RESUMO
OBJECTIVE: To develop an HPLC method for simultaneous determination of multiple-components in Hedyotis diffusa Willd. METHODS: The HPLC analysis was carried out on a C18 column (4.6 mm×250 mm, 5 μm) by gradient elution with acetoni-trile-water[both containing 0.1‰ (V/V) acetic acid] as mobile phase at a flow rate of 0.8 mL·min-1, the column temperature at 35°C, and the detection wavelength was set at 238 nm. External standard method and quantitative analysis of multi-components by single marker (QAMS) method were adopted for simultaneous determination of six components in Hedyotis diffusa Willd, respectively. RESULTS: The linear ranges for asperulosidic acid, quercetin-3-O-[2-O-(6-O-E-feruloyl)-β -D-glucopyranosyl]-β-D-glucopyrano-side, kaempferol-3-O-[2-O-(6-O-E-feruloyl)-β-Z) -gfucopyranosyl]-β-D-galactopyranoside, (E)-6-O-p-coumaroyl scandoside methyl ester, (E)-6-O-feruloyl scandoside methyl ester, (Z)-6-O-p-coumaroyl scandoside methyl ester were 2.34-93.50, 2.61-104.33, 0.67-26.69, 3.42-136.84, 0.65-26.07, and 1.10-44.17 μg·mL-1 (r<0.9993), respectively. The RSD values of precision, reproducibility, and sample stability were not more than 2.2%. The average recoveries of the six components were 99.8%-101.1% with RSDs not more than 1.2%. The P values of external standard method and QAMS by paired t-test were greater than 0.05. CONCLUSION: There is no significant difference in the content analysis results of the two methods, which can both used for simultaneous determination of the four iridoids and two flavonoids in Hedyotis diffusa Willd.
RESUMO
OBJECTIVE: To study the feasibility of quantitative analysis of multi-components by single-marker (QAMS) in determination of flavones in Shuanghuanglian preparations by comparing the results assayed by QAMS and external standard method (ESM). METHODS: The analysis was performed on a Xtimate™ C18 column (4.6 mm × 250 mm, 5 μm)with methanol and water containing 0.2% phosphoric acid as the mobile phase in gradient elution. The flow rate was set at 1.0 mL · min-1. The UV detector wavelength was 274 nm, and the column temperature was 30°C. RESULTS: Within the linear ranges, the analysis results showed no significant difference between QAMS and ESM. CONCLUSION: The QAMS method is credible and will be a new quality evaluation pattern for the determination of multiple flavones in Shuanghuanglian preparations.
