Параметри стандартизації трави Alfredia nivea KAR.&KIR.
The Kazakhstan flora is rich in promising poorly-studied plants, which are traditionally used in folk medicine, but their introduction into medical practice requires additional in-depth research using modern scientific methods. Alfredia nivea KAR&KIR of the Asteraceae family, which is us...
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| Опубліковано в: | Журнал органічної та фармацевтичної хімії |
|---|---|
| Дата: | 2023 |
| Том: | 21 |
| Випуск: | 1 |
| Сторінки: | 46-53 |
| ISSN: | 2518-1548 |
| Автори та афіліації: |
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| Автори: | , , , , , |
| Формат: | Стаття |
| Мова: | Англійська |
| Опубліковано: |
National University of Pharmacy
2023
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| Онлайн доступ: | https://ophcj.nuph.edu.ua/article/view/276004 |
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| Назва журналу: | Journal of Organic and Pharmaceutical Chemistry |
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Репозитарії
Journal of Organic and Pharmaceutical Chemistry| _version_ | 1874364446741102592 |
|---|---|
| author | Rustemkulov, Almat G. Gontova, Tetiana M. Makhatova, Balzhan G. Rustemkulova, Aisana E. Datkhayev, Ubaidilla M. Koshovyi, Oleh M. |
| author_facet | Rustemkulov, Almat G. Gontova, Tetiana M. Makhatova, Balzhan G. Rustemkulova, Aisana E. Datkhayev, Ubaidilla M. Koshovyi, Oleh M. |
| author_institution_txt_mv | [
{
"author": "Almat G. Rustemkulov",
"institution": "Asfendiyarov Kazakh National Medical University",
"orcid": ""
},
{
"author": "Tetiana M. Gontova",
"institution": "National University of Pharmacy of the Ministry of Health of Ukraine",
"orcid": ""
},
{
"author": "Balzhan G. Makhatova",
"institution": "Asfendiyarov Kazakh National Medical University",
"orcid": ""
},
{
"author": "Aisana E. Rustemkulova",
"institution": "Asfendiyarov Kazakh National Medical University",
"orcid": ""
},
{
"author": "Ubaidilla M. Datkhayev",
"institution": "Asfendiyarov Kazakh National Medical University",
"orcid": "0009-0009-8876-6927"
},
{
"author": "Oleh M. Koshovyi",
"institution": "National University of Pharmacy of the Ministry of Health of Ukraine",
"orcid": ""
}
] |
| author_orcid_str_mv | 0009-0009-8876-6927 |
| author_sort | Rustemkulov, Almat G. |
| baseUrl_str | https://ophcj.nuph.edu.ua/oai |
| collection | OJS |
| container_end_page | 53 |
| container_issue | 1 |
| container_start_page | 46 |
| container_title | Журнал органічної та фармацевтичної хімії |
| container_volume | 21 |
| datestamp_date | 2026-08-23T20:00:17Z |
| description | The Kazakhstan flora is rich in promising poorly-studied plants, which are traditionally used in folk medicine, but their introduction into medical practice requires additional in-depth research using modern scientific methods. Alfredia nivea KAR&KIR of the Asteraceae family, which is used in folk medicine as a neurotropic agent, is an interesting object for introduction into official medical and pharmaceutical practice.Aim. To create new medicines based on Alfredia nivea herb, it is necessary to develop methods for quality control of this raw material, therefore, the aim of the research was to determine the parameters for standardization of the Alfredia nivea KAR & KIR. herb.Materials and methods. The study objects were samples of the A. nivea herb collected in Kungei Alatau, 4.3 km southeast of the Karabulak village, Eastern Karabulak canyon, Almaty Region, Kazakhstan. The macroscopic and microscopic studies of the A. nivea herb were performed according to the methodology of the European pharmacopeia (EuPh) 2.8.23 “Microscopic examination of the medicinal plant raw material”. The macroscopic studies were performed using a magnifying glass and a MBS-9 binocular microscope, the microscopic studies were done using MS Microscopes 10 (oculars X5, X10, X15, lenses x10,x40), Micromed XS-4130 (oculars WF15X, lenses x40/0.65, x10/0.25) with a microphotonozzle (China). Identification of the main substances was carried out by the TLC method, testing and the quantitative determination of the flavonoid content were performed according to the EuPh methods.Results and discussion. Morphological and anatomical features of the A. nivea herb have been determined; on their basis Identifications A and B have been proposed; TLC Identification C of the main BAS of the raw material has been developed; indicators of purity tests have been determined. It has been proposed to carry out the quantitative determination by the content of flavonoids.Conclusions. The parameters of the A. nivea herb standardization have been determined on the basis of the following indicators: macroscopic and microscopic features, TLC identification of the main BAS of the raw material (hyperoside, rutin, quercetin and chlorogenic acid), related impurities (not more than 2 %), stems with a diameter of more than 20 mm (not more than 10 %), the loss on drying (not more than 13 %), the total ash (not more than 10 %) and at least 0.5 % flavonoids calculated with reference to rutin. |
| doi_str_mv | 10.24959/ophcj.23.276004 |
| first_indexed | 2025-07-23T04:43:16Z |
| format | Article |
| fulltext |
ISSN 2308-8303 (Print) / 2518-1548 (Online) 46
Original Research
http://ophcj.nuph.edu.ua
UDC 615.322:582.998
A. G. Rustemkulov2, T. M. Gontova1, B. G. Makhatova2, A. E. Rustemkulova2,
U. M. Datkhayev2, О. M. Коshovyi1
Standardization Parameters of Alfredia nivea KAR.&KIR Herb
1 National University of Pharmacy of the Ministry of Health of Ukraine,
53 Pushkinska str., 61002 Kharkiv, Ukraine
2 Asfendiyarov Kazakh National Medical University,
94 Tole bi str., 050000 Almaty, Kazakhstan
Abstract
The Kazakhstan flora is rich in promising poorly-studied plants, which are traditionally used in folk medicine, but their intro-
duction into medical practice requires additional in-depth research using modern scientific methods. Alfredia nivea KAR&KIR
of the Asteraceae family, which is used in folk medicine as a neurotropic agent, is an interesting object for introduction into
official medical and pharmaceutical practice.
