Organic Chemistry
IX. Distribution of electrolytes between trans
udates and serum. A. B. Ha s t i n g s, H . A. Sa l- v e s e n, J. Se n d r o y, and D. D. Va n Sl y k e (J. Gen.
Physiol., 1927, 8, 701— 711).— The electrolyte con
tent of serum transudates differs from that of the serum itself in a maimer explicable on the assumption of the existence of the Doiman membrane equilibrium between the two fluids. When serum and salt solutions approximating to oedema fluid in composition are equilibrated across a collodion membrane, the
distribution of electrolytes becomes similar to that between serum and oedema fluid in vivo.
W . O. Ke r m a c k.
Blood-sugar of cod, sculpin, and pollock during- asphyxia. M. L. Me n t e n (J. Biol. Chem., 1927, 72, 249— 253).— The normal fasting blood-sugar concentration of the above fish was from 0-014 to 0-04% ; both under normal and asphyxial conditions, however, this value was greatly influenced by the previous feeding. C. R. Ha r i n g t o n.
Micro-determination of sugar and reducing substances in blood. V. La p a (Bull. Soc. Chim.
biol., 1927, 9, 310— 323).— Treatment of blood or plasma with mercuric acetate in presence of sodium hydrogen carbonate precipitates, not only proteins, but also all nitrogenous components. The reducing power may then be determined by any of the ordinary micro-methods. The reducing power after ferment
ation is independent of the method used and varies from 3 to 6 mg. (as dextrose) per litre.
W . O. Ke r m a c k.
Determination of small quantities of sugar.
Application to blood and to other body-fluids.
A. Ba u d o u i n and J. Le w i n (Bull. Soc. Chim. biol., 1927, 9, 280— 309).— To 15 c.c. of the neutral solu
tion to be examined 1 c.c. of a solution containing 3-60 g. of mercuric iodide and 12 g. of sodium iodide per 100 c.c. and 1 c.c. of iV-sodium hydroxide solution are added. The mixture is heated for 3 min. on the boiling water-bath and then cooled, and there is then added 1 c.c. of a solution containing 1-783 g. of potass
ium iodate dissolved in sufficient 5% sulphuric acid to make the volume to 1 litre. The iodine liberated converts the mercury formed during the heating into mercuric iodide, and the excess of iodine is then titrated with sodium thiosulphate.
W . 0 . Ke r m a c k.
Blood-sugar. Fractionation of the reducing"
substances in blood-filtrates. B. Sj o l l e m a (Bio- chem. Z., 1927, 182, 453—459).— Blood-filtrates- obtained by the method of Folin and Wu contain, besides dextrose, an amount of reducing substance much greater than the content of reducing non
sugar substances such as uric acid. A method is- described for the determination of dextrose in presence of very small amounts of disaccharides (lactose, maltose). When such sugar solutions are shaken with animal charcoal and a little acetic acid is added,- dextrose remains in solution, whereas the disaccharide is almost completely adsorbed. If, in place of acetic acid, the solution is shaken with ether, the disaccharide remains in solution. P. W . Cl u t t e r b u c k.
Analytical method of establishing the 11 nitro
gen formula ” for blood-serum. M . La u d a t
(Bull. Soc. Chim. biol., 1927, 9, 137— 14S).— Detailed methods are described for establishing the “ nitrogen formula ” (cf. Widal and Laudat, Compt. rend. Soc.
Biol., 1926, 95, 1233), the determinations including:
those for total non-protein nitrogen, carbamide, uric acid, creatine, creatinine, ammonia, and amino- acids, and the undetermined nitrogen. Well-known methods, with certain modifications, are recom
mended, and this method of analysis has proved.
useful in studying the retention of nitrogenous
sub-BIOCHEMISTRY. 477
stances in renal impermeability. The amount of serum required for the complete analysis is not greater
than 15 c.c. A. Wo r m a l l.
Thiasine ; its structure and identification with ergothioneine. E. B . Ne w t o n, S. R. Be n e d i c t,
and H . D . Da k i n (J. Biol. Chem., 1927, 7 2 , 367— 373).
