HPLC法测定苦杏仁苷两种差向异构体的含量
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摘要
目的分离纯化L-苦杏仁苷对照品,建立苦杏仁药材中D-、L-苦杏仁苷的含量测定方法。方法采用YMC-Pack Ph高效液相色谱柱(250 mm×4.6 mm,5μm);乙腈-水(5∶95)为流动相;流速:1.00 mL·min~(-1);柱温:30℃;检测波长:207 nm。结果本实验分离纯化的L-苦杏仁苷对照品纯度为96.3%,D-、L-苦杏仁苷分别在12.118 0～363.540 0μg·mL~(-1)和1.521 6～45.648 0μg·mL~(-1)范围内具有良好的线性关系,相关系数(r)分别为0.999 8和0.999 9,平均加样回收率分别为98.86%(RSD=0.64%)和97.33%(RSD=0.87%)。结论本实验分离纯化的L-苦杏仁苷对照品纯度高,建立的高效液相色谱法(HPLC)简单可行,结果准确可靠,对D-、L-苦杏仁苷具有良好的分离效果,可用于苦杏仁药材中苦杏仁苷的质量控制,为中成药中苦杏仁苷的质量控制提供参考。
Objective To separate and purify L-amygdalin reference substance,and to establish a HPLC methodfor the content determination of D-amygdalin and L-amygdalin. Methods The YMC-Pack Ph chromatographiccolumn(250 mm×4.6 mm,5 μm) was used. The 5% acetonitrile solution was used as the mobile phase. The flowrate was 1.00 mL · min~(-1). The column oven was set at 30 ℃, and the detection wavelength was set at 207 nm.Results The purity of the L-amygdalin reference substance was 96.3%. Under the above chromatographicconditions,good linear relationships were shown between peak area and the concentration of D-amygdalin and L-amygdalin in the ranges of 12.118 0-363.540 0 μg · mL~(-1)and 1.5216-45.6480 μg · mL~(-1), respectively. Thecorrelation coefficients were 0.999 8 and 0.999 9,respectively. The average recoveries were 98.86%(RSD=0.64%)and 97.33%(RSD=0.87%),respectively. Conclusion The purity of the L-amygdalin reference substance obtainedin this experiment was high. The method is simple, feasible, and accurate with high specificity. It has goodseparation of D-and L-amygdalin,which can be used for the quality control of amygdalin in Armeniacae semenand provide reference for the quality control of amygdalin in Chinese patent medicines.
Objective To separate and purify L-amygdalin reference substance,and to establish a HPLC methodfor the content determination of D-amygdalin and L-amygdalin. Methods The YMC-Pack Ph chromatographiccolumn(250 mm×4.6 mm,5 μm) was used. The 5% acetonitrile solution was used as the mobile phase. The flowrate was 1.00 mL · min~(-1). The column oven was set at 30 ℃, and the detection wavelength was set at 207 nm.Results The purity of the L-amygdalin reference substance was 96.3%. Under the above chromatographicconditions,good linear relationships were shown between peak area and the concentration of D-amygdalin and L-amygdalin in the ranges of 12.118 0-363.540 0 μg · mL~(-1)and 1.5216-45.6480 μg · mL~(-1), respectively. Thecorrelation coefficients were 0.999 8 and 0.999 9,respectively. The average recoveries were 98.86%(RSD=0.64%)and 97.33%(RSD=0.87%),respectively. Conclusion The purity of the L-amygdalin reference substance obtainedin this experiment was high. The method is simple, feasible, and accurate with high specificity. It has goodseparation of D-and L-amygdalin,which can be used for the quality control of amygdalin in Armeniacae semenand provide reference for the quality control of amygdalin in Chinese patent medicines.
引文
[1]国家药典委员会.中华人民共和国药典(一部)[M].北京:中国医药科技出版社,2015:201-202.
[2]KWON H J,LEE J H,HONG S P.Improvement of the extraction efficiency of D-amygdalin from armeniacae semen powder through inactivating emulsin and suppressing the epimerization of D-amygdalin[J].Archives Pharmacal Research,2010,33(1):81-86.
[3]汤庆发,谢颖,陈飞龙,等.苦杏仁中苦杏仁苷的存在形式及其影响因素[J].中国实验方剂学杂志,2013,19(8):107-109.
[4]YU L S,YE H Z,ZHENG L L,et al.Determination of the epimerization rate constant of amygdalin by microemulsion electrokinetic chromatography[J].Electrophoresis,2011,32(2):218-222.
[5]吴军,屠鹏飞,赵玉英.补阳还五汤中苦杏仁苷的存在形式及其产生机制[J].中草药,2002,33(2):101-104.
[6]BOLARINWA I F,ORFILA C,MORGAN M R.Amygdalin content of seeds,kernels and food products commercially-available in the UK[J].Food Chemistry,2014,152:133-139.
[7]BOLARINWA I F,ORFILA C,MORGAN M R.Determination of amygdalin in apple seeds,fresh apples and processed apple juices[J].Food Chemistry,2015,170:437-442.
[8]宋学英,杨华.色谱柱的选择及应用[J].分析仪器,2016,48(6):84-88.
[9]刘易,龚莹,郭磊,等.中成药中氰苷及其差向异构体的成分分析[J].药物分析杂志,2016,36(6):1011-1019.
[2]KWON H J,LEE J H,HONG S P.Improvement of the extraction efficiency of D-amygdalin from armeniacae semen powder through inactivating emulsin and suppressing the epimerization of D-amygdalin[J].Archives Pharmacal Research,2010,33(1):81-86.
[3]汤庆发,谢颖,陈飞龙,等.苦杏仁中苦杏仁苷的存在形式及其影响因素[J].中国实验方剂学杂志,2013,19(8):107-109.
[4]YU L S,YE H Z,ZHENG L L,et al.Determination of the epimerization rate constant of amygdalin by microemulsion electrokinetic chromatography[J].Electrophoresis,2011,32(2):218-222.
[5]吴军,屠鹏飞,赵玉英.补阳还五汤中苦杏仁苷的存在形式及其产生机制[J].中草药,2002,33(2):101-104.
[6]BOLARINWA I F,ORFILA C,MORGAN M R.Amygdalin content of seeds,kernels and food products commercially-available in the UK[J].Food Chemistry,2014,152:133-139.
[7]BOLARINWA I F,ORFILA C,MORGAN M R.Determination of amygdalin in apple seeds,fresh apples and processed apple juices[J].Food Chemistry,2015,170:437-442.
[8]宋学英,杨华.色谱柱的选择及应用[J].分析仪器,2016,48(6):84-88.
[9]刘易,龚莹,郭磊,等.中成药中氰苷及其差向异构体的成分分析[J].药物分析杂志,2016,36(6):1011-1019.