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ROBININ, VIOLAQUERCETIN, MPRTICOLORIN, AND OSYRITRIN. 473

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XLYIIL-Robinin, VioZaquercetin, Myrticolorin, and

0 syyitrin.
GEORGEPERKIN,
By ARTHUR F.R.S.E.
Robinin.
INa previous communication (Trans., 1900, 77, 430), it was shown
t h a t the leaves of Robinia Psezcdacacia contain acacetin, a monomethyl
ether of apigenin, a n interesting point i n view of the discovery by
Zwenger and Dronke (Annalen Xuppl., 1861,1,263) t h a t the flowers of
the same plant contain a quercetin glucoside, robinin. AS these
flowers, however, are practically devoid of dyeing properties, it seemed
doubtful whether a quercetin compound was present, and reference to
the work of these chemists made i b probable this suspicion was well
founded. At the time of Zwenger and Dronke's investigation, the
obstinacy with which these colouring matters retain water of crystal-
lisation was not fully appreciated, and results accurate in themselves
frequently received a wrong interpretation. The analyses of their
" quercetin " were made with material dried at looo, a temperature at

which it is rarely rendered anhydrous; again, it was unlikely that

picric acid mould result from the action of nitric acid on quercetin.
Some difficulty wa8 at first experienced in procuring raw material
for this investigation, but ultimately this was overcome by the kind-
ness of Dr. J. van Rijn, of Maastricht, who was good enough to
superintend the gathering and drying of some quantity of the flowers.
For isolating robinin, Zwenger and Dronke digested the flowers with
boiling water, subsequently evaporating the extract and treating the
residue with alcohol. The following method is more rapid, and is suit-
able for dealing with small quantities of raw material.
The flowers were digested with ten times their weight of boiling
alcohol for 4 hours, the mixture strained through calico, and the
residue well pressed and again treated in a similar manner. The pale
green extract, which deposited a wax on cooling, was concentrated by
evaporation, poured into water, and the mixture extracted with ether,
the alcohol contained in the aqueous liquid being removed by distilla-
tion ; on standing overnight, this solution deposited crystals which
were collected and washed with a mixture of alcohol and chloroform
until the washings were colourless. The residue was then purified by
two o r three crystallisations from water with the aid of animal char-
coal, One hundred and ninety grams of the flowers gave 1.76 grams,
or 0.82 per cent., of robinin in the crude condition, this being reduced
approximately one-Ealf on purification.
As thus obtained, it consisted of extremely pale yellow needles sinter-
VOL. LXXXI. K K
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474 PERKIN : ROBININ, VTOLAQUERCETIN,

ing a t 190' and melting a t 196-19'7' (2. and D. 195O), and had the
general properties mentioned by these authors, When dried a t looo i
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0.1103 gave 0.3123 GO, and 0.0550 H,O. C = 52-49 ; H = 5.54.