—Thiasine (A., 1926, 421) gives the Pauly reaction, yields trimethylamine on warming with potassium hydroxide, and reduces permanganate in the cold;
the sulphur is readily removed by oxidation with ferric chloride, although it is stable towards alkali.
Re-determination of the sulphur content indicated 13-4%; the substance was therefore suspected to be identical with ergothioneine (cf. Tanret, A., 1909, i, 671), and the identity was confirmed by direct com
parison, and by the identity of the behaviour of the substance with that described by Barger and Ewins (J.C.S., 1911, 99, 2336) for ergothioneine.
The compound (which it is proposed to name thioneine) has been isolated also from human blood (cf. Eagles and Johnson, this vol., 369). C. R . Ha r i n g t o n.
Non-protein sulphur compounds of blood. I.
Sympectothion. G. Hu n t e r and B. A. Ea g l e s (J.
Biol. Chem., 1927, 7 2 , 123— 132).—Modifications are described in the method of isolation of the compound previously obtained by the authors (A., 1926, 85) from b lood ; this compound is now shown to contain sulphur, and is named sympectothion, the formula suggested being C18H320 6NgS2; it has [ajg8 +115°, the previously recorded value being in error. The possible relationship of the compound to the thiasine of Benedict and others (A., 1926, 421) is discussed (cf. preceding abstract). C. R . Ha r i n g t o n.
Non-protein sulphur compounds of blood. II.
Glutathione. G. Hu n t e r and B. A. Ea g l e s (J.
Biol. Chem., 1927, 7 2 , 133— 146).— A substance apparently identical %vith glutathione has been isolated from blood-corpuscles b y a modification of the original method of Hopkins (A., 1921, i, 635);
colorimetric determinations indicate the presence of about 100 mg. of this substance per 100 c.c. of red corpuscles. C. R. Ha r i n g t o n.
Measurement of hsemolysin. C . B . Co u l t e r
(J. Gen. Physiol., 1927, 10, 541— 544).— A method is described for the measurement of hsemolysin con
centration, the principle of which is to compare under standard conditions the percentage of haemolysis produced by a given amount of the unknown with that produced by varying amounts of a standard haanolysin solution. W . O. Ke r m a c k.
Protein associated with hsemolysin in rabbit- seruna and -plasma. C . B. Co u l t e r (J. Gen.
Physiol., 1927, 10, 545— 550).— The hsemolysin of immune serum is associated with the water-insoluble puglobulin. Its activity is retarded by the presence in the solution of fibrinogen. Hsemolysis and hsemag- glutinin appear to be distinct entities, since the activity of the former is destroyed, and that of the latter apparently increased by extraction with alcohol.
W . O. Ke r m a c k.
Haemolysis by the photo-sensitising action of hsematoporphyrin. R. Fa b r e and H. Si m o n n e t
(Compt. rend., 1927, 184, 707— 709).— Lecithin freed
from cholesterol by the method previously described (A., 1926,1283), when irradiated in presence of hsemato- porphyrin by means of a mercury-vapour lamp for 60 min., acquires haemolytic powers towards the red cells of the dog. Partial separation of the hsemo- lysing product has been effected by alcohol-ether fractionation. R. Br i g h t m a n.
Unsealed fibres. A. T. Ki n g (Biochem. J., 1927,
2 1 , 434— 436).— A method has been devised for the mechanical removal of the scales or outer layer of wool fibres in such a way as to leave the cortex intact.
It consists essentially in drawing the fibre across the edge of a suitable glass plate, or over a series of such plates. The effectiveness of the method is illustrated by photomicrographs of wool fibres in various stages of scale removal. S. S. Zi l v a.
Distribution of unsaturated fatty acids in tissues. II. Voluntary muscle of the ox. W . R.
Bl o o r (J. Biol. Chem., 1927, 7 2 , 327— 343).— Figures are given for the distribution of lipins in various voluntary muscles and in the heart-muscle of the ox.
