ABRUS OR

JEQUIRITY

POISON.

By Surgeons Warden And Waddell.

(Contd. from page 162.) The Chemical Nature

of

Abrus-Poison.

CHAPTER I. As it

found that the

was

abrus-seeds

are

due neither

poisonous properties to

bacteria

nor

of

to decom-

position products set free during bacterial growth, we then turned to chemistry, in the endeavour to isolate the poisonous principle, which from the preceding observations was evidently of a chemical nature. We have already seen that the seeds contain no volatile or fixed alkaloid or glucoside, and that the crystalline acid?abric acid, is inert, so that we are forced to conclude that the active principle of abrus-seeds must belong to an altogether different category from that which includes ordinary vegetable poisons, and that unusual means must therefore be resorted to for its detection and isolation.

Whilst we

filtering

observed that

the watery infusion of the seeds, opalescent haze speedily formed

an

exposure for a few minutes to the air; and removing this hazy precipitate by refiltration, a fresh precipitate, slightly less dense, in the clear filtrate

on

again ; and it was only after filtration and through 12 to 14 filter-papers that a permanently clear filtrate was obtained. The original filtrate gave with picric and nitric acids a copious white, flaky precipitate, showing it to be loaded with albumin, while this permanently clear filtrate gave no definite precipitate with these reagents. formed

re-filtration

Here, then, nature

by to

the

was some

substance of

an

albuminous

which had been removed from the infusion

repeated filiations, and a question arose as or not the resulting liquid still retained

whether

ABRUS OR

July, 1884.]

JEQUIRITY

toxic power undiminished. The three following experiments were made to elicit this point:?-

The

ts

Exp. XXXVI.?Ten minims of the fresh infusion, which had been passed through two folds of filter-paper, were in-

jected

into

chicken.

a

Death resulted in 18 hours.

Exp. XXXVII.?Of the

through a

filter-paper, 15 minims days. died on the 7th day.

14 folds of

chicken.

were

No apparent effect for four

food, pined,

and

Then it refused

Exp. XXXVIII.- A similar experiment the 10th

to the

last was,

Death occurred

day.

From these experiments it was therefore evident that the activity of the seeds was associated with this proteid material, which had been in great part removed by the repeated filiations (1). To isolate this proteid material so as to enable us to test it

by

from

directly

its

physiological action,

we

precipitated

absolute alcohol from the clear filtrate obtained watery infusion.

concentrated

a

It fell

as a

fine white, flocculent precipitate, which, collected filter, became of a light slaty hue on exposure

a

air.

About

solution

the

few

a

drops

While

precipitate,

on

a

similar

of sodic carbonate

with the effect of

added

were

dissolving chicken,

which

to

to

precipitate mixed and injected into

with 15 minims of distilled water a chicken caused death in 20 hours.

quantity,

on

of this moist

grains

1

thoroughly

injection into another

caused death inn hours, with the character-

istic

post-mortem appearances of abrus-poisoning. The details of these two experiments are given

below: Exp. XXXIX.?A

infusion

was

freshly-prepared concentrated watery through two folds of Swedish filter-paper,

filtered

and the clear filtrate treated

examination showed

extravasations

over

with absolute alcohol in excess.

copious fine, white, flaky precipitate immediately appeared, and was collected by filtration. The filtrate, which was clear and of a light straw colour, gave no precipitate with picric or nitric acids.

Of the moist albuminous residue on the filter* taken and

were

mixed

with 30 minims of

distilled water, in which the albumin appeared to dissolve sparingly. Half this quantity of liquid, 2. e , 15 minims, were

injected

into

a

It died in 20 hours with all the con-

chicken

gestive and minute hemorrhagic extravasations characteristic of

of the mixture of water

XL.?The

and the alcoholic

remaining portion precipitate had 2 minims

sodic carbonate added

to

it.

of concentrated

A clear solution was at once ob-

tained, the liquid becoming of a faintly yellowish tinge. on injection into a chicken caused death in 11 hours. (1) Schmidt showed

the

sub-

lungs

The intestines were

very markedly injected. The serum from the seat of wound showed different kinds,

one

few bacilli of

a

variety being small, thick-set,

less than the diameter of the nucleus of

length

a

with

a

red blood-

cell, and its breadth ^3 of its length, ends abruptly rounded off. The

of these formed small chains of two and three.

majority

was about l/z the diameter of the nucleus of a blood-corpuscle, with a slender body and square ends, not chain forming. This proved body was therefore without doubt the

red

active

of the

principle

seeds,

and it seemed to act

with greater rapidity in proportion to the completeness c f its solution at the time of administration. Form

in

which it

Exists

It exists in the seeds in

in the

water, from which solution it is not strong acids, by alkaline carbonates, It thus behaves like

sodium. It

is, however, precipitated,

Seeds.

form which is soluble in

a

a

precipitated by by chloride of

or

native animal albumin. globulin, from its

like

watery solution by a current of carbonic anhydride, but The re-dissolves as the carbonic acid evaporates. in soluble in this is obtained readily way precipitate

hazy precipitate which forms in exposure to air is apparently due to the carbonic acid contained in the air, for when the filtrate distilled

The

water.

the infusion

on

permanently clear. highly acid on account of the abric acid present; but the proteid does not exist in the seed in the form of an acid albuminate, for a large proportion of it is precipitated on boiling the infusion, and on neutralising its aqueous solution it is not is collected in

vacuo

it remains

The watery solution is

thrown down. Effect of Temperature on

the

Activity of the

Poison.

