DEVELOPMENT OF METHODS FOR IDENTIFICATION AND DETERMINATION OF BIOLOGICALLY ACTIVE SUBSTANCES IN SEDAVIT HERBAL TABLET PRODUCTS | International Journal of Medical Sciences And Clinical Research

DEVELOPMENT OF METHODS FOR IDENTIFICATION AND DETERMINATION OF BIOLOGICALLY ACTIVE SUBSTANCES IN SEDAVIT HERBAL TABLET PRODUCTS

International Journal of Medical Sciences And Clinical Research
HAC
inLibrary
Google Scholar
doi
 
CC BY f
58-66
0
To share
Salomov Shokhabbos Nozimjon ugli, ., & Djalalova Ozoda Kasimjanovna, . (2024). DEVELOPMENT OF METHODS FOR IDENTIFICATION AND DETERMINATION OF BIOLOGICALLY ACTIVE SUBSTANCES IN SEDAVIT HERBAL TABLET PRODUCTS. International Journal of Medical Sciences And Clinical Research, 4(09), 58–66. https://doi.org/10.37547/ijmscr/Volume04Issue09-09
0
Citations
Crossref
Сrossref
Scopus
Scopus

Abstract

Bioactive groups were identified and methods for their identification and determination in the herbal preparation Sedavit, tablets, were developed. Methods were developed for high-performance liquid chromatography (HPLC) for the identification of polyphenolic compounds and the quantitative determination of pyridoxine and nicotinamide; gas chromatography (GC) for the determination of isovaleric acid, spectrophotometry (SP) for the quantitative determination of flavonoids.


background image

Volume 04 Issue 09-2024

58


International Journal of Medical Sciences And Clinical Research
(ISSN

2771-2265)

VOLUME

04

ISSUE

09

P

AGES

:

58-66

OCLC

1121105677
















































Publisher:

Oscar Publishing Services

Servi

ABSTRACT

Bioactive groups were identified and methods for their identification and determination in the herbal preparation

Sedavit, tablets, were developed. Methods were developed for high-performance liquid chromatography (HPLC) for

the identification of polyphenolic compounds and the quantitative determination of pyridoxine and nicotinamide; gas

chromatography (GC) for the determination of isovaleric acid, spectrophotometry (SP) for the quantitative

determination of flavonoids.

KEYWORDS

Herbal medicines, Sedavit, identification, quantitative determination, control methods.

INTRODUCTION

Herbal medicines consist of a complex of substances

belonging to several chemical groups and under the

influence of technological processes or internal

enzymatic reactions, chemical transformations of

these substances can occur. The task of standardizing

medicinal plant raw materials and drugs based on them

Research Article

DEVELOPMENT OF METHODS FOR IDENTIFICATION AND
DETERMINATION OF BIOLOGICALLY ACTIVE SUBSTANCES IN SEDAVIT
HERBAL TABLET PRODUCTS

Submission Date:

Sep 20, 2024,

Accepted Date:

Sep 25, 2024,

Published Date:

Sep 30, 2024

Crossref doi:

https://doi.org/10.37547/ijmscr/Volume04Issue09-09


Salomov Shokhabbos Nozimjon ugli

Student of Andijan State Medical Institute, Uzbekistan

Djalalova Ozoda Kasimjanovna

PhD. Department of Pathophisiology, Andijan State Medical Institute, Uzbekistan




Journal

Website:

https://theusajournals.
com/index.php/ijmscr

Copyright:

Original

content from this work
may be used under the
terms of the creative
commons

attributes

4.0 licence.


background image

Volume 04 Issue 09-2024

59


International Journal of Medical Sciences And Clinical Research
(ISSN

2771-2265)

VOLUME

04

ISSUE

09

P

AGES

:

58-66

OCLC

1121105677
















































Publisher:

Oscar Publishing Services

Servi

is often complicated by the lack of accurate control

indicators and the existing indicators are not accepted

for testing qualitative and quantitative properties. The

main goal of our work is to develop a method for

standardizing finished pharmaceutical products (PPF)

on a plant basis, ensuring their effectiveness and safety

for consumers, ensuring the development of

appropriate technical parameters and quality control

methods, among other things, for the identification of

counterfeit drugs [1, 2]. Standardization includes the

following interrelated stages of pharmaceutical

product development:

Select individual compounds, biologically active

compounds (BAC) or markers to assess the quality of

FMPs;

select methods and develop techniques for the

identification and/or quantification of individual

compounds, BAS groups or markers in FMPs;

selection and substantiation of standardization

criteria taking into account the requirements of

pharmacopoeial monographs, data from scientific

literature

and

characteristics

of

production

technology.