The content of phospholipins, unsaponifiable matter, and cholesterol was highest in the most active muscles;
a similar relationship was not observed with respect to the glycerides. Cholesterol esters are often absent from muscle, and, when present, are found in small amounts only. Emphasis is laid on the relationship of the phospholipin content to the mitochondria of muscle-ceOs, and the probable role of the phos- pholipins as constant constituents of protoplasm is discussed. C. R . Ha r i n g t o n.
Ovary. X II. Fatty acids of lecithin from corpus luteum. M. C. Ha r t and F. W . He y l (J.
Biol. Chem., 1927, 7 2 ,395— 402; cf. this vol., 69,168).
— The lecithin-cadmium chloride compound obtained from the acetone extract of corpus luteum yielded, on hydrolysis, 49-4% of fatty acids, of which 17%
was palmitic, 26% stearic, 22% oleic, 26% linoleic, and 7% arachidonic acid; there was also obtained 2% of an acid, 02^ 3402, the presence of which had been previously observed in the neutral fat fraction, and for which the name ovarenic acid is now proposed.
The above figures are similar to those previously obtained for the neutral fat, which supports the hypo
thesis that a function of the corpus luteum is to main
tain a supply of the labile phospholipins which are required during pregnancy. C. R. Ha r i n g t o n.
Structure of the histone of the thymus. III.
Acid- and base-binding power after peptic digestion. K . Fe l i x and A. Ha r t e n e c k (Z. physiol.
Chem., 1927, 1 6 5 , 103— 120).— Peptic digestion of the histone of the thymus causes an equal increase in acidic and basic groups and an equal increase in free amino-nitrogen and in formaldehyde-titratable carboxyl groups. The acidic groups liberated appear to be entirely carboxyl groups; the basic non-amino- groups set free probably belong to the guanidine group of arginine; this hypothesis is supported by the low figure for the increase in the carboxyl groups titratable in alcohol compared with the increase in base-binding capacity. No ammonia could be detected. Pepsin, therefore, apparently splits acid- basic linkings. After preliminary digestion of the
histone with trypsin, pepsin caused no further increase iii acid- or base-binding power, but the nitrogen which could be methylated increased in amount.
C. R. Ha r i n g t o n.
Crystals of Charcot, Leyden, Bôttcher, and Neumann. F . W r e d e , F . Bo lt, and E . Bu c h (Z.
physiol. Chem., 1927, 1 6 5 , 155— 166).— The crystals described by the above workers (Compt. rend., 1853, 3 ; Arch. Anat. und Physiol., 1872, 5 4 , 324; ibid., 1865, 3 2 , 525; Arch. mikr. Anat., 1866, 2 , 508) were almost certainly those of spermine phosphate.
C. R. Ha r i n gt o n.
Glutathione. G. Hu n t e r and B. A. Ea g l e s (J.
Biol. Chem., 1927, 7 2 , 147— 166).— B y a method similar in principle to that of Hopkins (A., 1921, i, 635), there was isolated from yeast and from liver a sulphur-containing peptide which is regarded as glutathione ; the product from liver was contaminated with cystine, but was otherwise similar to that from yeast and from blood (cf. this vol., 477). The material agreed with that of Hopkins (loc. cit.) and with the compound synthesised by Stewart and Tunnicliffe (A., 1925, i, 795) in its optical rotation, but differed from it in possessing a higher content of nitrogen and a lower content of sulphur. The preparations contained total N 11-5— 12-0, amino-N 7-8— 7-9, S 8-85— 10-27%. On hydrolysis, the amino- nitrogen was increased by one half. The optical rotation after reduction was [a]0 -4 -7 4 ° ; it is thought that the completely reduced substance may be dextro
rotatory. On the basis of the above results, the authors deduce the presence of a third amino-acid ( ? serine) in ester combination in the glutathione molecule; the direct evidence adduced by Hopkins (loc. cit.) for the constitution of glutathione is ques
tioned on the ground of the present authors’ inability to effect a sharp separation of glutamic acid hydro
chloride from cystine dihydrochloride after hydrolysis of their products. C. R. Ha r i n g t o n.