The effect of temperature upon the activity of the poison is very marked. Boiling the watery infusion for

an

instant renders the poison

permanently

inert.

short exposure of the watery infusion to a temperature of about 7o?C. is sufficient to bring about And

a

permanent

loss of its

physiological properties.

That

temperature of much less than ioocC. will destroy the activity of the poison was found out while evapo-

a

abrus-poisoning. Exp.

to

not

A

about 3 grains

slight tendency

walls cf stcmach and bases of

The other form

made at the same time on another chicken. on

passing it injected into

post-mortem

187

towards their attachment to chest wall.

two

after

infusion,

same

serous

POISON.

that on filtering an albuminous filtrate contains less albumin, but no qualitative

occurs.?Poggeniforfs

Annalin for. 1865,

;r

?:

This

liquid change

rating

some of

the watery

distance above the

extract

at a

considerable

water-bath, 75?C. A temperature considerably below that necessary to coagulate albumin is sufficient to lower the activity of this poison.. Thus, exposure for three or four

ranged

from

65?

to

where the temperature

188

INDIAN

THE

MEDICAL GAZETTE.

temperature as 32?C. was found to very marked degree the activity of the alcoholic precipitate kept submerged in alcohol,?to

days

to so low

reduce in

Effect

a

a

That the loss of

plays a very important part in facilitating this decomposition, which occurs through heat. Although the exposure of the watery infusion for

destroy

following experiments also show that the effect of heat in destroying the activity of the watery infusion is not due to simple coagulation of the proteid, for it also occurs when coagulation is prevented by solution in an alkali or an acid. Thus, in the

in situ and put into a water-bath for the different periods as noted in the experiments, and the activity then tested

pre-

at one

?vacuum

one

precipitate forming. The injected into a chicken.

boiled

liquid

after

consistency

of an extract.

rub-

1

diied

being

20

was

in-

hours.

Of this l/i grain was and death resulted

chicken,

XLVII.?1^ grains of the same injected into another chicken. powder

dried and heate

Death occurred

were

in 20 hours.

Exp. XLVIII.?Another

[

as

before,

was

quantity

heated

to

of the

a

powder, dried

temperature

in

of ioo" for

Its colour had become slightly darker, and it gave faint burnt smell. Of this 4^ grains were injected into healthy chicken. On the day following it was manifestly

80 hours.

is had in the

oft"

the action of see

powder

Death occurred in

Exp.

afterwards

no

grain of the above

into another chicken.

temperature of loo?C. for three hours.

vacuo

wholly inert, although

temperature of ioo?C. for the inrush of air testi-

a

injected in the usual way into a iin 30 hours.

a

(Exp.

a

affected, refused food, and drooped. After three days it had improved considerably. In about eight days it had almost recovered, with the exception of extensive swelling of the injected leg, which rendered it lam? fpr $H>ut three weeks. Exp. XLIX.?Another quantity of the powder was simply

precipitation

occurs.

sealed up ed of air.

The watery infusion kept at a temperature of io6C. for several hours retained its activity -un-

diminished.

Exp. XLV.?One

a

LXVII) that the peptone thus formed possesses considerable activity. But on boiling for an instant it is rendered

at

broken,

Exp. XLVI.?Another quantity of the freshly-pounded seeds !was dried in vacuo in a similar manner and then exposed to

It was then dissolved in water and

behaviour of the peptone pepsin on abrin. We shall

Sprengel compleie

5

No effect.

point formed by

a a

the completeness of the vacuum. Half a grain of powder was rubbed up with a little water and injected hypodermically into a chicken. The animal manifested the usual sighs of abrus-poisoning and died in 26 hours.

No effect.

A further illustration of this

after

this crisp

Exp. XLIII.?The acetic acid solution of the alcoholic precipitate was boiled for two minutes and evaporated to the

injected.

minutes

It was then sealed off in situ and

water-bath

The tube was then

hour.

jected

was

a

attached to

was

20

fying j,to

Exp. XLII.?Two grains of the freshly-prepared moist alcoholic precipitate were dissolved in a small quantity of water, to which a few drops of sodic carbonate solution had This solution was boiled for two minutes withbeen added. any

extremity

had been made.

[introduced into

effect whatever.

out

The open

end.

pump and retained for about

bed up with a small quantity of water and boiled for an instant. Half this quantity was injected into a chicken. No

cooled

by hypodermic injection.

Exp. XLIV.?About 15 grains of the finely powdered seed was introduced into a piece of Bohemian glass tubing closed

cipitation occurs even on boiling; but the activity of the poison is lost. And the precipitate obtained by heating the watery infusion from 70? to ioo?C. may be re-dissolved by the aid of a small quantity of sodic carbonate and caustic soda; but this solution when injected is inactive. were

the

activity. following experiments the seeds were reduced to a very fine powder and placed in Bohemian glass tubes, which were attached to the vacuum These tubes, after their conof a Sprengel pump. tents were thoroughly dried, were then sealed off

The

Exp. XLI.?Three grains of the powdered seeds

of ioo?C. suffices to des-

In the

contrary increase, the activity of the fresh poison. The powdered seed also, on exposure to a similar temperature in the air, has its activity similarly reduced.

no

temperature

troy altogether activity of the poison, the dried be seeds may exposed to a temperature of ioo? for several hours without having their activity seriously deteriorated. But prolonged exposure to ioo?, even in the absence of air and moisture, serves also to

soluble in great part in distilled water with the acid of sodic carbonate and weak caustic soda solutions?reagents which do not destroy, but on the

of the sodic and acetic acid solutions

to a

the

readily

case

instant

an

activity is due to the proteid by the

the heat, and not to coagulation of alcohol, seems evident from the fact that the pre" cipitate, even after two weeks' contact with alcohol, was

of Dry Heat.