C,,H,2020requires C = 52.24 ; H = 5.54 per cent.
Zwenger and Dronke assign t o robinin at 100' the formula C,,H,oO,G
(C = 51.19 ; H = 5.1 l),and with this their analytical figures closely
agree. This discrepancy suggested t h a t our preparation might contain
a trace of free colouring matter which would raise the percentage com-
position. A n alcoholic solution of a second preparation was therefore
poured into ether (in which the colouring matter is soluble, but not
t h e glucoside) and the precipitated product collected, again crystallised
from water, and dried at 100' :
0*1131gave 0.2165 CO, and 0,0575 H20. C=52*20; H = 5 * 6 4 .
The glucoside was thus evidently homogeneous.
Determinations of the water of cry stallisation contained in the
glucoside dried at the ordinary temperature, employing two distinct
preparations, gave the following results :
1.6790, at looo, lost 0.2450 H,O. H20=1551.
1'6079, ), ,, 0.2606 H,O. H20=15.58.
--- C,3H420,0,8H,0requires H,O = 15-97 per cent,
C,3H420,(),7frH20 9 9 H,O= 15*11 3 9
Air-dried robinin thus crystallises with 8H,O. These results are
not in agreement with those of Zwenger and Dronke, who found 14-53
per cent. of water, agreeing with the amount requiFed for the formula
C,,Hao0,,,5~H20, or according t o the above, C,3~42020,7H,0.This
discrepancy is curious, although the lower figures these authors give
for robinin dried at 100' suggest the possibility t h a t their product still
contained some water of crystallisation,
hCO!rr?,p08itiO9?, with Acid,-The dried glucoside was boiled with dilute
sulphuric acid * for at least 2 hours, and after standing overnight, the
liberated colouring matter was collected, washed, and dried at 100' :
0.6110, at looo, gave 0.2330 colouring matter. Found, 38.13.
0.9804, ,, ,, 0.3'745 9, ,, 38.11).
la2266, ,) ,, 004650 7, ,, 37.92.
Zwenger and Dronke, on the other hand, found that air-dried
robinin gave 37.96 per cent. of '' quercetin " dried at loo', a n amount
considerably in excess of that given a.bove. Thus, air-dried robinin,
C,,H,20,0,SH,0, should give 33.70 per cent. of the colouring matter
C,,H,oO,,H,O, or but 31-70 per cent. of the anhydrous substance. In
their paper, they state that robinin is " extremely easily " decom-
* 750 C.C.of 1 per cent. solution for each gram of glucoside.
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MPRTfCOLORTN, AND OSYRITRIN. 475

posed by boiling dilute sulphuric or hydrochloric acids, but with
acid of 1 per cent. strength this was not found to be the case, for with
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a short digestion, 39-54 per cent. of “colouring matter ” was obtained,

a fact suggesting that it was contaminated with unaltered glucoside.
The CoZouring Mattey.- For analysis, the product was recrystallised
from dilute alcohol and dried at 160’ :
0.1122 gave 0.2589 CO, and 0-0355 H,O. C = 62.93 ; H = 3.51.
C,,H,,O, requires C = 62.93 ; H = 3.49 per cent.
Zmenger and Dronke found that this colouring matter, dried at
loo’, gave C=59*31, H=4.49, numbers in close agreement with those
required for the formula C:,,H,,O,,H,O (C= 59.2, H = 3’94).
Prepared as above, i t crystallised in slender, yellow needles melting
at 276-278’, readily soluble in boiling alcohol, and soluble in alkaline
solutions with a pale yellow colour, For additional proof that it was
not quercetin, comparative dyeing trials were carried out employing
woollen cloth mordanted with chromium, aluminium, tin, and iron.
Chromium. Aluminium, Tin. Iron.
Quercetin .....,....,...,...
Red-brown. Brown-orange- Bright Olive-black.
yellow. orange.
Robinin colouring matter Brownbyellow. Full golden- Lemon-yellow. Deep olive-
yellow. brown.
With mineral acids in the presence of acetic acid, it yielded crystal-
line compounds, and to confirm its molecular weight the sulphuric acid
derivative was analysed.
0&1556gave 0.2657 CO, and 0.0457 H20, C = 46.57 ; H = 3.26.
Cl,H,,06,H,S0, requires C = 46-87 ; H = 3.12 per cent.
Alcoholic potassium acetate yielded a monopotassium salt, but
owing to lack of material this was not fully investigated. The acetyl
derivative, prepared in the ueual manner, crystalliaed from methyl
alcohol in colourless needles :
0.1093 gave 0,2438 CO, and 0*0400H20. C = 60.83 ; H = 4a06.
C,,H,O,(C,H,O), requires c = 60.79 ; H = 3.96.per cent.
When fused with alkali, the colouring matter gave p-hgdroxybenxoic
acid (m. p. 208-210°) and phloroglucinol.
These facts, together with a comparative dyeing trial, conclusively
proved that the colouring matter derived from robinin is identical with
that isolated from the flowers of the Delphinum ConsoZida (Proc., 1900,
IS, 182). This similarity was corroborated by the peculiar behaviour
of their acetyl derivatives when heated ;thus, while some preparations
melted at 180--182O, others became liquid a t about 116O, resolidified
as the temperature rose, and finally melted at 180-182’. The colour-
K K 2
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476 PERKIN : ROBININ, VIOLAQUERCETIN,

ing matter is in reality hmpherol, first obtained by aordin (Diss.

