 50., 336 (1968j. (24Q) Schindler, A. E., Ratanasopa, V., Herrmann, W. L., Clin. Chem., 13, 186 (lU671 \ - - - . I .
( 2 5 9) Tang, C. IT., Tomlinson, R. H.,
hucl. Actw. Tech. Life Sci., Proc. Symp. Amsterdam, 427. 1967. Chem. Abstr.. 68, 93444 (1968). (26Q) Van Baelen, H., Hayiis, W., DeMoor, P., J . Clin. Endocr. Metab., 27, 1056 (1967). (27Q) Viktora, J. K., Bnitkal, .4.,Kolff, F. W.,Anal. Biochem., 23, 513 (1068). (28Q) Wallach, E. E., Touchstone, J. C., Clin. Chem., 13, 976 (1967). (29Q) \Veil-Malherbe, H., Bigelow, L. >I., Anal. Biochem., 22,321 (1968). (30Q) Wise, C. D., zbzd., 18, 94 (1967). (31Q) Yee, H. Y., Katz, E. S., Jenest, E. S., Clzn. Chem., 13, 220 (1967).
Organic Acids and Compounds (1R) Anton, A . H., Sayre, D. F., Clin. Chem., 13, 1017 (1967). (2R) Bjornesjo, K. B., Jarnulf, B Scand. J . Clin. Lab. Invest., 20, 39 (19;i). (3R) Camp, B. J., Farmer, L., Clin. Chem., 13, 501 (1967). (4R) Chibolm, J. J., Jr., Anal. Biochein., 22, 54 (1968). (5R) Domaak, G. F.. Schlicke. H. H.. ibid., p. 2i9. 16R) Dreyfus, P. AI., Dube, F-. E., Clin. Chim. Acta, 15, 525 (1967). (TR) Fornari, G., lIarchesi, N., Sorbini, C. .4.,Rass. Fisiopatol. Clin. Ter., 39, 423 (1967). Chem. Ahsty., 68, 112113 (1968). (8R) Free, A. H., Free, H. M.,Clin. Pathol. Serum Electrolutes. 104 11966). (9R) Gibbard, S., W a t k h , ’ P. ,J., Clih. Chim. dcta, 19, 511 (1968). (IOR) Gompertz, D., ibid., p. 477. (1lR) Hadjivassiliou, A . G., Rieder, S.V., ibid., p. 357. 112R! Hla-Pe. V.. .4im~-Than-Batu.Anal. Biochem., 20, 432 (1567). (13R) Jones, G. B., ibid., 21, 286 (1967). (14R) Kanter, S. L., Clin. Chem., 13, 406 (1967). (l5R) Iiawasaki, C., Ito, Y., Tanino, IC, Bitamins, 36, 430 (1967). Chem. Abstr., 68, 19441 (1968). (16R) Kehl, H., Clin. Chem., 13, 475 (1967). (17R) Lee, II., XLicl. Activ. Tech. Life Sci., Proc. Symp. Amsterdam 681 (1967). Chem. Abstr., 68, 66217 (1968). (9U) Ciuhandu. G , Rusu, V., 2. KEin. C h n . Klin. Biochem., 6,’204 (1968). (10111 Colehour. J. K.. ANAL.CHEM.. , 39., ‘ iiio (1967). ’ (11U) Collison, H. A., Rodkey, F. L., O’Keal, J. D., Clin. Chem., 14, 162 (1968). (12U) Comstock, E. G., Comstock, B. S., Ellison, K., ibid.,13, 1050 (1967). (13U) Curry, S. H., A N ~ LCHEM., . 40, 1251 (1968). (14C) Curry, A. S., Walker, G. W., Simpson, G. s., Analyst (London), 91, 742 (1966). (15U) Davis! J. R., Andelman, S. L., Arch. Envzron. Health, 15, 53 (1967). ( 1 6 r ) De Leeuw, R. J. M.,Hamelink, 11. L., Kreukniet, J., Visser, B. F., Douze, J. C. XI., Van Heijst, 4.N. P., Chem. Weekbl., 62, 591 (1966) (Dutch). Chem. Abstr., 66, 73078 (1967). (17U) Douglas, J. F., Kelley, T. F., Smith, X. B., Stockage, J. A., ANAL.CHEM., 39, 956 11967). 118U) Dunlop, >I., J . Clin. Pathol., 20, 300 (106i). (19U) Eisenberg, W. V., Tillson, A. H., J . Forensic Sci., 11, 529 (1966). (20U) Ewan, R. C., Baumann, C. A., Pone, A. L.. J . Aar. Food Chem., 16, 212 (‘1968). ’ (21Uj E’ischer, D. S.,Clin. Chem., 13, 6 (1967).
