found in the body, a possibility to which so far little consideration has been given.
The foregoing suggests the need for a far-reaching clarification of the optimum conditions for the determination of enzyme activity. Opposed to this is the necessity of making many determinations of enzyme activity when following the course of pathological developments.
The necessary compromise limits the accuracy of the determination, but this is permissible considering the extent of biological variation. The assay of enzyme activities has contributed to the elucidation of many phenomena and is a useful aid for clinical diagnosis, prognosis, and the control of progress and therapy.
8) R. E. Thiers and B. L. Vallee, Ann. N . Y. Acad. Sci. 75, 214 [1958].
9) H. D. Horn and F. H. Bruns, Verh. dtsch. Ges. inn. Med. 65, 407 [1959].
Table 7. Normal values for the enzyme activity in human serum
*, modified Staved Biicher units/ml.
p H 7.4; 2 5 ° C modified by 5.20)
Schmidt^'D Biicher units/ml.
p H 7.5; 2 5 ° C
23 0.15 ( < 0 . 0 6 - 0 . 5 3 ) a-Amylase Sugar estimation
8 Cholinesterase (ChE) Colorimetric
1 1
) u.moles acetylcholine/hr./
0.05 ml.; p H 7.4; 37°C
30 j 8 - 9 ( 7 - 1 0 )
1) J. A. Sibley and A. L. Lehninger, J. biol. Chemistry 777, 859 [1949].
2
* F. H. Bruns, Biochem. Z. 325, 156 [1954].
3
> G. Beisenherz, H. J. Boltze, Th. Biicher, R. Czok, K.-H. Garbade, E. Meyer-Ahrendt, E. and G. Pfleiderer, Z. Naturforsch. 8b, 555 [1953].
4
* U. Stave, Z. Kinderheilkunde 84, All [1958].
5) E. Schmidt, F. W. Schmidt and E. Wildhirt, Klin. Wschr. 36, 172 [1958].
6) E. Schmidt, F. W. Schmidt and E. Wildhirt, Klin. Wschr. 36, 280 [1958].
7
* E. Schmidt and F. W. Schmidt, unpublished.
8) M. Somogyi, Proc. Soc. exp. Biol. Med. 32, 538 [1934].
) R. Heinecker and H. Losse, Klin. Wschr. 33, 870 [1955].
1
4
* R. Ammon, Pfliigers Arch. ges. Physiol. Menschen Tiere 233, 486 [1933].
Table 7, Part I (cont.)
Enzyme M e t h o d Author Units N u m b e r of cases N o r m a l values SD
G lutamate-oxalo-acetate trans
aminase (GOT)
UV-Assay is) Wroblewski
16)
aminase (GOT) Colorimetric
1 7
Schmidt O'l) Bucher units/ml.
p H 7.5; 2 5 ° C
, modified Stave*) Bucher units/ml.
p H 7.4; 25°C
UV-Assay (Boehiingei) Lindner
21
) Wroblewski units/ml.
p H 7.4; 2 5 ° C
Henley 23) [xmoles Py/hr./ml.
p H 7.4; r o o m temperature
> DeRitis™) fxmoles Py/15 min./ml.
p H 7.4; 37°C 18 0.22 ( 0 . 1 1 - 0 . 3 6 ) G lutamate-pyruvate
transaminase (GPT)
UV-Assay 22) Wroblewski
22)
UV-Assay 22), modified Staved Bucher units/ml.
p H 7.2; 25°C 25 premature modified by 20)
Schmidt <><1) Bucher units/ml.
p H 7.5; 2 5 ° C
122 0.38 ( 0 . 1 6 - 0 . 8 8 ) G lutamate-pyruvate
transaminase (GPT)
UV-Assay (Boehringer) Lindner
21
) Wroblewski units/ml.
p H 7.4; 2 5 ° C 950 1 9 ( 6 - 5 4 )
Schmidt^'D Bucher units/ml.
p H 7.5; 25° C
80 < 0 . 0 6
Isocitric dehydrogenase ( I C D H )
UV-Assay 26) Wolfson 2^) mu.moles/hr./ml.
p H 7.5; 25°C modified by 20)
Schmidt*' modified by 29)
Wroblewski
29) Wroblewski units/ml.
p H 7.4; 2 4 - 2 7 ° C 161 470 ( 2 5 0 - 8 9 0 ) Lactic dehydrogenase
( L D H )
UV-Assay 3D Hsieh
32
) , modified Staved Biicher units/ml.
p H 7.6; 25°C 25 premature
° ) , modified Engelhardt-Gdlkel**)
17) N. E. Tonhazy, U. G. White and W. W. Umbreit, Arch. Biochem. Biophysics 28, 36 [1950].
