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目的建立同时测定地黄中葡萄糖、半乳糖和蜜二糖的RP-HPLC方法,为地黄药材的质量控制提供新的依据。方法地黄药材以水超声提取,再用1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化后,采用Diamonsil C18柱(150mm×4.6 mm,5μm)分离,流动相为乙腈和100 mmol/L醋酸铵水溶液(pH 5.5)进行梯度洗脱,体积流量1.0 mL/min,检测波长为245 nm,柱温为23.5℃,以内标法定量。结果葡萄糖、半乳糖和蜜二糖分别在90~900μg/mL(r=0.999 3)、90~900μg/mL(r=0.999 0)和27~270μg/mL(r=0.999 0)线性关系良好;3种糖的平均回收率均>98.7%(n=9),RSD<3.3%。结论该方法灵敏度高、分离效果好,可用于地黄药材的质量控制。
OBJECTIVE To establish a RP-HPLC method for simultaneous determination of glucose, galactose and melibi in Rehmannia glutinosa, providing a new basis for the quality control of Rehmannia glutinosa. Methods Rehmannia glutinosa extracts were extracted with water and then derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP). Diamonsil C18 column (150 mm × 4.6 mm, 5 μm) Acetonitrile and 100 mmol / L ammonium acetate aqueous solution (pH 5.5). The volume flow rate was 1.0 mL / min, the detection wavelength was 245 nm and the column temperature was 23.5 ℃. Results The linear relationship between glucose, galactose and melibiose was 90 ~ 900μg / mL (r = 0.999 3), 90 ~ 900μg / mL (r = 0.999 0) and 27 ~ 270μg / mL The average recoveries of all three kinds of sugars were> 98.7% (n = 9) with a RSD <3.3%. Conclusion The method has high sensitivity, good separation effect and can be used for the quality control of Rehmannia glutinosa.