(22U)-Fischl, J., Pinto, S., Gordon, C., ibid., 14, 371 (1968). (23C) Funk, K. F.. Die Pharmazie, 22, 241 (1967). Chem. Abstr., 67, 105814 (1967). (24C) Gassman, H. E., JVranne, L., Acta SOC.Med. Upsal., 72, 277 (1967). (25U) Girard, 11.L., Dreux, C., Paolaggi, F., Delattre, J., Ann. Biol. Clin. (Paris), 26, 401 (1968). (26U) Guidotti, G. G., Borghetti, A. F., Loreti, L., Bnal. Biochem., 17, 513 (19661. At. Absorption News(27U) Hessel, D. W., letter, 7, 55 (1968). (28U) Jones, D. W.,Gerber, L. P., Drell, IT., Clin. Chem., 13, 696 (1967). (29U) Kaempe, B., Acta Pharmacol. Tozicol., 25, 249 (1967). Chem. Abstr., 67, 71031 (1967). (30U) Kirk, P. L., J . Gas Chromatog., 5 , 11 (1967). (31U) Knowlton, Al., Goldbaum, L. R., Clin. Chem., 13, 683 (1967). (32U) Kondo, T., Kawashiro, I., Furuhashi, F., Shokuhin Eiseigaku Zasshi, 8 , 331 (1967). Chem. Abstr., 68, 75643 (1968).
(33U) Kopito, L., Shwachman, H., J . Lab. Clin. Med. 70, 326 (1967). (34U’r LeBlanc. A. E.. Can. J . Phvsiol. Pharmacol., 46, 665 (1968). (35U) llachata, G., Deut. 2. Gesmate Gerichtl. Med., 59, 181 (1967). Chem. Abstr., 67, 18407 (1967). (36U) >laddock, R. K., Jr., Bloomer, H. A,, Clin. Chem., 13, 333 (196i). (37U) McCann, H. G, Arch. Oral Biol., 13, 475 (1968). (38U) Nakaaki, K., Rodo Kagaku, 43, 713 (1967). (39U) Noirfalise, A . , Heusghem, C., De Vleeschhouwer, G. R., RIoerman, E., Ann. Biol. Clin. (Paris),26, 249 (1968). (40U) Ohkuma, S., Bunseki Kagakzc, Shinpo Sosetsu 162R (1966). Chem. r2bstr.. 68. 57230 119681. (41U) Oka,’ T., Keio J . Med., 16, 31 (1967). C h m . Bbstr., 67, 105850 (1967). 142U) Olesen. 0. V.. Scand. J . Clin. Lab. -Invest., 20,’109 (1967). (43U) Petrow, H. G., Strehlow, C. D., ANAL.CHEM..39. 265 (1967). (44U) Popa, L., Popa, L., Farmacia (Bucharest), 15,377 (1967).Chem. Abstr., 67, 97541 (1967). (45U) Raudonat, H. K., Deut. 2. Gesamte Gerichtl. Med., 59, 173 (1967). Chem. Abstr., 67,, 18406 (1967). 146U) Roach, 11.K., Creaven, P. J., Clin. Chim. Acta, 21, 275 (1968) (47C) Routh, J. I., Fhane, N. A., Arredondo E. G., Paul, IT. D., Clin. Chem., 14, 882 (1968). 148U) Sam\ahl, K., ASAL. CHEM., 39, 1480 (1967). (49U) Savory, J., Sunderman, F. W.,Jr., Roszel, K.”O., LIushak, P., Clin. Chem., 14, 132 (1968).