18) F. DeRitis, M. Coltorti and G. Giusti, Minerva med. [Torino] 47, 1 [1956].
19) G. A. Fleisher, K G. Wakim and N. P. Goldstein, Proc. Staff Meetings Mayo Clin. 32, 188 [1957].
2
0 ) E. Schmidt, F. W. Schmidt and E. Wildhirt, Klin. Wschr. 37, 1221 [1959].
2
24) H. Holzer, J. Haan and S. Schneider, Biochem. Z. 326, 457 [1955].
25) U. Gerlach, Klin. Wschr. 35, 1144 [1957].
2
6) S. K. Wolfson jr. and H. G. Williams-Ashman, Proc. Soc. exp. Biol. Med. 96, 231 [1957].
27) W. Kerppola, E. A. Nikkila and E. Pitkanen, Acta med. scand. 164, 357 [1959].
28) S. Ochoa, J. biol. Chemistry 174, 133 [1948].
29) F. Wroblewski and / . S. LaDue, Proc. Soc. exp. Biol. Med. 90, 210 [1955].
30) F. Kubowitz and P. Ott, Biochem. Z. 314, 94 [1943].
3D / . L. Strominger and O. H. Lowry, J. biol. Chemistry 213, 635 [1955].
32) K. M. Hsieh and H. T. Blumenthal, Proc. Soc. exp. Biol. Mee. 91, 626 [1956].
33) A. Engelhardt-Golkel, R. Lbbel, W. Seitz and 1. Woller, Klin. Wschr. 36, 462 [1958].
Table 7, Part I (cont.)
Enzyme Method Author Units Number of cases Normal values SE>
Leucine
^ , modified Engelhardt-Golkel^)
) Wroblewski units/ml.
p H 7.6; room temperature
67 2.9 ( 1 . 0 - 5 . 0 ) Phosphatase, alkaline Substrate: P-glyceroph.;
Buffer: veronal
<» 1 mg. phenol/30 min./lOO ml.
p H 9.3; 37.5°C
) m-moles p-nitroph./hr./
1 000 ml.
Phosphatase, acid Substrate: P-glyceroph.;
Buffer: acetate-veronal
Phosphohexose isomerase (PHI)
Fructose e s t .
4 5
) Bodanski
4
^ 25 u.g. fruct./30 min./ml.;
pH 7.4; 37°C
14 21 ( 8 - 4 0 ) ± 8
Fructose e s t .
4 6)
Bruns
47
) m m
3
F-6-P/hr./ml.
37° C
7 98 ( 8 6 - 1 0 9 ) Sorbitol dehydro
genase ( S D H )
| UV-Assay 24), modified by
4 8) Gerlach4
*) A E ^0 1/ m i n . / m l . pH 7.4; 24° C
16 < 1 . 0
| UV-Assay
4
»), i modified b y
7) Schmidt7
^ Biicher units/ml.
p H 7.5; 25° C
12 < 0 . 0 6
3 4
> G. A. Fleisher, H. R. Butt and K A. Huizenga, A n n . N . Y. Acad. Sci. 75, 363 [1958].
35) J. A. Goldbarg, E. Pineda and A. M. Rutemberg, Amer. J. clin. Pathol. 32, 571 [1959].
36) A. H. Mehler, A. Kornberg, S. Gisolla and 5. Ochoa, J. biol. Chemistry 174, 133 [1948].
37) A. Kornberg and B. L. Horecker in S. P. Colowick and N. O. Kaplan: Methods in Enzymology. Academic Press, N e w York 1955, Vol. I, p. 323.
38) O. Bodanski, J. biol. Chemistry 101, 93 [1933].
39) E. J. King and A. R. Armstrong, Canad. J. med. Assoc. 31, 376 [1934].
4
°) E. J. King and A. R. Armstrong, Canad. J. med. Assoc. 32, 379 [1935].
4
D C. Huggins and P. Talalay, J. biol. Chemistry 159, 399 [1945].
42
> O. A. Bessey, O. H. Lowry and / . Brock, J. biol. Chemistry 164, 321 [1946].
4
3 ) E. Kirberger and G. A. Martini, Dtsch. Arch. klin. Med. 197, 268 [1950].
44
> D. Remy and E. Gadermann, Dtsch. med. Wschr. 85, 1061 [I960].
4
5 ) O. Bodanski, Cancer 7, 1191 [1954].
4
6 ) F. H. Bruns and K. Hinsberg, Biochem. Z. 325, 532 [1954].
47
> F. H. Bruns and W. Jakob, Klin. Wschr. 32, 1041 [1954].
