ABSTRACT
Objective: To study the chemical compositions from the root tuber of Tetrastigma hemsleyanum and their anti-oxidative activities. Methods: The compounds were isolated by various column chromatography. Their structures were identified by spectroscopic methods including 1H-NMR, 13C-NMR, MS, HR-MS, etc. The anti-oxidative activities were evaluated by DPPH radical scavenging assay. Results: Fourteen compounds were isolated from the root tuber of T. hemsleyanum, they were identified as kaempferol (1), quercetin (2), salicylic acid (3), benzoic acid (4), oxyresveratrol (5), catechin (6), L-epicatechin (7), epigallocatechin (8), procyanidin B2 (9), procyanidin B1 (10), 3-O-caffeoylquinic acid (11), protocatechualdehyde (12), p-hydroxybenzoic acid (13), and protocatechuic acid (14). The anti-oxidative IC50 values of compounds 2, 8, 9, 10, and 12 were 14.15, 14.19, 15.99, 12.4, and 15.98 μmol/L, respectively, better than the positive control Vit C (IC50 value was 23.0 μmol/L). Conclusion: Compounds 4-12 are isolated from T. hemsleyanum for the first time. It has been demonstrated that the root tuber contain flavonoids and also is rich in organic acids. Moreover, compounds 2, 8, 9, 10, and 12 have the certain prospects in the development of natural anti-oxidative agents.
ABSTRACT
In this study, a rapid and sensitive analytical method was developed for the determination of 10 major compounds (procyanidin B1, catechin, procyanidin B2, rutin, isoquercitrin, kaempferol-3-O-rutinoside, astragalin, quercitrin, quercetin, and kaempferol) in Tetrastigma hemsleyanum by using ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-MS/MS) in multiple-reaction monitoring (MRM) mode. UPLC-MS/MS assay with negative ion mode was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 μm) with the mobile phase consisting of acetonitrile (A) and 0.1% aqueous formic acid (B) in gradient elution at a flow rate of 0.25 mL · min(-1) and the column temperature was set at 45 °C. Under the optimized chromatographic conditions, good separation for 10 target compounds were obtained including chiral isomer procyanidins B1 and B2 were completely separated within 8.5 min. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.996 6), the overall recoveries were ranged from 95.44%-110.40% with the RSD ranging from 2.37%-8.69%. It is the first report about simultaneous analysis of 10 major flavonoids components in Tetrastigma hemsleyanum by using UPLC-MS/MS method, which affords highly sensitive, specific, speedy and efficient method for quality control of Tetrastigma hemsleyanum
Subject(s)
Acetonitriles , Chromatography, High Pressure Liquid , Flavonoids , Chemistry , Kaempferols , Quercetin , Rutin , Tandem Mass Spectrometry , Vitaceae , ChemistryABSTRACT
A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) was developed for identification of multi-constituents and an analytical method was developed for simultaneously determining 4 major compounds (rutin, isoquercitrin, kaempferol-3-0-rutinoside, and astragalin) in Tetrastigma hemsleyanum Diels et Gilg. The HPLC-Q-TOF-MS assay was performed on a Welch Ultimate XB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% Formic acid (B) in gradient mode at a flow rate of 0.8 mL x min(-1). The column temperature was at 30 degrees C, and negative ion mode was used for TOF-MS. The UPLC-QqQ-MS assay was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% formic acid (B) in gradient mode at a flow rate of 0.25 mL x min(-1). The column temperature was at 45 degrees C, and MRM mode was used for QqQ-MS. Based on the retention time and MS spectra, 24 compounds were identified or tentatively characterized by comparing with reference substances or literatures. For quantitative the linear range of 4 detected compounds were good (r > 0.9966), and the overall recoveries ranged from 98.27% to 101.58%, with the RSD ranging from 3.15% to 5.88%. The results indicated that new approach conbined HPLC-Q-TOF-MS and UPLC-QqQ-MS was applicable in qualitative and quantitative quality control of Tetrastigma hemsleyanum.