Aim. To create new medicines based on Alfredia nivea herb, it is necessary to develop methods for quality control of this
raw material, therefore, the aim of the research was to determine the parameters for standardization of the Alfredia nivea
KAR & KIR. herb.
Materials and methods. The study objects were samples of the A. nivea herb collected in Kungei Alatau, 4.3 km southeast
of the Karabulak village, Eastern Karabulak canyon, Almaty Region, Kazakhstan. The macroscopic and microscopic studies of
the A. nivea herb were performed according to the methodology of the European pharmacopeia (EuPh) 2.8.23 “Microscopic
examination of the medicinal plant raw material”. The macroscopic studies were performed using a magnifying glass and a
MBS-9 binocular microscope, the microscopic studies were done using MS Microscopes 10 (oculars X5, X10, X15, lenses x10,
x40), Micromed XS-4130 (oculars WF15X, lenses x40/0.65, x10/0.25) with a microphotonozzle (China). Identification of the
main substances was carried out by the TLC method, testing and the quantitative determination of the flavonoid content
were performed according to the EuPh methods.
Results and discussion. Morphological and anatomical features of the A. nivea herb have been determined; on their basis
Identifications A and B have been proposed; TLC Identification C of the main BAS of the raw material has been developed;
indicators of purity tests have been determined. It has been proposed to carry out the quantitative determination by the
content of flavonoids.
Conclusions. The parameters of the A. nivea herb standardization have been determined on the basis of the following in-
dicators: macroscopic and microscopic features, TLC identification of the main BAS of the raw material (hyperoside, rutin,
quercetin and chlorogenic acid), related impurities (not more than 2 %), stems with a diameter of more than 20 mm (not
more than 10 %), the loss on drying (not more than 13 %), the total ash (not more than 10 %) and at least 0.5 % flavonoids
calculated with reference to rutin.
Keywords: Alfredia nivea; herb; standardization; quality control methods
А. Г. Рустемкулов2, Т. М. Гонтова1, Б. Г. Махатова2, А. Є. Рустемкулова2, У. М. Датхаєв2, О. М. Кошовий1
1 Національний фармацевтичний університет Міністерства охорони здоров’я України,
вул. Пушкінська, 53, м. Харків, 61002, Україна
2 Казахський національний медичний університет імені С. Д. Асфендіярова,
вул. Толе бі, 94, м. Алмати, 050000, Казахстан
Параметри стандартизації трави Alfredia nivea KAR.&KIR.
Анотація
Флора Казахстану багата на перспективні маловивчені рослини, які застосовують у народній медицині, але для їх впро-
вадження в медичну практику потрібні додаткові глибокі дослідження, за допомогою сучасних наукових методів.
Цікавим об’єктом для впровадження в офіцинальну медичну та фармацевтичну практику є Альфредія снігова (Alfredia
nivea KAR. & KIR.) родини Asteraceae, яку в народній медицині використовують як нейротропний засіб.
Мета. Для створення нових лікарських засобів на основі трави Альфредії снігової необхідно розробити методи контролю
якості цієї сировини, тому метою досліджень було визначити параметри стандартизації трави A. nivea KAR. & KIR.
ISSN 2308-8303 (Print) / 2518-1548 (Online) 47
Journal of Organic and Pharmaceutical Chemistry 2023, 21 (1)
Матеріали та методи. Об’єктами дослідження були зразки трави Альфредії снігової (A. nivea), заготовлені в Кунгей
Алатау, за 4,3 км південно-східніше від с. Карабулак, ущ. Східний Карабулак, Алматинської області, Казахстан. Макро- і мі-
кроскопічні дослідження трави Альфредії снігової виконували згідно з методикою Європейської фармакопеї (ЄФ) 2.8.23
«Мікроскопічне дослідження лікарської рослинної сировини». Для макроскопічних досліджень використовували лупу та
бінокулярний мікроскоп МБС-9, для мікроскопічних – мікроскопи МС 10 (окуляр х5, х10, х15, об’єктиви х10, x40), Micromed
XS-4130 (окуляр WF15X, об’єктиви х40/0,65, х10/0,25) з мікрофотонасадкою (КНР). Ідентифікували основні речовини мето-
дом ТШХ, випробування та кількісне визначення за вмістом флавоноїдів здійснювали відповідно до методик ЄФ.
