HPLC法测定雄蚕蛾中缬氨酸、异亮氨酸和亮氨酸的含量

Determination of valine,isoleucine and leucine in male silkworm moth by HPLC

目的建立HPLC法测定雄蚕蛾中缬氨酸、异亮氨酸和亮氨酸的含量。方法采用KromasilC18色谱柱(150mm×4.6mm,5μm),以乙腈-0.1mol·L^-1醋酸钠溶液(用醋酸调节pH值至6.5)(7∶93)为流动相A,以乙腈-水(4∶1)为流动相B,进行梯度洗脱;检测波长为254nm;柱温为43℃;流速为1.0mL·min^-1;异硫氰酸苯酯(PITC)柱前衍生化。结果雄蚕蛾中缬氨酸、异亮氨酸和亮氨酸各组分线性关系良好;平均回收率为99.29%~100.13%,RSD值为1.22%~1.45%。结论所建立的方法专属性强,重复性好,可用于雄蚕蛾药材的质量控制。

Objective To establish an HPLC method for the determination of the content of valine,isoleucine and leucine in male silkworm moth.Methods Kromasil C 18 chromatographic column(150 mm×4.6 mm,5μm)was used with acetonitrile-0.1 mol·L-1 sodium acetate(adjust pH value to 6.5 by acetic acid)(7∶93)as mobile phase A,and acetonitrile-water(4∶1)as mobile phase B in a gradient elution mode.The detection wavelength was 254 nm with the column temperature 43℃and the flow rate 1.0 mL·min^-1.Precolumn derivatization was performed by using phenylester isothiocyanate(PITC).Results The linear relationship was good.The average recoveries were 99.29%-100.13%,and the RSDs were 1.22%-1.45%.Conclusion The method has high reproducibility,which can be used in the quality control of male silkworm moth.