July, 1946
COMMUNICATIONS TO T H E EDITOR
1391
OXIDATION OF LIGNIN SULFONIC ACIDS BY lyst at atmospheric pressure. About one molar PERIODIC ACID equivalent of hydrogen was absorbed. The product obtained by drying the filtered solution from Sir: I t doubtless has occurred to niany investigators the frozen state was a white granular solid which showed ( c Y ) ? ~ D-8S.7O ( 6 , 1.0 in water), and had that the degradation of cellulose in wood by a n activity of about 750 units/mg. as compared periodic acid oxidation might provide a tnild and facile method of obtainini lignin, provided that with SO0 units/mg. for streptomycin. Dihydrostreptomycin trihydroehloride was con- ligneous substances are not attacked by the reverted to the trihelianthate as described for strep- agent. Wald, Ritchie and Purves reported' the toniycin.' A sample of the trihelianthate after isolation of lignin by the action of periodic acid, 14:e are moved therefore to make a preliminary recrystallization three times melted at 215-225' idee.), artivity about 400 units/mg. A n d . report on our study of periodic acid cisitlation of lignin sulfonic acids and other isolated lignins Calcd. for C21H41ILT,i)12(C14H16Na03S)3:C, ~ 50.4ti; H, d.79; N , 14.94. Found: C, 50.14; H, 5.P3; in progress in this Laboratory for more than 3 year. Periodic acid has been found to oxidize N, 15.08. Conversion of dihydrostreptomycin trihelian- lignin sulfonic acids, including samples scruputhate to the trihydrochloride as described for lously freed of carbohydrate material originating streptomycin' gave a white powder, m. p. 185- from the wood pulping process. The purification 190' (dcc.), ( c Y ) ? ~ D -S9.5' (c, 0.98 in water), processes employed to remove c:irboh?-drates are : sctivi ty about 730 units/nig. Potentiometric (a) icrrnentation with yeast; ( b j diffusion of iertitration of this sample gave an equivalent weight nieiitecl sulfite waste liquor in S orthup-Anson of ti!)O; calcd. niol. wt. (i93; ~ K '7.75. A Arrnd. type sintered glass diffusiun cells for sixty-two Calcd. for C21f141N~012.3HC1: C, 36.40; €1, fi.10; c1aj.s; (e) preparatioii of I ) a r i i t i r i ligno-sulfonate N,11.1.5. Found: C,30..50; H,G.21; N , 12.01. soluble i n 40To acetone-water solution, insoluble The presence of :L free or potential c:trbonyl in 70% acctone-mater wlution ; (d) dialysis clf group in streptomycin was denionstrated by the fermented sulfitc waste liquor against running formation of an oxime and semicarbazc~ne.~water for one hundred and sixty-eight hours2; Streptomycin was inactivated3 by hydroxylamine (e) purification by a quinoline extraction iiiethod, in xqueous pyricline solution a t pH 4. Dihydro- precipitation from quinoline solution of quinoliiistrc.1)tornycin is riot inactivated by hydroxylamine ium ligno-sulfonates by the addition of ctlicr : ~ i i t i uritler these contlitiotis, which is evidence that the re-solution of the quinolinium salts in aqucou:; carhonyl group in the streptobiosaiiiine moiety is arnnioniuin hydroside. Table I lists the various prel)arations .iiid the, the functicmal group which was reduced. Acid hyrlrolq-sis of dihydrostreptomycin yields strepti- equivalent weights nf lignin sulfonic acids pcr (line; hence, the reduction involves only the mole of periodate reduced. streptobiosamine moiety. 'rAIiLE I Dihydrostreptclm7:cin is not inactivated by % Slethoxyl content Equiv. w t irnole o f Preparation of ammoninrn salt periodate reduced cystcine ruder conditions* which cause the inacti1 8 ..-J vation of streptoriiyciri : ilius, it nplwars that a 1, 10.1 niechanisii; involving tlic rc~aclionof the carbonyl < 9.7 group with the ariiirio and/or mcrcapto groups of (1 13 0 cysteine is involvcd i n the activation. c 11.9 r>ihydrostreptoiiiyciii is not tlcgradecl to inalto16 when treated with alkali. It is apparent that preparations of low methox!-1 Tests by Dr. 1%.Robinson arid J'Ir. 0 . Graessle content contain extraneous periodate-reducing rjf the Merck Institute for Therapeutic Research substances, probably carhohytlrates ; but as rehave shown that single doses of 85 units of di- finement improves, the approach of the methoxyl hydrostrep trrmycin trihydrochloride as contrasted content to a limiting viilue is accoinpanied by ;I with 45 units of streptomycin trihydrochloride- similar constancy iii the periodate rquivalent calcium chlnritle tioithlc snlt were required to pro- weight. We believe s'iuip1t.s (d) anti ( e ) to be tcct o f t h e ?riicc agaiiist oiie lethal close of substantially free of carbohydrate materid. The Si1uz(171 t'llu si-/Ioftn:u Ile vi. equivalent weights of 52,; for preparations ( d ) K U U I R L. P E C K arid (e) correspond approximately to a ratio of CHARLES E. t f O P F I i I Y E , JR. two methoxyt groups for each linkage oxidizable l
