RP-HPLC测定正常人血清中的瓜氨酸
Determination of citrulline in human serum by reversed phase HPLC
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收稿日期： 年卷（期）页码：2020,35(06):-630-634
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：瓜氨酸;人血清;含量测定;高效液相色谱;柱前衍生化;2,4-二硝基氟苯
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中文摘要
目的采用反向HPLC法测定人血清中的瓜氨酸含量。方法以2,4-二硝基氟苯(DNFB)为衍生化试剂,柱前衍生;采用Amethyst C_(18)-H色谱柱(250 mm×4.6 mm,5μm),流动相A为乙腈-水溶液(1:1),流动相B为50 mmol·L(-1)磷酸二氢钾加1‰磷酸的缓冲盐溶液,梯度洗脱,检测波长360 nm,柱温30℃,流速1 mL·min(-1)磷酸二氢钾加1‰磷酸的缓冲盐溶液,梯度洗脱,检测波长360 nm,柱温30℃,流速1 mL·min(-1)。结果在该实验条件下,瓜氨酸在30 min内的分离度良好,0.25～100μg·mL(-1)。结果在该实验条件下,瓜氨酸在30 min内的分离度良好,0.25～100μg·mL(-1)瓜氨酸与峰面积呈良好的线性关系,精密度的RSD=1.98%,供试样品48 h内稳定,重复性的RSD=1.55%,平均加样回收率为87.45%,RSD=1.47%;在此方法基础上测定正常人血清中瓜氨酸的含量为33.08±6.46μmol·L(-1)瓜氨酸与峰面积呈良好的线性关系,精密度的RSD=1.98%,供试样品48 h内稳定,重复性的RSD=1.55%,平均加样回收率为87.45%,RSD=1.47%;在此方法基础上测定正常人血清中瓜氨酸的含量为33.08±6.46μmol·L(-1)。结论所用方法稳定可靠、高效简洁、灵敏度高、成本低。
参考文献
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