can be included without precipitation, if the titration is performed using the nitrate salt of tetren. CADMIUMIN PRESEXCE OF CALCIUJI, hlAGXESIUM, ALcivUNuhI, AND LAXTH.UKJR.I. Take a sample containing 0.20 mmole of cadmium ion, adjust the volume to 60 ml. with mater, and add 1 ml. of ammonia buffer, 1 gram of sodium tartrate, and 1 drop of mercury(I1)tetren. Titrate with 0.01M tetren. Barium can also be included without precipitation of barium sulfate, if the titration is performed using the nitrate salt of tetren.
COPPERAND ZINC
IN
PRESEXCE OF
CaLcIunr AND MAGSESIUM. Take a samp;l!. rnntaining 0.01 mmole of copper and ziiii ions, adjust the volume to 60 ml. with water, and add 1 ml. of acetate buffer. If necessary, adjust the pH to 5 with sodium hydroxide; add 1 drop of mercury(I1)-tetren and titrate the copper n-ith 0.01U tetren. Then add
3 nil. of ammonia buffer and titrate the zinc with 0.01ilI tetren. CADMIUM I N PRESENCE O F AIANG-4NESE. Take a sample containing 0.20 mmole each of cadmium and manganese(I1) ion, dilute to 60 ml. with distilled water, and add 1 drop of mercury(11)-tetren. Bubble the solution with tank nitrogen for a few minutes to exclude oxygen, add 1.5 ml. of ammonia buffer. and titrate with 0.01M tetren. LITERATURE CITED
(1) Bjerrum, J., Schwarzenbach, G., Silken, L. G. (compilers), “Stabilit; Constants of Metal Ion Complexes, Part [ &;, . k a l Society, London, Spec. Publ. 6 (1957). (2) Flaschka, H., Soliman, A., 2. anal. Chew 159, 30 (195(3) Latimer, \V, M.,‘ );‘Oxidation Potentials,” 2nd ed., Prentice Hall, New York, 1952. (4) Prue, J. E., Schwarzenbach, G., Helv.
Chini. Acta 33, 963, 974, 985, 995 (1950). ( 5 ) Reilley, C. N., Holloway, J. H. , J . Am. Chem. SOC.80, 2917 (1958). (6) Reilley, C. S., Schmid, R. W.,hsa~,. CHEX.30,953 (1958). ( 7 ) Ibzd , submitted. (8) Reilley, C. N., Schmid, R. W., J . Elisha Nztchell SOC.73, 279 (1957). (9) Reilley, C. S., Schmid, R. W., Lamson, D. IT., ASAL. CHEII. 30, 947 (1958). (10) Reilley, C. S . , Sheldon, AI. V., Chemzst Analyst 46,59 (1957). (11) Reilley, C. K., Sheldon, 11. V., Talanta 1, 127 (1958). (12) Schmid, R. IF-., Reilley, C. N., J . A m . Chem. SOC.78,5513 (1956). (13) Schwarzenbach, G., Ackermann, H., Heh f‘h7ni -4 $z 30, 1798 (1947). (14) S i u g ’ k $ , h. V.,J . Chem. SOC.1941, 433. RECEIVEDfor revierT May 12, 1958. Accepted July 28, 1958. Work supported by U. S. Air Force through the Office of Scientific Research of the Air Research and Development Command.
Comparative Phenolic Spot Tests Application to Some Air Particulate Fractions G. E. INGLETT’ and J. P. LODGE Robert A. Toft Sanitary Engineering Center, U. S. Public Health Service, Cincinnati 26, Ohio
Comparative spot tests utilizing four different reagents were applied to 34 phenols. Characteristic colors and lower detection limits were determined. The tests were further applied to {he weak acid fractions of benzene-soluble particulate matter from 1 1 cities. Samples from two cities yielded weak evidence of the presence of phenolic compounds. Clear evidence was obtained for the presence of flavonoids, presumably derived from pollen in the samples.