4
8 ) u. Gerlach. Klin. Wschr. 37, 93 [1959].
Table 7, Part 2
Enzyme Method Author Units Number of cases Normal values S D
Aconitase Citrate est., modified b y
4 9
) Beutler^o) 245 u.g. citrate/5 min. per assay or 0.085 umoles aconi-tate/5 min. per assay; 37°C
6 not detectable
Meister 54) [jimoles/hr./ml.
0.40 0.31
Arginase Kochakian
55
) not detectable
Aryl sulphatase Substrate: p-nitrophenyl-s u l p h a t e
5 6
)
0.005 M in 0.5 M acetate buffer
Huggins 56) 10 pig. p-nitrophenol in 10 ml./
lOhr./ml.
Dodgson 57) pig. phenol/hr./lOO ml.
2 0 - 2 4 ° C < 2 0
Quinine oxidase Spectrophotometric 58) Baier 59) A E ^ / I O min./4 ml. assay/ml. not detectable Cholesterol esterase Cholesterol est.
6
°) Turner*®) 1 % decrease in free
cholesterol/24 hr.; 37°C 20 3 3 - 4 4 (31 - 5 0 ) Creatine phospho
kinase (CPK) Creatine determination
6 1
) [jimoles creatine/hr./ml.
38°C
) [jig. deoxyribose/4 hr./ml., from 0.6% D N A solution (0.5 ml.)
Enolase ( E N ) UV-Assay 3), modified b y
5 , 2 0) Schmidt*^ Bucher units/ml.
pH 7 . 5 ; 2 5 ° C
15
UV-Assay 66) Merten
67
) [jtmoles/min./l 000 ml. 30 Esterases Substrate: p r o c a i n e
6
8 ) Substrate: phenyl-b e n z o a t e
6 9
)
Fischer
70
^ [jig./hr./ml.
Fructose-1 -phosphate aldolase ( P F A )
UV-Assay 7D Wolf
72
) sec./AEjff (2 ml. assay) p H o f the serum; 25° C U V - A s s a y 7 D ,
modified by
2 0
)
Schmidt
7
) Bucher units/ml.
p H 7.5; 2 5 ° C
18 Fumarase ( F U M ) Spectrophotometric 73) DeRitis
7
*) Wroblewski units/ml.
0.17 ( < 0 . 0 6 - 0 10.5
50 1 4 - 3 3
> 3,600
< 0 . 0 6 not detectable E. Racker, Biochim. biophysica Acta 4, 211 [1950].
E. Beutler and M. K. Y. Yeh, J. Lab. clin. Med. 54, 456 [1959].
F. B. Staub, O. Stephaneck and G. Acs, Biochemie [russ.] 22, 118 [1957].
O. Bodanski in: Enzymes in Health and Disease. T h o m a s , Springfield, Jll., 1960, p. 309.
E. Negelein and H. Widf, Biochem. Z. 293, 351 [1937].
A. Meister, Science [Washington] 106, 167 [1947].
C. D. Kochakian, E. H. Kentmann and E. E. Garber, Conf. of Metabolic Aspects, Transact. 17, 187 [1948].
C. Huggins and D. R. Smith, J. biol. Chemistry 170, 391 [1947].
K. S. Dodgson and B. Spencer, Biochem. J. 56, 2, XIII [1959].
H. Baier and K. Lang, Biochem. Z. 329, 381 [1957].
H. Baier, Klin. Wschr. 36, 569 [1958].
K. B. Turner, G. H. MacCormack and A. Richards, J. clin. Invest. 32, 801 [1953].
A. H. Ennor and H. Rosenberg, Biochem. J. 57, 203 [1954].
F. Schapira, J.-C. Drey fuss, G. Schapira and / . Demos, Rev. franc. Etud. clin. biol. 5, 990 [I960].
Y. Allfrey and A. E. Mirsky, J. genet. Physiol. 36, 221 [1952].
K. Schreier, A. Raspe and V. Heinke, Klin. Wschr. 33, 1096 [1955].
O. D. Kowlessar and R. K. McEvoy, J. clin. Invest. 35, 1325 [1956].
Th. Bucher in S. P. Colowick and N. O. Kaplan: Methods in Enzymology. Academic Press, N e w York 1955, Vol. I, p. 427.
