HPLC of Hypericum perforatum Herb Cell Suspension Reveals Attractive Medical Flavanone Glycosides Compounds that are Supported by DFT Study

Author's: Aziza M. Taj Al-Deen1*, Hossam H. Manaf2, Sameh A. Rizk3
Authors' Affiliations
1Biology Department, Faculty of Science, Sana’a University, Sana’a, Yemen.2Agricultural Botany Department, Faculty of Agriculture, Ain Shams University, Cairo, Egypt.3Chemistry Department, Faculty of Science, Ain Shams University, Cairo, Egypt.*CorrespondenceAziza M. Taj AldeenEmail:azizataj1@yahoo.com
Article Type: Research Article     Published: Jan. 11, 2022 Pages: 14-33
DOI:        Views 500       Downloads 0


In the present study, in vitro Hypericum perforatum leaves were used as a segment to induce callus culture. Callus morphogenesis was achieved using Murashige and Skoog’s culture media supplemented with different combinations of growth regulators. The best callus biomass was obtained with 0.22 mg/l (2,4-D) + 0.024 mg/l (Kin), and cell suspension culture succeeded in the production of new good circular cell biomass that was treated by three different stimulating compounds. The results showed that the best increase in the active ingredients was by chromone glycoside derivatives 2-5, as revealed via HPLC test of fractionation. The extracted H. perforatum L. was conveniently synthesized in 3 steps, from the commercially available Acetophloroglucine, which supported the extracted ingredients 2-5. The glucosyl moiety was introduced by stereoselective O-glycosylation with a per-O-benzylated glucosyl bromide donor, followed by the formation of the chalcone with debenzylation, which was carried out to afford 3 and 5 with overall yields of 64% and 73%, respectively. Finally, the density functional theory (DFT) was then applied to explore the structural and electronic characteristics of these materials. All the synthesized compounds have been characterized based on their 1H NMR, MS, and elemental analyses for the optimized precursors of the synthesized chromone glycosides.


In vitro,

Callus culture,

Density functional theory,

Synthesized chromone glycosides.

How to cite

Taj Al-Deen, A.M., Manaf, H.H., Rizk, S.A., 2022. HPLC of Hypericum perforatum Herb Cell Suspension Reveals Attractive Medical Flavanone Glycosides Compounds that are Supported by DFT Study. PSM Biol. Res., 7(1): 14-33.


Al- Mahweety, J.A.N., 2016. Phytochemical Studies on Medicinal Plants, Dracaenaceae resin, of Socotra Island, Yemen. PSM Biol. Res., 1(2): 62-65.

Ali, K., Shuaib, M., Ilyas, M., Hussain, F., Hussain, F., 2017. Medicinal Uses of Chemical Extracts from Withania somnifera and Its Antimicrobial Activity: A Mini-Review. PSM Microbiol., 2(1): 20-23.

Amin, R.A., Edris, S.N., 2017. Grape Seed Extract as Natural Antioxidant and Antibacterial in Minced Beef. PSM Biol. Res., 2(2): 89-96.

Asres, K., Seyoum, A., Veeresham, C., Bucar, F., Gibbons, S., 2005. Naturally derived anti-HIV agents. Phytother. Res., 19(7): 557-81.

Attia, S., Elgendy, A., Rizk, S., 2019. Efficient green synthesis of antioxidant azacoumarin dye bearing spiro-pyrrolidine for enhancing electro-optical properties of perovskite solar cells. J. Mol. Struct., 1184: 583-592.

Ayan, A.K., Çirak, C., KEVSEROĞLU, K., Sökmen, A., 2005. Effects of explant types and different concentrations of sucrose and phytoharmones on plant regeneration and hypericin content in Hypericum perforatum L. Turk. J. Agric. For., 29(3): 197-204.

Azab, M.E., Rizk, S.A., Mahmoud, N.F., 2016. Facile synthesis, characterization, and antimicrobial evaluation of novel heterocycles, schiff bases, and N-nucleosides bearing phthalazine moiety. Chem. Pharm. Bull., 64(5): 439-450.

Bakry, b.a., Sadak, M., El-Monem, A., 2020. Physiological Aspects of Tyrosine and Salicylic Acid on Morphological, Yield and Biochemical Constituents of Peanut Plants. Pak. J. Biol. Sci., 23: 375-384.

