分别采用饱和硫酸铵法和辛酸—硫酸铵法纯化兔抗酸性红73(acid red 73,AR73)免疫球蛋白G(IgG)。通过二喹啉甲酸(BCA)法、十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)试验、间接ELISA法分别测定IgG的浓度、纯度及效价。结果显示,饱和...分别采用饱和硫酸铵法和辛酸—硫酸铵法纯化兔抗酸性红73(acid red 73,AR73)免疫球蛋白G(IgG)。通过二喹啉甲酸(BCA)法、十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)试验、间接ELISA法分别测定IgG的浓度、纯度及效价。结果显示,饱和硫酸铵法所提IgG浓度为36.44mg/mL,最高效价达1.6×105;辛酸—硫酸铵法所提IgG浓度为15.79mg/mL,最高效价达3.2×105。提示,辛酸—硫酸铵法IgG纯度优于饱和硫酸铵法。展开更多
[Objectives]To identify stained Schisandrae Chinensis Fructus in Lifei Tablets with carmine and acid red 73 as target substances.[Methods]High performance liquid chromatography(HPLC)and liquid chromatography tandem-ma...[Objectives]To identify stained Schisandrae Chinensis Fructus in Lifei Tablets with carmine and acid red 73 as target substances.[Methods]High performance liquid chromatography(HPLC)and liquid chromatography tandem-mass spectrometry(LC-MS/MS)was used to analyze the 70%methanol extract of the drug.The HPLC was equipped with an Inertstain-C_(18)(250 mm×4.6 mm,5μm)chromatographic column.The mobile phase was acetonitrile-5 mmol/L ammonium acetate solution,gradient elution:volume flow rate of 1.0 mL/min,column temperature of 30℃,detection wavelength of 509 nm.LC-MS/MS was equipped with a Waters-C_(18)(2.1 mm×150 mm,1.7μm)chromatographic column,the mobile phase was methanol-0.1%formic acid solution(containing 5 mmol/L ammonium acetate),gradient elution;the volume flow rate was 0.3 mL/min;ESI ion source,positive ion mode,full scan of primary and secondary mass spectrometry.[Results]Carmine and acid red 73 showed good linear relationship in HPLC and LC-MS/MS,with r greater than 0.995.The sample recovery and RSD values of HPLC and LC-MS/MS met the requirements.The RSD of the results determined by HPLC and LC-MS/MS was not greater than 5.0%.Both methods had good compatibility and could be used for the examination of carmine and acid red 73 in Lifei Tablets.[Conclusions]The method is reliable and reproducible and can be used to identify stained Schisandrae Chinensis Fructus in Lifei Tablets.展开更多
文摘分别采用饱和硫酸铵法和辛酸—硫酸铵法纯化兔抗酸性红73(acid red 73,AR73)免疫球蛋白G(IgG)。通过二喹啉甲酸(BCA)法、十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)试验、间接ELISA法分别测定IgG的浓度、纯度及效价。结果显示,饱和硫酸铵法所提IgG浓度为36.44mg/mL,最高效价达1.6×105;辛酸—硫酸铵法所提IgG浓度为15.79mg/mL,最高效价达3.2×105。提示,辛酸—硫酸铵法IgG纯度优于饱和硫酸铵法。
基金Supported by Chongqing Pharmaceutical Research Project of Chongqing Drug Administration and Chongqing Science and Technology Bureau(Yu Yao Jian[2021]30)。
文摘[Objectives]To identify stained Schisandrae Chinensis Fructus in Lifei Tablets with carmine and acid red 73 as target substances.[Methods]High performance liquid chromatography(HPLC)and liquid chromatography tandem-mass spectrometry(LC-MS/MS)was used to analyze the 70%methanol extract of the drug.The HPLC was equipped with an Inertstain-C_(18)(250 mm×4.6 mm,5μm)chromatographic column.The mobile phase was acetonitrile-5 mmol/L ammonium acetate solution,gradient elution:volume flow rate of 1.0 mL/min,column temperature of 30℃,detection wavelength of 509 nm.LC-MS/MS was equipped with a Waters-C_(18)(2.1 mm×150 mm,1.7μm)chromatographic column,the mobile phase was methanol-0.1%formic acid solution(containing 5 mmol/L ammonium acetate),gradient elution;the volume flow rate was 0.3 mL/min;ESI ion source,positive ion mode,full scan of primary and secondary mass spectrometry.[Results]Carmine and acid red 73 showed good linear relationship in HPLC and LC-MS/MS,with r greater than 0.995.The sample recovery and RSD values of HPLC and LC-MS/MS met the requirements.The RSD of the results determined by HPLC and LC-MS/MS was not greater than 5.0%.Both methods had good compatibility and could be used for the examination of carmine and acid red 73 in Lifei Tablets.[Conclusions]The method is reliable and reproducible and can be used to identify stained Schisandrae Chinensis Fructus in Lifei Tablets.