Berne) by the decomposition of its monomethyl ether karnpheride, which
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is contained in galanga root (AZpinia oficinarum). This, as Kostan-

ecki suggests (Ber., 1901, 34,3723), has in all probability the con-
stitution

and may be considered as the connecting link between apigenin and

yuercetin. Kampherol, and not guercetin, is the colouring matter pro-
duced when robinin is hydrolysed with acid.
The Sugars.-The acid filtrates formed by the decomposition of
robinin were neutralised with barium carbonate, filtered, and
evaporated to a small bulk. The product yielded an osazone which
after three crystallisations from dilute alcohol was obtained as a
spongy mass of yellow needles sintering at 165' and melting at
178-180°. As a further treatment in this manner did not yield a
homogeneous substance, it was dissolved in alcohol, the solution
poured into ether, and well washed with water. On slow evaporation,
a small quantity of crystalline matter separated, and this was collected,
washed with ether, and recrystallised from alcohol. It melted at
204-205O and resembled glucosaxone.
The filtrate which contained the main bulk of the osazone, on
spontaneous evaporation, deposited crystals which were extracted with
benzene, washed with traces of ether, and recrystallised from dilute
alcohol, I n the preliminary notice (Proc., 1901, 17, 87), it was
considered probable that this substance was galac tosazone, but fer-
mentation experiments kindly carried out for me by Dr. Turnbull,
of the Leather Industries Department, did not corroborate this view.
Employing the sugar solution, as obtained in the above manner from
robinin, a s1igb.t fermentation did indeed occur, but the main bulk
was unattacked and gave an osazone meeting at 180-182°. This
was found to be identical with rhamnosaxone prepared from pure
rhamnose, thus harmonising with the. results of Zwenger and Dronke.
Whether the trace of the glucose derivative simultaneously isolated is
a n ingredient of >robininitself or is derived from 8 second glucoside OF
kampherol also present in minute quantity must, although it appears un-
likely, remain a. matter for conjecture. Very large quantities of r a m
material would be required to decide this point. According to these
views, the formula of robinin is consequently either C33H44020or
C33H42020, and its decomposition by acid may be represented as
follows :
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MYRTICOLORIN, AND OSYRITRIN. 477

C33H44020f QH20= C15H,oO6 -k 3C6H1406.

C33H42020 f 4 H 2 0= C15H1006
4- 2c6H1406 4- C6HI2O6.
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It is thus a most interesting glucoside, as it appears t o be the first

known substance of this class to contain three sugar nuclei. The
above equations require respectively a yield of 37.63 and 37.73 per
cent. of kampherol, whereas that actually obtained was 38-13,
38.19, and 37.92 per cent.
Dyeing sxperiments carried out in the usual manner with woollen
cloth showed that robinin is almost devoid of tinctorial properties ; this
was to be expected from the preliminary examination of the flowers.

The Identity of Osyritrin, Violaquercitrin and Myrticolorin.

Jn a previous communication (Trans., 1897, 71, 1133), it was shown
that the leaves of the CoZpoon compressurn contain a quercetin glucoside,
osyritrin, to which, dried at 130°, the formula C2'7H30017was assigned.
A t that time, the raw material available yielded little more than
1 gram of the pure substance, but subsequently, through the kindness
of Mr. R. H. Coaton, of Wellington, Cape of Good Hope, a larger
supply was obtained, and it became possible to investigate i t more fully.
Water of Crystallisation.-For determining the loss of weight which
the air-dried glucoside undergoes, it was exposed for about three
weeks over sulphuric acid in a desiccator. As the product on exposure
to the atmosphere rapidly assumed its original weight, the analysis
was performed indirectly in this manner.
1,3345, dried as above, gained 0.0770 H,O. Found 5.76.
C27H30017,2H20 requires H20= 5.44 per cent.
At 130°, employing a n oil-bath jacketed with amyl alcohol (b. p.
130-131"), it suffered a further loss of 0*0200H,O, or 1.49 per cent.
($H,O requires 1.43) and finally, at 160" (oil-bath jacketed with 'tur-
pentine), 0.0220 gram H20was evolved, or 1.67 per cent. ($H,O = 1.47).
Osyritrin, therefore, crystallises from water with 3 mols. of water of
crystallisation.
Anhydrous osyritrin is remarkably hygroscopic, thus, on exposure
for 1 hour in the air of the room, it completely regains its water of
crystallisation. On analysis :
0.1170 gave 0.2275 CO, and 0.0537 H20. C = 53.03 ; H = 5.09.
C27H28016requires C 53.28 ; H = 4.60 per cent.
=)

