高效液相色谱法测定大鼠血清中的高良姜素___

 

 

目的：建立测定大鼠血清中高良姜素的HPLC内标法。方法：SD大鼠腹腔注射给药，血清样品用乙酸乙酯萃取，以大黄素为内标；采用Shim-pack C₁₈柱（2.1mm×250 mm，5 μm），以A相：0.1%乙酸-水，B相：乙腈-甲醇-四氢呋喃（15∶40∶45）为流动相，梯度洗脱\[0 min时，A-B（61∶39）；20 min时，A-B（39∶61）；30 min时，A-B（0∶100）；45 min时，A-B（0∶100）]，流速0.2 mL·min^-1；检测波长为345 nm，柱温35 ℃。结果：高良姜素的线性范围为0.34~26.0 μg·mL^-1（r=0.9998）,最低定量限为0.34 μg·mL^-1，方法回收率为90.6%~111.9%，绝对回收率大于85%，日内和日间的RSD均小于6.3%；血清样品的稳定性符合要求。结论：所建立的大鼠血清中高良姜素的HPLC测定方法灵敏、准确、精密、稳定，适用于大鼠体内高良姜的药代动力学研究。

 

 

 

Objective:To develop an HPLC internal standard method for the determination of galangin in rat serum.Methods:The determination was performanced on Shim-pack C18 column（2.1 mm×250 mm，5 μm）with the mobile phase consisting of 0.1% acetic acid-water (A) and acetonitrile-methanol-tetrahydrofuran(15∶40∶45;B) with gradient elution mode at a flow rate of 0.2 mL·min-1.The following gradient procedure was used:0-20 min,39%→61%B;20-30 min,61%→100%B;30-45 min,100%→100%B.Detection wavelength was 345 nm,the column temperature was 35 ℃.The route of administration of rats was ip,serum samples were extracted with ethyl acetate,methyl parahydroxybenzoate was used as an internal standard.Results:The linear range of galangin was 0.34 to 26.0 μg·mL^-1（r=0.9998）with the lower limit of quantification for 0.34 μg·mL^-1.The method recovery was 90.6% to 111.9% and the absolute recovery was more than 85%.The intra-day RSD and inter-day RSD were less than 63%.Conclusion:The developed method for the quantification of galangin in rat serum is sensitive,accurate,precise,stable,and is suitable for the pharmacokinetic studies of galangal in rats.

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