HPLC法同时测定大黄药材中8个非蒽醌类成分的含量

Simultaneous Determination of 8 Non-anthraquinone Constituents in Rheum palmatum by HPLC

目的:建立同时测定大黄药材中8个非蒽醌类成分含量的方法。方法:采用高效液相色谱法。色谱柱为Symmetry C18,流动相为甲醇-0.1%磷酸水溶液(梯度洗脱),流速为1.0mL/min,柱温为30℃,检测波长为280nm,进样量为30μL。结果:没食子酸、儿茶素、表儿茶素、白藜芦醇4′-O-葡萄糖苷、表儿茶素没食子酸酯、白藜芦醇4′-O-β-D(- 6″-O-没食子酰)-葡萄糖苷、番泻苷A、4′-羟基苯基-2-丁酮-4′-O-β-D(- 2″-O-没食子酰-6″-O-对羟基桂皮酰)-葡萄糖苷检测进样量的线性范围分别为6.16~2 464 ng(r=0.999 9)、37.4~14 960 ng(r=0.999 9)、7.635~3 054 ng(r=0.999 7)、7.63~3 052 ng(r=0.999 9)、8.32~3 328 ng(r=0.999 9)、11.5~4 600 ng(r=0.999 9)、16.08~6 432 ng(r=0.999 9)、29.3~11 720 ng(r=0.999 9);定量限分别为3.48、4.30、6.40、4.40、3.39、2.87、8.40、4.95ng,检测限分别为2.32、2.58、2.40、2.64、2.26、1.23、4.20、2.97ng;精密度、稳定性、重复性试验的RSD均小于5%;加样回收率分别为94.32%~100.54%(RSD=2.78%,n=6)、91.15%~99.36%(RSD=3.72%,n=6)、92.16%~98.04%(RSD=2.39%,n=6)、93.41%~100.73%(RSD=3.17%,n=6)、93.89%~98.40%(RSD=1.99%,n=6)、92.61%~101.74%(RSD=3.71%,n=6)、92.66%~103.40%(RSD=3.76%,n=6)、95.45%~102.70%(RSD=3.06%,n=6)。结论:所建方法简便、准确、专属性强,可用于同时测定大黄药材中8个非蒽醌类成分的含量。

OBJECTIVE:To establish a method for simultaneous determination of 8 non-anthraquinone constituents in Rheum palmatum. METHODS:HPLC method was adopted. The determination was performed on Symmetry C18 column with mobile phase consisted of methanol-0.1% phosphoric acid(gradient elution)at the flow rate of 1.0 mL/min. The column temperature was 30 ℃ and detection wavelength was 280 nm. Sample size was 30 μL. RESULTS:The linear range of gallic acid,catechin,epicatechin, resveratrol 4′-O-glucopyranoside, epicatechin gallate, resveratrol 4′-O-β-D-(6″-O-galloyl)-glucopyranoside, sennoside A, 4′-hydroxyphenyl-2-butanone-4′-O-β-D-(2″-O-galloyl-6″-O-p-hydroxy cinnamyl)-glucopyranoside were 6.16-2 464 ng(r=0.999 9), 37.4-14 960 ng(r=0.999 9),7.635-3 054 ng(r=0.999 7),7.63-3 052 ng(r=0.999 9),8.32-3 328 ng(r=0.999 9),11.5-4 600 ng(r=0.999 9),16.08-6 432 ng(r=0.999 9),29.3-11 720 ng(r=0.999 9),respectively. The limits of quantitation were 3.48, 4.30,6.40,4.40,3.39,2.87,8.40 and 4.95 ng,respectively. The limits of detection were 2.32,2.58,2.40,2.64,2.26,1.23, 4.20,2.97 ng,respectively. RSDs of precision,stability and reproducibility tests were all lower than 5%. Recoveries were 94.32%-100.54%(RSD=2.78%,n=6),91.15%-99.36%(RSD=3.72%,n=6),92.16%-98.04%(RSD=2.39%,n=6),93.41%-100.73%(RSD=3.17%,n=6),93.89%-98.40%(RSD=1.99%,n=6),92.61%-101.74%(RSD=3.71%,n=6),92.66%-103.40%(RSD= 3.76%,n=6),95.45%-102.70%(RSD=3.06%,n=6),respectively. CONCLUSIONS:The established method is simple, accurate and specific,and can be used for the simultaneous determination of 8 non-anthraquinone constituents in R. palmatum.