| The chemical constituents and quality control of the fruit of Acanthopanax sessiliflorus (Rupr. et Maxim.) Seem.were studied in this paper. The details were listed as follows:1. Isolation and identification of the compoundsNine compounds were isolated from the 70% ethanol extract of the fruit of Acanthopanax sessiliflorus (Rupr. et Maxim.) Seem., which was fractionated on a macroporous resin column by 60% ethanol. They were identified as scopoletin (1), protocatechuic acid methyl ester (2), quercetin (3), acanthoside B (4), hyperin (5), acanthoside D (6), (-)-pinoresinol-4,4’-di-O-β-D-glucopyranoside (7), chiisanoside (8) and 22α-hydroxychiisanoside (9) by analysis of their spectroscopic data and by comparison with those reported on the literature. Compounds 2 and 7 were isolated from Araliaceae for the first time and compound 3 was obtained from Acanthopanax genus for the first time.2. The method was developed for determination of the content of total flavonoidsThe content of total flavonoids of the fruit of Acanthopanax sessiliflorus was determined by colorimetric method at 510 nm with rutin as reference. The standard curve of rutin was linear in the range of 0.0109-0.0654 mg·mL-1 (r=1.0000). The average recovery was 99.5%(RSD=3.0%). The contents of total flavonoids in the fruit and pericarp pulverize by general technique were 10.7 mg·g-1 (RSD=1.1%) and 12.1 mg·g-1 (RSD=1.2%), respectively; the content of total flavonoids in the fruit pulverize by nanotechnology was 22.2 mg·g-1 (RSD=2.9%).3. The extract technology was improved for determination of the content of total saponinsThe content of total saponins of the fruit of Acanthopanax sessiliflorus was determined by colorimetric method at 563 nm with ginsenoside Re as reference. The standard curve was linear in the range of 0.00787-0.0393 mg·mL-1 (r=0.9999) of ginsenoside Re. The average recovery was 101.5%(RSD=2.8%). The contents of total saponins in the fruit and pericarp pulverize by general technique were 9.4 mg·g-1 (RSD=1.1%) and 9.2 mg·g-1 (RSD=1.6%), respectively; the content of total saponins in the fruit pulverize by nanotechnology was 11.0 mg·g-1 (RSD=0.2%).4. The method was established for simultaneous determination of six active compounds.A simple, reliable and accurate method for simultaneous determination of protocatechuic acid (1), scopolin (2), (-)-pinoresinol-4,4’-di-O-β-D-glucopyranoside (3), acanthoside D (4), hyperin (5) and acanthoside B (6) in the fruit of Acanthopanax sessiliflorus was established by high performance liquid chromatography. The optimal chromatographic conditions were achieved on Diamasil C18 column with gradient elution of acetonitrile and 0.05% (v/v) phosphoric acid aqueous solution at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 210 nm. Good linearity was observed over the investigated concentration range of 2.08-41.6μg·mL-1 for compound 1,2.16-43.2μg·mL-1 for compound 2,2.94-58.8μg·mL-1 for compound 3,2.44-48.8μg·mL-1 for compound 4, 13.44-268.8μg·mL-1 for compound 5 and 1.04-20.8μg·mL-1 for compound 6 with the values of r2 higher than 0.999 for all analytes. The method provided good accuracy with recoveries in the range of 95.6-101.6% and good precision with RSD values of intra-day and inter-day precision less than 3.0%. The contents of compounds 3-6 of green fruit were 351.9±1.9μg·g-1,105.6±0.98μg·g-1，630.7±1.3μg·g-1 and 95.6±1.2μg·g-1, respectively; the contents of compounds 1-6 of mature fruit were 55.8±0.97μg·g-1,143.4±1.1μg·g-1,353.0±3.0μg·g-1,211.0±1.1μg·g-1,430.1±1.9μg·g-1 and 180.8±2.0μg·g-1. |
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