Результати та їх обговорення. Визначено морфологічні й анатомічні ознаки трави Альфредії снігової, на основі яких
запропоновано Ідентифікації А та В, розроблено ТШХ Ідентифікацію С основних БАР сировини, визначено показни-
ки випробувань на чистоту, а також запропоновано проводити кількісне визначення за вмістом флавоноїдів.
Висновки. Визначено параметри стандартизації трави Альфредії снігової за такими показниками: макро- та мікро-
скопічні ознаки, ТШХ ідентифікація основних БАР сировини (гіперозид, рутин, кверцетин та хлорогенова кислота),
сторонні домішки (не більше 2 %), наявність стебел діаметром понад 20 мм (не більше 10 %), втрата в масі під час
висушування (не більше 13 %), загальна зола (не більше 10 %) і не менше 0,5 % флавоноїдів у перерахунку на рутин.
Ключові слова: Альфредія снігова; Alfredia nivea; трава; стандартизація; методи контролю якості
Citation: Rustemkulov, A. G.; Gontova, T. M.; Makhatova, B. G.; Rustemkulova, A. E.; Datkhayev, U. M.; Коshovyi, О. M. Standardization
parameters of Alfredia nivea Kar. & Kir herb. Journal of Organic and Pharmaceutical Chemistry 2023, 21 (1), 46 – 53.
https://doi.org/10.24959/ophcj.23.276004
Received: 29 December 2022; Revised: 30 January 2023; Accepted: 10 February 2023
Copyright© 2023, A. G. Rustemkulov, T. M. Gontova, B. G. Makhatova, A. E. Rustemkulova, U. M. Datkhayev, О. M. Коshovyi.
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0).
Funding: The authors received no specific funding for this work.
Conflict of interests: The authors have no conflict of interests to declare.
■ Introduction
The Kazakhstan flora is rich in promising
poorly-studied plants, which are traditionally used
in folk medicine, but their introduction into me-
dical practice requires additional in-depth re-
search using modern scientific methods [1, 2].
Alfredia nivea KAR&KIR of the Asteraceae fami-
ly, which is used in folk medicine as a neurotro-
pic agent, is an interesting object for introduc-
tion into official medical and pharmaceutical
practice in Kazakhstan [3].
A. nivea is a perennial plant of 25 – 70 cm
height with leathery, oblong-lanceolate, pinnate-
ly lobed leaves. The venation is pinnate, the la-
teral veins at the ends turn into sharp yellowish
spines. The leaf blade is grayish-green on the
upper side, whitish-pubescent on the lower side.
The inflorescence is a spherical head of pink tubu-
lar flowers with a multi-row involucre. The plant
blooms in July, fruits ripen in August – Septem-
ber. It grows in the subalpine and alpine belts of
the mountains, along the stepped rocky slopes,
in spruce-fir forests. It is found in Tarbagatai,
Dzungarian, Trans-Ili and Küngöy Ala-Too Range
Alatau [4 – 6]. The plant’s raw material reserves
are sufficient for the needs of Kazakhstan.
Plants of the Alfredia genus are insufficient-
ly studied. Most of the studies are devoted to
A. cernua L. Thus, extracts of this species have
been shown to affect the central nervous system
and improve memory [7, 8]. It has also been found
that A. cernua extracts have nootropic properties,
contribute to the improvement of indicators of
orientation-exploratory behavior, the preserva-
tion of the passive escape reflex during hypoxic
shock, and the increase of physical performance
in mice [9, 10]. The most pronounced effect was
observed with the use of the A. cernua extract
obtained with 95 % ethanol [10, 11]. It has also
been shown that the extract of the A. nivea
above-ground part in 95 % ethanol in the dose of
100 mg kg–1 restores exploratory behavior and
reflex safety after hypoxic trauma [12]. Thus, the
A. nivea herb is a promising raw material for
creating new neurotropic medicines.
Previously, the standardization of the A. cer-
nua herb [13], the macroscopic and microscopic
study of the A. nivea herb were carried out, and
its diagnostic signs were determined [14]. These
data were taken into account when developing
a project of quality control methods for the A. ni-
vea herb.
Therefore, the aim of the research was to de-
termine the parameters of the standardization
of the A. nivea KAR. & KIR. herb.
■ Materials and methods
The study objects were samples of the A. nivea
herb collected in Kungei Alatau, 4.3 km south-
east of the Karabulak village, Eastern Karabulak
canyon, Almaty Region, Kazakhstan (1762 m abo-
ve the city, N 43°02’30.2”, E 078°34’16.0”) July
ISSN 2308-8303 (Print) / 2518-1548 (Online) 48
Журнал органічної та фармацевтичної хімії 2023, 21 (1)
06, 2021. The identity of the plant was determi-
ned by professor Tetiana M. Gontova, D. Sc., Na-
tional University of Pharmacy (Kharkiv, Ukrai-
ne) [4, 15]. Voucher specimens were deposited at
the School of Pharmacy, Asfendiyarova Kazakh Na-
tional Medical University (Almaty, Kazakhstan,
No. 432 – 435). The raw material was dried at room
temperature [16, 17] in a well-ventilated area for
ten days and stored in paper bags [18, 19].