Decomposed with dilute sulphuric acid:

0*8400gave 0.4137 C15H1007. Found 49.25.
C27H2,0,,requires Cl5HI0O7= 49.67 per cent.
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478 PEREIN : ROBININ, VIOLAQUERCETIN,

The osazone of the sugar melted at 204-2305', as previously found,

and was evidently glucosazone.
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Osyritrin dried a t 160' has thus the formula C27H28016, aqd at 130'
(C27H28016)2,H20, and not C27HR0017 as formerly suggested. Its hydro-
lysis with acid is analogous to that of rutin, which in a similar manner
gives quercetin and rhamnose (Schunck, Trans., 1888, 53, 264) :
C27H2,ol6 + 3H20 = +
C15Hlo07 2C6H120,.
C27H32016 4- 3H20 = C,,Hl00, 4- 2C6H#6.
Rutin.

Vio2ccquercitrim.
Mandelin (Jahresber., 1883, 1369) isolated this glucoside from the
flowers of the Viola tricolor uariensis, and assigned to it the formula
C,,H42024. It has been previously pointed out that this is more
correctly C27H,601,,as the true molecular weight of quercetin was not
known at that time.
For its preparation from the flowers, a method identical with that
employed for the preparation of osyritrin (Zoc. cit.) gave an excellent
result :
0.1071, dried at 130°, gave 0,2036 CO, arid 0.0510 H20. C = 51-84 ;
H = 5.29 per cent.
0.1144, dried at 160', gave 0.2230 CO, and 0-0535 H20. C = 53.16 ;
H = 5.19 per cent.
As Mandelin employed water for the isolation of his substance, i t
seems possible that the result he obtained was due to the contamination
of his product with a trace of quercetin.
Water of CvystaZZiscction.-When dried over sulphuric acid, 0.970 2
gained, on exposure to the atmosphere, OgO580 H,O. Found H 2 0 =
5.64 ; 2H20 requires 5-44 per cent.
1.0280, a t 130°, lost 0.070 H20. H 2 0= 6.80.
2.5H20 requires H,O = 6.79 per cent.
1.0280, a t 160°, lost 0.085 H20. H20= 8.26.
3H20 requires H20= 8.1 6 per cent.
Violaquercitrin thus crystallises with 3H20,
When dried at 160' and decomposed with acid, it gave 49.35 per
cent. of quercetin, which is in accordance with the following equa-
tion (49.67 per cent.) :
C27H2,0164- 3H20 = cl5Hl007 4- 2C6H120p
The osazone of the sugar melted at 204-205' and was evidently
glucosazone. Violaquercitrin melts at 186' when slowly and at 190°
when rapidly heated, and is undoubtedly identical with osyritrin.
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MYRTICOLORIN, AND OSYRITRIN. 479

Ngrticolorin.
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This quercetin glucoside was isolated by H. G. Smith (Trans., 1898,

73, 697) from the leaves of the Ezccalyptue mcworhynaha, who
assigned to it the formula C27H28016,
and represented its decomposition
with acid by the equation
C2!7H28016 f.3 H 2 0 = C,,H,,07 + 2c6H1206*