Let's look at each of these moments using the example

of the development process of a finished herbal

product, Sedavit tablets.

METHODS

The object of our study is the drug Sedavit, in tablet

form, whose active ingredient is a concentrated

extract containing 5 plant ingredients (valerian

rhizome, hawthorn fruit, St. John's wort, peppermint

leaves, hop cones), vitamins - nicotinamide (vitamin P)

and pyridoxine hydrochloride (vitamin B6).

The study used gas chromatography (GC) and high-

performance liquid chromatography (HPLC). GC

studies were performed

on an Agilent 6890 N” gas

chromatograph with a flame ionization detector

(Agilent, USA). For HPLC studies, an Agilent 1200 liquid

chromatograph with a diode array detector (Agilent,

USA) was used. Quantitative determination of total

flavonoids

was

performed

on

a Cary

100

spectrophotometer (Varian, Australia). The study used

standards including chlorogenic, caffeic, ferulic and

rosmarinic acids (Fluka), rutin, hypersaccharide,

luteolin, quercetin, apigenin (Fluka). The reagents used

for the study complied with the requirements of the

European Pharmacopoeia (EPh) and the State

Pharmacopoeia of Uzbekistan (SPU). The reaction

solutions

were

prepared

according

to

the

requirements of EP/SPU [3, 4].

RESULTS AND DISCUSSION

Isolation of individual compounds, groups of bioactive

substances or markers. Each plant component of the

complex extract contains a variety of bioactive

substances, the composition and proportion of which


background image

Volume 04 Issue 09-2024

60


International Journal of Medical Sciences And Clinical Research
(ISSN

2771-2265)

VOLUME

04

ISSUE

09

P

AGES

:

58-66

OCLC

1121105677
















































Publisher:

Oscar Publishing Services

Servi

may vary depending on the geographical and climatic

conditions of growth and cultivation, harvesting,

drying, etc. The complex extract is obtained by

simultaneous extraction of a mixture of five plant

materials. This complicates the development of a

control strategy that includes the qualitative or

quantitative determination of the bioactive substances

or markers inherent in each medicinal plant material

(MPM), since the extract will contain both substances

common to different MPMs and substances

characteristic of one MPM. In this regard, we

conducted a literature review on BAS of the studied

raw materials [5

7], thanks to which we selected

groups of substances whose presence and content

could be used for identification and quantitative

determination (evaluation results are presented in the

table).

Table 1. Biologically active substances of the studied types of raw materials

Raw materials

Compounds

Peppermint leaves

Essential oil (from 2.40 to 3.75%). Essential

oil basically consists of menthol (41-65%),

menthone and menthyl acetate. Carotene,

flavonoids:

apigenin,

hesperidin,

rutin,

chlorogenic, rosmarinic, ursolic and oleanolic

acids; microelements: copper, manganese,

strontium, etc.

St. John’s wort herb

Flavonoids: hypericin, hyperoside, rutin,

quercitrin, quercetin, chlorogenic, caffeic

acids,

tannins,

coumarins,

carotenoids,

vitamins C and PP

Hawthorn berries

Flavonoids: quercetin, hyperoside, rutin, etc.;

chlorogenic,

caffeic

acids,

glycosides,

carotenoids, tannins, fatty oils, saponins,

pectin substances, etc.


background image

Volume 04 Issue 09-2024

61


International Journal of Medical Sciences And Clinical Research
(ISSN

2771-2265)

VOLUME

04

ISSUE

09

P

AGES

:

58-66

OCLC

1121105677
















































Publisher:

Oscar Publishing Services

Servi

Rhizomes with roots of valerian

Essential oil (from 0.5 to 2%), the main part of

which is valerianic-borneol ester, various

organic acids: valerianic, isovaleric, formic,

acetic, etc., valepotriates, alkaloids: valerine,

chatinine, etc.

Hop cones

Bitter

substances

humulone,

lupulone,

humulenic

acid,

humulinone

isomers,

essential oil, flavonoids rutin, quercitrin,

tannins, etc.