Moisture

prevent the development of bacteria; and, after exposure for two weeks, the precipitate had become

completely inert.

[July, i?S4.

for -

a

hermetically,

similar

meat.

without

having

been dried and exhaust"

This-was subjected.to thc $5ime-degree of heat- and

period

Its colour

of tirae(t?is;j was

$$iV>ors)as

last

experi-

then of ? : light coffee colour, and it

ABRUS OR

Julv, 1884.] gave off a strong burnt smell.

into

chicken, This last a

Of this 3 grains

were

JEQUIRITY POISON.

injected

and no effect whatever ensued.

appears to show that the moisture contained within the seed is in itself suffi-

experiment

cient

to effect rapid decomposition. In the preparation of " sut's" the poison is exposed to the direct rays of the tropical sun to dry. Lard or other greasy matter, however, is always pre-

viously

poison, and apparently decomposition which occurs by

mixed with the

to hinder this

tends heat.

freshvery much less active than the 2 "sui" One seeds. ly powdered grains, weighing on being rubbed up with water and injected into a chicken, does not usually produce a fatal result till But

"

suis"

after the

are

whilst

lapse

of about

36 hours;

the fresh seed

produces

death in about 18

The Mode

of

Extracting

the

grain of hours (1).

Active

Principle?Abrin.

difficulty was experienced in obtainprinciple in a pure form, chiefly through susceptibility to heat and its association with

Considerable

ing

the active

its great abric acid and extractives, which cause the crude product to become of a dark, slaty hue on exposure to the air.

Of the

two

following methods of extraction, appeared to yield a product

the second is that which

less contaminated with extractives. The first method

was as

follows :?The seeds in

were

iron powder by pounding being removed to a considerable exThe powder was packed in a tent by winnowing. cotton wool and a layer of with percolator plugged cold distilled water, the with sand, and percolated process being hastened by connecting the receiver to an exhausting pump. The first portions of the percolate were turbid, but after exhaustion the liquid that passed through was bright. The dark sherrycoloured and strongly acid percolate was now mixed with a large excess of 60 per cent, alcohol, and the precipitate allowed to settle. The supernatant liquid was then siphoned off, and the precipitate collected on reduced

to a coarse

mortar, the

an

testae

paper-filter. After exposure to air for a few hours to vaporise the alcohol, the precipitate was scraped a

(1) The effect of heat in destroying the inflammation-exciting power of the seeds was determined as follows by M. Salomonsen (loc cit). He exposed small quantities of the watery infusion in sealed tubes to temperatures varying from 6o? to ioo?C in a water bath from x/t an hour to 1 hour, and then tested their inflammation-exciting power after rapid cooling. After % hour's exposure to 64? their activity was considerably weakened, not completely annulled. A freshly-prepared, very active infusion was warmed at 65? for one hour, and when afterwards dropped into the eye produced an evanescent hyperemia without any swelling or pus-formation. Exposure to 70? for

although

qpe-how completely destroyed this power.

1S9

off the filter and mixed with cold distilled water, the resulting mixture being of a slate-grey colour and very turbid.

Alcohol

again added,

was

the

precipitate

allowed to settle, collected on a filter, and again treated with water and alcohol. The principle obtained

by

this process while moist

which

rapidly deepened

on

dried under the receiver ,of

was

of

a

exposure an

dark-slate colour to

air-pump

air.

It

with

sulphu-

was

ric acid, powdered and percolated with chloroform (i), which removed some fatty matter and traces of abric

acid;

the chloroform

being

succeeded

by

absolute

alcohol, which dissolved traces of extractive. The principle thus purified was, when pulverized, of a dark fawn colour and somewhat The second method of

hygroscopic. separation was conducted :?The follows as pulverized seeds were first percolated with chloroform, which removed fatty matter, some abric acid, and a yellow colouring principle. The powder was then exposed to air to evaporate the chloroform, and re-percolated with 40 per cent, alcohol (to remove extractives), until the percolate was nearly colourless. The powder was again exposed to air, and, when free from alcohol, percolated with cold distilled water. The resulting percolate had only a slight yellow tinge, and was very faintly acid in reThe active principle was obtained from this ; action. percolate by precipitation with alcohol, and purified by repeated precipitation and solution in water, as already described, and subsequently dried over sulphuric acid. The precipitate, whilst contained in the liquid from which it had been precipitated, is perfect ly white, but on exposure to air it becomes, whilst drying, of a very light slate colour. To this proteid so obtained we have applied the It is much more active than an equal name ah in. weight of the powdered seed, as will be seen from the experiments which are detailed further on. The Physical Properties of Abrin.

When obtained in this solid of

purified form, abrin is an pale grey colour, and tasteless.

amorphous In thin layers, it is of a gummy translucent appearance and yellowish colour, like dried white of egg. Cold water readily dissolves it, with the exception of a few flocks, and the solution,, which is of a faintly yellow colour, froths on agitation. In glycerine it is also soluble. And it is precipitated from its aqueous and glycerine solutions by alcohol in white,curdy flakes, which become of a light slaty hue on exposure to air. (i) Chloroform acid than ether.

was

a

used

as

being a

better solvent of abric

,

THE INDIAN MEDICAL GAZETTE.