R. Merten and H. G. Solbach, Klin. Wschr. 39, 222 [1961].
R. Hazard, Presse med. 113 [1949].
S. Dan and / . Vajda, Kiserl. Orvostud. 1, 118 [1948].
A. Fischer, L. Perenyi and S. Rohmy, Acta med. Acad. Sci. hung. 12, 229 [1958].
F. Leuthardt, E. Testa and H. P. Wolf, Helv. chim. A c t a 36, 221 [1953].
H. P. Wolf, G. Forster and F. Leuthardt, Gastroenterologia 87, 172 [1957].
F. DeRitis, G. Giusti and M. Coltorti, J. infect. Diseases 101, 219 [1957].
F. DeRitis, M. Coltorti and G. Giusti, Clin. chim. A c t a 4, 213 [1959].
Table 7, Part 2 (cont.)
Enzyme | Method Author Units Number of cases Normal values SD
Glucose-6-phosphatase Pi est. 75) Koide
1
^ mg. P/hr./ml.
pH 6.5 ! 0.012
(^-Glucuronidase Substrate: phenol- i 1
phthalein-gluc.76) Fishman
76
^ [ig. phenolphth./hr./lOO ml.
pH 4.5; 38°C 16 | J 0 - 1 8 1
) [ig. <?-aminophenyl-gluc./
40 min./ml.
pH 7.4; 37.5°C
! not detectable
! 1 i
!
Glutathione
reductase (GR) UV-Assay78) Coenzyme: D P N H
Glyceraldehyde-3- | UV-Assay3), | Schmidt
6
'
1
) phosphate dehydro- modified by
5
'
2 0
>
genase (GAPDH) i Biicher units/ml.
Engelhardt-GdlkenV Biicher units/ml. 30 not detectable j
Hexokinase (HK)
Lipase Substrate: olive oil82) Goldstein
82
) u.moles/24 hr./ml. 180 |
Lipase
Substrate: olive oil;
manometric 83) Adams
83
) u.moles/20 hr./ml. 2 . 0 ( 1 . 5 - 2 . 5 ) Lipase
Substrate: olive oil;
thymolphthalein 84) TietzM ml. 0.05 N NaOH/6 hr./ml. 100 cf0.41 !
P 0.45 « 1 . 0 ) ! 5'-Nucleotidase Substrate: 5'-AMP85) Dixon 85) mg. phosphate/hr./lOO ml.
pH 7.5 j 0 - 1 . 6
5'-Nucleotidase
Substrate: 5'-AMP;
EDTA added 86) Young*
6
) mg. phosphate/hr./lOO ml.;
pH 7.5 30 0 . 3 - 3 . 2
Ornithine carbamyl- i transferase (OCT)
Citrulline est. 88) Brown **) u.moles citrulline/15 min./ml.;
37°C 70 0.22 ( 0 . 1 2 - 0 . 3 0 ) i
1
1
Phosphofructokinase UV-Assay 24) Merten*
1
^ (jimoles/min./l 000 ml. 30 not detectable
6-Phosphogluconic dehydrogenase (6-PG-DH)
UV-Assay 26) Wolfson 20) mu.moles/hr./ml.
25° C 16 cT
1 0 $
137 ( 8 0 - 1 8 8 ) 137 ( 4 0 - 2 3 8 ) 6-Phosphogluconic
dehydrogenase
(6-PG-DH) UV-Assay »9)
modified by 20)
Schmidt
61
^ Biicher units/ml.
pH 7.5; 25°C
17 < 0 . 0 6 Phosphoglycerate
kinase (PGK)
UV-Assay90) Engelhardt-GolkelM
Biicher units/ml.
room temperature
107 0.61 ± 0 . 1 7
Phosphoglycerate mutase (PGM)
UV-Assay 90 Merten
61)
[i.moles/min./l 000 ml. 30 not detectable Phosphohexose mutase Estimation of G-6-P92) Bodanski
92
) 100 u-moles G-6-P/4 hr./ml.
pH 7.6; 37°C
19 46 ( 1 9 - 8 4 ) ± 1 7
Phosphohexose mutase
Estimation of G-6-P + F-6-P93)
Noltmann
9
*) umoles G-6-P + F-6-P/
hr./ml.; 37°C
10 0.5 ( 0 . 1 - 1 . 4 ) Phosphorylase Estimation of Pi from
G-1-P94); modified
Merten
61
) fxmoles/min./l 000 ml. 30 not detectable
Polyphenol oxidase 95) Markowitz
96
) [jimoles 02/hr./ml. 10 3 . 9 ( 3 . 0 - 4 . 7 ) ± 0 . 7
H. Koide and T. Oda, Clin. chim. Acta 4, 554 [1959].
W. G. Fishman, B. Springer and R. Brunetti, J. biol. Chemistry 173, 449 [1948].
/. M. Arias, B. A. Lowry and /. M. London, J. clin. Invest. 37, 875 [1958].
