A five-herb containing traditional homemade medicine is extensively used to treat gout but has not been standardized for quercetin content. Therefore, the current study describes a reversed-phase liquid chromatographic method for quercetin determination in traditional herbal remedy. The elution was carried out using aqueous 2.0% acetic acid, acetonitrile and tetrahydrofuran (55:40:5, V/V/V) as mobile phase at flow rate of 0.8 mL/min and detection was performed using diode array detector operated at 370 nm. The response of the detector was linear in the range investigated (2.5-160.0 µg/mL) with R2 = 0.996. Results of recovery (98.26-103.22%, SD<5%), intraday accuracy and precision (94.68-104.08%, RSD<5%) and interday accuracy and precision (92.31-104.92%, RSD<5%) showed that the method was reliable, repeatable and reproducible, hence may be used for determination of quercetin in herbal remedy. The medicine contained 0.2425 mg/g quercetin. The molecular interactions of this marker compound were also studied against anti-gout and anti-inflammatory protein targets. Hence, the developed RP-HPLC method may be used to produce standardized anti-gout medicine for quercetin content. Moreover, the molecular interactions help in understanding underlying mechanism of action of this marker compound against gout.
Careri, M., Corradini, C., Elviri, L., Nicoletti, I., & Zagnoni, I. (2003). Direct HPLC analysis of quercetin and trans-resveratrol in red wine, grape, and winemaking byproducts. Journal of Agricultural and Food Chemistry, 51(18), 5226-5231.
Chen, X.Q., Xiao, J.B. (2010). RP-HPLC-DAD detrmination of flavonoids: separation of quercetin, luteolin and apigenin in Marchantia convoluta. Iranian Journal of Pharmaceutical Research, (3), 175-181.
Dmitrienko, S. G., Kudrinskaya, V. A., & Apyari, V. V. (2012). Methods of extraction, preconcentration, and determination of quercetin. Journal of Analytical Chemistry, 67(4), 299-311.
Duan, Y. (2014). Ultraviolet-visible spectrum characterizations of quercetin in aqueous ethanol solution with different pH values. Journal of Chemical and Pharmaceutical Research, 6(9), 236-240.
Ewais, E. A., Abd El-Maboud, M. M., Elhaw, M. H., & Haggag, M. I. (2016). Phytochemical studies on Lycium schweinfurthii var. schweinfurthii (Solanaceae) and Isolation of five Flavonoids from leaves. Journal of Medicinal Plant Studies, 4, 288-300.
Fasolo, D., Schwingel, L., Holzschuh, M., Bassani, V., & Teixeira, H. (2007). Validation of an isocratic LC method for determination of quercetin and methylquercetin in topical nanoemulsions. Journal of Pharmaceutical and Biomedical Analysis, 44(5), 1174-1177.
Ferrandiz, M. L., & Alcaraz, M. (1991). Anti-inflammatory activity and inhibition of arachidonic acid metabolism by flavonoids. Agents and Actions, 32(3), 283-288.
Goo, H. R., Choi, J. S., & Na, D. H. (2009). Simultaneous determination of quercetin and its glycosides from the leaves of Nelumbo nucifera by reversed-phase high-performance liquid chromatography. Archives of Pharmacal Research, 32(2), 201-206.
Ishii, K., Furuta, T., & Kasuya, Y. (2003). High-performance liquid chromatographic determination of quercetin in human plasma and urine utilizing solid-phase extraction and ultraviolet detection. Journal of Chromatography B, 794(1), 49-56.
Kalyaanamoorthy, S., Chen, Y.P.P. (2011). Structure-based drug design to augment hit discovery. Drug Discovery Today, 16(17-18), 831-839.
Kim, H. P., Mani, I., Iversen, L., & Ziboh, V. A. (1998). Effects of naturally-occurring flavonoids and biflavonoids on epidermal cyclooxygenase and lipoxygenase from guinea-pigs. Prostaglandins, Leukotrienes and Essential Fatty acids, 58(1), 17-24.