90

On boiling the watery solution of abrin it becomes opalescent, and a whitish flocculent deposit falls, and

supernatant fluid becomes

the This

collected

precipitate

in caustic

soda,

by

of

alight

Strong caustic potash gives rise to no precipitate. boiling this solution ammonia is evolved, and the liquid becomes of a somewhat darker hue.(i) On

brown colour-

filtration is partly soluble on neutraliza-

The Ultimate Composition

from which solution

The abrin which

precipitated by picric and nitric acids and alcohol, and the alcoholic precipitate is sparingly

tion it is

light

water.

dissolves in

excess

of the acid.

A

copiou;

the usual

compounds

white pre-

and the alkaline

acid albuminate.

dry heat, as has been already takes place much more slowly. position a

seen,

decom-

Carbon

a

chloroform appears

cipitate

to

watery solution of abi in throws the

addition of ether

hasten the subsidence of

acid in

a

pre-

produces

excess

a

1

>t

,,

ages

(I)

9-1620

pitate. a

white

chloride,

precipitate

and

insoluble in

excess.

ferrocyanide and perchloridc of iron do precipitate abrin from its watery solutions. Millon's reagent gives the usual purple-red proteid Potassium

not

reaction.

...

filled with dilute

hydrochloric

!>

>>

*9640 gram, 5622

spongy

platinum.

,,

to?

,,

following

percent-

?

,,

,,

,,

sulphur determination, with fusion mixture of 1-355? gram- abrin dried at 140*0., yielded '0376 grams Ba S04, equivalent to '3763 per cent, sulphur. One

The

ultimate

follows

percentage

of

abrin

therefore be stated

:?

..

...

...

...

43*5754

77652

N

10-2801

0

288599

S Ash

composition

the first method may (I)

C. II.

Dilute acetic acid gives a white precipitate soluble in large excess of acid. Tannic acid gives a copious white curdy preci-

give

...

per cent, of ash.

(II) 9-3370

of the

excess.

silver nitrate,

(II)

:?

as

mercuric

give?

100

43*5754

(I) 10 2801 per cent. Nitrogen. (II) 10-5283 Two ash determinations gave the

prepared by

of lead,

...

being

76^4 which is equivalent

Glacial phosphoric acid and met aphosphate of soda give a white precipitate which is soluble in large'

acetate

...

"3402 gram, abrin gavt:

acid.

Basic

?

in parts per

and the ammonium chloride converted into the

(I) (II)

or

with alcohol.

Very dilute hydrochloric or nitric precipitate which is extremely soluble

1258 gram.' IisO. *1508

?

platinum-ammonium salt, yielded on ignition following amounts of spongy :? platinum

soluble in dilute alcohol.

but

?

43*S235 77964 77652 determinations by the soda-lime

...

process, the bulbs

dilute one, a very large excess of alcohol requires to be added to effect precipitation, as abrin is slightly

precipitate,

expressed

...

a

no

'

of the double salt the

decidedly acid reaction. It gives the following reactions. Absolute alcohol readily precipitates abrin from If the solution is its aqueous or glycerine solutions.

down

acid

combus-

on

double

The concentrated aqueous solution of abrin has

a

-3442

?

Hydrogen Two nitrogen

acid,

Its Chemical Reactions.

Ether agitated with

results

the

(I)

neutralizing this amber-coloured filtrate no precipitate is thrown down, showing that the proteid is With

?

These results

On

not an

sulphuric

over

:?

(II) -2143

by tannin, alcohoi and ether; carbonates give a slight precipitate.

the first described

by

-1790 grain, gave -286S CO.. and

(I)

is thrown down

cipitate

been dried

Abrin.

of

following cupric oxide and oxygen in an open tube, precautions being adopted for nitrogenous

tion with

cloudiness with distilled water; and weak solutions of acetic acid throw down a flocky precipitate, which

obtained

was

method, after having for 48 hours, yielded

The filtrate, which is of a amber colour and of acid reaction, gives a slight

soluble in distilled

[Jui.v, 1884.

...

'3574 9-1620 loo'cooo

(II) 43*8235 7 7964 10-5283 28-1574

Mean.

43*6994 7*7808 10-4024

285105

-

-3574

9*337?

9-2495

99-6426

(1) Salomonsen {!oc .cit) also being led to conclude that the local action of the seeds on the conjunctiva was not due to bacteria, endeavoured to isolate the poison by chemical means, and adopted the ordinary method for extracting a pro,teid-? similar to that made use of by us 5 and he found that the dried " soluble in water or glycerine and a alcoholic precipitate was few drops of this watery or glycerine solution brought into the conjunctival sac of a rabbit calls forth an inflammation with all the clinical and anatomical characters of abruS'ophtbalmia."

.

W/

ABRUS OR

July, 1884.J On

abrin in the hot-air bath

drying

i4o?C. it

at

POISON.

JEQUIRITY

plant-albumin the proteid held in solution

As

1.

t9t

juices or extracts, which is precipitated by acetic acid, but which coagulates when heated.

lost 5*2540 per cent

in aqueous plant

the

not

H3 O. Deducting ash and water, percentage would be as follows :?

mean

C.

...

...

...

...

H.

...

...

...

...

N. O. S.

...

...

...

...

...

...

...

...

...

...

5CS241 8-0422

...

...

As

2.

plant-fibrin

12-0878 286313 '4146

which is insoluble in

As

3.

The abrin dried

prepared by the second method was first sulphuric acid and then in a hot-air bath

over

i4o?C.

