THE PROTEIDS OF THE KIDNEY BEAN

THE PKOTEIDS OF THE KIDNEY BEAN. 757 complex iron, molybdenum, tungsten, and chromium molecules and their union to form a new compound, i. e., the ...
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757

T H E PKOTEIDS O F T H E KIDNEY B E A N .

complex iron, molybdenum, tungsten, and chromium molecules and their union to form a new compound, i. e . , the passing of a heterogeneous system of nlolecules of like elements to a hornogeneous system in the melted condition and again to a heterogeneous system in the resultant alloy ; that these properties indicate a condition in these alloys analogous to an aqueous solution of a salt converted by cold to the solid state. 3. The high melting-point of the element in combination with the iron and its chemical affinity for iron is especially favorable to the separation of alloys from the cooling solution, and is the cause of the heterogeneity of these compounds. This separation of salts, i. e . , segregation, draws the analogy between these alloys and solutions closely. XI. COMPOSITION A.

Alloys.

B.

OF

ALLOYS. C.

D.

E.

...... 0.2979 0.1665 ...... ...... ...... ...... ..................... 0.4646 0.1696 ...... 0.0702 ........................... .............. 0.6626 0.7942 0.5251 0.7832 0.8799 ........ 0.0031 0.0030 0.0004 0.00zs 0.0029 ............. 0.0074 0.0066 0.0014 0.0207 0.0337 . 0.0017 o.00~0 ) o.m,5 { 0.0184 0.0041 0.0082 0.0056 .. 0.0273 0.0277

Molybdenum. Tungsten.. Chromium Iron.. Phosphorus Silicon Combined carbon.. Graphitic carbon.

THE PROTEID5 OF THE KIDNEY BEAN. (PHASEOLUSVULGARIS.) BY

T H O M A S B. OSBORNE. (Confinuedjronrpage 7 1 2 . )

Another trial was made by treating 400 grams 6f bean-meal, previously exhausted by benzine, with one per cent. sodium chloride solution, dialyzing the extract for twenty-four hours. and .filtering off the precipitated phaseolin. T h e clear filtrate after standing over night, deposited a considerable quantity of proteid, but the solution with this deposit was returned to the dialyzer and left for two days longer, when it was filtered, the precipitate washed with water, alcohol, and ether, dried over sulphuric acid, and six grams of preparation 36 obtained.

T H O M A S B. OSBORNE.

PHASELIN, PREPARATION 36.

Carbon ........................ 50.44 Hydrogen ..................... 7.14 Nitrogen ...................... 14.51 Sulphur ....................... 0.46 Oxygen. ........................... Ash ........................... 1.94

Ash-free.

j1.41 7.28

14.59 0.47

26.25

..,.

__

100.00

T h e filtrate from preparation 36 was then dialyzed into distilled water which was renewed every twenty-four hours for several days. After a week the solution was filtered and the precipitate washed with water, alcohol, and ether, and dried over sulphuric acid. I t weighed I .60 grams, 3 7 . PHASELIN, PREPARATION 37.

Carbon ......... Hydrogen ...... Nitrogen Sulphur > Oxygen t ” ” . ’ Ash

.......

14.52

Ash-free. 52.19 7.24 14.79

....

25.78

1.72

....

I.

11.

Average.

51.38 7.25

51.22

51.30

14.52

....

............

.... 1.83

6.99 .

I

.

.

1.61

7~12

100.00

One more preparation, 3 8 , was made in the same manner as 37, and had the following composition : PHASELIN, PREPARATION 38.

........................

Carbon 50.20 Hydrogen ..................... 7.07 Nitrogen ...................... 14.02 Sulphur ....................... 0.50 Oxygen. ........................... Ash ........................... 2.06

Ash-free 51.27

7.22 14.32 0.51

26.68

....

100.00

Further dialysis in distilled water, of the filtrate from preparation 37, gave no more precipitate. T h e solution was therefore saturated with ammoniun sulphate, and the precipitate so produced filtered off and dissolved in distilled water. This solution which now had a volunie of about 400 cc. was dialyzed for some

T H E PROTEIDS O F T H E K I D N E Y B E A N .

