HPLC法同时测定黄芪药材中10种黄酮类成分的含量

Simultaneous Determination of 10 Flavonoids in Astragalus membranaceus by HPLC

目的:建立同时测定黄芪药材中10种黄酮类成分含量的方法。方法:采用高效液相色谱法。色谱柱为Agilent SB-C_(18),流动相为乙腈-0.3%甲酸溶液(梯度洗脱),流速为1.0 mL/min,检测波长为254 nm,柱温为35℃,进样量为10μL。结果:毛蕊异黄酮-7-O-葡萄糖苷、异槲皮苷、染料木苷、刺芒柄花苷、毛蕊异黄酮、槲皮素、染料木素、山柰酚、异鼠李素和芒炳花素检测进样量线性范围分别为0.030 29~1.514 5μg(r=0.999 4)、0.015 00~0.7 500μg(r=0.999 5)、0.007 39~0.369 5μg(r=0.999 1)、0.12011~6.005 5μg(r=0.999 8)、0.038 36~1.918μg(r=0.999 9)、0.029 89~1.494 5μg(r=0.999 5)、0.007 04~0.352μg(r=0.9994)、0.016 83~0.841 5μg(r=0.999 5)、0.004 54~0.227μg(r=0.999 9)、0.013 36~0.668μg(r=0.999 9);精密度、稳定性、重复性试验的RSD<2.0%;加样回收率分别为99.55%~100.45%(RSD=0.36%,n=6)、99.34%~101.00%(RSD=0.59%,n=6)、98.05%~100.36%(RSD=1.27%,n=6)、99.73%~100.13%(RSD=0.18%,n=6)、99.70%~100.30%(RSD=0.22%,n=6)、99.67%~103.27%(RSD=1.37%,n=6)、98.13%~104.41%(RSD=2.37%,n=6)、96.35%~100.06%(RSD=1.46%,n=6)、99.47%~101.13%(RSD=0.60%,n=6)、99.70%~100.06%(RSD=0.15%,n=6)。结论:该方法操作简便,精密度、稳定性、重复性好,可用于黄芪药材中10种黄酮类成分含量的同时测定。

OBJECTIVE： To establish a method for simultaneous determination of 10 flavonoids in Astragalus membranaceus. METHODS： HPLC method was adopted. The determination was performed on Agilent SB-C18 column with mobile phase consisted of acetonitrile-0.3% formic acid （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm, and the column temperature was 35 ℃. The sample size was 10 μL. RESULTS： The linear ranges of calycosin-7-O-glucoside, isoquercitrin, genistin, ononin, calycosin, quercetin, genistein, kaempferol, isorhamnetin and formononetion were 0.030 29-1.514 5 μg（r=0.999 4）,0.015 00-0.7 500 μg（r=0.999 5）,0.007 39-0.369 5 μg（r=0.999 1）,0.120 11-6.005 5 μg（r=0.999 8）,0.038 36-1.918 μg（r=0.999 9）,0.029 89-1.494 5 μg（r=0.999 5）,0.007 04-0.352 μg（r=0.999 4）,0.016 83-0.841 5 μg（r=0.999 5）,0.004 54-0.227 μg（r=0.999 9）, 0.013 36-0.668 μg（r=0.999 9）, respectively. RSDs of precision, stability and reproducibility tests were all lower than 2.0%. The recoveries were 99.55%-100.45%（RSD=0.36%,n=6）,99.34%-101.00%（RSD=0.59%,n=6）, 98.05%-100.36%（RSD=1.27%,n=6）, 99.73%-100.13%（RSD=0.18%,n=6）,99.70%-100.30%（RSD=0.22%,n=6）,99.67%-103.27%（RSD=1.37%,n=6）, 98.13%-104.41%（RSD=2.37%,n=6）,96.35%-100.06%（RSD=1.46%,n=6）, 99.47%-101.13%（RSD=0.60%,n=6）, 99.70%-100.06%（RSD=0.15%,n=6）, respectively. CONCLUSIONS： This method is convenient, sensitive, stable and reproducible, can be used for simultaneous determination of 10 flavonoids in A. membranaceus.