| Alpinia officinarum Hance is one of important medicinal plants growing mainly in Southern China. It is a traditional chinese medicine which has been recorded in China Pharmacopoeia for relieving stomachache, treating colds, invigorating the circulatory system, and reducing swellings. Its major chemical constituents include volatile oil, diaryheptanoid and flavonoids. Pharmacological experiments indicate that this plant has anti-ulcer, ant-diarrhea, antioxidant, anti-inflammatory antibacterial functions.16 samples were collected from different habitats, which were used for the studies on the chemical components, the method of assessment and antibacterial activities.The experiments show that the chloroform and acetoacetate extractions had remarkable antibacterial activities.The plants of Alpinia officinarum Hance(9 kg) were extracted with 80% aqueous alcohol to give an alcoholic extract, which was extracted with chloroform and acetoacetate to afford active extractions. Eight compounds were isolated from the chloroform and acetoacetate fractions by silica gel, Sephadex LH-20 and Polyacylamine chromatography and they were identified as follow by chemicalreactions and spectral analysis:galangin, kaempferide-4’-methylether, galangin-3-methylether, protocatechuic acid, quercetin, kaempferol,β-sitosterol,daucosterol.The antibacterial tests in vitro indicate that 4 of these compounds(galangin, protocatechuic acid, quercetin and kaempferol) could effectively inhibit the growth of the chosen bacteria. Among the 4 compounds, protocatechuic acid and quercetin have antibacterial activity against both Gram-positive(Staphylococcus aureus and Bacillus subtilis)and Gram-negative(Escherichia coli and Pseudomonas aeruginosa);galangin and kaempferol have antibacterial activity against Gram-negative(Escherichia coli and Pseudomonas aeruginosa).A RP-HPLC method was developed for the determination of protocatechuic acid in Alpinia officinarum Hance.The separation was performed on a Diamonsil-C18 column (200 mm×4.6 mm,5μm) with the mobile phase consisting of methanol-0.05%phosphoric acid (12:88,v/v) at a flow rate of 1.0 mL·min-1.The detection wavelength was set at 258 nm and the column temperature was set at 35℃. The calibration curve was linear over the range of 0.902-4.432 μg·mL-1 (r= 0.9998). The average recoveries were 99.2% (RSD= 1.8%, n=9). The method is simple, accurate and reproducible, and suitable for the quality control of Alpinia officinarum Hance.A RP-HPLC method was developed for simultaneous determination of 3 flavonoids in Alpinia officinarum Hance.The separation was performed on a Promosil-C18 column (250 mm×4.6 mm,5μm) with a gradient of acetontrile and 0.05%phosphoric acid (v/v) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 264 nm and the column temperature was set at 30℃. The calibration curves of galangin, kaempferide-4’-methylether and galangin-3-methylether were linear in the range of 12.18-21.8μg·mL-1 (r= 0.9999),4.44-44.4μg·mL-1 (r= 0.9999) and 2.53-25.32μg·mL-1 (r= 0.9998) respectively. The mean recoveries (n= 9) of them were 99.2%(RSD= 1.6%),100.5%(RSD= 1.8%) and 100.4%(RSD= 1.5%) respectively. The method is simple, accurate and reliable, which is helpful to control the quality of Alpinia officinarum Hance.The antibacterial tests indicate essential oil of Alpinia officinarum Hance could effectively inhibit the growth of the chosen bacteria,so we establish a GC method for simultaneous determination of a-pinene, P-pinene, eucalyptol and a-terpineolin in volatile fractions of Alpinia officinarum Hance. The essential oil was extracted by steam distillation. Capillary column DB-1(30 m×0.25 mm×0.25μm ) was used;The detector was FID;Inlet temperature was 230℃;The detector temperature was 250℃.Temperature programs was 50℃(hold 2min) programmed to 130℃(hold 3 min) at 8℃·min-1; Carrying gas was nitrogen(1.2 mL·min-1),split ratio was 10:1; Injection volumn was 1μL. The linear ranges for a-pinene,β-pinene, eucalyptol and a-terpineolin were 0.009-0.090 mg·mL-1 (r= 0.9998),0.009-0.091 mg·mL-1 (r= 0.9998),0.0604-0.604 mg·mL-1 (r= 0.9997),0.0374-0.374 mg·mL-1 (r= 0.9995),respectively; The average recoveries (n= 9) of a-pinene,β-pinene, eucalyptol and a-terpineolin were 96.2%(RSD= 0.8%),96.7%(RSD= 1.1%),98.7%(RSD= 1.1%),96.7%(RSD= 2.2%). The developed method is simple, quick and accurate, it can be used for the quality control of Alpinia officinarum Hance. |
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