Muñoz MJA, Morgan MJE, Trujillo GM

Language: Spanish
References: 14
Page: 91-102
PDF size: 195.81 Kb.

ABSTRACT

Introduction: calendula officinalis extracts are used as natural raw material in a wide range of pharmaceutical and cosmetic preparations; however, there are no official methods for quality control of these extracts.
Objective: to validate an HPLC-based analytical method for quantification total quercetin in glycolic and hydroalcoholic extracts of Calendula officinalis.
Methods: to quantify total quercetin content in the matrices, it was necessary to hydrolyze flavonoid glycosides under optimal conditions. The chromatographic separation was performed on a C-18 SiliaChrom 4.6x150mm 5µm column, adapted to a SiliaChrom 5µm C-18 4.6x10mm precolumn, with UV detection at 370nm. The gradient elution was performed with a mobile phase consisting of methanol (MeOH) and phosphoric acid (H₃PO₄) (0.08% w/v). The quantification was performed through the external standard method and comparison with quercetin reference standard.
Results: the studied method selectivity against extract components and degradation products under acid/basic hydrolysis, oxidation and light exposure conditions showed no signals that interfere with the quercetin quantification. It was statistically proved that the method is linear from 1.0 to 5.0mg/mL. Intermediate precision expressed as a variation coefficient was 1.8 and 1.74% and the recovery percentage was 102.15 and 101.32%, for glycolic and hydroalcoholic extracts, respectively.
Conclusions: the suggested methodology meets the quality parameters required for quantifying total quercetin, which makes it a useful tool for quality control of C. officinalis extracts.

REFERENCES

1. Attard A, Cuschieri A. In vitro immunomodulatory activity of various extracts of Maltese plants from the Asteraceae family. J. Med. Plant. Res. 2009;3(6):457-461.

2. Radioza SA, Iurchak LD. Antimicrobial activity of Calendula L. plants. Mikrobiol. Z. 2009; 69:21-25.

3. Chandran PK, Kuttan G, Kuttan R. Anti-inflammatory activity of flower extract of C. officinalis Linn. and its possible mechanism of action. Ind.J.Exp.Biol. 2009;47:113-120.

4. Leach MJ. C. officinalis and wound healing: A systematic review. Wounds 2008; 20(8):236-243.

5. Ćetković GS, Djilas SM, Ćanadanović-Brunet JM, Tumbas VT. Antioxidant properties of marigold extracts. Food Res.Int. 2004;37:643-650.

6. Guevara HA, Luengas PE, Garavito G. Revisión documental de los productos naturales legalmente autorizados para su mercadeo en Colombia. Colombia Médica. 2010;41(2):129-140.

7. INVIMA. Datos de Productos, Fitoterapéutico, Caléndula. [Citado el 13 de marzo de 2014] Disponible en: https://www.invima.gov.co

8. Olszewska M. Quantitative HPLC analysis of flavonoids and chlorogenic acid in the leaves and inflorescences of Prunus serotina EHRH. Acta Chromatographica. 2007;19:253-269.

9. D’Mello PM, Joshi UJ, Shetgiri PP, Dasgupta TK, Darji AK. A Simple HPLC Method for Quantitation of Quercetin in Herbal Extracts. Journal of AOAC International. 2011;94(1):100-105.

10. Rajalakshmi PV, Senthil KK. Direct HPLC analysis of quercetin in exudates of abutilon indicum (linn) Malvaceae. J.of Pharm.Sci. and Tech. 2009;1(2):80-83.

11. Menghinello P, Cucchiarini L, Palma F., Agostini D, Dachà M, Stocchi V. Simultaneous analysis of flavonoid aglycones in natural products using an RP-HPLC method. J. Liq. Chrom. & Rel. Technol. 1999;22(19):3007–3018.

12. The United States Pharmacopoeial Convention. The United States Pharmacopeia. 35 Edit.. Baltimore, 2012;1457-1460.

13. AOAC. Guidelines for Single Laboratory Validation of Chemical Methods for Dietary Supplements and Botanicals. 2012;22. [Citado el 13 de marzo de 2014] . Disponible en: http://www.eoma.aoac.org/app_k.pdf

14. FDA. Guidelines for the Validation of Chemical Methods for the FDA Foods Program. Version 1.0, 2012;26. [Citado el 13 de marzo de 2014]. Disponible en: http://www.fda.gov/downloads/ScienceResearch/FieldScience/UCM298730.pdf