According to literature data [5

7], the BAS

composition of four MPMs (hawthorn berries, St.

John's wort, peppermint leaves, hop cones) included

phenolic compounds, which were selected based on

their qualitative and quantitative properties. To

demonstrate the presence of valerian rhizomes in

MPMs, isovaleric acid was chosen as a marker.

Selection of criteria and methods for the identification

and/or quantification of individual compounds, groups

of biologically active substances or markers in the

finished medicinal product. When choosing a control

method, it is necessary to take into account the

selectivity, sensitivity of the method, and suitability for

quality control in the medicinal plant raw material -

extract - concentrate - finished product chain. Highly

selective chromatographic control methods best meet

these requirements and are suitable for the

standardization of herbal preparations [8]. Liquid

chromatography is proposed for the identification and

quantification

of

the

synthetic

components

nicotinamide (vitamin P) and pyridoxine hydrochloride

(vitamin B6). The conditions for conducting the

analysis allow for simultaneous identification and

quantitative determination. Liquid chromatography

was chosen for the determination of phenolic

compounds

in

pharmaceutical

products.

The

substances were identified by comparing the retention

times of the peaks in the chromatograms of the test

solutions with those of the standards. During the

development of the method, samples of chlorogenic,

caffeic,

ferulic

and

rosmarinic

acids,

rutin,

hypersaccharide, luteolin, quercetin and apigenin were


background image

Volume 04 Issue 09-2024

62


International Journal of Medical Sciences And Clinical Research
(ISSN

2771-2265)

VOLUME

04

ISSUE

09

P

AGES

:

58-66

OCLC

1121105677
















































Publisher:

Oscar Publishing Services

Servi

used as standards. For routine control, chlorogenic,

rosmarinic, caffeic and quercetin acids were chosen as

the most specific representatives of the biologically

active substances of the plant components contained

in the drug. Chromatography was performed on a

liquid chromatograph with a UV detector using a C18

chromatographic column; mobile phase A: 0.6 g/l

sodium dihydrogen phosphate monohydrate solution

with pH 2.5 (using phosphoric acid), mobile phase B:

acetonitrile; mobile phase flow rate - 1.0 ml/min;

detection at 330 nm. Typical chromatograms of the

comparison solution and the test solution are shown in

Figures 1.1 and 1.2. Gas chromatography was used to

determine isovaleric acid in the rhizomes and roots of

Valerian in FPP. The determination was carried out by

comparing the retention time of the main peak in the

chromatogram of the test solution with the retention

time of the isovaleric acid peak, a substance

characteristic of the rhizome and root of valerian. The

chromatography was carried out on a gas

chromatograph with a flame ionization detector, using

a 60 m capillary column with an internal diameter of

0.53 mm, the stationary phase was macrogol 20000 P

with a layer thickness of 1 μm; the carrier gas was

helium for chromatography P, the carrier gas flow rate

was 5 ml/min, the flow distribution was 1:1. Typical

chromatograms of isovaleric acid comparison solution

and Sedavit tablet test solution are shown in Figures 2.1

and 2.2.

Figure 1.1 Typical chromatogram of the reference solution obtained during the identification of

phenolic compounds


background image

Volume 04 Issue 09-2024

63


International Journal of Medical Sciences And Clinical Research
(ISSN

2771-2265)

VOLUME

04

ISSUE

09

P

AGES

:

58-66

OCLC

1121105677
















































Publisher:

Oscar Publishing Services

Servi

Figure 1.2 Typical chromatogram of the test solution of Sedavit, tablets, obtained during the

identification of phenolic compounds

When selecting quantitative determination criteria, the

content of biologically active substances in plant raw

materials, semi-finished products and technological

features should be taken into account. The

requirements should reflect the actual level of

biologically active substances, which characterizes the

effectiveness of the

Figure 2.1 Typical chromatogram of the comparison solution of isovaleric acid obtained during

its identification


background image

Volume 04 Issue 09-2024

64


International Journal of Medical Sciences And Clinical Research
(ISSN

2771-2265)

VOLUME

04

ISSUE

09

P

AGES

:

58-66

OCLC

1121105677
















































Publisher:

