反相高效液相色谱法同时测定生物转化液中的反式茴脑、茴香醛和茴香酸

建立了同时测定生物转化液中反式茴脑、茴香醛和茴香酸3种化合物的反相高效液相色谱(RP-HPLC)分析方法。采用Kromasil-100A C18色谱柱(250 mm×4.6 mm×5μm),流动相为V(乙腈)∶V(水)∶V(冰醋酸)=70∶30∶0.02,等度洗脱,流速0.8 mL/min,检测波长为260 nm,进样量5μL,柱温室温,15 min内3种物质得到了较好的分离。该方法中各标准品质量浓度与色谱峰面积线性关系良好,具有较好的精确度和重现性。反式茴脑、茴香醛和茴香酸线性范围分别为4.080～2.652×102 mg/L(R2=0.9999)、4.580～64.12 mg/L(R2=0.9997)、3.780～52.92 mg/L(R2=0.9993);平均加标回收率分别为100.34%、101.18%、100.31%;相对标准偏差RSD分别为0.92%、1.75%、0.53%。用于实际样品测定时,RSD均小于1.1%。该方法简便、快速、准确,能同时定量分析反式茴脑生物转化液等复杂体系中的反式茴脑、茴香醛和茴香酸。

Analysis of Trans-anethole,Anisaldehyde and Anisic Acid in Biotransformation Liquids by Reversed Phase High Performance Liquid Chromatography

A reversed phase high performance liquid chromatography method for the analysis of trans-anethole,anisaldehyde and anisic acid in biotransformation liquids was developed.The separation was performed on Kromasil-100A C18 column(250 mm×4.6 mm×5 μm) using V(acetonitrile)∶ V(water)∶ V(acetic acid)=70∶ 30∶ 0.02 as the mobile phase.The flow rate was 0.8 mL/min.The detection wave length was 260 nm.The injection volume was 5 μL,and the column temperature was room temperature.Three compounds were clearly separated in 15 minute.The method showed good linear relationship,precision and repeatability.The linearity was obtained within the range of 4.080~2.652×102 mg/L for trans-anethole(R2=0.999 9),4.580~64.12 mg/L for anisaldehyde(R2=0.999 7),and 3.780~52.92 mg/L for anisic acid(R2=0.999 3).The average recoveries of trans-anethole,anisaldehyde and anisic acid were 100.34%,101.18%,and 100.31% respectively.The relative standard deviations were 0.92%,1.75%,and 0.53% respectively.The relative standard deviations of practical sample were less than 1.1%.The method was found to be simple,rapid,sensitive and accurate for the determination of trans-anethole,anisaldehyde and anisic acid in biotransformation liquids.