高效液相色谱法测定肉松中的淀粉含量

Determination of Starch Content in Meat Floss by High Performance Liquid Chromatography

建立超声辅助双酶水解法联合高效液相色谱测定肉松制品中淀粉含量的检测方法。采用超声辅助双酶水解法水解肉松中的淀粉后,利用高效液相色谱法对水解液中的葡萄糖含量进行测定,并换算为淀粉含量,采用C18色谱柱(4.6 mm×250 mm,5 μm),以乙腈-20 mmol/L乙酸铵溶液(22∶78,V/V)为流动相,以245 nm为分析波长,葡萄糖在5~50 mg/L质量浓度范围内线性良好,相关系数r大于0.999。本实验采用耐高温α-淀粉酶与淀粉葡萄糖苷酶酶解淀粉,该方法仪器检出限为0.4 μg/m L,方法检出限为0.16%。通过肉松水解液加标验证,平均回收率为88.0%~96.6%,相对标准偏差小于5%(n=6)。与国家标准方法相比,该方法重复性好、回收率高,适用于各种肉松中淀粉含量的测定。

A high performance liquid chromatographic(HPLC)with ultrasonic-assisted two-step enzymatic pretreatment for the determination of the starch content in meat products was developed.After pretreatment,the sample was separated on a C18 column(4.6 mm×250 mm,5μm),using acetonitrile-20 mmol/L ammonium acetate(22:78,V/V)as mobile phase at a flow rate of 1.0 mL/min,and detected by an ultraviolet detector at a wavelength of 245 nm.The calibration curve was linear in the range of 5–50 mg/L with a correlation coefficient(r)of 0.999.The average recoveries of spiked hydrolysate were between 88.0%and 96.6%with relative standard deviations(RSDs)of less than 5%(n=6).The limit of detection(LOD)was 0.4μg/mL,and the limit of quantitation(LOQ)of the method was 0.16%by usingα-amylase and amyloglucosidase.With the advantages of high repeatability and recovery over the national standard methods,this method is suitable for the determination of starch in meat products.