Oscar Publishing Services

Servi

Figure 2.2 Typical chromatogram of the test solution of Sedavit, tablets, obtained during the

identification of isovaleric acid

finished medicinal product. In this case, if the finished

medicinal product contains a negligible amount of

biologically active substances or markers, the

specification should only include identification

requirements and during quantitative testing the

amount of biologically active substances should be

determined; in this case, tests can be carried out, for

example, by spectrophotometric methods. Sedavit

tablets from the UK contain a complex extract

consisting of five medicinal plant raw materials. Its

therapeutic effect is determined by a complex of

biologically active substances isolated from plant raw

materials. In this regard, when quantitatively

Figure 3 Differential electronic absorption spectra of the aluminum chloride complex with

flavonoids of the studied Sedavit solution, tablets (1) and standard rutin solution (2)

analyzing active substances, it is unreasonable to

determine

each

component

individually

by

chromatography and then calculate the total peak

area. For these purposes, it is sufficient to use

spectrophotometric methods to determine the total

optical density of the components determined under

the analytical conditions [9]. When choosing a

standard sample for recalculating the active substance


background image

Volume 04 Issue 09-2024

65


International Journal of Medical Sciences And Clinical Research
(ISSN

2771-2265)

VOLUME

04

ISSUE

09

P

AGES

:

58-66

OCLC

1121105677
















































Publisher:

Oscar Publishing Services

Servi

content, we took into account its availability for

routine control, the inclusion of the selected substance

in the determined compound group and the need to

use conversion factors [9]. French rutin was chosen as

the standard, which under analytical conditions gave a

maximum absorbance at 415 nm and did not show a

reliable difference in spectrum compared to the

indicator of the analyzed solution. For routine control,

it is proposed to use the absorption index value of rutin

with aluminium chloride, which simplifies and reduces

the cost of the method for quantitative determination

of flavonoids, a typical group for medicinal plant raw

materials used. The optical density of the test solution

was determined after the formation of a colored

complex of flavonoids extracted with ethyl acetate and

aluminium chloride. The initial solution (ethyl acetate

extract) without adding aluminium chloride reagent

was used as a compensation solution, which avoids the

influence of the accompanying substances extracted

with ethyl acetate on the analytical results. In parallel,

the optical density of the rutin standard sample

solution with aluminium chloride was measured using

the initial rutin standard solution without adding

aluminium chloride reagent as a compensation

solution. Typical differential absorption spectra of the

aluminium chloride complex with flavonoids of the test

sample (complex extract) and the rutin standard are

shown in Figure 3.

CONCLUSION

As a result of this work, a herbal medicine

standardization algorithm was proposed using Sedavit

tablets as an example, including conditions for

selecting bioactive groups and markers, identification

and quantitative determination methods, qualitative

and quantitative methods, and quantitative criteria for

drug evaluation.

REFERENCES

1.

Myagkiе

lеkarstvеnniе

formi:

uchеbno

-

mеtodichеskoе posobiе / V. F. Dzyuba [i dr.] pod

rеd. V. F. Dzyubi. –

Voronеj : Izd

-vo VGU 2015.

108

s.

2.

Nikolaеv, N. P. Tеxnologiya mazеy / N. P. Nikolaеv.

Moskva: MORION, 1998.

S. 250

261.

3.

Slivkin, A. I. Kontroli kachеstva ekstеmporalinix

lеkarstvеnnix form / A. I. Slivkin [i dr.] pod rеd. A. I.

Slivkina.

Voronеj: Izd

-vo VGU 2003.

S. 21

28, 46.

4.

Validasiya analitichеskix mеto

dov / A. P.

Arzamassеv [i dr.] // Farmasiya. –

2006.

№ 4. –

S.

8

12.

5.

Sur, S. V. Mеtodologiya osеnki kachеstva

rastitеlinix lеkarstvеnnix srеdstv na osnovanii

rеzulitatov, poluchеnnix s pomoshiyu sovrеmеnnix

analitichеskix

mеtodov

/

S.

V.

Sur

//

Farmas

еvtichеskiy jurnal. –

2002.

№ 6. –

S. 64

71.

6.

Guidelines for the assessment of herbal medicines

/ WHO Expert Committee on Specification for

Pharmaceutical Preparation.

Thirty-fourth


background image

Volume 04 Issue 09-2024

66


International Journal of Medical Sciences And Clinical Research
(ISSN

2771-2265)

VOLUME

04

ISSUE

09

P

AGES

:

58-66

OCLC

1121105677
















































Publisher:

Oscar Publishing Services

Servi

Report.

