Glycine and lysis hydrochloride assay in human coagulation factor Ⅷ by pre-column derivation
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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。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.
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Key words:
- glycine /
- lysis hydrochloride /
- human coagulation factor Ⅷ /
- pre-column derivation /
- HPLC
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[1] 国家药典委员会. 中华人民共和国药典(三部):2015年版[S]. 北京:中国医药科技出版社,2015:272. [2] 孙东坡,胡一桥. 蛋白质冷冻干燥制品中的保护剂及其保护机制[J]. 药学进展,2003,27(4):202-205. [3] 朱敖兰,杨洁. 生物制品冻干保护剂及其保护机理的研究进展[J]. 喀什师范学院学报,2007,28(6):46-50. [4] 高莉萍,李萍,姚佳佳. 高效液相色谱法测定人凝血因子Ⅷ制品中甘氨酸的含量[J]. 药学实践杂志,2016,34(1):59-61. [5] 孙向红,张建华. HPLC法测定盐酸赖氨酸的含量[J]. 齐鲁药事,2010(6):345-346. [6] 国家药典委员会. 中华人民共和国药典(四部):2015年版[S]. 北京:中国医药科技出版社,2015:235. -
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