柱前衍生化反相高效液相色谱法测定蝉蜕中3种氨基酸含量
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Determination of Three Amino Acids in Cicada Slough by Pre- column Derivatization Reversed- phase High- performance Liquid Chromatography
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    摘要:

    目的 建立柱前衍生化反相高效液相色谱法测定蝉蜕中天冬氨酸、酪氨酸和缬氨酸3种氨基酸含量的方法。方法 采用酸水解法制备氨基酸样品溶液,以异硫氰酸苯酯柱前衍生化后进行高效液相色谱分析。采用Kromasil C18色谱柱(250 mm×4.6 mm),梯度洗脱,流动相A为0.1 mol/L醋酸钠缓冲液- 乙腈(93∶7),流动相B为乙腈- 水(4∶1),流速1.0 mL/min,柱温为43 ℃,检测波长254 nm,进样量20 μL。结果 氨基酸衍生溶液在36 h内保持稳定,3种氨基酸在线性范围内峰面积与浓度的相关系数均大于0.999 0,加样回收率(n=6)分别为95.62%、101.26%和100.00%。结论 所建立的检测方法稳定可靠,精密度和重复性良好,可用于蝉蜕中天冬氨酸、酪氨酸、缬氨酸含量测定。

    Abstract:

    Objective To establish the method for determining the content of aspartic acid, tyrosine, and valine in cicada slough by pre- column derivatization reversed- phase high- performance liquid chromatography (HPLC). Methods Acid hydrolyzation was performed to prepare amino acid sample solution, and HPLC was performed after pre- column derivatization with phenyl isothiocyanate. HPLC was performed on a Kromasil C18 column (250 mm × 4.6 mm) with mobile phase A of 0.1 mol/L sodium acetate buffer solution- acetonitrile (93∶7) and mobile phase B of acetonitrile- water (4∶1) for gradient elution at a flow rate of 1.0 mL/min, a column temperature of 43 ℃, a wavelength of 254 nm, and a sample size of 20 μL. Results The amino acid derivative solution remained stable within 36 hours. The correlation coefficient between peak area and concentration for these three amino acids within the linear range was higher than 0.999 0, and the recovery rates (n=6) were 95.62%, 101.26%, and 100.00%, respectively. Conclusion The method established is stable and reliable and has good accuracy and reproducibility. It can be used for the determination of aspartic acid, tyrosine, and valine in cicada slough.

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张 楠,刘劲松,王国凯,郑 娟,刘松照.柱前衍生化反相高效液相色谱法测定蝉蜕中3种氨基酸含量[J].安徽中医药大学学报,2017,36(2):80-82

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  • 在线发布日期: 2017-04-07