Benavente-García, O., Castillo, J., Marin, F.R., Ortuño, A., Del Río, J.A., 1997. Uses and Properties of Citrus Flavonoids. J. Agric. Food Chem., 45(12): 4505-4515.

Bito, T. et al., 2002. Flavonoids differentially regulate IFN gamma-induced ICAM-1 expression in human keratinocytes: molecular mechanisms of action. FEBS Lett., 520(1-3): 145-52.

Burgess, J.R., Andrade, J.E., 2006. Antioxidant effects of citrus flavonoid consumption, Potential health benefits of Citrus (ACS Symp. Ser. 936), pp. 161-174.

Caristi, C., Bellocco, E., Gargiulli, C., Toscano, G., Leuzzi, U., 2006. Flavone-di-C-glycosides in citrus juices from Southern Italy. Food Chem., 95(3): 431-437.

Caristi, C. et al., 2003. Flavonoids detection by HPLC-DAD-MS-MS in lemon juices from Sicilian cultivars. J. Agric. Food Chem., 51(12): 3528-34.

Choi, J.-S., Choi, Y.-J., Park, S.-H., Kang, J.-S., Kang, Y.-H., 2004. Flavones mitigate tumor necrosis factor-α-induced adhesion molecule upregulation in cultured human endothelial cells: role of nuclear factor-κB. The J. Nutr., 134(5): 1013-1019.

Cushnie, T.P., Lamb, A.J., 2005. Antimicrobial activity of flavonoids. Int. J. Antimicrob. Agents, 26(5): 343-56.

De Jong, W.H., Borm, P.J.A., 2008. Drug delivery and nanoparticles:applications and hazards. Int. J. Nanomed., 3(2): 133-149.

El-Hashash, M.A., Darwish, K.M., Rizk, S.A., El-Bassiouny, F.A., 2011. The uses of 2-ethoxy-(4H)-3, 1-benzoxazin-4-one in the synthesis of some quinazolinone derivatives of antimicrobial activity. Pharm., 4(7): 1032-1051.

El-Hashash, M.A., Rizk, S.A., Atta-Allah, S.R., 2015. Synthesis and regioselective reaction of some unsymmetrical heterocyclic chalcone derivatives and spiro heterocyclic compounds as antibacterial agents. Molec., 20(12): 22069-22083.

El‐Hashash, M.A., Rizk, S.A., 2017. One‐pot Synthesis of Novel Spirooxindoles as Antibacterial and Antioxidant Agents. J. Heterocycl. Chem., 54(3): 1776-1784.

Elkholy, A.E., Rizk, S.A., Rashad, A.M., 2019. Enhancing lubricating oil properties using novel quinazolinone derivatives: DFT study and molecular dynamics simulation. J. Mol. Struct., 1175: 788-796.

Erlund, I., 2004. Review of the flavonoids quercetin, hesperetin, and naringenin. Dietary sources, bioactivities, bioavailability, and epidemiology. Nutr. Res., 24(10): 851-874.

Gattuso, G., Barreca, D., Gargiulli, C., Leuzzi, U., Caristi, C., 2007. Flavonoid composition of citrus juices. Molec., 12(8): 1641-1673.

Gattuso, G. et al., 2006. Flavonoid glycosides in bergamot juice (Citrus bergamia Risso). J. Agric. Food Chem., 54(11): 3929-3935.

Gil-Izquierdo, A., Gil, M.I., Ferreres, F., 2002. Effect of processing techniques at industrial scale on orange juice antioxidant and beneficial health compounds. J. Agric. Food Chem., 50(18): 5107-5114.

Gil-Izquierdo, A., Riquelme, M.T., Porras, I., Ferreres, F., 2004. Effect of the rootstock and interstock grafted in lemon tree (Citrus limon (L.) Burm.) on the flavonoid content of lemon juice. J. Agric. Food Chem., 52(2): 324-31.

Gross, M., 2004. Flavonoids and Cardiovascular Disease. Pharm. Biol., 42(sup1): 21-35.

Hara, H. et al., 2011. Inhibitory effects of chalcone glycosides isolated from Brassica rapa L. ‘hidabeni’ and their synthetic derivatives on LPS-induced NO production in microglia. Bioorg. Med. Chem., 19(18): 5559-68.