The macroscopic and microscopic studies of
the raw material were performed according to the
methodology described in the European Pharma-
copeia (EuPh) 2.8.23 “Microscopic examination
of the medicinal plant raw material” [18, 20].
The macroscopic studies were conducted using a
magnifying glass and a MBS-9 binocular micro-
scope. The study of the anatomical structure of
the A. nivea herb was carried out on samples of
the whole and cut raw material according to the
requirements of the EuPh. The herb was fixed
in a mixture of 96 % ethanol R – glycerin R – pu-
rified water R (1:1:1). The structure of stems
and leaves was studied on transverse sections.
The epidermal of the organs was considered from
the surface according to generally accepted me-
thods [18, 21]. The raw material was crushed ac-
cording to the requirements of the EuPh mono-
graph 2.9.12 “Sit analysis” and clarified with the
help of chloral hydrate R [18, 20]. The studies
of transverse and longitudinal sections, epider-
mis and preparations from the surface were car-
ried out using MS Microscopes 10 (oculars X5,
X10, X15, lenses x10, x40), Micromed XS-4130
(oculars WF15X, lenses x40/0.65, x10/0.25) with
a microphotonozzle (China). The results of the
study were recorded using the Canon IXUS 220
HS camera.
To develop the identification method for the
main biologically active substances (BAS) in the
A. nivea herb, the TLC method was used [18, 22].
Merck Silica Gel F254 plates were used for chro-
matography. Purity of solvents used for the pre-
paration of chromatographic systems were of che-
mically pure and analytical grades. The ratio of
solvents indicated by numbers were taken in vo-
lumetric units.
The content of related impurities (2.8.2), the
loss on drying (2.2.32), the total ash content
(2.4.16) were determined according to the re-
quirements of the EuPh [18, 23, 24].
Assay of flavonoids. The assay of flavonoids
was determined in accordance with the metho-
dology of the EuPh by the spectrophotometric me-
thod calculated with reference to rutin [18, 25].
About 5.0 g of Alfredia herb (accurate weight)
crushed to the size of particles passing through
a sieve with a diameter of 2 mm was placed in a
200 mL ground joint flask, 50 mL of 70 % etha-
nol solution was added. The flask was connected
to a condenser and heated on a water bath for
30 min, periodically shaken to wash off partic-
les of the raw material from the walls. The mix-
ture was cooled and filtered through cotton wool,
so that the particles of the raw material did not
get on the filter. The cotton wool was transferred
to the ground joint flask for extraction, and the
new portion of the extractant was added. The ex-
traction was repeated twice under the conditions
described above, and the extract was filtered in
the same flask. The combined extract was evap-
orated up to ¼ of the initial volume. The evapo-
rated extract was quantitatively transferred to
a 50.0 mL measuring flask, cooled and was di-
luted with 70 % ethanol solution to the volume
(Solution A).
2.0 mL of Solution A was placed into a measu-
ring 25 mL flask, 2.0 mL of 3 % aluminum chloride
solution in 96 % ethanol R was added, the volume
was diluted with 70 % ethanol solution and mixed.
In 30 min, the solution was filtered through
the paper filter, throwing away the first portions
of the filtrate, and the optical density was meas-
ured on a Specol 1500 spectrophotometer (Swit-
zerland) at a wavelength of 410 nm in a 10 mm
cuvette. Reference solution contained 2.0 mL of
Solution A, diluted with 70 % ethanol to the vol-
ume in a 25 mL flask.
In parallel, experiment with a Standard solu-
tion of rutin (pharmacopoeial standard) was car-
ried out in the same conditions. 1.0 mL of 3 % alu-
minum chloride solution was added to 1.0 mL of
the Standard solution of rutin, and the volume was
diluted to 25.0 mL with 70 % ethanol. As a referen-
ce, 1.0 mL of the Standard solution of rutin diluted
with 70 % ethanol to the volume of 25 mL was used.
The content of flavonoids in the raw material
was calculated with reference to rutin (%) by the
following equation [28, 29]:
X, % �
A × a × × × × ×1 0 50 1 25 100 100 ,
A × a × × × × – w0 1 25 2 50 (100 )
where А1 – is the optical density of the Test solu-
tion;
А0 – is the optical density of the Standard solu-
tion of rutin;
а1 – is the accurate weight of raw material, g;
а0 – is the accurate weight of rutin, g;
w – is the loss on drying, %.
ISSN 2308-8303 (Print) / 2518-1548 (Online) 49
Journal of Organic and Pharmaceutical Chemistry 2023, 21 (1)
Preparation of the standard solution of rutin.
About 0.01 g (accurate weight) of rutin (pharma-
copoeial standard FS 42-2508-87) dried at 135 oC
to the constant mass, was placed into a 25.0 mL
measuring flask, dissolved in 96 % ethanol, di-
luted with the same solvent to the volume and
mixed.
Preparation of a 3 % aluminum chloride solu-
tion in 96 % ethanol. According to the State stan-
dard 3759-85, 3.0 g of aluminum chloride (chemi-
cally pure) was dissolved in 96 % ethanol in a
100.0 mL measuring flask, diluted with the same
solvent to the volume and mixed.