759

days, first in river water, and afterwards in distilled water, but only a trace of substance separated. This was filtered off, and the perfectly clear solution gave the following reactions : Saturation with sodium chloride produced no precipitate until acetic acid was added. Acetic acid in the absence of salt gave no precipitate. Nitric acid gave a turbidity when added in considerable quantity, and the addition of soine sodium chloride produced no further precipitation. Copper sulphate gave no precipitate. This solution when heated became turbid at 57" and particulate at 63". T h e entire solution was therefore heated to 70" in a waterbath which did not exceed this temperature, and after two hours, filtered from the separated coagulum. This was then washed with distilled water, alcohol, absolute alcohol, and ether, and dried over sulphuric acid. It weighed 0.48 gram. Before drying, this coagulum dissolved readily in two-tenths per cent. hydrochloric acid, and in dilute potash solution, and gave a violet reaction with copper sulphate and caustic potash. T h e filtrate from this coagulnm gave a further small coagulum when heated at i o " , for some time longer. This was filtered off, and after treating it in the same way as the first coagulum, was added to it. T h e total amount of coagulum, preparation 39, amounted to 0.63 gram, and after drying at I IO" was found to contain, ash-free, 15.23 per cent. of nitrogen. The filtrate from 39 was theii dialyzed into alcohol, and the solution thereby concentrated. On adding an equal volume of strong alcohol, the proteid was precipitated. This was filtered off, washed with absolute alcohol and ether, and dried over sulpliuric acid. It then weighed 0.72 gram which shows that the proteids had been almost wholly precipitated by dialysis and coagulation. This substance gave a nearly clear solution with distilled water, not made clearer by adding a few drops of sodium chloride solution. With caustic potash and copper sulphate, a pink color was developed which had a distinct violet tinge, and was by no means so red as that given by pure proteoses and peptones. T h e aqueous solution heated to 85" gave a flocculent coagulum which apparently represented most of the substance. From

760

THOMAS I3. OSHORNE.

this, it would appear that true proteoses are present in extremely small amounts. Dried at I IO', this preparation, 40, contained, ash-free, 13.60 per cent. of nitrogen. Being thrown down by strong alcohol, it could scarcely be pure, and the nitrogen determination is only of value, as indicating that the coagulun: mainly consisted of proteid. Of these preparations, 2 j , 2 7 , and 3 z 1 are unquestionably mixtures of phaseolin and phaselin. Excluding these three arid also 3s and 40, evidently impure, the remaining agree fairly well, as is shown by the following table : SVMXiK\7 OF

ANALYSESO F PHASE1,IN 26

28.

Carbon. . . . . . . . . . . . H ydroge t i . . . . . . . . . Nitrogen. ..... 14.57

51.57 6.92 14.48

........ _-

27.03 ___

Sulphur Oxygen

! J

100.00

34.

Carbon ....... 51.38 Hydrogen .... 6.91 Nitrogen ..... 14.67 Oxygen

\

....

.... .... 14.81

.... __ .....

KIDNEYB E A N .

30.

31

51.59 6.71 14.84 26.87 ___

51.98 6.82 14.53 26.68 __

100.00

100.00

3.- marl!. that of pliaseliii. Makiiig liberal :illo.ivancc for impurities arid iricoiiiplett tlr>,iiig,they reprcsent about t n o per cent. of phaselin. ,j,'I'weiity graiiis of t)caii-nical xvcrc extracted :IS coiiipleiel!. a s possil)!e lvitli two tciitlis per cent. potash lvatcr. Tlic n.ashed and air-(lry residue weiglietl I I . ? 7 grains, a n t i coiitaiiietl o , C j i per cent. or 0 . 1 0 2 6 graiii of iiitroge~i.eciual to 0.61 I graiii of (water-free) proteid insoluble i n alkali, or to per cent. of the liieal. T h e alkali-soIuliIe prpteid aiiiouiited therefore to (23.65-.,;.oh2 0 , j g ) 20.6 per ceiit. oi the i i i c u l . 6 . On page 642 is recorded tlint the proteid insoluble i n salt solution, h i t dissolved b y ttw-tenths per ceiit. potash, I S , coritairied sixteen per cent. of iiitrogeii and accordingly h a d nearly the coinposition of phaseolili. I t is, therefore, probable that the proteid uridissolvecl 11). salt-solution. i s phaseolin. 0 1 1 this asbuiiiption, the kidne!- bean exaniiriecl, coiitains about 2 I .5

