毛脉酸模不同生长发育期HPLC色谱指纹图谱研究

采用HPLC-DAD方法,梯度洗脱,对10批不同生长发育期毛脉酸模根样品进行主成分分析。色谱条件为:Planetsil C18分析柱(5μm, 200 mm×4.6 mm),预柱Phenomenex ODS-C18(4×3.0 mm ID),柱温40℃,流动相A为甲醇;流动相B为水(磷酸调pH值为2.0),流动相A从30%甲醇到100%甲醇,时间为0~50 min,检测波长254 nm。10批毛脉酸模样品得到的色谱指纹图谱有27个共有峰,其特征峰指纹图谱可分为Ⅰ, Ⅱ, Ⅲ三组:第Ⅰ组包括0~17 min(1~14号峰),第Ⅱ组包括17~35 min(15~24号峰),第Ⅲ组包括35~50 min(25~27号峰)。HPLC-DAD方法分析毛脉酸模主成分,方法准确可靠,其本身具有多成分同时定性的优势;27个共有峰的出峰先后顺序及相对含量极具特征性、专属性,重复性好,形成了毛脉酸模特有的HPLC色谱指纹图谱,可作为毛脉酸模内在质量评价、鉴定及其最佳采收期确定提供科学依据。

Study on HPLC fingerprining of Rumex gmelini at different development stages

HPLC method was applied for analysis of main constituents of 10 lots different growing period samples. The HPLC column, mobile phase elution mode (isocratic or gradient) and gradient program were optimized in order to obtain high quality HPLC profile. The HPLC system consisted of Waters 600 pump and a 996 photodiode-array detector (DAD). HPLC analysis was performed on a Planetsil C18 column (5 μm, 200 mm×4. 6 mm) and Phenomenex HPLC Guard Cartridge System ODS-C18(4×3. 0 mm ID) with the mixture of methanol (A) and H2O (acidified to pH 2. 0 with phosphoric acid) (B) as mobile phase in gradient mode. Concentrations of A 30%, 100%during 0, 50min, column temperature 40℃, detection wavelength 254 nm. The HPLC chromatographic fingerprinting of main constituents showing 27 common peaks was established from 10 lots of samples. These peaks were divided into 3 groups: from 0 to 17 min, 14 peaks (including peak 1~14);from 17 to 35min, 10 peaks (including peak 15~24);from 35 to 50 min, 3 peaks (including peak 25~27). This method with high specificity and good reproducibility can provide scientific basis for determining the best collecting time, quality evaluation and identification of Rumex gmelini.