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建立HPLC同时测定山豆根中7种生物碱(金雀花碱、氧化苦参碱、氧化槐果碱、N-甲基金雀花碱、槐醇、苦参碱和槐果碱)及3种黄酮(三叶豆紫檀苷、芒柄花素和马卡因)含量的方法。采用Welch XtimateTMC18(4.6 mm×250 mm,5μm)色谱柱;流动相为乙腈(A)-0.01 mol·L-1醋酸铵(氨水调p H 8.0)溶液(B)进行梯度洗脱:0~20 min,4%~14%A;20~30 min,14%~25%A;30~45 min,25%~40%A;45~65 min,40%~55%A;65~75 min,55%A;流速1.0 m L·min-1,柱温30℃,检测波长225 nm。方法学验证结果显示,10种成分的分离度良好,且在各自浓度范围内呈现出良好的线性关系,r≥0.999 7,平均回收率在98.60%~102.6%,RSD为0.60%~3.7%(n=6),该方法简便、准确,重复性良好。含量测定结果显示,7种生物碱和3种黄酮成分在不同样品中的含量具有显著性差异。由此表明,不同批次的山豆根药材,其内在质量存在较大差异。该法可用于山豆根药材的多成分同步测定,同时可以为山豆根的全面质量评价和质量控制提供科学依据。
Simultaneous determination of seven alkaloids (semen camptothecum, oxymatrine, oxysophocarpine, N-methylxedine, sophyl alcohol, matrine and sophocarpine) and three Flavonoids (Trefoil glycosides, monascus and macacaine) content of the method. The gradient elution was carried out on a Welch XtimateTMC18 (4.6 mm × 250 mm, 5 μm) column with a mobile phase of acetonitrile-A (0.01 mol·L-1) ammonium acetate solution (B) min, 4% -14% A, 20-30 min, 14-25% A, 30-45 min, 25-40% A, 45-65 min, 40-55% A, 65-75 min, 55% A; flow rate 1.0 m L · min-1, column temperature 30 ℃, detection wavelength 225 nm. The methodological validation showed that the separation of the 10 components was good and showed a good linear relationship in the range of their concentration, with r≥0.9997, the average recoveries ranged from 98.60% to 102.6%, and the RSDs ranged from 0.60% to 3.7% n = 6), the method is simple, accurate, reproducible. The content of seven alkaloids and three flavonoids in different samples showed significant differences. This shows that different batches of Radix herbal medicine, there is a big difference in its intrinsic quality. The method can be used for the simultaneous determination of multi-components of Radix et Rhizoma and can provide a scientific basis for the comprehensive quality evaluation and quality control of Radix isatidis.