本研開發用高效液相層析法，分析芫花藥材中之六種類黃酮成分genkwanin 5-O-β-D-primeveroside (1),genkwanin 5-O-β-D-glucoside (2), genkwanin (3), potassium apigenin 7-O-βD-glucuronate (4), apigenin (5) 及tiliroside (6)，並進一步以此高效液相層析法調查市售芫花藥材中此六種成分之含量。本實驗之高效液相層析法係採用C18逆向層析管柱，移動相為乙-1.0% 醋酸溶液，利用線性梯沖提，檢測波長為254 nm，得到良好分析結果。其線性迴歸方程式及相關係數(r)分別為： 1 ：Y = 39.17X – 0.21 (r= 0.9999)；2： Y = 27.78X – 0.03 (r = 0.9994)；3 ： Y=23.59X + 0.66 (r = 0.9999)；4 ：Y = 38.18X + 0.65 (r =0.9999)；5： Y = 23.39X + 0.43 (r = 0.9999) ； 6 ： Y=34.74X + 0.15 (r =0.9999)，均呈良好線性關係。此六種類黃酮成分之添加回收率試驗結果為100.9%~106.1% ；同日內及異日間相對標準偏差試驗，同日內為0.49~3.46 %，異日間為0.52~4.61 % ，顯示此高效液相層析法用於芫花藥材之分析其效果好。市售芫花藥材16 種，利用上述高效液相層法，分析此六種類黃酮成分的含量，結果1~6 的含分別如下： 1 ： 0.13~0.40% ； 2 ： 0.02~0.12% ； 3 ： 0.24~0.56% ； 4 ： 0.16~1.19% ； 5 ： 0.30~0.73% ； 6 ：0.15~0.57% 。

In order to evaluate the quality of Daphnis Genkwae Flos, a high performance liquid chromatographic method (HPLC) was developed.
Six flavonoid constituents from Daphnis Genkwae Flos, genkwanin 5-O-β-D-primeveroside (1), genkwanin 5-O-β-D-glucoside (2),
genkwanin (3), potassium apigenin 7-O-β-D-glucuronate (4), apigenin (5) and tiliroside (6) determined by this simple, rapid and accurate
method.
Samples were analyzed using a Cosmosil 5C18-AR reversed phase column by gradient elution with varied proportion of 1.0 % (v/v)
acetic acid and acetonitrile as mobile phase at 254 nm. Sulfamethoxypyridazine was used as the internal standard.
Regression equations revealed the linear relationship (correlation coefficients: 0.9994~0.9999) between the peak-area ratios flavonoid
to internal standard and concentrations of flavonoids. The recoveries of six flavonoids were between 100.9~106.1%. The relative standard
deviations of six flavonoids ranged between 0.49~3.46% (intraday) and 0.52~4.61% (interday). The contents of the six flavonoids in sixteen
crude drugs of Daphnis Genkwae Flos were 1 0.13~0.40%, 2 0.02~0.12%, 3 0.24~0.56%, 4 0.16~1.19%, 5 0.30~0.73% and 6
0.15~0.57%, respectively.