Pharmaceutical Chemistry Journal
4, 1998
MEDICINAL PLANTS
CHEMICAL COMPOSITION AND PHARMACOLOGICAL
ACTIVITY OF AN AQUEOUS EXTRACT FROM SEA
BUCKTHORN WASTE PRODUCTS
IC A. Dychko, z E. V. Kulagina, 1 V. V. Khasanov, 1 E. N. Tveryakovafl
G. L. Ryzhovafl V. G. Paslainskii, 2 and N. V. Gribel '2
Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 32, No. 4, pp. 3 2 - 34, April, 19989
Original article submitted January 17, 1995.
As is known, the fruits of sea buckthorn are used for the
production of oil and juice. The pharmacological activity of
preparations based on the buckthorn oil is due to the biologi-
cally active substances, including carotinoids, sterols, etc.,
present in the fruits [ 1 - 4 ] .
The existing technologies of buckthorn processing are
based on displacement of the buckthorn oil by vegetable oils
(diffusion technology) [5, 6], or on the extraction with lique-
fied gases (CO2, freon-12) under pressure ['7-9] 9 Biologi-
cally active compounds also can be isolated from buckthorn
by extraction with organic solvents (petroleum ether, pen-
tane, hexane, methylene chloride) [1, 4, 10].
The processing of buckthorn leads to a large amount of
primary waste product still containing a combination of bio-
logically active substances. The chemical composition and
pharmacological activity of concentrates obtained from the
buckthom waste products upon extraction with petroleum ether,
hexane, and methylene chloride were studied in [ 10, 11].
The purpose of this work was to study the chemical com-
position and pharmacological activity of an aqueous extract
from the waste products ofbuckthorn processing by freon ex-
traction ("Altaivitaminy" Production Association, Biisk, Rus-
sia). The aqueous extract was obtained using a wasteless
ecologically safe technology developed for plant raw material
processing, providing maximum yield and preservation of
biologically active compounds.
Because the therapeutic action ofbuckthorn is now com-
monly attributed to the presence of carotinoids, we have de-
termined their content in the aqueous extract by a conven-
tional method [ 12].
I Tomsk State University, Tomsk, Russia.
2 Tomsk Institute of Pharmacology, Russian Academy of Medical Sciences,
Tomsk, Russia.
204
Vol. 32, No.
AS is known, many multivitamin preparations contain,
besides vitamins, carbohydrates, micro-, and macroelements.
Therefore, it was also of interest to analyze the aqueous buck-
thorn extract for these components.
EXPERIMENTAL CHEMICAL PART
The total content of carbohydrates was measured using
aphenol-sulfuric acid procedure with spectrophotometric
detection [13]. The component concentrations were deter-
mined with the aid of a calibration plot constructed by meas-
uring standard glucose solutions.
The water-soluble vitamins in the aqueous buckthom ex-
tract were determined by reversed-phase HPLC. The analysis
was performed on a Milikhrom-1 chromatograph ("Nauchpri-
bor" Production Association, Orel, Russia). The sample
preparation procedure included deproteinization, precipita-
tion of proteins with acetic acid, and concentration in ion-ex-
change cells as described in [14]. The concentrating cells
were of the Diapack-Sulfo and Diapack-Ta types ("BioKhim-
Mak" company, Moscow). The chromatographic separation
was performed at room temperature on a column (80
2 mm) filled with a Separon SGX RP-S sorbent (Elsico com-
pany, Moscow). The sample fractions were detected with a
9 UV detector operated at 230 rim. The mobile phases were
0.1 M sodium acetate (A) and an A - acetonitrile 1 : 1 mix-
ture (B); the gradient elution was effected using an A + B
mixture with the B component content varying from 20 to
80%.
The fat-soluble vitamins were extracted with hexane
from the initial aqueous buckthorn extract. The fractions ex-
tracted were combined, dried, and evaporated to reduce the
volume to 500 ~tl. The chromatographic analysis was per-
formed on the Milikhrom-1 chromatograph using a column
0091-150X/98/3204-0204520.00 9 1999 Kluwer Academic/Plenum Publishers
 Pharmacological activity of an Aqueous Extract from Sea Buckthorn Waste Products 205

