标 题:HPLC法测定二氢杨梅素含量的操作规程
起草单位:长沙上禾生物科技有限公司
目 的:HPLC法测定二氢杨梅素含量的操作规程
责任人:QC员。
内 容:
1、 试剂
1.1 甲醇(HPLC级)
1.2 蒸馏水(AR级)
1.3冰醋酸(HPLC级)
2、 对照品
二氢杨梅素(购自中国药品生物制品检定所)
3、 仪器和用具
3.1 电子天平(1/100000)
3.2 玻璃仪器:100ml容量瓶
3.3 安捷伦1220 HPLC
4、液相条件
4.1流动相:甲醇:水:冰醋酸=50:50:1
4.2检测波长:292nm
4.3流速:1ml/min
4.4柱温:30℃
5 、 操作步骤
5.1 对照品溶液的制备 精密称取在110℃干燥至恒重的二氢杨梅素对照品10mg,置50ml容量瓶中,加50%甲醇适量,超声处理10min使溶解,放冷,加50%甲醇至刻度,摇匀,备用。
5.2样品溶液制备 精密称取在110℃干燥至恒重的二氢杨梅素样品10mg,置50ml容量瓶中,加50%甲醇适量,超声处理10min使溶解,放冷,加50%甲醇至刻度,摇匀,备用。
6、检测
取对照品溶液和样品溶液0.22μm针孔过滤器过滤,进样20µl,根据峰面积计算样品含量。
含量按以下公式计算:
A2 x C1 x T
R=----------------- x 100
A1 x C2
C1: 对照品的参考浓度,mg/ml
C2: 样品的参考浓度,mg/ml
A1: 对照品溶液的色谱主峰值面积
A2: 样品溶液的色谱主峰值面积
T: 对照品的含量
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