柱前衍生-HPLC法测定天龙泉糯米调味酒中18种氨基酸

Determination of 18 amino acids in Tianlongquan seasoning glutinous rice wine by HPLC with pre-column derivatization

为满足企业对糯米调味酒精细化生产的品控需求,该研究利用反相色谱柱建立了测定天龙泉糯米调味酒中18种氨基酸的柱前衍生-高效液相色谱(HPLC)方法。通过优化洗脱梯度,使待测糯米调味酒的18种氨基酸色谱峰实现有效分离,确定最优色谱条件,进而建立了定量方法并对其进行方法学考察。结果表明,优化后的色谱条件为以异硫氰酸苯酯作为衍生试剂,乙腈∶水(4∶1,V/V)为流动相A,0.019 mol/L乙酸钠溶液(pH=6.2)为流动相B,梯度洗脱,柱温28℃,流速1 mL/min,检测波长245 nm。在各氨基酸标准曲线的线性范围内相关系数(R^(2))均≥0.998 7,平均加标回收率为92.38%~104.86%,相对标准偏差(RSD)≤4.45%,重复性、精密度试验结果RSD均<5%。该方法操作简单,准确高效,可定量测定天龙泉糯米调味酒及相关产品中各氨基酸的含量。

In order to meet the qualty control requrements of accurately production of seasoning glutinous rice wme.a method for quantitative deter mimation of18 amino acids in Tianlongquan seasoning glutinous rice wime by high performance liquid chromatography(HPL)with pre-column deriva tization was established basing on reverse phase column.By optimizing the elution gradient,the targeted chromatographic peaks of 18 amino acids ir the wime were efiectively separated,the optimal chromatographic conditions were detemmined,and then the quantitation method was developed fol lowed by corresponding methodology investigation.The results showed that the optimized chromatographic conditions were as folows.phenyl isoth iocyamate as derivative reagent,acetonitrile:water(4:1.V/V)as mobile phase A.0.019 moll sodium acetate solution(pH=6.2)as mobile phase B.gra dient elution,column temperature 28℃,fow rate 1 mlmin,detection wavelength 245 nm,in the linear range ofthe amino acid standard curves.the correlation coefficient(R^(2))>0.9987,the average standard recovery was 92.389%-104.86%6,the relative standard deviation(RSD)was less than 4.45%and the RD of repeatability and precision tests was less than 596.The method was simple.accurate and eficient.and could be used to quantitativel letermine the contents of amno acids in Tianlongauan seasoning glutmous rice wine and related products.