苍耳子水提取物UPLC和HPLC特征图谱的比较研究

Comparison on characteristic chromatograms of water extract of Fructus Xanthii with UPLC and HPLC

目的通过建立并比较苍耳子水提物的超高效液相色谱(UPLC)和常规高效液相色谱(HPLC)的特征化学图谱,筛选苍耳子药材品质控制的优化方案。方法分别建立苍耳子药材水提物的UPLC和HPLC特征化学图谱分析方法,比较2种方法的分离能力、分离效率以及检测灵敏度等。UPLC检测色谱条件如下:色谱柱为Agilent SB-C18柱(50 mm×2.1 mm,1.8μm);柱温为35℃;流动相A为甲醇,B为0.1%(体积分数,下同)H_3PO_4(p H=2.3),梯度洗脱;流速为0.4 m L/min;检测波长为220 nm。HPLC检测采用Waters Symmetry C_(18)柱(250 mm×4.6 mm,5μm);柱温为35℃;流动相A为甲醇,B为0.1%(体积分数)H_3PO_4(p H=2.3),梯度洗脱;流速为1.0 m L/min;检测波长为220 nm。结果分别建立了27批不同产地苍耳子药材的UPLC和HPLC特征图谱,共检测到16个共有色谱峰,指认了其中10个色谱峰的化学归属。结果表明,UPLC法可大大缩短苍耳子药材水提取物的分析时间,且检测灵敏度也有显著提高,说明UPLC法在分离能力和分离效率方面明显优于HPLC法。结论与HPLC法相比,UPLC法既达到了分析的高通量,得到更高的分析灵敏度;又大大提高了分析速度,减少了有机溶剂的消耗。UPLC法测定中药材苍耳子水提取物的特征图谱可为其质量控制提供更为高效快速的检测方法。

Objective To establish a more feasible quality control method for Fructus Xanthii water extract by comparing the characteristic chromatograms obtained with ultra performance liquid chromatography （UPLC） and conventional HPLC. Methods UPLC and HPLC characteristic chromatogram of water extract of Fructus Xanthii were established respectively, and then their chromatographic behaviors were compared such as their peak capacity, resolution and sensitivity, and so on. UPLC separation was carried out on an waters symmetry C18 column （50 mm×2.1 mm,1.8μm） with the mobile phase consisting of methanol and water containing 0.1% phosphoric acid （ pH - 2.3） at a flow rate of 0.4 mL/min, and the detective wavelength was at 220 nm and the column temperature was at 35 ℃. HPLC separation was performed on a waters symmetry C18 column （250 min×4.6 mm, 5 μm）.The mobile phase was consisted of methanol and water containing 0.1% phosphoric acid （pH = 2.3） with a flow rate of 1.0 mL/min, and the detective wavelength was set at 220 nm and column temperature was at 35 ℃. Results The established characteristicchromatogram of Fructus Xanthii had 16 characteristic common peaks, of which 10 constituents were identified by comparison with the chromatographic behaviors and UV spectra of reference substances. Compared with HPLC fingerprint, the UPLC fingerprint showed obvious advantages in peak shape, sensitivity, and separation time. Conclusion In consideration of the mentioned advantages along with higher analytical sensitivity, less injection volume and solvent consumption, UPLC characteristic chromatogram might be proposed for the quality evaluation method of Fructus Xanthii.