(80 x 2 mm) filled with a Sitasorb SPH-NH2 sorbent (Elsico
company, Moscow). The solvents were hexane (A) and a
20% hexane + 80% chloroform mixture (B); the change from
eluent A to B was performed 9 min after the beginning of
separation. The UV detection was performed at 260 urn.
The quantitative determination of both vitamin types was
based on the use of an external standard [15]. The standard
compounds were vitamins (DAB, Ph. Eur.) purchased from
H o f f m a n - La Roche company (Switzerland).
The micro- and macroelement contents in the aqueous
buckthom extract were determined by the method of radioac-
tivation analysis.
The results of analyses are presented in Table I.
E X P E R I M E N T A L BIOLOGICAL PART
Preliminary biological tests showed that peroral admini-
stration o f the dry buckthorn extract to mice at doses in the
range 50 - 5000 mg / kg neither led to a loss of the test ani-
mals nor produced noticeable changes in their general state
and behavior.
Experiments on the model of acute stress (induced by
18-h hanging of female mice by a skin fold) showed that a
5-day peroral administration of the buckthorn extract at a
dose of 100 or 200 m g / k g produced a pronounced antiulcer-
ous effect: the average amount of ulceration damage in the
stomach reliably (p < 0.05) decreased from 12.2 + 2.3 in the
male mice) having an initial weight of I0 g. The test animals
received the buckthorn extract daily over a time period of
1 month. Upon elapse of this time, the weight coefficients
were determined for the internal organs (thymus, spleen,
liver, pancreas, heart, and front tibial muscle) as the ratios of
the organ to body weights (mg/g). An analysis of these data
indicated that the weight gain of animals in the test groups
was on the average 1 g higher than that in the control group.
The group receiving the buckthom extract at a dose of
200 mg / kg showed an increase in the weight coefficients for
thymus, pancreas, and liver as compared to the control group.
The average 1-month weight gain in the control group was
14.6 and 12.7 g for males and females, respectively. In the
group receiving the buckthom extract at a dose of 50 mg / kg,
these increments were 17.2 and 13.7g, and the animals
treated at a dose of 200 m g / k g showed a weight gain of 15.8
and 14.2 g, respectively. Note that the l-month weight gain
T A B L E 1. C h e m i c a l C o m p o s i t i o n o f an A q u e o u s Extract f r o m the W a s t e
P r o d u c t o f B t m k t h o r n P r o c e s s i n g a n d the C o m p o s i t i o n s o f s o m e C o m m e r c i a l
Multivitamin Preparations
Dry
Component buckthom Revit, Supradyn, ** U n i c a p , **
extract ** m g mg mg
(0.6 g)*
Vitamin C 13.9 m g 35.0 150.0 50.0

control to 6.7 + 1.1 and 3.0 + 0.5, respectively. Administra- Vitamin B t 0.55 m g 1.0 2.0 2.5

tion of the doses 50 and 400 m g / k g was much less effective, Vitamin B2 89 I.tg 1.0 5.0 2.5
Vitamin B 6 8.56 m g - 10.0 0.5
reducing the amount of damage in the stomach to only 8.8 +
V i t a m i n Bt2 - - 2 . 0 x 10 - 3 5 . 0 x 10 - 3
1.2 and 9.3 + 2.5, respectively.
Vitamin P 6.0 lag - 50.0 -
Experiments on the hypoxic hypoxia model (sodium ni- V i t a m i n PP 5 lag - - -
troprusside, 25 m g / k g ) showed that administration of the Vitamin A l lag*** 0.86 1.0 1.5
buckthom extract at a dose of 200 m g / kg led to an increase Vitamin EA l lag - 10.0 -
in the survival lifetime of the test mice from 13.5 + 0.6 min in Vitamin D 55 lag 2.5 - 12.5

the control to 15.7 + 0.3 min (p < 0.05). Lower doses (50 and Vitamin D 2 - - 12.5 -

100 m g / k g ) did not significantly affect the survival of test Vitamin H - - 0.25 -
F o l i c acid - - 1.0 -
animals.
Carbohydrates 29.0 m g - - -
The effect of the buckthorn extract administered at the
Ca 0.66 - 51.3 40.0
doses 50, t00, 200 or 400 m g / k g on the work capacity was wt.%
studied in mice on the model of forced swimming under load Mg 1.7 x 10 - 2 - 21.2 6.0
(10% of the body weight). Here, the drug exhibited a maxi- wt.%
mum activity at a dose of 50 mg / kg. During the first swim, Fe 1.4 x 10 - 2 - 10.0 10.0

the mice receiving the above drug dose showed only some wt.%
Mn - - 0.5 1.0
tendency to increase in the work capacity (130+23 sec
P - - 23.8 -
against 102 + 17 sec in the control). In the repeated test (after
Cu 1.7 x 10 - 3 - 1.0 1.0
a 1-h rest), animals in the test group floated on the surface for wt.%
a much longer time as compared to that in the control group Zn - - 0.5 -
(119 + 17 sec against 65 + 9 sec). Note that other doses of the Mo - - 0.1 -

buckthorn extract did not increase the work capacity in test I - - - 0.15
animals. K - - - 5.0

We have performed a special series of experiments to T h e r a p e u t i c dose for adults.

study the effect of the buckthom extract (50 and 200 m g / k g ) 1 Drajee.
C a l c u l a t e d for carotinoids.
on the weight variation in test animals (mongrel male and fe-
 206
o f males in both control and test groups was higher than that
of females.
Experimental data obtained on the chemical composition
and pharmacological activity o f the aqueous extract from the
waste products o f b u c k t h o m processing, and a comparison o f
these data with characteristics o f the known multivitamin
preparations (Hexavit, Undevit, Supradyn, Unicap) suggest
that the aqueous buckthom extract is not inferior to these
drugs and even exceeds them in some respects.
Thus, the aqueous extract from the waste products o f
buckthorn processing can be used both as a multivitamin ad-
ditive to meals and as independent multivitamin preparation.
The fact that this product is obtained from a waste product
using on ecologically safe medium (water) is a contribution
to more rational use o f the natural resources.
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