Isolation of glutathione. F. G. Ho p k i n s (J.
Biol. Chem., 1927, 7 2 ,185— 187).— Hunter and Eagles (cf. preceding abstract) have omitted to discuss the evidence in favour of the accepted constitution of glutathione which is afforded b y the synthesis of the compound by Stewart and Tunnicliffe (A., 1925, i, 795) ; the sulphur in glutathione is very much more labile than that in cystine ; loss of sulphur at some stage in the isolation may therefore explain the results of Hunter and Eagles. C. R. Ha r i n g t o n.
Colorimetric determination of cystine and glutathione. G. Hu n t e r and B. A. Ea g l e s (J.
Biol. Chem., 1927, 72, 177— 183).— The method of Folin and Looney (A., 1922, ii, 539) for the colori
metric determination of cystine has been modified by the substitution of sodium hydroxide for sodium carbonate as the necessary alkali ; the modified method is applicable also to the determination of glutathione, which yields 0-464 times the depth of colour given by the same weight of cystine. C. R . Ha r i n g t o n.
Cystine in liver. G. Hu n t e r and B. A . Ea g l e s
(J. Biol. Chem., 1927, 7 2 , 167— 175).— Preparations of glutathione obtained from liver showed a high content of sulphur and a high specific rotation ; this was found to be due to contamination with free
cystine, which must therefore occur as such in the liver. The possible relationship of this observation to the physiological synthesis of taurocholic acid, and to the occurrence of cystinuria, is discussed.
C. R. Ha r i n g t o n.
Micro-volumetric determination of sulphur in biological fluids. B. Po h o r e c k a- Le l e s z (Bull.
Soc. Chim. biol., 1927, 9 , 263— 276).— Sulphate is precipitated as benzidine sulphate which is titrated in boiling distilled water, in presence of phenol-red, with 0-02/Y-sodium hydroxide solution. The method may be applied for the determination of sulphate in body-fiuids, only 0-5 c.c. of urine or 5 c.c. of serum being required, and also for the determination of total sulphur in such fluids if the sulphur is first oxidised to sulphate. W. O. Ke r m a c k.
Normal excretion of zinc in urine and faeces of man. K. R. Dr i n k e r, J. W. Fe h n e l, and M.
Ma r s h (J. Biol. Chem., 1927, 72, 375— 383).— The average daily excretion of zinc by the adult human is 0-89 mg. in the urine and 9-8 mg. in the faeces.
Ingestion of a single meal rich in zinc causes an immediate and great increase in the faecal excretion of zinc, which may persist for 2— 3 days; the urinary excretion of zinc is not perceptibly affected.
C . R. Ha r i n g t o n.
Micro-determination of ammonia in urine. J.
W e b e r and W . Kr a n e (Z . physiol. Chem., 1927,165,
45— 52).— The ammonia is precipitated with Nessler’s reagent or with sodium cobaltinitrite and alcoiol;
the ammonia is liberated from the precipitate, in the first case with sodium hydroxide and potassium sulphide, and in the second case with sodium hydroxide alone, steam-distilled into excess of standard acid, and determined by titration. The method is applic
able to quantities of ammonia of 0-5 mg. and upwards.
C. R . Ha r i n g t o n.
Determination of oxalic acid in urine by means of the rocking-extraction method of Widmark. C.
G. Ho l m b e r g (Biochem. Z ., 1927,182, 463— 469).—
Widmark’s method (Skand. Arch. Physiol., 1926, 84, 61) for the determination of urinary benzoic acid is adapted to the determination of oxalic acid in urine.
P. W . Cl u t t e r b u c k.
Effect of light on uroporphyrin. B. T. S q u i r e s
(Biochem. J., 1927, 2 1 , 437— 440).— Over the pn range 6-2—8-2, there is a differential fading which reaches a maximum at about pa 7-3. The rate of the general fading o f uroporphyrin solutions is greater in the case of those containing crude uroporphyrin
than in those containing the purified compound.
