超高效合相色谱法拆分及测定板蓝根中(R,S)-告依春对映异构体含量

Separation and Quantification of(R,S)-Goitrin Enantiomers in Isatidis Radix by Ultra-Performance Convergence Chromatography

目的建立拆分板蓝根中(R,S)-告依春对映异构体及测定其含量的超高效合相色谱(UPC^(2))法。方法色谱柱为Trefoil^(TM) AMY1柱(150 mm×3.0 mm,2.5μm),流动相为超临界CO_(2)-甲醇(梯度洗脱),流速为1.2 mL/min,检测波长为245 nm,柱温为45℃,动态背压为1800 psi,进样量为2μL。结果R-告依春、S-告依春能达到较好分离,质量浓度分别在2.01~64.32μg/mL(r=0.9999,n=6)和1.01~32.38μg/mL(r=1.0000,n=6)范围内与峰面积线性关系良好;检测限均为0.09μg/mL,定量限分别为0.46μg/mL和0.44μg/mL;精密度、稳定性、重复性试验结果的RSD均小于3.0%;平均加样回收率分别为100.97%和101.26%,RSD分别为1.73%和2.08%(n=6)。结论该方法稳定性、重复性均较好,结果准确可靠,可用于拆分板蓝根中(R,S)-告依春对映异构体及测定其含量。

Objective To establish an ultra-performance convergence chromatography(UPC^(2))method for the separation and quantification of(R,S)-goitrin enantiomers in Isatidis Radix.Methods The chromatographic column was Trefoil^(TM) AMY1 column(150 mm×3.0 mm,2.5μm),the mobile phase was supercritical CO_(2)-methanol(gradient elution),the flow rate was 1.2 mL/min,the detection wavelength was 245 nm,the column temperature was 45℃,the dynamic back pressure was 1800 psi,and the injection volume was 2μL.Results R-goitrin and S-goitrin could be separated well,and the linear ranges of R-goitrin and Sgoitrin were 2.01-64.32μg/mL(r=0.9999,n=6)and 1.01-32.38μg/mL(r=1.0000,n=6),respectively.The limits of detection(LOD)of R-goitrin and S-goitrin were 0.09μg/mL,and the limits of quantitation(LOQ)of R-goitrin and Sgoitrin were 0.46μg/mL and 0.44μg/mL,respectively.The RSDs of precision,stability and repeatability tests were less than3.0%.The average recoveries of R-goitrin and S-goitrin were 100.97%and 101.26%with RSDs of 1.73%and 2.08%(n=6),respectively.Conclusion The method is stable,reproducible,accurate and reliable,which can be used for the separation and quantification of(R,S)-goitrin enantiomers in Isatidis Radix.