Validated HPTLC method for the simultaneous quantification of diterpenoids in Vitex trifolia L.

Abstract

Vitex trifolia L. is an important Indian medicinal plant with diverse pharmacological properties. In a recent study, we reported the isolation and antitubercular activity evaluation of three new diterpenoids from its leaves; here we have developed a validated rapid, simple, precise, and accurate high-performance TLC method for the simultaneous quantification of isolated diterpenoids in V. trifolia. Diterpenoids, 6α,7α-diacetoxy-13-hydroxy-8(9),14-labdadien (A), 13-hydroxy-5(10),14-halimadien-6-one (B), and 9-hydroxy-13(14)-labden-16,15-olide (C) were separated on silica gel 60F254 high-performance TLC plates using chloroform/acetone (98:2, v/v) as mobile phase. The quantitation of diterpenoids was carried out using densitometric reflection/absorption mode at 610 nm after postchromatographic derivatization using a vanillin/sulfuric acid reagent. A precise and accurate quantification can be performed for compounds A and B in the linear working concentration range of 333-1000 ng/band and for C in the range of 670-2000 ng/band with good correlations (r = 0.9984, 0.9991, and 0.9994, respectively). The method was validated for peak purity, precision, accuracy, robustness, LOD, and LOQ, as per the ICH guidelines. The method reported here is simple, reproducible and may be applied for the quantitative analysis of the above diterpenoids in the leaves of V. trifolia.

Simultaneous quantitative analysis of 11 constituents in Viticis Fructus by HPLC‐HRMS and HPLC‐DAD combined with chemometric methods

“…The batch of VTLF with the lowest content of chlorogenic acid (3) still had a higher amount than the VTF batch with the highest content of chlorogenic acid (3). The contents of the main effective component (casticin (11)) in VTF and VTLF were also quite different. The casticin (11) content in VTF was significantly lower than that in VTLF.…”

“…According to the results, the contents of these components in VTLF were generally higher than those in VTF. Obvious differences in the contents of chlorogenic acid (3) and casticin (11) were observed between different Viticis Fructus samples. The average amounts of chlorogenic acid (3) in VTF and VTLF were 0.119 and 1.877 mg/g, respectively.…”

“…(1), 4-hydroxybenzoic acid (2), chlorogenic acid (3), vanillic acid (4), isoorientin ( 5), orientin (6), agnuside (7), 3,4-dicaffeoylquinic acid (8), 3,5-dicaffeoylquinic acid (9), 4,5-dicaffeoylquinic acid (10), and casticin (11)…”

“…The peaks were identified by comparing the retention times of chromatographic peaks and the corresponding MS and MS/MS spectrograms of each chromatographic peak with references. 22,[29][30][31][32][33] Peaks 1-11 were 3,4-dihydroxybenzoic acid (1), 4-hydroxybenzoic acid (2), chlorogenic acid (3), vanillic acid (4), isoorientin ( 5), orientin (6), agnuside (7), 3,4-dicaffeoylquinic acid (8), 3,5-dicaffeoylquinic acid (9), 4,5-dicaffeoylquinic acid (10), and casticin (11). Among them, 3,4-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid were identified and quantified in Viticis Fructus for the first time.…”

Traditional Medicinal Uses, Phytochemistry, Biological Properties, and Health Applications of Vitex sp.

“…Two new labdane diterpenoids, such as 6 α ,7 α -diacetoxy-13-hydroxy-8(9),14-labdadien ( 60 ), and 9-hydroxy-13(14)-labden-15,16-olide ( 61 ), were found in the leaf extract of V . trifolia L. Compound 60 was found to be active against Mycobacterium tuberculosis, with an MIC value of 100 μg/mL [ 93 ].…”