Smith at first considered the sugar thus produced (Zoc. cit.) to be

galactose, but more reoently, in a private communication, he informs
me that this is not the case, as he has obtained from it glucosazone,
(m. p. 204-205O). It is worthy of note t h a t he (Zoa. cit.) calls atten-
tion in his paper to the remarkable similarity of myrticolorip to
osyrit rin.
The raw material employed mas some commercial myrticolorin, for
which I am indebted to the kindness of Mr. Smith. After purifica-
tion, a sample dried a t 160' gave C = 53.03 and H = 5-09 per cent,,
and on decomposition with acid 49.25 per cent. of quercetin :
Driedoversulphuric acid,on exposure to air it gained H20= 5.76 percent.
9, ,, and heated a t 130' it lost H20= 1.49 per cent,
Dried at 130° and heated at 160° it lost H 2 0 =1.67 per cent,
Myrticolorin, therefore, crystallises from water with 3H20, and this
may be fractionally removed by methods identical with those employed
in the case of osyritrin. As the melting points, general reactions, and
dyeing properties also agree, there can be no doubt that the azlbstances am
identical, moreover, it was previously shown (Trans., 1899,75,433) that
osyritrin, violaquercitrin, and myrticolorin give, by means of potassium
acetate, monopotassium derivatives (found K = 6-21). There is no
doubt, therefore, t h a t these also must be regarded as one of the same
compound, represented by the formula C27H27OI6 (K = 6.03 per
cent.).
Certain glucosides, as ruberythric acid and the purpurin glucoside
contained in madder are decomposed during the dyeing operation by
means of the mordant, which combines with the alizarin or purpurin
thus liberated. Such, however, I 6nd is not the case with the known
glucosides of the quercetin group, which are dyestuffs of themselves,
and give shades differing considerably in most cases from those yielded
by the colouring matters from which they are derived.* A simple
experiment with Persian berries clearly illustrates this point. This
dye contains glucosides of rhamnetin, rhamnazin, and quercetin, and
* Nietzki (" Chemistry of Organic Dye-stuffs," 1892, 255) considers it probable
that in dyeing the quercitrin splits up, and that the shades obtained are due to
the formation of quercetin lakes.
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480 PERMAN: THE INFLUENCE OF SALTS ON THE

there is also present a ferment which at about 40' in the presence of

water hydrolyses these compounds ; if, therefore, the dye-bath be
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raised slowly t o the boiling point, this change occurs, and the resulting
shade is due to the free colouring matters and not to the glucosides. On
the other hand, if the berries be plunged into boiling water, the activity
of the ferment is a t once destroyed; and the tinctorial property of this
extract is now due to the glucosides as the shade indicates. This
difference is very similar to t h a t shown between quercetin and its
glucosides, which gave the following results with woollen cloth mor-
danted in the usual manner :
Chromium. Aluminium. Tin. Iron.
Quercitrin... Full brown-yellow. Full golden-yellow. Lemou-yellow. Deep olive.
Quercetin ... Red-brown. Brown-orange, in- Bright orange. Olive-black.
clining to red.
Osyritrin ... Brown-yellow. Full golden-yellow. Lemon-yellow. Dull brown.
Rntin. ........ Brown-yellow. Full golden-yellow. Lemon-yellow. Dull brown.
It is interesting to observe that rutin and osyritrin have identical
tinctorial properties, which points to the fact that the two sugar nuclei
of each (in the one case rhamnose and the other dextrose) are similarly
attached to the quercetin residue. As regards their actual disposition,
satisfactory proof is a t present wanting, but it is highly probable t h a t
one a t least is attached to the catechol group, as the dyeing properties
suggest the absence of o-hydroxyl groups. Their behaviour with
potassium acetate is a n indication that they contain intact the hydr-
oxyl of the pyrone ring, for it is found t h a t galangin, by this method,
forms a monopotassium salt. The properties as a whole would harmonise
closely with those of a compound containing both sugar nuclei attached
t o the catechol group, but further evidence is needed before this conclu-
sion can be adopted.
RESEARCHLABORATORY,
CLOTHWORKERB'
DYEINGDEPARTMENT,
Y ORKSHIRE COLLEGE.