Laughton, M. J., Evans, P. J., Moroney, M. A., Hoult, J. R. S., & Halliwell, B. (1991). Inhibition of mammalian 5-lipoxygenase and cyclo-oxygenase by flavonoids and phenolic dietary additives: relationship to antioxidant activity and to iron ion-reducing ability. Biochemical Pharmacology, 42(9), 1673-1681.
Li, S., Han, Q., Qiao, C., Song, J., Lung Cheng, C., & Xu, H. (2008). Chemical markers for the quality control of herbal medicines: an overview. Chinese medicine, 3(1), 1-16.
Lin, C. M., Chen, C. S., Chen, C. T., Liang, Y. C., & Lin, J. K. (2002). Molecular modeling of flavonoids that inhibits xanthine oxidase. Biochemical and Biophysical Research Communications, 294(1), 167-172.
Lin, S., Zhang, G., Liao, Y., Pan, J., Gong, D. (2015). Dietary flavonoids as xanthine oxidase inhibitors: Structure–affinity and structure–activity relationships. Journal of Agricultural and Food Chemistry, 63(35), 7784-7794
Liu, H. P., Shi, X. F., Zhang, Y. C., Li, Z. X., Zhang, L., & Wang, Z. Y. (2011). Quantitative analysis of quercetin in Euphorbia helioscopia L by RP-HPLC. Cell Biochemistry and Biophysics, 61(1), 59-64.
Meng, X. Y., Zhang, H. X., Mezei, M., & Cui, M. (2011). Molecular docking: a powerful approach for structure-based drug discovery. Current Computer-aided Drug design, 7(2), 146-157. 12
Nessa, F., Ismail, Z., & Mohamed, N. (2010). Xanthine oxidase inhibitory activities of extracts and flavonoids of the leaves of Blumea balsamifera. Pharmaceutical Biology, 48(12), 1405-1412.
Nile, S. H., Nile, A. S., Keum, Y. S., & Sharma, K. (2017). Utilization of quercetin and quercetin glycosides from onion (Allium cepa L.) solid waste as an antioxidant, urease and xanthine oxidase inhibitors. Food Chemistry, 235, 119-126.
Phani, C. R., Vinaykumar, C., Rao, K. U., & Sindhuja, G. (2010). Quantitative analysis of quercetin in natural sources by RP-HPLC. International Journal of Research in Pharmaceutical and Biomedical Sciences, 1(1), 19-22.
Rasoulzadeh, F., Jabary, H. N., Naseri, A., & Rashidi, M. R. (2009). Fluorescence quenching study of quercetin interaction with bovine milk xanthine oxidase. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 72(1), 190-193.
Rathee, P., Chaudhary, H., Rathee, S., Rathee, D., Kumar, V., & Kohli, K. (2009). Mechanism of action of flavonoids as anti-inflammatory agents: a review. Inflammation & Allergy-Drug Targets, 8(3), 229-235.
Savic, I. M., Nikolic, V. D., Savic, I. M., Nikolic, L. B., & Stankovic, M. Z. (2013). Development and validation of a new RP-HPLC method for determination of quercetin in green tea. Journal of Analytical Chemistry, 68(10), 906-911.
Shaukat, A., Hussain, K., Bukhari, N.I., Shehzadi, N., Naheed, S., Saghir, F., Iftikhar, S, Javed, O. (2020). In vitro anti-gout and anti-inflammatory activity of traditionally used polyherbal anti-gout remedy. International Journal of Biosciences, 16(5), 327-335.
Shivashankar, S., Murali, A., & Sangeetha, M. K. (2019). Molecular interaction of phytochemicals with snake venom: Phytochemicals of Andrographis paniculata inhibits phospholipase A2 of Russell's viper (Daboia russelli). Biocatalysis and Agricultural Biotechnology, 18, 101058.
Sladkovsky, R., Solich, P., Opletal, L. (2001). Simultaneous determination of quercetin, kaempferol and (E)-cinnamic acid in vegetative organs of Schisandra chinensis Baill. by HPLC. Journal of Pharmaceutical and Biomedical Analysis, 24(5-6), 1049-1054.
Yao, L. H., Jiang, Y. M., Shi, J., Tomas-Barberan, F. A., Datta, N., Singanusong, R., & Chen, S. S. (2004). Flavonoids in food and their health benefits. Plant Foods for Human Nutrition, 59(3), 113-122.