Geneva: World Health Organization,

1996.

(WHO Technical Report Series, No. 863).

7.

Farmasеvtichеskaya ensiklopеdiya Ukraini / Golova

rеd. radi V. P. Chеrnix. –

2-

е izd. –

K.: «MORІON»,

2010.

1632 s.

8.

Sur, S. V. Problеmi i pеrspеktivi razrabotki i

vnеdrеniya sovrеmеnnix lеkarstvеnnix srеdstv

rastitеlinogo proisxojdеniya / S. V. Sur,

O. M.

Grisеnko // Lіki Ukraїni. –

2002.

№ 4. –

S. 47

49.

9.

Rastitеliniе lеkarstvеnniе srеdstva / N. P.

Maksyutina [i dr.] // K.: Zdorov’ya, 1985. –

280 s.

10.

Muraviеva, D. A. Farmakognoziya / D. A. Muraviеva.

Moskva.: Mеdisina, 1981. –

656 s.

References

Myagkiе lеkarstvеnniе formi: uchеbno-mеtodichеskoе posobiе / V. F. Dzyuba [i dr.] pod rеd. V. F. Dzyubi. – Voronеj : Izd-vo VGU 2015. – 108 s.

Nikolaеv, N. P. Tеxnologiya mazеy / N. P. Nikolaеv. – Moskva: MORION, 1998. – S. 250–261.

Slivkin, A. I. Kontroli kachеstva ekstеmporalinix lеkarstvеnnix form / A. I. Slivkin [i dr.] pod rеd. A. I. Slivkina. – Voronеj: Izd-vo VGU 2003. – S. 21–28, 46.

Validasiya analitichеskix mеtodov / A. P. Arzamassеv [i dr.] // Farmasiya. – 2006. – № 4. – S. 8–12.

Sur, S. V. Mеtodologiya osеnki kachеstva rastitеlinix lеkarstvеnnix srеdstv na osnovanii rеzulitatov, poluchеnnix s pomoshiyu sovrеmеnnix analitichеskix mеtodov / S. V. Sur // Farmasеvtichеskiy jurnal. – 2002. – № 6. – S. 64 – 71.

Guidelines for the assessment of herbal medicines / WHO Expert Committee on Specification for Pharmaceutical Preparation. – Thirty-fourth Report. – Geneva: World Health Organization, 1996. – (WHO Technical Report Series, No. 863).

Farmasеvtichеskaya ensiklopеdiya Ukraini / Golova rеd. radi V. P. Chеrnix. –2-е izd. – K.: «MORІON», 2010. – 1632 s.

Sur, S. V. Problеmi i pеrspеktivi razrabotki i vnеdrеniya sovrеmеnnix lеkarstvеnnix srеdstv rastitеlinogo proisxojdеniya / S. V. Sur, O. M. Grisеnko // Lіki Ukraїni. – 2002. – № 4. – S. 47–49.

Rastitеliniе lеkarstvеnniе srеdstva / N. P. Maksyutina [i dr.] // K.: Zdorov’ya, 1985. – 280 s.

Muraviеva, D. A. Farmakognoziya / D. A. Muraviеva. – Moskva.: Mеdisina, 1981. – 656 s.

inLibrary — это научная электронная библиотека inConference - научно-практические конференции inScience - Журнал Общество и инновации UACD - Антикоррупционный дайджест Узбекистана UZDA - Ассоциации стоматологов Узбекистана АСТ - Архитектура, строительство, транспорт Open Journal System - Престиж вашего журнала в международных базах данных inDesigner - Разработка сайта - создание сайтов под ключ в веб студии Iqtisodiy taraqqiyot va tahlil - ilmiy elektron jurnali yuridik va jismoniy shaxslarning in-Academy - Innovative Academy RSC MENC LEGIS - Адвокатское бюро SPORT-SCIENCE - Актуальные проблемы спортивной науки GLOTEC - Внедрение цифровых технологий в организации MuviPoisk - Смотрите фильмы онлайн, большая коллекция, новинки кинопроката SMARTY - Увеличение продаж вашей компании ELECARS - Электромобили в Ташкенте, Узбекистане CHINA MOTORS - Купи автомобиль своей мечты! PROKAT24 - Прокат и аренда строительных инструментов