柱前衍生化HPLC测定人纤维蛋白粘合剂中盐酸精氨酸和亮氨酸的含量
Determination of two amino acids in human fibrin sealant kit by pre-column derivatization HPLC
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摘要: 目的:建立HPLC法测定人纤维蛋白粘合剂中盐酸精氨酸和亮氨酸的含量。方法:采用Shim-pack CLC-ODS色谱柱,柱前衍生化,流动相A为50%乙腈,流动相B为醋酸盐缓冲液;检测波长362 nm;用外标法检测。结果:本色谱条件下2种氨基酸衍生物能良好地分离,衍生化试剂对氨基酸分析无干扰。平均回收率:盐酸精氨酸为102.8%,RSD为0.83%;亮氨酸为103.8%,RSD为0.92%。结论:本方法准确,方便,费用低,可用于人纤维蛋白粘合剂中2种氨基酸含量的控制。Abstract: Objective :To set up a method for determination of arginine and leucine in human fibrin sealant kit. Methods :A Shim-pack CLC-ODS column was used after pre-column derivatization with 2,4-dinitrobenzene fluoride,with the mobile phase of 50% acetonitrile and 0.05mol/L acetate buffer,and the UV detection wavelength was 362 nm.The content was analyzed by external standard method. Results :These two amino acid derivatives can be well separated under the chromatographic conditions.The average recovery was 102.8%(RSD=0.83%) for Arg,and 103.8%(RSD=0.92%) for Leu. Conclusion :The method is accurate,convenient,low-cost and suitable for determination of the two amino-acids in human fibrin sealant kit.
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Key words:
- arginine hydrochloride /
- leucine /
- HPLC /
- derivation /
- 2,4-dinitrofluorobenzene
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