■ Results and discussion
Source. The raw material is a dried herb,
which includes whole or cut leaves, flowers and
stems of A. nivea KAR. & KIR. (Asteraceae).
The herb was collected in summer, the below ground
parts were removed, and then the soil was dried
to obtain the intact form. Alternatively, the herb
was sliced while fresh, and dried immediately to
obtain the sliced form of the A. nivea herb.
Based on the results of the macroscopic ana-
lysis of the A. nivea herb [14], such macroscopic
signs, which served as the basis for the develop-
ment of Identification A, were determined.
The stem is usually unbranched in the lower
part, slightly branched in the upper part, rough,
slightly ribbed, 20 – 60 cm long, 5 mm in diameter.
It is externally grayish-green, tomentose-pubes-
cent. The texture is hard. Rosette leaves are
15 – 35 cm long and 2.5 – 6.0 cm wide; stem leaves
are short-petiolate or sessile, 5.0 – 7.0 cm long and
1.5 – 2.5 cm wide, leathery, linear-lanceolate and
lanceolate, pinnatipartite, with a pinnate vena-
tion with prominent lignified veins and long sharp
ends; non-tomentose and green on the adaxial
surface and grayish green, abundant tomentose-
pubescent on the abaxial surface. The petiole of
the rosette leaf is elongated-triangular in out-
line with elongated “wings” directed upwards at
an angle of 45 degrees, narrow at the ends.
The inflorescence is a spherical drooping flo-
wer head up to 6.0 cm in diameter with a 3 – 5-row,
tiled involucre made of hard lanceolate, pointed
leaflets with filmy fring-rowed outgrowths on the
sides. The tubular pink flowers are 5-tooth, thin,
up to 3.0 cm in length; numerous sepals are mo-
dified to thin long hairs; stamens and the pis-
til protrude from the corolla tube; the cypsela
is up to 6.0 mm, pale yellowish with variegated
brown spots and stripes on the surface.
Based on the results of the microscopic ana-
lysis of the A. nivea herb [14], such microscopic
signs, which served as the basis for the develop-
ment of Identification B, were determined.
The stem and leaves are tomentose-pubescent
with covering and rarely glandular trichomes; co-
vering trichomes are of three types – long filamen-
tous hairs with a 2-cell base, large multicellular
wide lumen hairs with collapsed cells; and multi-
cellular narrow lumen hairs with collapsed cells;
glandular hairs are of 2 types – 1-headed with a
1-celled stalk, and headed with a multicellular
stalk and a multicellular head with collapsed cells.
The stem is rounded with slightly protruding ribs.
The cells of the epidermis are covered with a layer
of a folded cuticle; prosenchymatous cells are
straight-walled with slightly thickened walls and
simple straight pores. The central cylinder is of
transitional type. The collenchyma is angular and
lacunar; parenchyma cells are collenchymatous;
the endoderm is distinct; open collateral bundles
are with a well-developed sclerenchyma, xylem
vessels are porous and spiral.
Leaf is dorsiventral; the cells of the upper and
lower epidermis above the veins are prosenchy-
matous, straight-walled, with slightly thickened
walls and straight pores; between the veins the
cells of the upper epidermis are 5 – 6-angle, iso-
diametric and on the lower – parenchymal, si-
nuous-walled, with slightly thickened walls; the
stomata are frequent only on the lower epider-
mis. The stomatal apparatus is anomocytic.
The adaxial surface of the leaf is rarely pu-
bescent with thin long hairs, and abaxial – den-
sely pubescent with filamentous hairs with a 2-cell
base and less often multicellular hairs with col-
lapsed cells. The cells of the outer and inner epi-
dermis of the involucre leaflets are elongated,
slightly thickened and straight-walled with light
brown contents. The inner epidermis is covered
with filamentous trichromes. The cells of the pe-
tal epidermis are prosenchymatous, thin-walled
or heavily thick-walled, straight-walled with co-
lorless, rarely brown contents; the epidermis is
covered with a weakly expressed folding cuticle;
vessels are thin spiral. The style of the pistil is
densely covered with short conical hairs. Pollen
grains are rounded, yellow-brown, the structure
of ectine is slightly spiny.
The powdered raw material. The powder
is yellowish-green in color, examined under a mic-
roscope using Chloral hydrate solution R. The pow-
der shows the following diagnostic signs: frag-
ments of epidermis (Figure 1, A) composed of cells
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Журнал органічної та фармацевтичної хімії 2023, 21 (1)
A B C
D E F G
H
I J
K L M
Figure 1. Microscopic features of the powder
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Journal of Organic and Pharmaceutical Chemistry 2023, 21 (1)
with prosenchymatous straight-walled and slightly
thick-walled cells with simple straight pores (Fi-
gure 1, B); cells with light brown contents (Figu-
re 1, C); stomata of the anomocytic type (Figure 1, A);
fragments of the angular and lacunar collenchy-
ma (Figure 1, D); collenchymatous cells of the pa-
renchyma (Figure 1, E); open collateral bundles
(Figure 1, F); sclerenchyma (Figure 1, G), porous
and spiral vessels; long filamentous hairs with a
2-cell base, large multicellular wide lumen hairs
with collapsed cells (Figure 1, H), and multicellu-
lar narrow lumen hairs with collapsed cells (Fi-
gure 1, I); glandular hairs of 2 types – 1-headed
with a 1-celled stalk (Figure 1, J), and headed
with a multicellular stalk and a multicellular head
with collapsed cells (Figure 1. K); styles with short
conical hairs (Figure 1, L); rounded, yellow-brown
pollen grains with slightly spiny ectine (Figure 1, M).