The fading of crude uroporphyrin is in part associated with the presence of oxygen; this is not the case with pure uroporphyrin. As uroporphyrin solutions fade, there is no’ change in the positions of the absorption bands, which simply fade away. Within the above range of pa, the appearance of “ Kalilichtreaktion ” band described by Schumm at 461 ¡xu. could not be detected. On exposure of crude or purified uropor
phyrin solutions in quartz tubes to ultra-violet light, a general fading irrespective of pa is obtained.
S. S. Zi l v a.
Iron in nutrition. IV. Correction of nutri
tional anaemia with ash of plants and animal
BIOCHEMISTRY. 479
tissues and with soluble iron salts. E . B. Ha r t,
C. A. El v e j h e m, J. Wa d d e l l, and R. C. He r r in
(J. Biol. Chem., 1927, 72, 299— 320; cf. A., 1925, i, 1354).— The nutritional anaemia which occurs in young rats on a diet of whole milk and ferric oxide can be cured by the addition of the ash, or an alcoholic extract, of lettuce or cabbage; spleen marrow and an alcoholic extract of maize were efficient supple
ments, but the ash of these substances was not so.
Pure ferrous sulphate had no effect, although a com
mercial sample of the salt cured the anaemia. The supplementing factor is thus apparently of an inor
ganic character, but is not connected with the solu
bility of the iron compounds administered (cf. Mitchell and Schmidt, A., 1926, 1269). C. R . Ha r i n g t o n.
Action of mineral waters on carbohydrate metabolism. Experiments [on diabetics]. 0 .
Ka u f f m a n n-Co s l a, R . Zo r k e n d o r f e r, and W . Zo r k e n d o r f e r (Bull. Soc. Chim. biol., 1927, 9 ,
174— 202).— 1000— 1500 G. per diem of the natural mineral waters of Carlsbad and Marienbad, of which the principal constituent is sodium sulphate, bring about the recovery of mild and moderately severe
diabetics. A. W o r m a l l .
Oxidative processes in the living cell. A.
Op a r i n (Biochem. Z., 1927, 1 8 2 , 155— 179).— An investigation of the aerobic oxidation of glycine under the influence of the plant respiratory chromogen chlorogenic acid, a didepside of caffeic and quinic acids (A., 1922, i, 309). It is concluded that in the living cell the chromogen is oxidised by molecular oxygen and phenoloxydase to a pigment, and that the pigment is then reduced by oxido-reductase to the original chromogen, the necessary hydrogen being derived from water, whilst the hydroxyl remains for the actual oxidation. The oxidation and reduc
tion of the chromogen are normally in equilibrium.
A high oxygen uptake by the living cell, following chemical or mechanical injury, leads to deep-seated oxidation of the respiratory chromogen, with the formation of an inactive pigment, and oxidation within the cell ceases. J. Pr y d e.
Metabolism of tissues growing in vitro. I.
Ammonia and urea production by kidney. B. E.
Ho l m e s and E. Wa t c h o r n (Biochem. J., 1927,
21, 325— 334).— Growing embryonic rat kidney-tissue produces considerable amounts of ammonia and urea in contrast to “ resting ” embryonic kidney-tissue, i.e., living but non-proliferating tissue, which produces neither ammonia nor urea. Preliminary experiments with brain-tissue indicate the probable existence of urease in the rat brain. S. S. Zi l v a.
Brain metabolism. IV. Carbohydrate meta
bolism of brain-tissue of depancreatised cats.
B.-E. Ho l m e s and E. G. Ho l m e s (Biochem. J., 1927, 21, 412— 418).— The lactic acid values of the brain fall and rise with the blood-sugar. Hyperglycsemia brought about by the administration of anaesthetics is also accompanied by an increased brain lactic acid content. Similarly, the hvperglycaemia of depancreatised cats is responsible for a correspondingly higher value of resting lactic acid in the brain of the cat. The brain-tissue of diabetics, like that of normal
animals, is capable of converting dextrose into lactic acid and of removing lactic acid under aerobic con
ditions. The lactic acid of the brain is therefore formed from the dextrose supplied by the blood.
