HPLC法测定L-丙氨酸异丙酯盐酸盐中L-丙氨酸的含量

Determination of L-alanine in L-alanine Isopropyl Ester Hydrochloride by HPLC

建立测定L-丙氨酸异丙酯盐酸盐中L-丙氨酸含量的反相高效液相色谱法。采用十八烷基硅烷键合硅胶为填充剂(Waters XBridge shield C18,4.6 mm×250 mm,5μm)的色谱柱,流动相为乙腈-0.02 mol/L辛烷磺酸钠(用磷酸调节pH至2.5)(V∶V=25∶75),流速0.5 mL/min,检测波长210 nm,柱温30℃。定量限为0.72μg/mL(相当于样品浓度的0.024%),检测限为0.24μg/mL。L-丙氨酸在0.72~225μg/mL浓度范围内,线性方程为Y=619.7733X-157.8515,r=1.0000>0.999。回收率为100.8%~104.9%(RSD<10.0%,n=9)。该方法操作简便、专属性强、灵敏度高,可有效检出L-丙氨酸异丙酯盐酸盐中杂质L-丙氨酸的含量。

A method for determining the L-alanine in L-alanine isopropyl ester hydrochloride by HPLC was established.The chromatography separation was performed on a Waters XBridge shield C18(4.6 mm×250 mm,5 μm).The mobile phase was acetonitrile-0.02 mol/L sodium octane sulfonate(pH 2.5 with phosphoric acid)(V∶V=25∶75),the flow rate was 0.5 mL/min,column temperature was 30 ℃,the detection wavelength was 210 nm.The quantitative limit was 0.72 μg/mL(corresponding to a percentage of sample concentration 0.024%),and the detection limit was 0.24 ug/mL.The caliblation curve for L-alanine was linear in the concentration range of 0.72 to225 μg/mL(Y=619.773 3X-157.851 5,r=1.000 00.999).The recoveries were 100.8%~104.9%(RSD10.0%,n=9).This method was simple,specific and sensitive,and could be used to effectively detect the content of L-alanine impurity in L-alanine isopropyl ester hydrochloride.