Identification C of BAS of the A. nivea herb
was developed using the TLC method with stan-
dards of hyperoside, quercetin, rutin and chloro-
genic acid in the system: ethylacetate R – aqua R –
formic acid R – acetic acid anhydrous R (72:14:7:7).
For this analysis, 300 mg of the Alfredia herb
was extracted with 3 mL of methanol for 30 min
at 40 oC on a water bath with ultrasound irra-
diation. 15 μL of the extract was applied to the
chromatogram. The standards were prepared by
dissolving 1 mg of the standards in 10 mL of pure
methanol. After spraying the plate with diphenyl-
borinic acid 2-aminoethyl ester R, macrogol 400 R
the zones at the level of quercetin, hyperoside,
rutin and chlorogenic acid were detected in UV
light [28].
A photo of the sequence of zones on the chro-
matogram of the A. nivea the test extract and
the reference solutions are presented in Figure 2.
The main substances found in the TLC chroma-
togram are quercetin, chlorogenic acid, rutin and
hyperoside. Other additional fluorescent zones can
be found on the chromatogram of the test extract,
but they do not play an important role when iden-
tifying the raw material. Also, using the chroma-
togram obtained in the same conditions, after
spraying the plate with the Drangendorff rea-
gent [29], it was determined that there were no
alkaloids in the raw material.
In the lower part of the chromatogram of the
reference solutions, a yellow fluorescent zone
corresponding to rutin, as well as the blue fluo-
rescent zone corresponding to chlorogenic acid,
and the yellow fluorescent zone corresponding to
hyperoside, were found. The yellow fluorescent
zone corresponding to quercetin was detected
in the upper part of the chromatogram. On the
chromatogram of the test extract, the intense
fluorescent yellow zone at the level of the refer-
ence solution of rutin was detected. At the level
of the reference solution of chlorogenic acid the
intense blue fluorescent zone was also deter-
mined.
Since the zones of rutin and quercetin are most
clearly identified and have a significant area du-
ring the TLC analysis, it has been noticed that
they dominate in the raw material, therefore, the
quantitative determination of BAS in the A. ni-
vea herb should be carried out by the assay of fla-
vonoids calculated with reference to rutin adapt-
ing the methodology of the EuPh [18].
The results of determining related impuri-
ties, indicators “loss on drying during drying”,
“total ash” and the quantitative content of flavo-
noids are given in Table.
The tests were proposed to be carried out ac-
cording to the following indicators: related im-
purities (2.8.2), namely, stems with a diameter
of more than 20 mm – not more than 10 %; other
impurities – not more than 2 %; the loss on dry-
ing (2.2.32) – not more than 13.0 % (1.000 g of
the raw material crushed into a powder at a tem-
perature of 105oC for 2 h); the total ash (2.4.16) –
not more than 10 %.
1 2 3
Figure 2. The TLC chromatogram of A. nivea phenolic compounds:
1. the standards of quercetin, chlorogenic acid, rutin;
2. hyperoside; 3. the A. nivea herb extract
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Журнал органічної та фармацевтичної хімії 2023, 21 (1)
The assay was proposed to be carried out by
the content of flavonoids – at least 0.5 % calcula-
ted with reference to rutin (C27N30O16; M.M. 610.5)
in the dry raw material.
■ Conclusions
The parameters of the A. nivea herb stan-
dardization have been determined on the basis
of the following indicators: macroscopic and mic-
roscopic features, TLC identification of the main
BAS of the raw material (hyperoside, rutin, quer-
cetin and chlorogenic acid), related impurities
(not more than 2 %), stems with a diameter of
more than 20 mm (not more than 10 %), the loss
on drying (not more than 13 %), the total ash
(not more than 10 %) and at least 0.5 % flavo-
noids calculated with reference to rutin.
■ References
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Indicators Standardization
Samples of the raw material
1 2 3 4 5 6
Description According to the parameters
developed + + + + + +
Identification A. External signs –
macroscopy
According to the parameters
developed + + + + + +
Identification B.
Anatomical signs – microscopy
According to the parameters
developed + + + + + +
Identification C. TLC method According to the parameters
developed + + + + + +
Impurities
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than 20 mm not more than 10 % 8.7 6.2 9.6 7.1 8.5 8.0
Related impurities – not more
than 2 % 1.5 1.8 1.0 1.5 1.0 0.5
Loss on drying Not more than 13.0 % 8.7 9.3 7.2 9.7 8.5 11.0
Total ash Not more than 10 % 7.2 6.6 8.6 5.3 8.2 8.9
The content of flavonoids
calculated with reference to rutin Not less than 0.5 % 0.74 0.69 0.63 0.76 0.60 0.58
Note: «+» – meets the requirements
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Journal of Organic and Pharmaceutical Chemistry 2023, 21 (1)
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Information about the authors:
Almat G. Rustemkulov, Postgraduate Student of the “Pharmaceutical Production Technology” Program,
Asfendiyarov Kazakh National Medical University; https://orcid.org/0000-0002-1792-9399; e-mail: arustemkulov@gmail.com.
