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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