S. S. Zi l v a.
p a of muscles of marine animals. K . F u r t j - s a w a and P. M . T. K e r r i d g e (J . Marine Biol. Assoc., 1927, 1 4 , 657— 659).— The average paof the muscles of various marine animals was 7-06 in the resting condition and 6-33 in rigor; these figures are in close agreement with those obtained for the muscles of the frog and the cat. C. R. H a r i n g t o n .
P o s t-m o r tem changes in the free sugar, glycogen, phosphates, and lactic acid in m am malian muscle. W . W . Su i t so n and J. J. R.
Ma c l e o d (Trans. R oy. Soc. Canada, 1926, [iii], 2 0 ,
V, 371— 375).— Experiments have been carried out to determine the total reducing substances, the free sugar, and the glycogen content of extracts of the liver and muscle of standard white rats, previously starved for 24 hrs. The free sugar, which accounts for a small percentage only of the total reducing value, is much lower in the liver extracts of insulin- treated rats than with the normals. Insulin treat
ment causes little, if any, change in the glycogen content or total reducing value. W ith muscle- extract, on the contrary, there is no reduction in free sugar following insulin. Changes taking place in the glycogen, free sugar, lactic acid, and soluble phosphorus of frozen rabbit muscle, after various short periods of thawing, indicate that some polymerised carbohydrate (perhaps a lower dextrin) must be formed from the disappearing glycogen. A. Wo r m a l l.
Acid formation in thiocyanate rigor in frog’s muscle. G. E. Se l t e r (Z. physiol. Chem., 1927,
1 6 5 , 18— 27).— In thiocyanate rigor in frog’s muscle there is a marked increase in lactic acid and no con
stant change in the phosphoric acid. The phenomenon is therefore probably due to increased acidity.
C. R. Ha r i n g t o n.
Significance of ions in muscular function.
IX . Influence of different anions on lactic acid formation and phosphoric acid exchange in minced muscle. G. E . Se l t e r (Z . physiol. Chem., 1927,1 6 5 ,1— 17).— The effects of additions of various sodium salts to minced muscle on the formation of lactic acid and the amount of free phosphoric acid vary greatly (they may, indeed, be in the reverse direction) according to the concentration of salt employed. A salt which increases the rate of dis
appearance of phosphoric acid does not necessarily diminish the formation of lactic acid. Complete inhibition of lactic acid formation is obtained only in presence of those anions (fluoride and oxalate) which entirely suppress the degradation of lactacid- ogen. In view of these results, it is unsafe to deduce the behaviour of the lactic acid, under any given conditions, from the observed behaviour of the phosphoric acid. C. R . Ha r i n g t o n.
Degradation of glycogen in muscle. H . v o n Eu l e r, R . Ni l s s o n, and B. Ja n s s o n (Z. physiol.
Chem., 1927, 1 6 5 , 121— 129).— The decolorisation of methylene-blue b y muscle is influenced to an equal extent by sodium and potassium phosphates. The
rate of decolorisation was not definitely affected by the state of nutrition of the animal; the process of reduction is accelerated to a slight extent by addition of small concentrations of glycogen; it is inhibited by high concentrations of glycogen and by all con
centrations of dextrose; in the case of yeast, the reduction is not accelerated by glycogen.
C. R. Ha r i n g t o n.
Carbohydrate utilisation. II. Rate of dis
appearance of carbohydrates from the blood.
J. G. Re i n h o l d and W . G. Ka r r (J. Biol. Chern., 1927, 72, 345— 365).— Oral administration to rabbits of galactose, maltose, sucrose, starch, lactose, and
J. G. Re i n h o l d and W . G. Ka r r (J. Biol. Chern., 1927, 72, 345— 365).— Oral administration to rabbits of galactose, maltose, sucrose, starch, lactose, and