柱前衍生化法同时测定人凝血因子Ⅷ中甘氨酸和盐酸赖氨酸的含量 |
投稿时间:2017-11-22 修订日期:2018-04-18 点此下载全文 |
引用本文:姚佳佳,闫晨.柱前衍生化法同时测定人凝血因子Ⅷ中甘氨酸和盐酸赖氨酸的含量[J].药学实践杂志,2018,36(4):362~364,384 |
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中文摘要:目的 建立一种柱前衍生-高效液相色谱法同时测定人凝血因子Ⅷ中2种氨基酸含量的方法。方法 以2,4-二硝基氟苯(DNFB)为衍生剂,采用C18色谱柱,流动相A为水,B为乙腈,C为含4 mmol/L三乙胺的0.05 mol/L醋酸钠缓冲液(pH=6.4),进行梯度洗脱,流速1.0 ml/min,检测波长360 nm。结果 甘氨酸的线性范围为2.98~14.90 g/L (r=0.999 9),盐酸赖氨酸线性范围为1.195~5.975 g/L(r=0.999 9),甘氨酸和盐酸赖氨酸的平均回收率分别为99.1%和98.2%。结论 此方法检测结果准确,可用于人凝血因子Ⅷ中甘氨酸和盐酸赖氨酸的含量测定。 |
中文关键词:甘氨酸 盐酸赖氨酸 人凝血因子Ⅷ 柱前衍生化 高效液相色谱 |
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Glycine and lysis hydrochloride assay in human coagulation factor Ⅷ by pre-column derivation |
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Abstract:Objective To establish a method of analysis for glycine and lysis hydrochloride in human coagulation factor Ⅷ by pre-column derivation HPLC. Methods The sample was derivatized with 2,4-dinitrofluorobenzene(DNFB). HPLC was performed on a C18 column with gradient elution. The mobile phase A was water, B was acetonitrile and C was 0.05 mol/L sodium acetate buffer containing 4 mmol/L trimethylamine. The flow rate was 1.0 ml/min and detection wavelength was 360nm. Result There was a linear range for glycine between 2.980 g/L~14.90 g/L (r=0.999 9) and lysis hydrochloride between 1.195 g/L~5.975 g/L (r=0.999 9). The average recovery rate of glycine and lysis hydrochloride was 99.1% and 98.2%. Conclusion The established method is accurate and can be used to determine the contents of glycine and lysis hydrochloride in human coagulation factor Ⅷ. |
keywords:glycine lysis hydrochloride human coagulation factor Ⅷ pre-column derivation HPLC |
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