?'

>>

,,

C.

(II)

...

...

...

42-330?

4i'35?3

H....

...

...

70032

6-9674

determination with soda-lime, gave :? ?5458 gram, abrin gave '3276 gram, spongy platinum, which is equivalent to? One

nitrogen

8-6502

per cent of

Nitrogen.

Two ash determinations gave?

(I) 11'9359 Per cent ?f ash. (II) 12-2449 The ultimate percentage composition of abrin prepared by the second method may therefore be ?

stated

follows

as

(ID 418503

(I) ...

H.

...

...

N.

...

...

O. and S. Ash

42'33??

...

According to

...

...

...

6*9674 8-6502 29-8075

7-0032

12-2449

u'9359

6'985 3 8*6502 30*1840 12-0904

100*0000

precipitated from by heat, but is precipi-

not

plant-gelatin, which animal-gelatin.

this classification abrin

to

either the 2nd

or

is gene-

cannot

belong

4th category, and its being pre-

cipitable by acetic acid prevents its being regarded plant-albumin," whilst its being precipitated from its aqueous solution on boiling prevents its being So that abrin is altogether classed as plant-casein. excluded from a place in this unduly restricted classification. If, instead of" not precipitated by acetic "

as

acid,"

we

abrin

read

"

soluble in

would obtain

albumins,"

to

a

which

excess

of acetic

place amongst class

it

seems

acid," then

"

the

plant naturally to

belong. Abrin

Mean. 42*0901

and is

rally similar to

:

C.

the constituent which dissolves

water

The last class is

4.

,t

(I)

plant-casehi in cold

its aqueous solution tated by acetic acid.

On combustion with oxide of copper the results were obtained :? following (I) *1862 gram, gave *2872 CO? and *1174 gram. II? O. '2687 ,, (II) "1751 *1105 ), These results give the following percentages :?

at

and aqueous

water

ammonia..

ioo-oooo

.

seeds

of

constituent

the

form of to

cannot certainly be plant-casein, for casein,

considered which is

be identical with alkali-albuminates

cipitated

from its aqueous solutions

insoluble in distilled

ties,

not

of

a

on

as

now

being

a

believed

(1), is not preboiling, and is

Abrin has the properderived-albumin, but of a nativewater.

(animal) such as that described by Foster (2), being soluble in water and not precipitated by very dilute acids, by carbonates of the alkalis, or by sodium chloride, coagulated by heating If in solution to a temperature of about /o'C. dried at 4o?C, the resulting mass is of a pale yellow colour, easily friable, tasteless, inodorous, and albumin

"

the

Deducting ash, would be

as

follows

mean

percentage

:?

C.

...

...

...

...

II.

...

...

...

...

N.

...

..

...

...

...

...

...

O. and S.

composition 47*8674 7*949o

98273 34*3593

soluble." The ash

phosphoric reaction,

found

was

acid.

It

was

and contained

The Place

of

to

a

contain very

decided

large amount of faintly alkaline in a

trace of

iron.

Abrin amongst the Proteids.

The classification of in

use

is that of

vegetable proteids at present Liebig(i). He distinguishes :?

(1) On the Nitrogenous Food-stuffs of the dom.?Ann, P/iarm.) XXX/X, 129.

Vegetable King-

The

typical native albumin is egg-albumin. In following table is given the percentage composition of egg-albumin, together with that of pea-albumin, and a few other vegetable albumins for comparison with the percentage composition of abrin

the

which is also appended (1) (2)

:?

Foster's Physiology, Ibid, p. 702.

p.

708, 4th ecln.

THE INDIAN MEDICAL GAZETTE.

192

Table

Comparing Percentage Composition of certain Vegetable-albumins with Egg-albumin and Abrin. C.

H.

N.

Ash.

[July, 1884.

Exp. LII.?A small

of the filtered watery infusion

quantity

applied to the eye of a rabbit produced on the following day intensely acute inflammation of conjunctiva, with hemorrhagic iritis,

extravasations and

the formation of a

ultimately

and

corneal ulcer.

Egg-albumin (i)

Pea-albumin (2) Gourd-seed vitellin^) Para-nut vitellin (4).. Wheat-albumin (5).. (6) ? Rye Almond ,, (7) Potato (8) , ..

.. ..

Abrin

53'98 52*45 51*36 52*43

7'5i 6'8i

22*34

7*58 7'I2

I7"86

22*66 2I*8o

53-74

7*11

53*7' 55*96 53'8r 50*82

7*77 7*53

15*85

23*46 22*67 22*76

i8'io

13*75

7-32

2*3

0*98

i*4

0*89 1*12

8-S

0*41 Dedctd.

2*8*63

8*04

i*93 0.80 ?*54 ?*55 1*04 o*77

large The

following experiments show that principle?abrin is more abundant in the in the heart-wood of the Exp. LIII.?X

oz.

stem was reduced to

a

stems and roots :

of the mixed rind of the roots and

coarse

powder

and mixed with water and

macerated with occasional trituration in

It will be

proximates somewhat and the

in the

vegetable foregoing

composition of abrin ap" closely to that of egg-albumin

that the

seen

albumins.

