柱前衍生高效液相色谱法测定胎盘多肽注射液中的氨基酸  

Determination of amino acids in Placenta Polypeptide Injection by high performance liquid chromatography with pre-column AQC derivatization

作　　者：黄莉[1] 吴小曼[2] 纪宇[2] 王玉[1,2] 

机构地区：[1]中国药科大学,江苏南京210009 [2]江苏省食品药品检验所,江苏南京210008

出　　处：《中国生化药物杂志》2012年第4期373-376,380,共5页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的建立柱前衍生-高效液相色谱法测定胎盘多肽注射液中游离氨基酸和水解后总氨基酸的方法。方法以α-氨基丁酸为内标,以6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯(AQC)为柱前衍生化试剂,采用KromasilC18(250 mm×4.6 mm,5μm)色谱柱,140 mmol/L醋酸铵缓冲液(pH 5.10)和57%乙腈溶液为流动相进行梯度洗脱,检测波长为248 nm,同时测定胎盘多肽注射液中21种游离氨基酸和水解后氨基酸含量。结果该方法重现性好,精密度高,各氨基酸浓度在0.005～0.50 mmol/L范围内线性关系良好;各氨基酸检出限≤1.57 ng。结论建立的AQC柱前衍生-高效液相色谱法可用于检测胎盘多肽注射液中氨基酸的含量,测定的结果为胎盘多肽注射液的质量研究和评价提供了依据。Purpose To establish a pre-column derivatization method with HPLC for the determination of amino acids in Placenta Polypeptide Injection.Methods The sample was mixed with 2-amino butyric acid as the internal standard,derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate（AQC） and analyzed on a Kromasil C18 column（250 mm×4.6 mm,5 μm）,ammonium acetate solution（pH 5.10） and acetonitrile solution as mobile phase,using gradient elution with detection at 248 nm.Results All 21 amino acid derivatives were separated well when an optimized elution condition was applied.Consequently,good reproducibility,high sensitivity and high precision of the method were achieved.This method had a good linearity when the amino acid concentration w as set at the range of 0.005-0.50 mmol/L,and the detector limit of each amino acids was lower than 0.5 pmol.Conclusion The method could be used for determination of content of amino acids in Placenta Polypeptide Injection.

关 键 词：胎盘多肽注射液 AQC 柱前衍生 氨基酸 

分 类 号：R927.2[医药卫生—药学]