(50U) Sereda, G. A., Gig. Tr. Prof. Zabol., 12, 54 (1968). Chem. Abstr., 68, 102481 (1968). (51U) Serra, A. S., Rev. Port. Farm., 17, 448 (1967). Chem. Abstr.., 68., 112068 11968). ’ (52U) Simon, R. K., Christian, G. D., Purdy, W.C., Am. J . Clin. Pathol., 49, 207 (1968). (53U) Singer, L., Armstrong, W. D., ANAL. CHEM., 40, 613 (1968). 154U) Skinner, R. F., J . ForensicSci., 12. 230 (1967). ’ (55U) Sprung, W. D., Reihe, 15, 745 (1966). Chem. Abstr., 67, 88193 (1967). (56U) Stone, L. R., J . Ass. Ofic. Anal. Chem., SO, 1361 (1967). (57U) Sunshine, I., Rlaes, R., Faracci, R., Clin. Chem., 14, 595 (1968). (58U) Tewari, S. N., Mikrochim. Acta, 2 , 390 (1968). Chem. Abstr., 68, 93421 (1968). (59U) Thompson, H. L., Decker, R. J., Am. J . Clin. Pathol., 49, 103 (1968). (60U) Truffert, L., Girard-Wallon, C., Arch. Mal. Prof., Med. Trav., Secur. SOC.,29, 23 (1968). Chem. Abstr., 69, 602 (1968). (61U) Vidic, E., Z . Klin. Chem., 5, 143 (1967). Chem. Abstr., 67, 71039 (1967). (62U) Watkinson, J. H., Symp. Selenium Biomed., Int. Symp., 1st; Oiegon State Univ. 97 (1966) (Pub. 1967). Chem. Abstr., 68, 36527 (1968). (63U) Wawschinek, O., Paletta, B., Beyer, IT., Arch. Toxikol., 23, 52 (1967). Chem. Abstr., 68, 19433 (1968). (64U) Weber, K., Deut. 2. Ges. Gerichtl. M e d . , 57, 410 (1966). Chem. Abslr., 67, 114271 (1967). (65U) Wieczorek, H., Mikrochim Acta, 4, 802 (1968). I
(66U) Winsten, S., Brody, D., Clin. Chem., 13, 589 (1967). (67U) Yoshimura, H., Oguri, K., Tsukamoto, H., Chem. Pharm. Bull. (Tokyo), 14, 1286 (1966). Chem. Abstr., 66, 73111 119671. (68U)-Zsoldos, T., Csevari, S., Toth, A,, Nagy, L., Med. Radiol., 11, 56 (1966). Chem. Abstr., 66, 73185 (1967).
.
Vitamins
(1V) Contractor, S. F., Shane, B. Clin. Chim. Acta, 21, 71 (1968). (2V) Drujan, B. D., Castillon, R., Guerrero, E., Anal. Biochem., 23, 44 (1968). (3V) Fabianek, J., DeFilippi, J., Rickards, T., Herp, A. Clin. Chem., 14,456 (1968). (4V) Frodyma, M.lI.,Lieu, V. T., ANAL. CHEM.,39, 814 (1967). (5V) Hansen, L. G., Warwick, TV. J., Am. J . Clin. Pathol., SO, 525 (1968). (6V) Kahan, J., Scand. J . Clin. Lab. Inuest., 18,679 (1866). (7V) lIcLaren, D. S., Read, W. \V. C., Awdeh, Z. L., Tchalian, lf,,Methods Biochem. Anal., 15 1 (1967). (8V) Michaelsson, d., Michaelbson, LI., Scand. J . Clin. Lab. Invest., 20. 97 (1967). 19V) Ostlund. S. G. S..Biorlin. G.. Odontol Revy, 17, 208 (1966). 6’hem: Abstr., 66, 62537 (1967). (10V) Pelletier, O., J. Lab. Clin. Med., 72, 674 (1968). (1lV) Sherman, B. S., Clin. Chem., 13, 1039 (1967). (l2V) Yamane, Y . , JIotomiya, IC., Yamada, Y., Bitamins, 36, 203 (1967). Chem. .Ibstr., 67, 97542 (1967).
Coatings M. N. Swann, M. l . Adams, and G . G. Esposito, U.S. Army Coating and Chemical laboratory, Aberdeen Proving Ground, Md.
T
contributions t o the analysis of coating materials as selected by the reviewers since the previous summary (1 19) are contained in this biennial review. The period covered extends from December 1966 through December 1968, although a few significant foreign publications, located by abstracts, may predate this coverage. I n this attempt t o be selective, we hope that valuable publications have not been omitted. Other reviews were made nithin this period (89,117, 124).
