(322) Shibsta, S., Furukswa, M., Goto, K., Anal. Chim. Acta. 53,369 (1971). (323) Shiga, T., Shiga, K., Kuroda, M., Anal. Biochem., 44,291 (1971). (324) Shimaki, N., Ikeda, K., Hsmaguchi, K., J. Bwchem. (Tokyo), 68, 795 (1970). (325) Shmukler, H. W., J. Chromaiogr. Sn’., 8,581 (1970). (326) Siigur, J., Hsldna, U., Reakts. Sposobnost Org. Soedzn, 7, 197 (1970); Org.Reactiv., 7 , 8 7 (1970). (327) Singh, P. R., Sahai, R., J. Indian Chem. SOC.,46,945 (1969). (328) Skancke, A., Sksncke, P. N., Acta Chem. Scand., 24,23 (1970). (329) Smith, D. J., J. Ass. Ofic.Anal. Chem., 53, 116 (1970). (330) Smith, E., Sharkey, M. F., Levine, J., ibid., 54, 609 (1971). (331) Smolinski, S., Deja, I., Tetrahedron, 27, 1409 (1971). (332) Sokolova, R. S., Belkova, E. V., Opt. Spektrosk., 30, 562 (1970); Opt. Spectrosc., 30, 303 (1970). (333) Soni, R. N., Gupta, K. L., Collect. Czech. Chem. Commun., 36,2371 (1971). (334) Soonike, E. R., Haldna, U. L., Reakts. Sposobnost Org. Soedin, 6 , 1133 (1969); Org. Reactiv. ( U S S R ) , 6, 490 (1969). (335) Ssekaalo, H., Anal. Chim. Acta, 51, 503 (1970). (336) Stefanovich, V., Ceprini, M. Q., J. Pharm. Sci., 60,781 (1971). (337) Stein, T. P., Reid, T. W., Fahrney, D., Anal. Biochem., 41, 360 (1971). (338) Stern, E. S., Timmons, C. J., “Gillam and Stern’s Introduction to Electronic Absorption Spectroscopy in
Organic Chemistn,” Third Ed., Edward Arnold, London, 1970. (339) Stevens, H. N. E., Stevens, M. F. G., J. Chem. Soc. C, 1970,765. (340) Steward, 0. W.,. Dziedzic, J. E., Johnson, J. S., Frohliger, J. O., J. Org. Chem., 36,3480 (1971). (341) Stewart, R. G., Anal. Bwchem., 35,
321 (1970). (342) Strickland, E. , H., Horwitz, J., Billups, C., Biochemzstry, 9,4915 (1970).
(343) Stromberg, L. E., Widmark, G., J . Chromutogr., 47,27 (1970). (344) Sukhova, T. G., Temkin, 0. N., Flid, R. M., Zh. Neorg. Khim., 15, 1849 (1970); Russ. J. Inqrg. Chem., 15, 949 (1970). (345) Sundbom, M., Acta Chem. Scand., 25,487 (1971). (346) Suzuki, H., J. Amer. Oil Chem. SOC., 47, 273 (1970). (347) Swihart, D. L., Mason, W. R., Inorg. Chem., 9,1749 (1970). (348) Talwar, U. B., Haldar, B. C., Anal. Chim. Acta, 51,53 (1970). (349) Tamres, M., Grundnes, J., J. Amer. Chem. SOC.,93,801 (1971), (350) Tanaka, K., Takagi, N., Anal. Chim. Acta, 48,357 (1969). (351) Tillman, J., Whymark, D. W., Analyst (Lonabn),96,689 (1971). (352) Tishchenko, V. G., Fetisova, M. M., Opt. Spektrosk., 30, 23 (1970); Opt. Spectrosc., 30, 12 (1970). (353) Topham, R. W., Gaylor, J. L., J. Bwl. Chem., 245,2319 (1970). (354) Treinin, A,, Wilf, J., J. Phys. Chem., 74,4131 (1970). (355) Tselenskii I. V., Kosmynina, A. S., Dronov, V. d., Shokhor, I. N., Reakts. S p o s o h s t Org. Soedin, 7, 50 (1970), Org.Reactiv. 7 , 2 0 (1970). (356) Tsuji, d.,Seiki, T., J. Polymer Sei., Part B., 8,817 (1970). (357) Tutt, D. E., Schwartz, M. A., ANAL.CHEM.,43,338 (1971). (358) Tyler, N. W., Jr., Becker, R. S., J . Amer. Chem. SOC.,92, 1289 (1970). (359) Umeyama, H., Nagai, T., Nogami, H., Chem. Pharm. Bull., 19,1714 (1971). (360) Umeyama, H., Nagai, T., Nogami, H., Oguma, T., ibid., p 412. (361) Vasil’ev, V. P., Zaitseva, G. A., Zh. Neorg. Khim., 15, 1016 (1970); Russ. J . Inorg. Chem., 15, 518 (1970). (362) Vierling, F., Bull. SOC.Chzm. Fr., 1971, 25. (363) Vinogradov, I. P., Dodonova, N. Ya., Opt. Spektrosk., 30, 27 (1970); Opt. Spectrosc., 30, 14 (1970). (364) Wagner, A. J., Moeller, T., J. Inorg. Nucl. Chem., 33,1307 (1971).