Zhu, J. X., Wang, Y., Kong, L. D., Yang, C., & Zhang, X. (2004). Effects of Biota orientalis extract and its flavonoid constituents, quercetin and rutin on serum uric acid levels in oxonate-induced mice and xanthine dehydrogenase and xanthine oxidase activities in mouse liver. Journal of Ethnopharmacology, 93(1), 133-140.
Careri, M., Corradini, C., Elviri, L., Nicoletti, I., & Zagnoni, I. (2003). Direct HPLC analysis of quercetin and trans-resveratrol in red wine, grape, and winemaking byproducts. Journal of Agricultural and Food Chemistry, 51(18), 5226-5231.
Chen, X.Q., Xiao, J.B. (2010). RP-HPLC-DAD detrmination of flavonoids: separation of quercetin, luteolin and apigenin in Marchantia convoluta. Iranian Journal of Pharmaceutical Research, (3), 175-181.
Dmitrienko, S. G., Kudrinskaya, V. A., & Apyari, V. V. (2012). Methods of extraction, preconcentration, and determination of quercetin. Journal of Analytical Chemistry, 67(4), 299-311.
Duan, Y. (2014). Ultraviolet-visible spectrum characterizations of quercetin in aqueous ethanol solution with different pH values. Journal of Chemical and Pharmaceutical Research, 6(9), 236-240.
Ewais, E. A., Abd El-Maboud, M. M., Elhaw, M. H., & Haggag, M. I. (2016). Phytochemical studies on Lycium schweinfurthii var. schweinfurthii (Solanaceae) and Isolation of five Flavonoids from leaves. Journal of Medicinal Plant Studies, 4, 288-300.
Fasolo, D., Schwingel, L., Holzschuh, M., Bassani, V., & Teixeira, H. (2007). Validation of an isocratic LC method for determination of quercetin and methylquercetin in topical nanoemulsions. Journal of Pharmaceutical and Biomedical Analysis, 44(5), 1174-1177.
Ferrandiz, M. L., & Alcaraz, M. (1991). Anti-inflammatory activity and inhibition of arachidonic acid metabolism by flavonoids. Agents and Actions, 32(3), 283-288.
Goo, H. R., Choi, J. S., & Na, D. H. (2009). Simultaneous determination of quercetin and its glycosides from the leaves of Nelumbo nucifera by reversed-phase high-performance liquid chromatography. Archives of Pharmacal Research, 32(2), 201-206.
Ishii, K., Furuta, T., & Kasuya, Y. (2003). High-performance liquid chromatographic determination of quercetin in human plasma and urine utilizing solid-phase extraction and ultraviolet detection. Journal of Chromatography B, 794(1), 49-56.
Kalyaanamoorthy, S., Chen, Y.P.P. (2011). Structure-based drug design to augment hit discovery. Drug Discovery Today, 16(17-18), 831-839.
Kim, H. P., Mani, I., Iversen, L., & Ziboh, V. A. (1998). Effects of naturally-occurring flavonoids and biflavonoids on epidermal cyclooxygenase and lipoxygenase from guinea-pigs. Prostaglandins, Leukotrienes and Essential Fatty acids, 58(1), 17-24.
Laughton, M. J., Evans, P. J., Moroney, M. A., Hoult, J. R. S., & Halliwell, B. (1991). Inhibition of mammalian 5-lipoxygenase and cyclo-oxygenase by flavonoids and phenolic dietary additives: relationship to antioxidant activity and to iron ion-reducing ability. Biochemical Pharmacology, 42(9), 1673-1681.
Li, S., Han, Q., Qiao, C., Song, J., Lung Cheng, C., & Xu, H. (2008). Chemical markers for the quality control of herbal medicines: an overview. Chinese medicine, 3(1), 1-16.
Lin, C. M., Chen, C. S., Chen, C. T., Liang, Y. C., & Lin, J. K. (2002). Molecular modeling of flavonoids that inhibits xanthine oxidase. Biochemical and Biophysical Research Communications, 294(1), 167-172.