Heakal, F.E.-T., Attia, S., Rizk, S., Abou Essa, M., Elkholy, A., 2017. Synthesis, characterization and computational chemical study of novel pyrazole derivatives as anticorrosion and antiscalant agents. J. Mol. Struct., 1147: 714-724.

Hirano, R. et al., 2001. Antioxidant ability of various flavonoids against DPPH radicals and LDL oxidation. J. Nutr. Sci. Vitaminol., 47(5): 357-362.

Hussain, F. et al., 2016. Antibacterial Activities of Methanolic Extracts of Datura inoxia. PSM Microbiol., 1(1): 33-35.

Hussein, M.A. et al., 2018. Synthesis, molecular docking and insecticidal activity evaluation of chromones of date palm pits extract against Culex pipiens (Diptera: Culicidae). Int. J. Mosq. Res, 5(4): 22-32.

Iqbal, M.N., Ashraf, A., 2019. Withania somnifera: Can it be a Therapeutic Alternative for Microbial Diseases in an Era of Progressive Antibiotic Resistance? Int. J. Nanotechnol. Allied Sci., 3(1): 16-18.

Jaddoa, K., Ibrahim, B., 2014. Role of yeast extract in the growth of black seed callus and production of secondary metabolism compounds. Iraqi J. Agric. Sci., 45(7): 685-691.

Kalim, M., Hussain, F., Ali, H., Ahmad, I., Iqbal, M.N., 2016. Antifungal Activities of Methanolic Extracts of Datura inoxia. PSM Biol. Res., 1(2): 70-73.

Kim, H.P., Son, K.H., Chang, H.W., Kang, S.S., 2004. Anti-inflammatory plant flavonoids and cellular action mechanisms. J. Pharmacol. Sci., 96(3): 229-45.

Knekt, P. et al., 2002. Flavonoid intake and risk of chronic diseases. The Am. J. Clin. Nutr., 76(3): 560-568.

Kris-Etherton, P.M. et al., 2004. Bioactive compounds in nutrition and health-research methodologies for establishing biological function: the antioxidant and anti-inflammatory effects of flavonoids on atherosclerosis. Annu. Rev. Nutr., 24: 511-538.

Lotito, S.B., Frei, B., 2006. Dietary flavonoids attenuate tumor necrosis factor α-induced adhesion molecule expression in human aortic endothelial cells: structure-function relationships and activity after first pass metabolism. J. Biol. Chem., 281(48): 37102-37110.

Materska, M., Perucka, I., 2005. Antioxidant activity of the main phenolic compounds isolated from hot pepper fruit (Capsicum annuum L). J. Agric. Food Chem., 53(5): 1750-6.

Meignanalakshmi, S., Vinoth Kumar, S., Deepika, J., Farida Begum, I., 2013. Evaluation of antibacterial activity of methanol extract of leaves of Adhatoda vasica on mastitis pathogens. Hygela J. Drugs Med., 5: 1-4.

Middleton, E., Jr., Kandaswami, C., 1992. Effects of flavonoids on immune and inflammatory cell functions. Biochem. Pharmacol., 43(6): 1167-79.

Minato, K. et al., 2003. Lemon flavonoid, eriocitrin, suppresses exercise-induced oxidative damage in rat liver. Life Sci., 72(14): 1609-16.

Miyake, Y., 2006. Characteristics of Flavonoids in Niihime Fruit-a New Sour Citrus Fruit. Food Sci. Technol. Res., 12: 186-193.

Miyake, Y., Yamamoto, K., Morimitsu, Y., Osawa, T., 1997. Isolation of C-glucosylflavone from lemon peel and antioxidative activity of flavonoid compounds in lemon fruit. J. Agric. Food Chem., 45(12): 4619-4623.

Nichenametla, S.N., Taruscio, T.G., Barney, D.L., Exon, J.H., 2006. A review of the effects and mechanisms of polyphenolics in cancer. Crit. Rev. Food Sci. Nutr., 46(2): 161-83.

Nijveldt, R.J. et al., 2001. Flavonoids: a review of probable mechanisms of action and potential applications. Am. J. Clin. Nutr., 74(4): 418-25.

