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COMMUNICATIONS TO THE EDITOR
Vol. 82
hin~kiflavone,~ sotetsuflavone pentaethyl ether produces 2,4-diethoxy-6-hydroxyacetophenone,p ethoxybenzoic acid, and a phenolic acid (V), m.p. 257-258" (found: C, 63.80; HI G.02. Calcd. for CzoHz?07: C, 64.16;H, 5.92). V is not identical, by admixture, with a formerly reported acid (VI)'j, m.p. 258-259O, similarly obtained from sciadopitysin triethyl ether, and gives a monoketodicarboxylic acid (VII), m.p. 245-249' (dec.) (found: C, 55.79; H, 4.25. Calcd. for CllHloOs: C, 55.40; H, 4.23) on oxidation with alkaline potassium permanganate solution. VI1 was identified with a synthetic sample, which was newly prepared by the mild oxidation of a condensation product (VIII),6 through admixtures of itself and of its 2,4-dinitrophenylhydrazone, CliH140&J4, m.p. 239241".
ceed very rapidly, in a second or two the systern approaches closely the state of equilibrium if the viscosity of the solution is not too high. A polystyrene endowed with the -CH(Ph) -, Li+ end-groups has been prepared recently by polymerizing styrene in T H F with lithium naphthalene. The initially fluid solution becomes gradually more and more viscous and a gel is formed after a day or two if the concentration is sufficiently high. The following experiment proved that this phenomenon is not due to a slow polymerization. A T H F solution of polystyrene polymerized by lithium naphthalene was divided into three portions. By addition of water the organolithium compound was destroyed in the first portion shortly after its preparation, and at that time the solution was quite fluid. The second portion was kept at room ternperature for 16 hours and then, when it was viscous, water was added. Finally, the last portion was kept for 40 hours and it was very viscous when the organolithium compound was destroyed by addition of water. iZfter precipitation of the respective polymers it was shown that the polymerization was conipleted in each portion, and the intrinsic viscosities of the resulting polymers were identical, [q] = 0.130 in units g./100 cc. The slowness of the association is astonishing and suggests that the process requires activation energy. This indeed seems to be the case. -4 solution of polystyrene polymerized by lithium naphthalene I I was divided into two portions. One was stored a t -SO", while the second was introduced into a This degradative evidence leads to the assign- viscosimeter ( a falling weight type). The time of ment of structure IV for sotetsuflavone. There- falling slowly increased, being initially 9 sec., then fore, a ketoflavone obtained by the hydrolysis of 20 sec. after 12 hours, 40 sec. after 24 hours, and sotetsuflavone must be represented by formula ( I X ) eventually 250 sec. after 48 hours. At that time instead of 11. the solution was very viscous, while the stored We thank Dr. Kariyone for his encouragement solution still remained fluid in spite of its low tenithroughout this work, which was carried out with perature. On the third day the stored solution the financial support of a Grant-in-Aid for Scientific was brought up to room temperature and then Research from the Ministry of Education (Japan). introduced into the same viscosimeter. Its initial viscosity corresponded again to 9 sec., and over a (4) N. Kawano and Y . Fukui, THISJOURNAL, 81, 6331 (195q) ( 5 ) pi. Kawano and H Miura, Yakrcgaku Zass/zz, 1 9 , 14FS (lq50) period of two days the viscosity increased to about PHARMACEUTICAL FACULTY 250 sec., the rate of increase being similar to that UVIVERSITYOF NAGASAKI l T KAYAVO observed ~ ~ ~ ~ previously. SAGASAKI, JAPAX MITSUTO\*AMADA An explanation of this phenoliicrion is proposed. RECEI~E DECEXBER D 15, 1959 The polymeric ends might exist in 2 forms, e.g., as covalent C-Li bonds and as ion pairs. Since each form represents a stable configuration, transformaASSOCIATION OF POLYMERS THAT REQUIRES tion of one form into the other must require activaACTIVATION ENERGY' tion energy. If the initially