Tetiana M. Gontova, D.Sc. in Pharmacy, Professor of the Pharmacognosy Department, National University of Pharmacy of the Ministry
of Health of Ukraine; https://orcid.org/0000-0003-3941-9127; e-mail: tetianaviola@ukr.net.
Balzhan G. Makhatova, Postgraduate Student, Associate Professor of Engineering Disciplines and Good Practices Department,
Asfendiyarov Kazakh National Medical University; https://orcid.org/0000-0002-0206-0416; e-mail: makhatova.b@kaznmu.kz.
Aisana E. Rustemkulova, Postgraduate Student of the “Pharmaceutical Production Technology” Program,
Asfendiyarov Kazakh National Medical University; https://orcid.org/0000-0003-2363-7576; e-mail: rustemkulova.a@gmail.com.
Ubaidilla M. Datkhaev, D.Sc. in Pharmacy, Professor, Vice-Rector for Corporate Development,
Asfendiyarov Kazakh National Medical University; https://orcid.org/0000-0002-2322-220X.
Oleh M. Koshovyi (corresponding author), D.Sc. in Pharmacy, Professor of the Pharmacognosy Department,
National University of Pharmacy of the Ministry of Health of Ukraine; https://orcid.org/0000-0001-9545-8548;
e-mail for correspondence: oleh.koshovyi@gmail.com.
|
| id | oai:ojs.journals.uran.ua:article-276004 |
| institution | Journal of Organic and Pharmaceutical Chemistry |
| issn | 2518-1548 |
| keywords_txt_mv | |
| language | English |
| last_indexed | 2026-08-24T01:00:32Z |
| publishDate | 2023 |
| publisher | National University of Pharmacy |
| record_format | ojs |
| resource_txt_mv | ophcjnupheduua/54/035918d20ad5d78327de8f1d00431254.pdf |
| spelling | oai:ojs.journals.uran.ua:article-2760042026-08-23T20:00:17Z Standardization Parameters of Alfredia nivea KAR.&KIR Herb Параметри стандартизації трави Alfredia nivea KAR.&KIR. Rustemkulov, Almat G. Gontova, Tetiana M. Makhatova, Balzhan G. Rustemkulova, Aisana E. Datkhayev, Ubaidilla M. Koshovyi, Oleh M. Альфредія снігова Alfredia nivea трава стандартизація методи контролю якості Alfredia nivea herb standardization quality control methods The Kazakhstan flora is rich in promising poorly-studied plants, which are traditionally used in folk medicine, but their introduction into medical practice requires additional in-depth research using modern scientific methods. Alfredia nivea KAR&KIR of the Asteraceae family, which is used in folk medicine as a neurotropic agent, is an interesting object for introduction into official medical and pharmaceutical practice.Aim. To create new medicines based on Alfredia nivea herb, it is necessary to develop methods for quality control of this raw material, therefore, the aim of the research was to determine the parameters for standardization of the Alfredia nivea KAR & KIR. herb.Materials and methods. The study objects were samples of the A. nivea herb collected in Kungei Alatau, 4.3 km southeast of the Karabulak village, Eastern Karabulak canyon, Almaty Region, Kazakhstan. The macroscopic and microscopic studies of the A. nivea herb were performed according to the methodology of the European pharmacopeia (EuPh) 2.8.23 “Microscopic examination of the medicinal plant raw material”. The macroscopic studies were performed using a magnifying glass and a MBS-9 binocular microscope, the microscopic studies were done using MS Microscopes 10 (oculars X5, X10, X15, lenses x10,x40), Micromed XS-4130 (oculars WF15X, lenses x40/0.65, x10/0.25) with a microphotonozzle (China). Identification of the main substances was carried out by the TLC method, testing and the quantitative determination of the flavonoid content were performed according to the EuPh methods.Results and discussion. Morphological and anatomical features of the A. nivea herb have been determined; on their basis Identifications A and B have been proposed; TLC Identification C of the main BAS of the raw material has been developed; indicators of purity tests have been determined. It has been proposed to carry out the quantitative determination by the content of flavonoids.Conclusions. The parameters of the A. nivea herb standardization have been determined on the basis of the following indicators: macroscopic and microscopic features, TLC identification of the main BAS of the raw material (hyperoside, rutin, quercetin and chlorogenic acid), related impurities (not more than 2 %), stems with a diameter of more than 20 mm (not more than 10 %), the loss on drying (not more than 13 %), the total ash (not more than 10 %) and at least 0.5 % flavonoids calculated with reference to rutin. Флора Казахстану багата на перспективні маловивчені рослини, які застосовують у народній медицині, але для їх впровадження в медичну практику потрібні додаткові глибокі дослідження, за допомогою сучасних наукових методів. Цікавим об’єктом для впровадження в офіцинальну медичну та фармацевтичну практику є Альфредія снігова (Alfredia nivea KAR. & KIR.) родини Asteraceae, яку в народній медицині використовують як нейротропний засіб.