摘要
目的:建立OPA-FMOC柱前衍生-HPLC法测定脾氨肽口服液中17种游离氨基酸的含量。方法:采用Capcell PAK C18MG色谱柱,以含10 mmol.L-1磷酸氢二钠和10 mmol.L-1硼酸钠的缓冲溶液(pH8.2)为流动相A;以乙腈-甲醇-水(45∶45∶10)为流动相B,梯度洗脱,流速1.5 mL.min-1,柱温40℃,自动在线衍生,检测波长338 nm/262 nm。结果:在一定范围内17种氨基酸的溶液浓度与其峰面积线性关系良好(r>0.999 0),各氨基酸的平均回收率均在93.52%～112.12%之间,重复性RSD为1.0%(n=6)。结论:本方法准确、灵敏、简便、专属性强、重现性好,适用于脾氨肽口服液中游离氨基酸的含量测定。
Objective： To develop an HPLC method for the determination of 17 kinds of free amino acids in oral solution of spleen amino peptide by pre-column OPA-FMOC derivatization. Methods： The separation was performed on Capcell PAK C18 MG column（gradient elution）with column temperature at 40 ℃. Using 10 mmol·L^-1 Na2HPO4 and 10 mmol·L^-1 Na2B4O7 buffer solution （pH 8.2） as mobile phase A and acetonitrile-methanol-water （45∶45∶10） as mobile phase B. Flow rate was 1.5 mL？min-1. OPA and FMOC were used as the on-line automatic derivation reagent. The detection wavelength was set at 338 nm/262 nm. Results： All calibration curves of seventeen amino acids were linear （r〉0.999 0）. The average recoveries were 93.52%~112.12%. The method precision （RSD） was in the range of 1.0% （n=6）. Conclusion： The method is accurate, sensitive and simple, and proved to be suitable for the quantification of each amino acid in oral solution of spleen amino peptide.
出处
《中国新药杂志》
CAS
CSCD
北大核心
2013年第11期1255-1258,1268,共5页
Chinese Journal of New Drugs
基金
国家"重大新药创制"科技重大专项(2012ZX09301002-001-019)
教育部高等学校博士学科点专项科研基金新教师基金(20101106120032)
关键词
高效液相色谱法
柱前衍生
游离氨基酸
脾氨肽口服液
HPLC
pre-column derivatization
free amino acids
spleen amino peptide oral solution