T H E PKOTEIDS OF THE K I D N E Y B E A N .

763

per cent. of phaseolin, and about two per cent. of pliaselin. 7 . T h e foregoing data are suiiiiiiarized as follow: T h e " white medium field bean " contains approximately :

.............................

Phaselin, salt-soluble.. 2 per cent. Phaseolin, salt-soluble .............................. 15 " Phaseolin, salt-insoluble, alkali-solulile ............. 3.5 Phaseolin, insoluble in salt and in & per cent. alkali .t "

..

Total protei&

................................

23.j

''

Schulze, Steiger and Maxwell have stated' that ten per cent. of the nitrogen of the seeds of the horse bean ( Vicia f k b a ) the vetch and the pea, exists in non-proteid form. Should such prove to be the case with the kidney bean, then its total proteids would be about twenty-one per cent. iristead of 23.5 per cent. COXCLLJSION. T h e kidney bean contains two globulins characterized by great solubility in very dilute saline solutions, and by yielding precipitates with acids which are soluble in sodium chloride solutions. One of these globulins, pltaseoliiz, probably forms about twenty per cent. of the seed, and has the followin,G- coniposition, which is the average of analyses of twenty-four different preparations : PHASEOLIN. Carbon Hytlrogen ................................ Nitrogen ................................. Sulphur Oxygen ..................................

...................................

..................................

52.58 6.S.4

16.47 0.56 23.55 100.00

This is the proteid described by Ritthausen in 1884, to which he assigned very nearly the same composition as that above giveri. T h e other proteid, phaseliin, is much more soluble, remaining in solution after the phaseolin has separated. I t is slowly coagulated by heat at temperatures varying with the amount of salts present and the rapidity of heating. I t is precipitated by acids, on prolonged dialysis yields insoluble or albuminate niodifications, and has inore nearly the properties of a globulin 1 V~nucl~ S fsa t m i e x , 3gV306.

.\ .S;\FETY

764

.\TTACHMBNT F O R R I D E K S

than of any other recognized class of proteitis. It has ari unusually lo\\ iiitrogrii a i d high os\.geii content as shown by the subjoined ayerage of the aiialyses of eleven different preparatioiis. 12:r.\si 26.24 100.00

I n addition to these two globulins, the extracts were found to contain an extremely small amount of proteose. ?+fAY. 1$3 .

.

~. .

A SAFETY ATTACHMENT FOR RIDERS. H\

CHAS.

E.

I’:\IIKEH.

1’II.c

\ C d Ailyllrt 3 3 . 18u4

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chnierit for manipulating the weighiiig rider of balances was introduced by T’erbeck and Peckholdt. T h e arrangement, devised by the writer, and here described, appears to have sonie atl~atitagesin simplicity, lightness, and adaptability Tt consists of a piece of sheet brass cut i n tlie shape shown i n 6 , Fig. I , to which is soldered a bit of hair-spring froiii a watch, a ; and of a light glass rod, nz, 1 Fig. 3 $ secured i n a position parallel to the beam behind and slightly above it. The piece of brass, 6. is bent, .is shoivii i i i Figs. z atid 3, to forin a sleeve upon the rod, c, together with a prong in apposition to the hook 1))- which the rider is usually lifted, 1 2 , Fig. z ; and an extension, c , back under the rod, m ,which thus limits the rotation of the sleeve and prong Fig. 1. I f in one direction. I,I L‘ T h e operation is readily understood ; the usual hook arrangement being converted into forceps

OME years since, an

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