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[摘要]
目的 建立HPLC-DAD法同时测定益肺清化颗粒中苦杏仁苷、齐墩果酸、熊果酸、仙鹤草酚B、甘草苷、毛蕊异黄酮葡萄糖苷、鲁斯可皂苷元、没食子酸、补骨脂素和紫菀酮的方法。方法 采用反相液相色谱法,色谱柱为Waters Symmetry-C18(250 mm×4.6 mm,5.0μm);流动相为KH2PO4缓冲溶液(KH2PO4 1.0 g,加水1 000 mL使溶解,磷酸调节pH值至3.5)-(乙腈-甲醇1∶1),梯度洗脱,体积流量0.8 mL/min;柱温40℃。结果 苦杏仁苷、齐墩果酸、熊果酸、仙鹤草酚B、甘草苷、毛蕊异黄酮葡萄糖苷、鲁斯可皂苷元、没食子酸、补骨脂素和紫菀酮10种成分能够达到良好分离;其线性范围分别为0.2~2.0 μg/mL(r=0.999 5)、0.4~4.0 μg/mL(r=0.999 4)、0.3~3.0 μg/mL(r=0.999 6)、0.1~1.0 μg/mL(r=0.999 3)、0.5~5.0 μg/mL(r=0.999 1)、0.15~1.50 μg/mL(r=0.999 2)、0.25~2.50 μg/mL(r=0.999 5)、0.6~6.0 μg/mL(r=0.999 1)、0.45~4.50 μg/mL(r=0.999 3)、0.12~1.20 μg/mL(r=0.999 4),平均加样回收率分别为98.7%、98.0%、99.6%、98.0%、99.1%、99.4%、98.8%、101.1%、100.6%、101.7%,RSD分别为0.7%、1.3%、1.4%、1.4%、1.1%、0.8%、0.9%、0.5%、0.5%、0.8%。10批次供试品中苦杏仁苷、齐墩果酸、熊果酸、仙鹤草酚B、甘草苷、毛蕊异黄酮葡萄糖苷、鲁斯可皂苷元、没食子酸、补骨脂素和紫菀酮质量浓度分别为0.104~0.123、0.600~0.621、0.501~0.523、0.100~0.121、0.103~0.122、0.231~0.260、0.043~0.065、0.055~0.069、0.061~0.079、0.031~0.043 mg/g。结论 本方法操作简便,结果准确可靠,可用于益肺清化颗粒的质量控制。
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[Abstract]
Objective To establish an HPLC-DAD method for the simultaneous determination of amygdalin, oleanolic acid, ursolic acid, agrimol B, liquiritin, calycosin-7-glucoside, ruscogenin, gallic acid, psoralen, and shionone in Yifei Qinghua Granules (YQG). Methods The chromatographic separation was achieved on an Waters Symmetry-C18 (250 mm×4.6 mm, 5.0 μm) column with mobile phase consisted of (1.0 g potassium phosphate monobasic in 1 000 mL water, the pH value was adjusted to 3.5 with phosphate)-(acetonitrile-methanol 1:1) for gradient elution, at the flow rate of 0.8 mL/min; The column temperature was 40℃. Results The linear ranges of amygdalin, oleanolic acid, ursolic acid, agrimol B, liquiritin, calycosin-7-glucoside, ruscogenin, gallic acid, psoralen, and shionone were 0.2-2.0 μg/mL (r=0.999 4), 0.4-4.0 μg/mL (r=0.999 4), 0.3-3.0 μg/mL (r=0.999 6), 0.1-1.0 μg/mL (r=0.999 3), 0.5-5.0 μg/mL (r=0.999 1), 0.15-1.50 μg/mL (r=0.999 2), 0.25-2.50 μg/mL (r=0.999 5), 0.6-6.0 μg/mL (r=0.999 1), 0.45-4.50 μg/mL (r=0.999 3), and 0.12-1.20 μg/mL (r=0.999 4), respectively. These ten components were well resolved. Their average recoveries (n=6) respectively were 98.7% (RSD=0.7%), 98.0% (RSD=1.3%), 99.6% (RSD=1.4%), 98.0% (RSD=1.4%), 99.1% (RSD=1.1%), 99.4% (RSD=0.8%), 98.8% (RSD=0.9%), 101.1% (RSD=0.5%), 100.6% (RSD=0.5%), and 101.7% (RSD=0.8%). The content of ten batches of the amygdalin, oleanolic acid, ursolic acid, agrimol B, liquiritin, calycosin-7-glucoside, ruscogenin, gallic acid, psoralen, and shionone was 0.104-0.123, 0.600-0.621, 0.501-0.523, 0.100-0.121, 0.103-0.122, 0.231-0.260, 0.043-0.065, 0.055-0.069, 0.061-0.079, 0.031-0.043 mg/g, respectively. Conclusion The method is accurate, sensitive, credible, and repeatable, which can be applied to the quality control of YQG.
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