(365) Wahbi, A. M., Farghaly, A. M., J. Pharm. Phurmacol., 22, 848 (1970). (366) Walborg, E. F., Jr., Kondo, L. E., Anal. Bwchem., 37,320 (1970). (367) Wallace, J. E., Biggs, J. D., J. Phurm. hi.,60, 1346 (1971). (368) Waters, T. N., Wright, P. E., J. Imrg. Nucl. Chem., 33, 359 (1971). (369) Watkin, J E , Miller, R. A., Anal. Bzochem., 34,424 (1970). (370) Watson, F. H., Jr., Armstrong, A. T., McGlynn, S. P., Theor. Chim. Acta, 16,75 (1970). (371) Weiss, K. G., Boltz, D. F., Anal. Chim. Acta, 55,77 (1971). (372) Welling, P. G., Lee, K. P., Khanna, U., Wagner, J. G., J. Pharm. Sci., 59, 1621 (1970). (373) Wetters, J. H., Uglum, K. L., ANAL. CHEM.,42,335 (1970). (374) Wladislaw, B., Viertler, H., Demant, E. B., J. Chem. SOC.B , 1971,565. (375) Wojtowicz, E. J., J. Pharm. Sei., 59,240 (1970). (376) Wojtowicz, P. F., Diliberto, J. J., ibid., 60, 1385 (1971). (377) Wong, J., Angel], C. A,, Appl. Spectrosc. Rev., 4 , 9 7 (1970). (378) Wood, P. M., Crane, F. L., Anal. Biochem., 36, 127 (1970). (379) Woodhouse, E. J., Norris, T. H.‘ Imrg. Chem., 10, 614 (1971). (380) Woods, A. H., O’Bar, P. R., Science, 167, 179 (1970). (381) Woodson, A. L., J. Ass. Ofic. Anal. Chem., 53,819 (1970). (382) Worm, K., Acta Pharmucol. Tozicol., 27, 439 (1969). (383) Worman, J. J., Pool, G. L., Jensen, W. P., J. Chem. Educ., 47,709 (1970). (384) Wu, W. H., Chin, T. F., Lach, J. L., J . Pharm. Scz., 59,1234 (1970). (385) Wiinsch, G., Umland, F., 2. Anal. Chem., 250,248 (1970). (386) Yoshida, Z., Kobayaahi, T., Theor. Chim. Acta, 20,216 (1971). (387) Zalewski, R. I., Dunn, G. E., Cun. J. Chem., 48,2538 (1970). (388) Zholnin, A. V., Bobyrenko, Yu. Ya., Zh. Neorg. Khim., 16, 372 (1971); Russ. J . mnurg. Chem., 16, 194 (1971).
Use of Enzymes in Analytical Chemistry Myer M. Fishman and Howard F. Schiff, Department o f Chemistry, The City College, City Uniyersity of New York, New York, N.Y. 10037
T
in analytical chemistry is not a new concept. Nearly 40 years ago, the phosphatases were among the first enzymes to find widespread use as diagnostic tools. Since then, needless to say, enzymes have become increasingly important and extensively used in clinical chemistry, in the food and pharmaceutical industries, and as diagnostic tools in medicine. I n all these cases, enzymic analysis has included not only the analyses carried out with the aid of enzymes, but also the assay of enzyme activity itself in organic and biological fluids. HE USE OF ENZYMES
Enzyme techniques have found increasing favor for a variety of reasons. Careful studies on the preparation, purification, and properties of enzymes have led to new methods of assay. The ready availability of highly purified and crystalline enzymes has facilitated their use as analytical reagents. Studies on enzymes which have concerned theniselves with optimal conditions of pH and temperature, cofactor concentration, and the effect of inhibitors have done much to delineate the course of an enzymatic reaction. I n turn, these have enabled many investigators to
establish methods of measuring either the enzyme or its substrate with a high degree of specificity and reproducibility Some 25 years ago, Warburg developed a method of enzymatic analysis based on the absorption of light by the coenzymes nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP). The method was rapidly expanded to include a wide variety of dehydrogenases which are pyridine-nucleotide dependent. Thus, the enzymatic conversion of lactate to pyruvate was accompanied by a reduction of KAD to NADH and a
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