標 題:HPLC法測定二氫楊梅素含量的操作規程
起草單位:長沙上禾生物科技有限公司
目 的:HPLC法測定二氫楊梅素含量的操作規程
責任人:QC員。
內 容:
1、 試劑
1.1 甲醇(HPLC級)
1.2 蒸餾水(AR級)
1.3冰醋酸(HPLC級)
2、 對照品
二氫楊梅素(購自中國藥品生物制品檢定所)
3、 儀器和用具
3.1 電子天平(1/100000)
3.2 玻璃儀器:100ml容量瓶
3.3 安捷倫1220 HPLC
4、液相條件
4.1流動相:甲醇:水:冰醋酸=50:50:1
4.2檢測波長:292nm
4.3流速:1ml/min
4.4柱溫:30℃
5 、 操作步驟
5.1 對照品溶液的制備 精密稱取在110℃乾燥至恆重的二氫楊梅素對照品10mg,置50ml容量瓶中,加50%甲醇適量,超聲處理10min使溶解,放冷,加50%甲醇至刻度,搖勻,備用。
5.2樣品溶液制備 精密稱取在110℃乾燥至恆重的二氫楊梅素樣品10mg,置50ml容量瓶中,加50%甲醇適量,超聲處理10min使溶解,放冷,加50%甲醇至刻度,搖勻,備用。
6、檢測
取對照品溶液和樣品溶液0.22μm針孔過濾器過濾,進樣20µl,根據峰面積計算樣品含量。
含量按以下公式計算:
A2 x C1 x T
R=----------------- x 100
A1 x C2
C1: 對照品的參考濃度,mg/ml
C2: 樣品的參考濃度,mg/ml
A1: 對照品溶液的色譜主峰值面積
A2: 樣品溶液的色譜主峰值面積
T: 對照品的含量
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