It is to be noted that

tabulated

analysis of these latter, the albumin was obtained by coagulation of the cold queous extract by boiling. The best proof that abrin is nothing more nor less than the albumin of Abrus precatorius is had in the fact that the albumin of the roots and stems of that plant is possessed of the identical poisonous prowbieh characterize the albumin of the seeds, is evident from the following experiments :?

perties as

The fresh of a

February, concentrated

same

stems, gathered in the month reduced to a coarse powder, and

roots and were

watery infusion made after the when operating with the seeds.

cold,

manner

as

This infusion

on

being

colour and

filtered

was

acid reaction.

of

pale straw yielded with fairly copious a

slightly nitric and picric acids in the cold a white precipitate, and also on boiling. With alcohol a white precipitate was thrown down, which was slightly soluble in distilled water, and more readily in caustic soda

or

sodic carbonate.

sion and the alcoholic

precipitate

It

Both the infuon

injection

sub"

cutaneously produced death with the usual appearof abrus-poisoning. The infusion also, when ocular to the of a rabbit, induced conjunctiva applied acute abrus-ophthalmia, going on to partial sloughing of the cornea.

ances

Exp. L.?Of the concentrated infusion of pounded abrus root

25 minims were subcutaneously injected into a Death occurred in 50 hours with the usual symp-

and stems,

chicken.

toms and post-mortem appearances of abrus-poisoning. Exp. LI.?Two grains of the moist alcoholic precipitate

dissolved in

20

minims of water and injected hypodermically

produced death in iS'hours. (1) Theile, Kopp's Jahresb., 1867, 774. (2) Ruling. Ann. Pltarm., 58., 306. (3) J. Barbieri, Jour. Prakt. Chem. [2] 18., 102-16. (4) Weyi, PJluger's Archiv. Bd, 12. (5) D umas & Cahours, N. Ann. Chim. Phys. 6,409. (6) Jones, Ann. Pharm, 40, 66. (7) Gmelin, Handbk. Org. CktmVol. XII,p. 427. (I) Ruling, let tit,

hours.

the active

rind than

a

mortar

three

for

resulting infusion was filtered, and of the clear minims were injected subcutaneously into a chicken.

The

filtrate 30 Death occurred in 17 hours. Exp. LIV.?A similar quantity of the clear filtrate

jected

into another chicken.

was

in-

Death occurred in 18^.'

Exp. LV.?% oz. of the mixed heart-wood of the roots and was cut into small, thin slices and pounded in a mortar,

stems

then macerated with occasional trituration for three hours and

filtered.

Of this clear filtrate 30 minims were injected into a Death occurred in 43 hours, with the usual appear-

chicken. ances

of abrus-poiscning.

Exp. LVI.?The another

chicken,

foregoing experiment

and death took

place

was

repeated

on

in 47 hours.

This discovery that the roots and stems contain large quantities of the deadly principle?abrin, is of great practical importance, for it is a common practice amongst the natives of this country to chew the roots for the sake of their contained sweetish principleThe differences between of abrus

root

true

precatorius

can

liquorice root and the be readily determined

examination.

by microscopic

These differences

are

following description of abrus root Fluckiger's Pliarmacographia (p. 165 ): Description.?The root is long,woody,tortuous, and branching. It usually occurs cut up into short lengths. The thickest pieces are of about the diameter of a man's finger, but most of it is much more slender. The cortical layer is extremely thin and of a light brown hue. The woody part breaks across with a decidedly fibrous fracture, exhibiting a light yellow interior. It has a faint, somewhat disagreeable odour, and a bitterish acrid flavour, leaving a faintly sweet aftei-taste.

indicated in the taken

from

"

Microscopic Structure?On tion the bark exhibits loaded with brown the middle

layer are

of

zone

some

a

layers

transverse of cork

sec-

cells,

matter, and then, within of the bark, a comparatively thick

coloring

sclerenchymatous

tissue.

Strong

liber fibres

scattered through the interior of the cortical tissue,

but are 7iot distributed so

form wedge-shaped rays liquorice root. In the latter the selerenchyme (thick-walled cells) is wanting. These differences are sufficient to distinguish the two roots. such

as are

met

as

ivith in true

to

ABRUS OR

July, 18S4.]

to

CHAPTER II. In

have been

abrin

seeking to ascertain the manner active principle?abrin, exercises its

POISON.

JEQUIRITVT

in which this Table showing the Percentage Composition of Vegetable toxic action on Enzymes as compared 7oith that of Abrin.

the animal economy, we are at once led to consider or not it may behave as a chemical ferment. Its proteid nature, as well as the destruction of its exposure of its aqueous solution for temperature of ioo?C.; the consider-

by a

able interval which elapses between the administration of what ought to be an overwhelming dose and the

development of symptoms ; its harmlessness when given by the mouth or injected per rectum, all suggest the probability of its acting in this way. The "

term

has hitherto

ferment

two groups of agents,

in

contrasted with that of

analysed?is

:?

whether

toxic power an instant to

193

been

applied

to

or Emulsin, Synaptase (i). Diastase (2)

41*59 48-63

..

Papayatin (3)

Remarks.

H.

C.

11

"64 36-56 4*8 39'i6

6'g6 7*34

1

25

22 "o

7*34

a trace

4

14

..

whether got from tree-sap

Abrin by

first

Abrin by second

47*86

method

But the term

special

I2'o8

So'82

method

a

tact"

set of

48*63

8*04

9-82 34*35

7'94

9*04

'41

Ditto

12'09

do.

