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目的:建立柱前衍生化高效液相色谱法同时测定猪胆粉结合胆酸中甘氨酸和牛磺酸含量。方法:采用10%氢氧化钠溶液对猪胆粉中的结合胆酸进行碱水解,得到游离氨基酸,再用2,4-二硝基氟苯衍生化制备样品。采用Kromasil(KR100-5C18色谱柱(250mm×4.6mm,5μm);流动相:0.05mol.L-1醋酸钠缓冲盐(醋酸调pH至5.5)和甲醇(75:25);流速:1.0mL.min-1;柱温:35℃。检测波长:360nm。结果:甘氨酸与牛磺酸能达到基线分离,空白无干扰,线性良好。结论:该方法简便、快捷、重复性好,能准确测定猪胆粉结合胆酸中甘氨酸及牛磺酸的含量,为猪胆粉的质量控制提供了理论依据。
OBJECTIVE: To establish a method for the simultaneous determination of glycine and taurine in porcine bile powder and cholic acid by precolumn derivatization high performance liquid chromatography. Methods: 10% sodium hydroxide solution was used to hydrolyze bound bile acids in porcine bile powder to obtain free amino acids. Then, 2,4-dinitrofluorobenzene was derivatized to prepare samples. Kromasil (KR100-5C18 column (250 mm × 4.6 mm, 5 μm) was used; mobile phase: 0.05 mol·L-1 sodium acetate buffer (adjusted to pH 5.5 with acetic acid) and methanol (75:25); min-1, column temperature: 35 ℃, detection wavelength: 360 nm.Results: Glycine and taurine could achieve baseline separation with no interference in blank and good linearity.Conclusion: The method is simple, rapid and reproducible, Bile powder combined with bile acid glycine and taurine content for the quality control of porridge powder provides a theoretical basis.