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Objective:To develop an HPLC method for the simultaneous determination of oleanic acid and ursolic acid in Salvia plebeia R. Br. from different origins. Methods:The column was Agilent-C18 (250 mm × 4. 6 mm,5μm) with the temperature of 25℃, the mobile phase was methanol-water-glacial acetic acid-triethylamine (90 ∶10 ∶0. 03∶0. 06) at the flow rate of 1. 0 ml·min-1 ,and the detection wavelength was 210 nm. Results: The calibration curves were linear within the range of 7. 50-75. 00 μg · ml-1 for oleanic acid(r=0. 999 7)and 5. 00-50. 00 μg · ml-1 for ursolic acid(r=0. 999 7). The average recovery of oleanic acid and ursolic acid was 100. 15%(RSD=1. 14%,n=6)and 99. 40%(RSD=1. 67%,n=6), respectively. Conclusion: The method is simple and accurate, which is suitable for the determination of oleanic acid and ursolic acid in Salvia plebeia R. Br. from different origins.
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Objective To explore the effect of oleanic acid on key enzyme activity in insulin-resistant HepG2 cells. Methods The HepG2 cells were divided into normal control,model control,metformin,and oleanic acid groups.Glycogen content in insulin-resistant HepG2 cell model were detected by hepatic glycogen test kit upon treatment with oleanic acid.Activities of glucokinase ( GK) ,phosphoenolpyruvate carboxylase kinase (PEPCK),and glucose-6-phosphatase (G-6-Pase) were assayed by the glucose 6-phosphate dehydrogenase coupling colorimetric, lactate dehydrogenase coupling colorimetric and ammonium molybdate constant phosphorus methods. Results The oleanic acid enhanced glucose consumption,lowered the activity of G-6-Pase and PEPCK by 54.8% and 18.8%,respectively,and increased the activity of GK and glycogen content in also insulin-resistant HepG2 cells by 100.6% and 98.6%,respectively. Conclusion Aqueous extracts of shirako play a role in lowering PEPCK and G-6-Pase activities and inhibiting glucogenesis, resulting in the reduction of endogenous glucose in the cell. In addition,it can augment the activity of GK,accelerate the process of glucolysis,increase the glycogen content,and alleviate insulin resistance of HepG2.
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Objective: To identify the saponins in the rhizomes of Anemone davidii by ultra performance liquid chromatography coupled with time-of-fight mass spectrometry (UFLC/Q-TOF-MS/MS). Methods: The separation was performed on UPLC Welch C18 column (100 mm × 2.1 mm, 1.7 μm), with a mobile phase using 0.1% acetonitrile (A) and water containing 0.1% formic acid (B) for gradient elution. Q-TOF/MS and electrospray ion (ESI) source were applied for the analysis under the negative ion mode, and the running time was 40.25 min. Using target compound screening method, the structures of monitored chemical constituents were identified by retention time, exact relative molecular mass, and cleavage fragments of MS/MS. Results: Fifty-two triterpenoids were separated and identified from the methanol extract of A. davidii, 47 triterpenoids were identified for the first time from A. davidii among which nine pairs of structural isomers were included. Conclusion: UPLC/Q-TOF-MS/MS method can identify the main chemical constituents from A. davidii rapidly and accurately, which develops a new strategy for identification of the chemical constituents in A. davidii.
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Objective: To establish an HPLC-ESI-MS method for quickly identifying the chemical constituents in the extracts of Akebiae Fructus. Methods: The main saponin components in the extracts of Akebiae Fructus were detected with the HPLC-ESI-MS in negative ion mode. These components were further analyzed by MS2 and MS3 spectra, and by comparing with the corresponding reference substances and literature data. Results: Seventeen saponins in the extracts of Akebiae Fructus were well separated in one run. Conclusion: The new method is accurate and rapid. It could be used to identify the main chemical constituents in the extracts of Akebiae Fructus and be suitable for the quality control of Akebiae Fructus.