formed bond does not Sii. associate while the other does, then the observed It is known that some groups, if present in association would show an apparent activation polymeric molecules, may cause their reversible energy. This explanation is supported by another association in solution. Hydrogen bonding, di- observation, namely, deepening of the color of the pole interactions3 or formation of clusters in poly- solution (from a bright red to a dark brown) with meric salts4 are responsible for these phenomena. increase in the viscosity. Thus, the solution kept For example, polystyrene endowed with the a t -80" remained bright red and fluid; the color N a + end groups changed only after i t was brought to room tempera-COO-, N a + or -CHzO-, associates in T H F solution, while presence of the ture and then left for two days. Alternatively it -CH (Ph) - Na+ end-groups does not produce any is possible t h a t the activation energy is required detectable association? These associations pro- for the reaction 2 Li+ -+ \-C, Li, (1) Research supported by Quartermaster Contract DA19-129-QMC-} -, Li+, and it is hoped that future experi1297, and by the National Science Foundation, G 5914. inents will permit distinction between these alterna( 2 ) Q. A. Trementozzi, R. F. Steincr and P . Doty, THISJ O U R N A L , tives. 1 4 , 2070 (1952). ( 2 ) 11 Urr>d\ n H Kiihard. a d 31 S L h d r C ( I i i n l I ~ l i r s l v y ( 3 ) E . I?. Evans and I€ 11.Spurlin, ibicl , 1 2 , 4750 (1050)
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(4) C . R . Singleterry, J . AJWV.Oil C h e w SCC.,32, 446 (1935).
( I c n d o i a ) , 1-173 (1928)
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March 20, 1960
COMMUNICATIONS TO THE EDITOR
1507
Whatever is the explanation, it seems that this is the first observed case of an association (other
in the general reaction a product identical with the aldahexoses and in the secondary reaction of hexoses than coagulation of electrically charged colloidaI gave a product identical with glucose. A solution particles) that requires an activation energy. of alkaline phosphatase containing 30 pg. N per DEPART~XENT OF CKEMISTRY K. s. DA4 ml. after 2 hr. a t 100’ with 1 N HC1 was found to contain 38 pg. glucose as determined by the reducSTATEUNIVERIITYCOLLEGEOF FORESTRY AT SYRACUSE UNIVERSITY M. FELD ing sugar method as determined by the cysteine SYRACUSE 10, NEW YoRK M. SZWARC reaction on the untreated material. The hydroRECEIVED JANUARY 19, 1960 lysate was found to contain approximately 25 pg. glucose as determined with glucose oxidase.1° The ninhydrin-phosphomolybdate positive GLUCOSE, A CONSTITUENT OF ALKALINE material was found to migrate as a cation a t PH PHOSPHATASE’ Sir: 8 and to be unstable in more alkaline solutions. Alkaline phosphatase of swine kidney, found in Other reactions of the compound were an immediate the ribonucleoprotein particles of the microsomes2 reaction with K3FeCNe and FeCl3 (blue) and an imand released as an active fragment by proteolysis,2r3 mediate reaction with K3FeCN6 alone (blue). has been isolated in an apparently homogeneous Both these reactions were destroyed by previous condition by application of the procedures pre- treatment with traces of cupric ion with exposure viously describedS plus ion-exchange chromatog- to air. In a study of model compounds, pkenolic raphy with Ecteola c e l l ~ l o s e . ~In a typical puri- compounds appear to have been eliminated but fication 2,000,000 units activity2 with a specific similar reactions have been observed with tri- and activity of over 100,000 units per mg. total N tetra-substituted pyrimidines; from the absorb(micro-Kjeldahl) was placed on a column of Ec- ancy in the ultraviolet, from studies of the model teola 5 X 100 cm. and, after thorough washing with pyrimidines and from analogy with the proposed water, was eluted with a gradient of barium acetate structure for vicine,’l i t is suspected that the matea t pH 9 varying from 0.01 to 0.05 iZI in 10 liters of rial may be a diamino-5-hydroxypyrimidine atsolution. The active material was eluted sym- tached, in the active material, to the glucose by a metrically near 0.03 M and was concentrated by glycosidic linkage a t the 5-hydroxy position. the barium procedure3 to yield about 1,500,000 (10) “Glucostat,” Worthington Biochemical Corp , Freehold, h-.J. (11) A. Bendich and G. C. Clements, Biochirn. Btophys. Acta, 12, units material with a specific activity of 295,000 to 462 (1953). 