H. D. Horn and F. H. Bruns, Biochem. Z. 331, 58 [1958].
E. Racker in S. P. Colowick and N. O. Kaplan: Methods in Enzymology. Academic Press, N e w York 1955, Vol. II, p. 722.
C. Manso and F. Wroblewski, J. clin. Invest. 37, 214 [1958].
8 1
) / . Trautschold, Dissertation, Universitat Munchen, 1956.
N. P. Goldstein and J. H. Roe, J. Lab. clin. Med. 28, 1368 [1943].
D. H. Adams and V. P. Whittaker, Biochem. J. 44, 62 [1949].
N. W. Tietz, T. Borden and H. D. Stepleton, Amer. J. clin. Pathol. 31, 148 [1959].
Th. F. Dixon and M. Purdom, J. clin. Pathol. 7, 341 [1954].
J. Young, Ann. N . Y. Acad. Sci. 75, 357 [1958].
H. Reichard and P. Reichard, J. Lab. clin. Med. 52, 709 [1958].
R. W. Brown and S. Grisolia, J. Lab. clin. Med. 54, 617 [1959].
B. L. Horecker and P. Z. Smirniotis in S. P. Colowick and TV. O. Kaplan: Methods in Enzymology. Academic Press, N e w York 1955, Vol. I, p. 323.
Th. Biicher in S. P. Colowick and N. O. Kaplan: Methods in Enzymology. Academic Press, N e w York 1955, Vol. I, p. 415.
E. W. Sutherland, T. Pasternak and C. F. Cori, J. biol. Chemistry 181, 153 [1949].
O. Bodanski, Cancer 10, 859 [1957].
E. Noltmann and F. H. Bruns, Hoppe-Seylers Z. physiol. Chem. 313, 194 [1958].
G. T. Cori, B. Illingworth and P. J. Keller in S. P. Colowick and N. O. Kaplan: Methods in Enzymology. Academic Press, N e w York 1955, Vol. I., p.200.
C. G. Holmberg and C. B. Lowell, Acta chem. scand. 5, 476 [1951].
H. Markowitz, C. J. Gubler, J. P. Mahoney, G. E. Cartwright and M. W. Wintrobe, J. clin. Invest. 34, 1498 [1955].
Table 7, Part 2 (cont.)
Enzyme M e t h o d A u t h o r Units N u m b e r of cases N o r m a l values SD
Pyruvic kinase (PK) UV-Assay 3), modified by5>20)
Schmidt*^ Bucher units/ml.
p H 7.5; 2 5 ° C
19 0 . 9 0 ( 0 . 2 1 - 1 . 9 0 ) U V - A s s a y 9 7 ) van
Rymenant
9
®
Wroblewski units/ml. 20 62.3 ( 3 2 . 8 - 1 0 3 . 3 ) Ribose-5-phosphate
isomerase
Ru-5-P est. 99) BrunsW) (i.moles/hr./ml.
37° C
21 3.5 ( 2 . 0 - 5 . 4 ) Succinic dehydro
genase measurement at 400 m(i.ioo) Red. of K3F e ( C N )6, Engelhardt-Gdlkel**)
Bucher units/ml. not detectable
Transketolase S-7-P est. ioi) Bruns [xmoles/hr./ml.;
37°C
15 0.049 ( 0 . 0 2 3 - 0 . 0 8 5 )
Tributyrinase Manometric Hauss 102) mm3 CO2/hr./0.04 ml. 29 88.5 ± 1 7 . 0
Triosephosphate isomerase (TIM)
UV-Assay 103) Merten
61
^ u.moles/min./l 000 ml. 30 42.8 ± 1 5 . 0
Tripeptidase Substrate: Gly-gly-gly 104) Fleisher
3
*) [xmoles/hr./ml. 39 3.88 ± 0 . 2 5
97) E. Negelein quoted by F. Kubowitz and P. Ott, Biochem. Z. 317, 193 [1944].
98) M. van Rymenant and / . Robert, Cancer 72, 1087 [1959].
99) F. H. Bruns, Biochem. Z. 327, 523 [1956].
io°) E. C. Slater and W. D. Bonner, Biochem. J. 52, 185 [1952].
i°i) F. H. Bruns, H. Dunwald and E. Noltmann, Biochem. Z. 330, 497 [1958].
102) w. Hauss and H. /. Leppelmann, Klin. Wschr. 35, 65 [1957].
1 Q
3 ) G. Beisenherz in S. P. Colowick and TV. O. Kaplan: Methods in Enzymology. Academic Press, N e w York 1955, Vol. I, p. 387.
i°
4
> G. A. Fleisher, Arch. Biochem. 61, 119 [1956].