目的 建立岩黄连培养细胞的高效液相色谱法(HPLC)指纹图谱质量控制方法,测定主要生物碱的含量。方法 采用高效液相色谱法,色谱柱为Dikma Diamonsil C18柱(4.6 mm×250 mm, 5 μm),流动相为乙腈和水体积比为40∶60,每升流动相加3.4 g磷酸二氢钾和1.7 g SDS,检测波长345 nm,流速1 mL·min-1,柱温25 ℃。结果 在线性范围内,脱氢卡维丁和小檗碱的浓度与色谱峰面积呈良好的线形关系,回归方程分别为Y=3 645.2X+47.61(r=0.998 9)和Y=4 172.7X+73.52(r=0.999 2),加样回收率分别为97.9％和98.2％,RSD均在3％以内。结论 该方法准确、可靠,能有效分离和测定岩黄连培养细胞中脱氢卡维丁和小檗碱含量,所建立的指纹图谱相似度评价和方法学考察结果良好,可用于质量控制和定量分析。
Abstract
ObjectiveTo establish a method of HPLC fingerprint to determine the contents of primary alkaloids in Corydalis saxicola Bunting cells. Methods The determination was performed on a Dikma Diamonsil C18 column (4.6 mm×250 mm,5 μm). The mobile phase was acetonitrile and water (40∶60) supplemented with 3.4 g·L-1 KH2PO4 and 1.7 g·L-1 SDS. The chromatogram was monitored at 345 nm. The flow rate was 1 mL·min-1, and the column temperature was kept at 25 ℃. Results The linear regression equations for dehydrocavidine and berberine were Y=3 645.2X+47.61 (r=0.998 9) and Y=4 172.7X+73.52 (r=0.999 2), and the average recoveries were 97.9％ and 98.2％, respectively. The intra- and inter-day RSDs of these alkaloids were lower than 3%. Conclusion An effective method with good precision was established for the determination of the alkaloids in cultured C. saxicola cells. The Results of resemblances evaluation of the fingerprints were satisfactory. This method can be used as the quality and quantity control of C. saxicola cells.
关键词
高效液相色谱法 /
脱氢卡维丁 /
岩黄连 /
指纹图谱
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Key words
HPLC /
dehydrocavidine /
Corydalis saxicola Bunting /
fingerprint
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中图分类号：
R284.1
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参考文献
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