P
constitute an interesting portion of the organic constituents of atmospheric particulate matter. During a preliminary study of the paper chromatographic separation of this class of compounds, it was felt desirable to select a generally applicable spot reagent for the development of the resulting chromatograms. The lower limit of identification of four different reagents was determined for a series of phenols. The first of these reagents, 4-aminoantipyrine ( I ) , has been used previously for the quantitative estimation of phenol. but the HENOLS
present report is believed to be the first record of its use in a phenol spot test. Diazotized sulfanilic acid has been used for many years for detecting phenols ( 4 ) . It will also couple n ith various imidazoles to produce colored derivatives. Formaldehydesulfuric acid (w) is a sensitive test for phenols as !vel1 as for aromatic hydrocarbons (3). lloreover, i t reacts with phenols a t room temperature, whereas some aromatic hydrocarbons require elevated temperatures for positive tests. The Millon reagent (6,7 ) has also been applied previously to various phenols. There are numerous other spot tests Tvhich could be used, but only these four reagents were employed for the comparative observations reported here. I n the light of these tests, spot test analysis was applied to the weak acid fraction taken from the particulate matter of several iimerican cities (6) and to the alcoholic extracts of defatted samples taken from St. Louis County, Xssouri, during the high pollen incident months of August and September. EXPERIMENTAL
4-Aminoantipyrine
Present address, Corn Products Refining Co., P. 0 . Box 345, h g o , Ill. 1
248
*
ANALYTICAL CHEMISTRY
(UP).
RE^-
Smmonium chloride buffer. pH 10; 395 alcoholic 4-aminoantiGEXTS.
pyiine; aiid 2% aqueous potassium ferricyanide prepared fresh each time. METHOD. A drop of the unknown solution is spotted on a white porcelaiii spot plate, and one drop each of ammonium chloride, 4-aminoantipyrine. and potassium ferricyanide solutions are added successively. The colored spot is comuared n-ith a yellow reference spot of the reagents alone. Diazotized Sulfanilic Acid (DSA).
Five milliliters of 5% sodium nitrite, 5 ml. of sulfanilic acid in 9 S hydrochloric acid, and 15 mi. of butyl alcohol are placed in a 25ml. graduate and shaken. T h e butyl alcohol layer is separated and placed in a dropping bottle. d 0 . 2 5 S sodium carbonate solution is also required. METHOD. A drop of the unknown solution i s spotted on a white porcelain spot plate. followed by drops of diazotized sulfanilic acid in butyl alcohol and sodium carbonate solution. The colored spot develops rapidly. REAGEXTS.
Formaldehyde-Sulfuric Acid (FSA).
REAGESTS.I n 10 ml. of concentrated sulfuric acid, 0.20 ml. of 37% formaldehyde is mixed. METHOD. The unknown solution is spotted on a white porcelain spot plate follon ed by one drop of formaldehydesulfuric acid reagent. Color develops immediately for a positive test.
REAA 10-gram sample of mei-
Millon’s Reagent (MR). GESTS.
cury is dissolved in 14.1 ml. of con-
centrated nitric acid (specific grari t y 1.42) a n d subsequently diluted n-ith 20 ml. of distilled n-ater. METHOD.A drop of the u n k i m m solution is 100 T e 40 Br 10 > 100 2. 2,6-Di-te~t-butylphenol >lo0 Ye 20 Ye 10 Re 10 3. 2,6-Di-tert-butyl-4> ... 100 methylphenol > 100 l e 100 Br 10 4. 3,4-Diniet hylphenol 20 >100 Ye 10 Br 1 0 ’ Or 5 . 2,4-Dimethylphenol Ye 10 Br 10 >IO0 Pi 10 Or 6. 2,6-Dimethylphenol Ye 10 Ye Or 50 10 Or 10 7. 2,5-Dimethylphenol Te 10 Or 10 Te 10 Pu 10 8. 3,5-Dimet hplphenol Vi 10 1-e 10 T e 10 Or 5 9. p-tert-But ylphenol Pi 20 Re 10 Ye 10 Gr 50 10. p-Benz ylougphenol Te 50 Te 100 Te 50 Br 20 11. p-Benz ylphenol Te 50 Ye 40 Te 100 Re 10 12. p-Pheny lphenol Or 20 Re 50 B1 10 Pi 5 13. a-Tocophenol 50 > 100 > 100 Ye 10 Ye 14. 6-Hgdroxy-2,2,5,7,%pent a50 met hylchroman >I00 > 100 Gr 10 l e 15. 5-Hgdroxy-2,4,6,7-t et ra\-e 5 met hplcoumaran > 100 Te 20 Re 10 ~. 