"

implies the possession of qualities?ferments being substances ihe property of exciting chemical "

ferment

"

And in

(4).

fruit. Dried at 140*. or

which, although nearly allied which possess action, nevertheless belong to changes in matters with which they

origin and mode of essentially different categories. The organised or formed ferments of which yeast is the type are independent organisms, with powers of growth and reproduction, and the transformations which constitute their special characteristics as ferments are inseparably associated with the nutritive operations of these

Composition varied according to

come

the exercise of this

into

con-

action, the

ferment itself appears to be undestroyed, it being "asserted that in the fermentive proctexcited by soluble ferments, the amount of ferment at the end of the process is the

the

beginning" (5). might possibly of consist ordinary pea-albumin plus some soluble organisms. The ferment power cannot be separated from the ferment organism by any method or by any ferment which so reacted upon pea-albumin as to solvent. The soluble ferments, on the other hand, render it poisonous, the following experiments were undertaken. First of all the bland leguminous matter pass freely into solution in water; their action is the common pea (pis?im) was injected without any of dissociated from the life of the gland-cells which produced them, and they are wholly devoid of the poisoning symptoms developing :? Exp. LVII.?The solid matter of one pea deprived of its Kuhne desigpower of growth and reproduction. cuticle was rubbed up with 20 minims distilled water and innates these soluble ferments enzymes(i)." Examples jected hypodermically into thigh of chicken. No effect, of chemical or soluble ferments are emulsin, papayaExr. LVIII.?The above experiment was repeated on tin, diastase of malt, myrosin, ptyalin, trypsin, &c. another chicken with no effect. Then a very small quantity of a weak infusion of It is to this latter class?the so-called enzymes was rubbed up with the bland abrus-seed would abrin and leguminous belong (eV in, (vfirj, ferment)?that if it proved to be a ferment. The mode of. extract- matter of the common pea, and injected, but death ing chemical ferments from vegetables is identical did not occur sooner under these circumstances than with that which we have employed for the isolation in the control experiments, where a similar quantity of the weak abrus infusion had been given by itself, of abrin. But it is a similar process also which serves as the following experiments show :? to extract albumin from seeds or other vegetable tissues :?emulsin may, in fact, be called sweet-almond albumin; papayatin, papaya albumin ; diastase, wheat

barley

albumin ; and

abrin, abrus albumin

; and so

on.

Exp. LIX.

composition

of the known

vegetable

enzymes has not yet been completely worked out. In the following table the published composition of thiee of these ferments?the

only

ones

(i) \V.( Roberts, Free, Roy, Soc32,

at

not

abrin

One-eighth grain

of abrus-sced was rubbed up minims of distilled water and injected into a chicken. Death occurred after 6 days. Exp. LX.?The foregoing experiment was repeated in

with about

20

another chicken.

The chemical

or

"

"

or

same as

To ascertain whether

which appear p 145.

Death occurred in

Exp. LXI.?With

grains

of bland

^jth of pea-albumin were an

a

5^ days. grain of abrus-seed,

rubbed up

along

with

(1) Bull, Ann. rh. Pharmac., LXIX, 145. (2) C. Kranch, Bied. Caitr., 1879, p. 122. (3) A. Wurtz, Compt. Rendns, 90, 1379 (1S81). (4) McKendrick, Physiology, p. 49. (5) Ibid\ P- 54.

1

yz 20

minims of distilled water and injected. 6

Death took

place

in

days. Exp. LXII.?The

another chicken.

The

most

power of

foregoing experiment days.

was

repeated

on

Death occurred in 7 common

property of ferments is their Direct or starch.

decomposing proteids

It has been asserted that certain

ferments, although harmless when taken by the mouth, become fatally active

when

injected subcutaneously. Thus Rossbach is reported(i) to state that after the intravesolution of papayatin the blood nous injection of a contains in a few hours enormous quantities of micrococci and biscuit-shaped bacteria in lively movement, and death speedily results with all the symptoms This property attributed to papayatin of septicemia. a comparison of rendered papayatin with abrin highly desirable.

is

a

ferment obtained from the juice of

It presents

all

the characters of

a

strong

digestive ferment, resembling pepsin and that secreted by carnivorous plants. Placed in contact with moist fibrin in slightly acid, neutral, or slightly alkaline solutions, it dissolves large quantities of that substance the fibrin, first softening, then disintegrating without swelling, and, becoming dissolved, leaves a residue of dys-peptone (2). Indian cooks are aware of this property of papaya juice, and use it for rendering tough tender.

The

following experiments juice.

were

made with the

fresh papaya

LXIII.?An eighth of an ounce of the fresh juice was mixed with an equal quantity of water, well rubbed up in The clear filtrate gave a copious a mortar and filtered. pre-

cipitate filtrate

20

minims

were

picric injected hypodermically

Exp. LXIV.?A similar amount was

ly

into another chicken. EXP. LXV.?A

smeared

over

From

regards

Of this clear into the

thigh

large

injected

the above true

subcutaneous-

amount

of the fresh

experiments Rossbach's

fection of the blood, resembling in all essential resThe unfiltered saliva produces pects septiccemia. part where injected(i). But, as he parotid saliva was innocuous, it would appear likely that the toxic properties of mixed saliva were due to some poison generated during the growth of the non-specific bacteria which infest the saliva once it has entered the buccal cavity. The subcutaneous injection of small quantities of Pkpsix also seems (Exp. LXIX) to be un-

gangrene of the found pure

attended

juice

we

was

freely

No result.

must

conclude

statement may

be

as

injection, the subcutaneous injectolerably large quantities is unattended by

intravenous

toxic results.

they necessarily produce poisoning

symptoms when introduced hypodermically. Destructibility of the active properties by exposure to a temperature of ioo?C. is a quality common to almost all forms of albuminous

special properties, the substance is

a

and does not.