Nogata, Y. et al., 2006. Flavonoid composition of fruit tissues of citrus species. Biosci., Biotechnol., Biochem., 70(1): 178-192.

Oyama, K.-i., Kondo, T., 2004. Total synthesis of apigenin 7, 4′-di-O-β-glucopyranoside, a component of blue flower pigment of Salvia patens, and seven chiral analogues. Tetrahedron., 60(9): 2025-2034.

Pauli, G.F., Junior, P., 1995. Phenolic glycosides from Adonis aleppica. Phytochem., 38(5): 1245-1250.

Pretto, F.R., Santarém, E.R., 2000. Callus formation and plant regeneration from Hypericum perforatum leaves. Plant Cell Tissue Organ Cult., 62(2): 107-113.

Rabie, K.A., El-Aal, A., Mona, S., Manaf, H., 2010. Enhanced silymarin accumulation as influence of medium composition in cell suspension cultures of Silybum marianum (l.) gaertn. J. Plant Produc., 1(2): 319-332.

Rizk, S.A., Abdelwahab, S.S., Elrazaz, E., 2019. Synthesis and QSAR study of some novel heterocyclic derivatives as in vitro cytotoxic agents. J. Heterocycl. Chem., 56(2): 443-449.

Rizk, S.A., El-Naggar, A.M., El-Badawy, A.A., 2018a. Synthesis, spectroscopic characterization and computational chemical study of 5-cyano-2-thiouracil derivatives as potential antimicrobial agents. J. Mol. Struct., 1155: 720-733.

Rizk, S.A., El‐Hashash, M.A., El‐Badawy, A.A., 2017. Ultrasonic and Grinding Aptitudes of One‐Pot Synthesis of 5‐(4‐Chlorophenyl)‐7‐(3, 4‐Dimethyl Phenyl)‐2‐oxo‐2H‐Pyrano [2, 3‐b] Pyridine Derivatives as Antibacterial Agents. J. Heterocycl. Chem., 54(3): 2003-2011.

Rizk, S.A., Elsayed, G.A., El-Hashash, M.A., 2018b. One-pot synthesis, spectroscopic characterization and DFT study of novel 8-azacoumarin derivatives as eco-friendly insecticidal agents. J. Iran. Chem. Soc., 15(9): 2093-2105.

Sánchez‐Moreno, C., Plaza, L., de Ancos, B., Cano, M.P., 2003. Quantitative bioactive compounds assessment and their relative contribution to the antioxidant capacity of commercial orange juices. J. Sci. Food and Agric., 83(5): 430-439.

Scalbert, A., Manach, C., Morand, C., Rémésy, C., Jiménez, L., 2005. Dietary polyphenols and the prevention of diseases. Crit. Rev. Food Sci. Nutr., 45(4): 287-306.

Shimoi, K., Saka, N., Kaji, K., Nozawa, R., Kinae, N., 2000. Metabolic fate of luteolin and its functional activity at focal site. BioFactors, 12(1-4): 181-6.

Steinberg, D., 1989. Modifications of low-density lipoprotein that increase its atherogenicity. N. Engl. J. Med., 320: 915-924.

Takaya, Y., Kondo, Y., Furukawa, T., Niwa, M., 2003. Antioxidant constituents of radish sprout (Kaiware-daikon), Raphanus sativus L. J. Agric. Food Chem., 51(27): 8061-6.

Tijburg, L.B., Mattern, T., Folts, J.D., Weisgerber, U.M., Katan, M.B., 1997. Tea flavonoids and cardiovascular disease: a review. Crit. Rev. Food Sci. Nutr., 37(8): 771-85.

Walker, T.S., Bais, H.P., Vivanco, J.M., 2002. Jasmonic acid-induced hypericin production in cell suspension cultures of Hypericum perforatum L.(St. John’s wort). Phytochem., 60(3): 289-293.

Wightman, J.D., 2003. Red Berries and Their Health Benefits, Nutraceutical Beverages. ACS Symposium Series. Am. Chem. Soc., pp. 123-132.

Xi, Q., Lai, W., Cui, Y., Wu, H., Zhao, T., 2019. Effect of yeast extract on seedling growth promotion and soil improvement in afforestation in a semiarid chestnut soil area. Forests, 10(1): 76.