310,000 on the basis of total N. RechromatogDEPARTMEXT OF BIOCHEMISTRY raphy on Ecteola-cellulose, on Deae-cellulose4 FRANCIS BINKLEY or on D0wex-2~~ paper electrophoresis and paper EMORYUNIVERSITY ATLANTA 22, GEORGIA chromatography (ethanol-1 M ammonium aceRECETVED JANUARY 18, 1960 tate, 70-30) revealed no dissociation of absorbancy a t 280 rnp from activity. The material was free of peptidase and diesterase activity when tested unACID (CF,)?POH AND T H E diluted, amino acids were without effect on the T H E PHOSPHINOUS DIPHOSPHOXANE (CF,),POP(CF,,)zI activity and at no time was i t possible to demon- Sir: strate dialyzable cofactors other than magnesium We have recently isolated the new-type comion. The absorbancy in the ultraviolet was characteristic of protein with a maximum a t 278 and a pounds (CF3)zPOH and (CF3)?POP(CF3)zas stable minimum a t 250 mu. However, in the course of liquids contrasting with the apparently complete treatment with dilute acid (0.1 to 1.0 M a t 100”) instability of the corresponding hydrocarbon dethe absorbancy was found to increase remarkably rivatives. Evidently the highly electronegative and, a t the end of 2 hr., the absorbancy a t 278 was CF3 groups lower the power of phosphorus lonenearly tripled. There was a parallel release of pair electrons to bond either H + or (CF3)2P+cornreducing of ninhydrin reactive material,6 ing from 0. Thus these new (CF3)zPcompounds and of material reacting with pho~phomolybdate.~ do not undergo the rearrangements RzPOH -+ H 0 Paper chromatography (propanol-water, 80-20) ~ probably separated a phosphomolybdate and ninhydrin reac- RzPOH and RzPOPRz + R z P - P R which tive material from a ninhydrin negative but aniline represent the first stages of decomposition when R hydrogen phthalate positive8 (brown color) mate- is a hydrocarbon group. Synthesis and Characterization of the Dirial with the same Rfas glucose. The untreated reaction 2(CF3)2PI Agzmaterial in the cysteine methods of Discheg gave phosphoxane.-The COS -+ COZ 2AgI (CF3)2POP(CF3)2(room (1) These studies were supported by grants from t h e U. S. Public temperature, repeated shaking with fresh silver Health Service. Detailed studies of t h e purification, t h e effects of amlno acids and of divalent metal ions will be described b y C. Lea carbonate) gave yields above 79y0. The unused (2) F. Binkley, J. Davenport and F. Eastall, Biochem. Biophys. Re(CFJ2PI (1%) was converted by AgCl to the easily
+
Corn.. . 1.. 206 (1959). . . (3) F. Binkley, V. Alexander, F. E . Bell and C. Lea, J . Bioi. Chem., 228. 559 (1957). (4) E. A. Peterson and H. A. Sober, THIS JOWRNAL, 78, 751 (1956). (5) N. Nelson, J . Bid. Chern., 163, 375 (1944). (6) S. Moore and W. H . Stein, ibid., 176, 367 (1948). ( 7 ) 0. Folin and V. Ciocalteu, ibid., 73, 627 (1927). (8) S. M. Partridge, Nalure, 164, 443 (1949). (9) Z. Dische, in D. Glick, “Methods of Biochemical Analysis,” Vol. 2 , Interscience Publishers, Inc., New York, N. Y., 1935, p. 313 8.
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(1) This research was supported b y t h e United States Air Force under Contract A F 33(616)-5435 (Subcontract No. 1) monitored b y the Materials Laboratory, Wright Air Development Center, WrightPatterson Air Force Base, Ohia. (2) G. M. Kosolapoff, “Organophosphorus Compounds,” John Wiley and Sons, Inc., New York, N. Y., 1950, p. 144. We also have found t h a t reactions expected t o form (CHs)xPOH give nearly quantitative yields of (CH3)sPH and (CH8)zPOOH; and attempts t o make (CHs)2POP(CH3)2also give products suzgesting dis.xoportionation.
