高效液相色谱法同时测定青娥丸中5种黄酮类成分的含量

Simultaneous Content Determination of Five Flavonoids Chemical Components in Qing'e Pills by HPLC

目的:建立同时测定青娥丸中陆地锦苷、紫云英苷、异鼠李素、山奈酚及槲皮素含量的方法。方法:采用高效液相色谱法,色谱柱为Kinetex-C_(18)柱(4.6 mm×250 mm,5μm),以甲醇-1%磷酸为流动相,线性梯度洗脱,流速为1.0 ml/min,检测波长为360 nm,柱温为30℃。结果:陆地锦苷、紫云英苷、异鼠李素、山奈酚及槲皮素检测的线性范围分别为1.312~131.200μg/ml(r=0.999 9)、1.566~155.600μg/ml(r=0.999 7)、1.097~109.700μg/ml(r=0.999 9)、0.607~60.700μg/ml(r=0.999 9)及0.763~76.300μg/ml(r=0.999 6);定量限分别为0.13、0.15、0.11、0.30及0.35μg/ml;检测限分别为0.05、0.05、0.04、0.10及0.15μg/ml;平均加样回收率分别为94.1%(RSD=2.1%,n=6)、96.1%(RSD=2.1%,n=6)、96.5%(RSD=2.5%,n=6)、95.1%(RSD=2.3%,n=6)及96.3%(RSD=2.3%,n=6)。结论:该方法简便、准确,适用于青娥丸中陆地锦苷、紫云英苷、异鼠李素、山奈酚及槲皮素含量的测定。

OBJECTIVE： To establish the method for glucosidonin,astragalin,isorhamnetin,kaempferol and quercetin in Qing＇e pills. METHODS： HPLC method was adopted. The separation was performed on Kinetex-C18（ 4. 6 mm × 250 mm,5 μm） column with mobile phase consisting of methanol-1% phosphoric acetic solution,at the flow rate of 1. 0 ml/min. The detection wavelength was set at 360 nm. The column temperature was 30 ℃. RESULTS：The linear ranges of glucosidonin,astragalin,isorhamnetin,kaempferol and quercetin were respectively 1. 312-131. 200 μg/ml（ r = 0. 999 9）,1. 566-155. 600 μg/ml（ r = 0. 999 7）,1. 097-109. 700 μg/ml（ r = 0. 999 9）,0. 607-60. 700 μg/ml（ r = 0. 999 9） and 0. 763-76. 300 μg/ml（ r = 0. 999 6）; the limit of quantitation were respectively0. 13 μg/ml,0. 15 μg/ml,0. 11 μg/ml,0. 30 μg/ml and 0. 35 μg/ml; and the detection limits were respectively0. 05 μg/ml,0. 05 μg/ml,0. 04 μg/ml,0. 10 μg/ml and 0. 15 μg/ml; the average recoveries were respectively94. 1%（ RSD = 2. 1%,n = 6）,96. 1%（ RSD = 2. 1%,n = 6）,96. 5%（ RSD = 2. 5%,n = 6）,95. 1%（ RSD = 2. 3%,n = 6） and 96. 3%（ RSD = 2. 3%,n = 6）. CONCLUSIONS： The method is simple and accurate,and can be used for content determination of simultaneous determination of glucosidonin, astragalin, isorhamnetin, kaempferol and quercetin in Qing＇e pills.