柱前衍生HPLC法测定脾氨肽口服液中17种游离氨基酸  

Determination of 17 kinds of free amino acids in oral solution of spleen amino peptide by HPLC with pre-column derivatization

作　　者：山广志[1] 左利民[1] 余立[2] 张彤[2] 张博文 

机构地区：[1]中国医学科学院医药生物技术研究所,北京100050 [2]北京市药品检验所,北京100035 [3]北京第一生物化学药业有限公司,北京100029

出　　处：《中国新药杂志》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)

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.

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 

分 类 号：R927.2[医药卫生—药学]