HPLC法测定L-肌肽中的游离氨基酸的含量  

Determination of contents of the free amino acids in L-carnosine by HPLC

作　　者：韩木南[1] 陈晓辉[1] 纪晓影[1] 其其格[1] 毕开顺[1] 

机构地区：[1]沈阳药科大学药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2010年第4期303-305,共3页Journal of Shenyang Pharmaceutical University

摘　　要：目的采用柱前衍生高效液相色谱法对L-肌肽原料药中的游离氨基酸进行测定。方法采用异硫氰酸苯酯(phenyl isothiocyanate,PITC)柱前衍生高效液相色谱法,使L-肌肽中游离的β-丙氨酸和L-组氨酸衍生物达到完全分离,并测定其含量。色谱柱为DiamonsilTM C18(200 mm×4.6 mm,5μm),流动相为甲醇-0.05 mol.L-1醋酸钠溶液(体积比为13∶87),流速为1.0 mL.min-1,检测波长为254 nm,柱温为35℃。结果HPLC法测定β-丙氨酸和L-组氨酸的线性范围分别为2.01～20.10 mg.L-1(r=0.999 7)和2.00～20.00 mg.L-1(r=0.999 3);平均回收率分别为110.6%和98.2%。结论HPLC法无需专业的氨基酸分析仪,可用于L-肌肽中游离氨基酸的检查。Objective To establish an HPLC method with precolumn derivatization for determining the contents of two free amino acids in L-carnosine.Methods Using phenyl isothiocyanate（PITC）as the derivating reagent,the contents of β-alanine and histidine in L-carnosine were determined,which were completely separated.The column was DiamonslTM C18（200 mm×4.6 mm,5 μm）;the mobile phase was methanol-0.05 mol·L^-1 sodium acetate solution（V∶V=13∶87）;the flow rate was 1.0 mL·min^-1;the detection wavelength was set at 254 nm and the column temperature was 35 ℃.Results The liner ranges of β-alanine and histidine were 2.01-20.10 mg·L^-1（r=0.999 7）and 2.00-20.00 mg·L^-1（r=0.999 3）,respectively;their average recoveries were 110.6% and 98.2%.Conclusions The method does not need the special amino acid analyzer,and it is efficient,sensitive and accurate for determination of the free amino acids in L-carnosine.

关 键 词：L-肌肽 氨基酸 高效液相色谱法 

分 类 号：R917[医药卫生—药物分析学]