用柱前衍生RP-HPLC法测定脑蛋白水解物注射液中牛磺酸的含量

Determination of taurine in cerebroprotein hydrolysate injection by RP-HPLC with precolumn derivatization

目的：建立测定脑蛋白水解物注射液中牛磺酸含量的柱前衍生RP—HPLC法。方法：在碱性条件下，氨基酸与异硫氰酸苯酯反应，生成异硫氰酸苯一氨基酸的衍生物．采用Diamonsil C18色谱柱（250mm×4．6mmm，5μm），用二元梯度洗脱方式．流动相A为0．1mol／L乙酸钠-乙腈（93：7）．流动相B为乙腈-水（80：20），检测波长254nm。结果：牛磺酸在0．01-0．07mg／mL浓度范围内呈良好的线性关系（r=0．9999），精密度良好，低、中、高3种浓度（0．019、0．024、0．029mg／mL）的加样回收率分别为（97．0±0．5）％、（98．8±0．20）％、（100．0±1．6）％（n=3）。结论：本法结果准确，重复性好。

Objective： To develop a RP-HPLC method with phenylisothiocyanate precolumn derivation for determining the content of taurine in cerebroprotein hydrolysate injection. Methods： In alkaline condition, reaction between amino acids and phenylisothiocyanate formed a derivant of phenylisothiocyanate-amino acid. For RP-HPLC, the Diamonsil C18 column （250 mm×4.6 mm,5 μm） and binary gradient elution were used. The mobile phase A was 0. 1 mol/L sodium acetate solutionacetonitrile （93 ： 7） and the mobile phase B was acetonitrile-water（80 ： 20）. The detecting wavelength was 254 nm. Results： The concentrations of taurine showed good linearity within the range of 0.01-0.07 mg/mL（r=0. 999 9）. High precision was demonstrated by this method, The recovery rates of 0.019,0.024 and 0.029 mg/mL were （97.0±0.5）%,（98.8＋0.20） and （100. 0±1. 6）%, respectively （n=3）, Conclusion： This method is accurate and reproducible.