中国中药杂志

2014, v.39(08) 1463-1467

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柱前衍生化RP-HPLC测定鸡内金中16种氨基酸的含量
Determination of amino acids in Galli Gigerii Endothelium Corneum by HPLC with pre-column derivatization

梁琨;张丹;史辑;胡泓;修彦凤;贾天柱;
LIANG Kun;ZHANG Dan;SHI Ji;HU Hong;XIU Yan-feng;JIA Tian-zhu;Shanghai University of Traditional Chinese Medicine;Liaoning University of Traditional Chinese Medicine;

摘要(Abstract):

采用酸水解法制备水解氨基酸样品溶液,以异硫氰酸苯酯(PITC)柱前衍生化后进行HPLC分析。采用Ultimate Prime C18色谱柱(4.6 mm×250 mm,5μm);流动相A 0.1 mol·L-1醋酸钠溶液(调pH至6.5)-乙腈(93∶7),流动相B乙腈-水(8∶2);流速1.0 mL·min-1;柱温40℃;检测波长254 nm;梯度洗脱。氨基酸衍生溶液在36 h内保持稳定,16种氨基酸线性相关系数均大于0.999 5,加样回收率在98.01%~101.8%。该方法操作简便,精密度和重复性良好,可用于鸡内金中16种水解氨基酸含量的测定。
Hydrolytic amino acids were extracted by acid hydrolysis method,then derivatized with phenyl isothiocyanate( PITC). And the samples were analysed by HPLC on an Ultimate Prime C18( 4. 6 mm ×250 mm,5 μm) column with gradient elution of 0. 1 mol·L- 1sodium acetate buffer solution( adjusted to pH 6. 5)-acetonitrile( 93∶ 7)( A) and acetonitrile-water( 8∶ 2)( B) at a flow rate of 1. 0 mL·min- 1. Column temperature was 40 ℃ and the detected wavelength was 254 nm. Amino acids derivative solution remained stable in 36 hours. The response was linear for 16 amino acids with a correlation coefficient r > 0. 999 5. The average recoveries were 98. 01%-101. 8%. The method is reliable with good accuracy and repeatability,which is useful for the determination of amino acids in Galli Gigerii Endothelium Corneum.

关键词(KeyWords): 鸡内金;氨基酸;衍生化;异硫氰酸苯酯;高效液相色谱
Galli Gigerii Endothelium Corneum;amino acids;derivatization;phenyl isothiocyanate(PITC);HPLC

Abstract:

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基金项目(Foundation): 国家药品标准科研项目;; 上海市自然科学基金项目(13ZR1442000)

作者(Author): 梁琨;张丹;史辑;胡泓;修彦凤;贾天柱;
LIANG Kun;ZHANG Dan;SHI Ji;HU Hong;XIU Yan-feng;JIA Tian-zhu;Shanghai University of Traditional Chinese Medicine;Liaoning University of Traditional Chinese Medicine;

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