16. Catechol Te 2 Br a Pu 1 Re 10 17. Carvacrol 20 Te Or 1 Br 30 Or 10 18. Orcinol Te 1 11 10 Ye 0 Ye 1 19. Resorcinol Ye 1 Br 1 Or 1 11 10 20. p-terl-Butylcatechol Te 2 Br a Te 10 Pu 1 21. p-Octylphenol Ye 20 > 10 > 10 Br 5 22. p-Nonylphenol 20 > 10 > 10 Br 5 Ye 23. 2,6-DiisopropyIphenol 1 Te Te >10 Ye Re 1 5 24. o-Cresol Ye 10 Te Re 1 1 Re 1 25. na-Cresol Or 10 Or 1 Ye 1 Re 1 26. p-Cresol Te 10 Br 1 B1 1 Re 2 27. Thymol Ye 20 Pi 3 Te Br 1 28. Gu‘aiacol Pi 2 Ye 2 Vi 1 Or 1 ~. _. > 10 >io B; i Ye 20 29. p-Cumylphenol 30. Phenol 10 Pu 0 5 Te 0 5 Pu 1 Br 31. Arbutin 1 1-e 1 Ye 5 Ye 5 Pi 32. Rutin G\-g 1 Pi 2 Ye 0 5 Gr 5 33. Quercetin Y& 1 Or 2 Ye 0 5 Te 5 Gno 1 Or 2 Ye 0 5 Te 5 34. Quercitrin a 4-Aminoantipyrine reagent. Diazotized sulfanilic acid. c Formaldehyde-sulfuric acid. d Millon reagent. e B1, blue; Br, bron-n; Gn, green; Gy, gray; Or, orange; Pi, pink; Pu, purple; Re, red: Vi, violet; Ye, yellow. f Limit of detection in micrograms. 0 Color with ammonium chloride (pH 10) reagent alone. ~
A comparatire spot test study of a large number of phenols n.as made (Table I). An estensive examination of these data is not attempted, but it can be seen that the hindered phenols, 1, 2, 3, 14, 15, and 23, do not readily lend themselves to these tests. Similar analysis Tvas applied to the weak acid fraction taken from the benzene extraction from 11 American cities (Table 11). The formaldehydesulfuric acid reagent reveals the presence of considerable amounts of reactive materials. Becauqe of the nonspecificity of this reagent, it cannot be said conclusively t h a t these reactive materials are phenols. Positive reactions of samples 9 and 11 with both 4-aminoantipyrine and diazotized sulfanilic acid reagents seem to substantiate the presence of phenols. These tests were complicated b y the colored unknon-n solution, and, therefore, reference spots were required for each test. The more complex, highly polar phenols are not removed by the defatting operation and remain in the matrix of the residual particulate matter. Therefor?, ten residues from the defatted samples from St. Louis County were extracted for 6 hours nith 95% ethyl alcohol. The yellow-colored extracts w r e concentrated to a volume of 5.0 ml. Spot test analysis gave a positive test with formaldeliyde-siilfuric acid and diazotized sulfanilic acid reagents. The former reagent gave straw-colored reactions. and the latter, orange. -411 g a r e barely detectable colors n i t h 10 pl. of test solution. These color reactions logically may be attributed, a t least in part. to flayonoids derived from pollen.
Table II. Spot Test Analysis of W e a k Acid Fractions .4APa DS.\a FSA* llRn __Weak Acid Fraction Colorc p1 d Colorc p1.d Colop p1.d Color. pLd I . Cincinnati Br 1 2. Houston Br 1 3 Los Angeles Br 5 4. h’ew Orleans Br 6 5. New York Br 1
SUMMARY
A comparative spot test detection of 34 phenols has been made with 4aminoantipyrine, diazotized sulfanilic acid, formaldehyde-sulfuric acid, and Millon’s reagent. When these reagents were applied to the analysis of \Teak acid fractions of collected particulate air pollutants, some evidence indicated the existence of phenols. More conclusive, honrever, appears to be the presence of flavonoids derived from pollen present in the alcoholic extracts of defatted air particulate matter.
11. Detroit Br-Ye 1 Light Or 1 Br 0 5 . 30-ml. sample taken from n-eak acid fraction. Taken fiom weak acid fraction in 5 ml. of chloroform. c Br, brown; Re, red; Ye, yellow; Or, orange. Lack of color, negative test. d Smallest volume in microliters from which noticeable color could be obtained
LITERATURE CITED
(1) Emerson, E., J . Org. Chem 8 , 417 (1943). ( 2 ) Feigl, F., “Spot Tests,” Vol. 11, p. 104, Elsevier, Kew York, 1954. (3) Zbid., p. 107. (4) Zbid., p. 108. (5) Zbid., p. 134.
(6) Tabor, E. C., Hauser, T. R., Lodge, J. P., Burttschell, R. H., A X . . 4 . Arch. Znd. Health 17, 58-63 (1958). (7) Vaubel, W., Z. angew. Chem. 13, 1127 (1900).
RECEIT-ED for review April 7, 1958. Accepted October 7, 1058.
VOL. 31, NO. 2, FEBRUARY 1959
249
