酶解反应-柱前衍生-高效液相色谱法测定高淀粉类食品中的牛磺酸
HPLC Determinination of Taurine in High-Starch Food by Pre-column Derivatization after Enzymolysis
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摘要: 5.000g高淀粉类食品中加入不少于1 mg的高温α-淀粉酶(37.2 U·mg-1)于60 ℃酶解反应20 min,所得酶解产物经丹磺酰氯在50 ℃下进行衍生化,所得牛磺酸的衍生物用高效液相色谱法分析。选用Diamonsil C18色谱柱(250 mm×4.6 mm,5 μm)为固定相,以pH 7.2乙酸盐缓冲溶液与乙腈按体积比75比25混合所得溶液为流动相进行淋洗,于波长247 nm处进行检测。牛磺酸的色谱峰面积与其质量浓度在90 mg·L-1以内呈线性关系,检出限(3S/N)为0.3 mg·kg-1。在4个浓度水平下做加标回收试验,酶解反应后衍生所得回收率在95.3%~101%之间,测定值的相对标准偏差(n=6)在1.1%~1.5%之间。Abstract: Sample of high starch food (5.0 g) was enzymolized with not less than 1 mg of high-temperature-amylase (37.2 U·mg-1) at 60 ℃ for 20 min, and the product obtained was derivatized with Dansyl-Cl at 50 ℃. The derivate of taurine was used for HPLC analysis, using Diamonsil C18 chromatographic column (250 mm×4.6 mm, 5 μm) as stationary phase and mixture of acetate buffer solution of pH 7.2 and acetonitrile (75+25) as mobile phase. Linear relationship was found between values of peak area determined at wavelength of 247 nm and mass concentration of taurine in the range within 90 mg·L-1, with values of detection limit (3S/N) of 0.3 mg·kg-1. Tests for recovery were performed by standard addition method at 4 concentration levels, giving values of recovery in the range of 95.3%-101%. Values of RSD (n=6) found were in the range of 1.1%-1.5%.