Мета. Для створення нових лікарських засобів на основі трави Альфредії снігової необхідно розробити методи контролю якості цієї сировини, тому метою досліджень було визначити параметри стандартизації трави A. nivea KAR. & KIR.Матеріали та методи. Об’єктами дослідження були зразки трави Альфредії снігової (A. nivea), заготовлені в Кунгей Алатау, за 4,3 км південно-східніше від с. Карабулак, ущ. Східний Карабулак, Алматинської області, Казахстан. Макро- і мікроскопічні дослідження трави Альфредії снігової виконували згідно з методикою Європейської фармакопеї (ЄФ) 2.8.23 «Мікроскопічне дослідження лікарської рослинної сировини». Для макроскопічних досліджень використовували лупу та бінокулярний мікроскоп МБС-9, для мікроскопічних – мікроскопи МС 10 (окуляр х5, х10, х15, об’єктиви х10, x40), Micromed XS-4130 (окуляр WF15X, об’єктиви х40/0,65, х10/0,25) з мікрофотонасадкою (КНР). Ідентифікували основні речовини методом ТШХ, випробування та кількісне визначення за вмістом флавоноїдів здійснювали відповідно до методик ЄФ.Результати та їх обговорення. Визначено морфологічні й анатомічні ознаки трави Альфредії снігової, на основі яких запропоновано ідентифікації А та В, розроблено ТШХ ідентифікацію С основних БАР сировини, визначено показники випробувань на чистоту, а також запропоновано проводити кількісне визначення за вмістом флавоноїдів.Висновки. Визначено параметри стандартизації трави Альфредії снігової за такими показниками: макро- та мікроскопічні ознаки, ТШХ ідентифікація основних БАР сировини (гіперозид, рутин, кверцетин та хлорогенова кислота), сторонні домішки (не більше 2 %), наявність стебел діаметром понад 20 мм (не більше 10 %), втрата в масі під час висушування (не більше 13 %), загальна зола (не більше 10 %) і не менше 0,5 % флавоноїдів у перерахунку на рутин. National University of Pharmacy 2023-06-03 Article Article application/pdf https://ophcj.nuph.edu.ua/article/view/276004 10.24959/ophcj.23.276004 Journal of Organic and Pharmaceutical Chemistry; Vol. 21 No. 1 (2023); 46-53 Журнал органической и фармацевтической химии; Том 21 № 1 (2023); 46-53 Журнал органічної та фармацевтичної хімії; Том 21 № 1 (2023); 46-53 2518-1548 2308-8303 en https://ophcj.nuph.edu.ua/article/view/276004/273040 Copyright (c) 2023 Almat G. Rustemkulov, Tetiana M. Gontova, Balzhan G. Makhatova, Aisana E. Rustemkulova, Ubaidilla M. Datkhayev, Oleh M. Koshovyi http://creativecommons.org/licenses/by/4.0 |
| spellingShingle | Альфредія снігова Alfredia nivea трава стандартизація методи контролю якості Rustemkulov, Almat G. Gontova, Tetiana M. Makhatova, Balzhan G. Rustemkulova, Aisana E. Datkhayev, Ubaidilla M. Koshovyi, Oleh M. Параметри стандартизації трави Alfredia nivea KAR.&KIR. |
| title | Параметри стандартизації трави Alfredia nivea KAR.&KIR. |
| title_alt | Standardization Parameters of Alfredia nivea KAR.&KIR Herb |
| title_full | Параметри стандартизації трави Alfredia nivea KAR.&KIR. |
| title_fullStr | Параметри стандартизації трави Alfredia nivea KAR.&KIR. |
| title_full_unstemmed | Параметри стандартизації трави Alfredia nivea KAR.&KIR. |
| title_short | Параметри стандартизації трави Alfredia nivea KAR.&KIR. |
| title_sort | параметри стандартизації трави alfredia nivea kar.&kir. |
| topic | Альфредія снігова Alfredia nivea трава стандартизація методи контролю якості |
| topic_facet | Альфредія снігова Alfredia nivea трава стандартизація методи контролю якості Alfredia nivea herb standardization quality control methods |
| url | https://ophcj.nuph.edu.ua/article/view/276004 |
| work_keys_str_mv | AT rustemkulovalmatg standardizationparametersofalfredianiveakarampkirherb AT gontovatetianam standardizationparametersofalfredianiveakarampkirherb AT makhatovabalzhang standardizationparametersofalfredianiveakarampkirherb AT rustemkulovaaisanae standardizationparametersofalfredianiveakarampkirherb AT datkhayevubaidillam standardizationparametersofalfredianiveakarampkirherb AT koshovyiolehm standardizationparametersofalfredianiveakarampkirherb AT rustemkulovalmatg parametristandartizacíítravialfredianiveakarampkir AT gontovatetianam parametristandartizacíítravialfredianiveakarampkir AT makhatovabalzhang parametristandartizacíítravialfredianiveakarampkir AT rustemkulovaaisanae parametristandartizacíítravialfredianiveakarampkir AT datkhayevubaidillam parametristandartizacíítravialfredianiveakarampkir AT koshovyiolehm parametristandartizacíítravialfredianiveakarampkir |