Lin, S., Zhang, G., Liao, Y., Pan, J., Gong, D. (2015). Dietary flavonoids as xanthine oxidase inhibitors: Structure–affinity and structure–activity relationships. Journal of Agricultural and Food Chemistry, 63(35), 7784-7794
Liu, H. P., Shi, X. F., Zhang, Y. C., Li, Z. X., Zhang, L., & Wang, Z. Y. (2011). Quantitative analysis of quercetin in Euphorbia helioscopia L by RP-HPLC. Cell Biochemistry and Biophysics, 61(1), 59-64.
Meng, X. Y., Zhang, H. X., Mezei, M., & Cui, M. (2011). Molecular docking: a powerful approach for structure-based drug discovery. Current Computer-aided Drug design, 7(2), 146-157. 12
Nessa, F., Ismail, Z., & Mohamed, N. (2010). Xanthine oxidase inhibitory activities of extracts and flavonoids of the leaves of Blumea balsamifera. Pharmaceutical Biology, 48(12), 1405-1412.
Nile, S. H., Nile, A. S., Keum, Y. S., & Sharma, K. (2017). Utilization of quercetin and quercetin glycosides from onion (Allium cepa L.) solid waste as an antioxidant, urease and xanthine oxidase inhibitors. Food Chemistry, 235, 119-126.
Phani, C. R., Vinaykumar, C., Rao, K. U., & Sindhuja, G. (2010). Quantitative analysis of quercetin in natural sources by RP-HPLC. International Journal of Research in Pharmaceutical and Biomedical Sciences, 1(1), 19-22.
Rasoulzadeh, F., Jabary, H. N., Naseri, A., & Rashidi, M. R. (2009). Fluorescence quenching study of quercetin interaction with bovine milk xanthine oxidase. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 72(1), 190-193.
Rathee, P., Chaudhary, H., Rathee, S., Rathee, D., Kumar, V., & Kohli, K. (2009). Mechanism of action of flavonoids as anti-inflammatory agents: a review. Inflammation & Allergy-Drug Targets, 8(3), 229-235.
Savic, I. M., Nikolic, V. D., Savic, I. M., Nikolic, L. B., & Stankovic, M. Z. (2013). Development and validation of a new RP-HPLC method for determination of quercetin in green tea. Journal of Analytical Chemistry, 68(10), 906-911.
Shaukat, A., Hussain, K., Bukhari, N.I., Shehzadi, N., Naheed, S., Saghir, F., Iftikhar, S, Javed, O. (2020). In vitro anti-gout and anti-inflammatory activity of traditionally used polyherbal anti-gout remedy. International Journal of Biosciences, 16(5), 327-335.
Shivashankar, S., Murali, A., & Sangeetha, M. K. (2019). Molecular interaction of phytochemicals with snake venom: Phytochemicals of Andrographis paniculata inhibits phospholipase A2 of Russell's viper (Daboia russelli). Biocatalysis and Agricultural Biotechnology, 18, 101058.
Sladkovsky, R., Solich, P., Opletal, L. (2001). Simultaneous determination of quercetin, kaempferol and (E)-cinnamic acid in vegetative organs of Schisandra chinensis Baill. by HPLC. Journal of Pharmaceutical and Biomedical Analysis, 24(5-6), 1049-1054.
Yao, L. H., Jiang, Y. M., Shi, J., Tomas-Barberan, F. A., Datta, N., Singanusong, R., & Chen, S. S. (2004). Flavonoids in food and their health benefits. Plant Foods for Human Nutrition, 59(3), 113-122.
Zhu, J. X., Wang, Y., Kong, L. D., Yang, C., & Zhang, X. (2004). Effects of Biota orientalis extract and its flavonoid constituents, quercetin and rutin on serum uric acid levels in oxonate-induced mice and xanthine dehydrogenase and xanthine oxidase activities in mouse liver. Journal of Ethnopharmacology, 93(1), 133-140.
Shaukat, A., & Hussain, K. (2022). Quercetin Based Standardization Of Polyherbal Anti-Gout Remedy And İts Molecular Docking Study Against Anti-Gout And Anti-İnflammatory Protein Targets. Fabad Eczacılık Bilimler Dergisi, 47(3), 317-330. https://doi.org/10.55262/fabadeczacilik.1085825