matter n

possessed

ecessarily imply

of

that

ferment.

of the symptoms even after the subcutaneous injection of what ought to be an overwhelming dose is not apparently the result of The

relatively

slow

onset

fermentative change or leavening action, but due to the fact that abrin is a colloid body and

any

seems

readily

not

XL)

by

absorbed.

that when the

the addition occurs

jection,

circulation,

It has been

already

is rendered

proteid

of sodic

more

where the

carbonate

or

And in

seen

more

(Exp.

soluble

caustic soda,

intravenous in-

rapidly. poison gains speedy

death occurs with much

access to

the

greater rapidity

than under any other conditions. Exi\ LXVI.?Two grains of the moist alcoholic precipitate were dissolved in about 20 minims distilled water, and the solution which was faintly acid neutralized with sodic carbonate

quantity of one per cent, solution of comadded, and the whole injected into the pectoral Death took place in 10 hours. vein of a chicken. no find direct proof that abrin possesses We thus

mon

any

A small

salt was

of the

the soluble and

of the term

characterize and

pepsin,

4

ferment,' when applied such as abrin, is calculated unorganized poison,

as

to an

the

properties which specially ferments?papayatin, ptyalin, use

to convey an

(1) Lancet, Vol. 1, 1882, p. 280. (2) Wurtz and Bouchet, Conipt. rcndus, 89, p. 425 (1879.)

constitutional effects, so be regarded as a property of soluble

any marked

by

cannot

solution.

No result.

the ocular conjunctiva of a rabbit.

that, however tion of

acids.

No result whatever.

of a chicken.

porous plates, so as to exclude bacteria, and beneath the skin of rabbits, does not produce any local gangrenous changes, but a general in-

death

Exp.

with alcohol and nitric and

Vulpian

through injected

ferments that

the fruit and trunk of Carica papayj, an exceedingly common fruit-tree in India, by precipitation with alcohol.

M.M. Pasteur and

by

toxic power when injected under the skin. M. Griffini has lately repeated their experiments, and finds that ordinary mixed saliva, when filtered

that it

Papayatin contrasted with Abrin.

Papayatin

Saliva is asserted to possess

experiment showed that abrin possesses no amylolytic power, or peptonising action on egg-albumin.

meat

[July, T8S4.

THE INDIAN MEDICAL GAZETTE.

194

altogether

erroneous

impression

(1) Archiv. per It Scienzt Medichc,

t. v. 1882.

as

to

its

'?

/

EWART ON SIMPLE AND SLOUGHING DYSENTERY.

July, 1884.]

195

r':

these

true nature, it seems of this

i

term

with

undesirable

highly regard

abrin.

to

prefer to designate abrin simply of a proteid nature. Action

Pepsin

of

to

make

use

ed,

therefore the chemical poison We

as a

as

experiments

the albumin

in

pepsine resulting filtrate probably due to

on Abrin.

a

a

true was

peptone

not

was not

presented

to

really form-

the action of

coagulated form?the failure of the to

give

a

precipitate

on

boiling being

the conversion of the abrin into

an

acid albuminate by the action of the hydrochloric harmlessness of And the acid albuminate of abrin, as we have acid. abrin when taken by the mouth, it is interesting to is seen, poisonous. note the results of the following experiments, in which The intestinal juices appear to have the property abrin was digested with pepsin outside the body, and of rendering the poison inert, for a large quantity of the resulting product, apparently a peptone, then inabrin may be given per rectum with no other effect than jected into chickens. The pepsin used in these exthe production of acute colitis. periments was found to be fairly well up to the As

bearing

the

on

comparative

British

Pharmacopoeia standard of activity, where two grains of pepsin are stated to dissolve 100 grains of coagulated egg-albumin. Exp. LXVII.?Six grains of the freshly-prepared moist abrin from which the alcohol had been entirely driven off were dissolved in about]'%

oz.

of distilled

water.

To this were

added three grains of pepsin with 6 minims of dilute hydrochloric acid, and the whole digested for six hours at a temperature of 38? Cent. This solution was then filtered, the filtrate

being

a clear, limpid, and almost colourless fluid, which gave scanty white precipitate with absolute alcohol, and on boiling deposited no precipitate, showing that the proteid had apparent-

a

ly become coverted into a peptone. Of this peptone solution 25 minims, representing about 11/2 grains of moist abrin, were injected into a healthy chicken. The chicken presented no symptoms till the following day. It died 53 hours from the time of administration of the injection with all the usual symptoms of abrus-poisoning. Exp. LXVIII.?The previous experiment was repeated on another chicken, which died 60 hours after the administration of the

injection.

abrin-peptone proved fatal in both these experiments, it became necessary to ascertain what the effect of a hypodermic injection of pepsin by itself might be. In the following experiment twice the amount of pepsin which had been used in the two foregoing experiments was injected subcutaneously into a chicken without any positive result. As the

Exp.

LXIX.?Three

hydrochloric

of

grains

minims of distilled water, acid had been

to

pepsin

dissolved

in

25

which three minims of dilute

added,

were

injected

into

a

chicken.

No result.

From these

peptone

of

experiments abrus-albumin

it would appear that the is poisonous, although

very much less active than abrin itself.

It becomes

reconcile these results with the I acknowledged innocuousness of the seeds when taken by the mouth. For on introduction into the stomach therefore difficult

to

a peptone will be formed, and this absorbed ought to prove poisonous

foregoing experiments.

But it is

readily becoming according to the just possible that in