快速分析紫雲膏指標成分之高效液相層析方法   全文下載

A Rapid, Simple High Performance Liquid Chromatography Method for the Determination of Traditional Chinese Medicine Ointment Shiunko

本研究之目的在於開發出一種快速、簡單、靈敏的HPLC方法供紫雲膏中之指標成分之定量分析用，這些指標成分包括紫草中的紫草素、丙醯紫草素、β, β-二甲基丙烯紫草素、異丁基紫草素、去氧紫草素和異戊醯紫草素以及當歸中的阿魏酸等。紫雲膏檢品以乙?抽提後，以逆相層析管柱（C18, ThermoQuest, 250 × 4.6 mm i.d.），於室溫下分析。沖溶條件如次，移動相，乙?47%和pH 2.56 醋酸水溶液53%；流速，每分鐘1.0 mL進行，檢定波長，UV 280 nm；內部標準品，diclofenac。整個分析時間為35分鐘內。分析方法確認方面，在特定濃度範圍內（紫草素0.5-10μg／mL、丙醯紫草素0.5-10μg／mL、β, β-二甲基丙烯紫草素0.35 -7μg／mL、異丁基紫草素0.4-8μg／mL、去氧紫草素0.255-5.1μg／mL、異戊醯紫草素0.25-5μg／mL以及阿魏酸0.175-3.5μg／mL），其分析之相對標準差和相對誤差均小於7%；而且每個指標成分在此設定濃度範圍下其濃度與得到之相關面積比均有良好的線性關係，顯示此分析方法可應用於定量分析。這些指標成分的偵測極限至少可達0.05 μg／mL。"

A rapid, simple and sensitive reversed-phase HPLC method with Ultraviolet (UV) absorbance measurement at 280 nm was applied for the simultaneous quantitative determination of marker substances in the traditional Chinese herbal ointment Shiunko. Shiunko consists of Lithospermum radix containing shikonin, acetylshikonin, β,β-dimethylacrylshikonin, isobutylshikonin, deoxyshikonin, and isovalerylshikonin, as well as Angelica radix containing ferulic acid. Specimens of Shiunko ointment were extracted with acetonitrile, separated on a reversed-phase C18 (ThermoQuest, 250 × 4.6 mm i.d.) column at ambient temperature, with a solvent system consisting of 47% acetonitrile and 53% acetic acid (v/v, pH 2.56) at a flow rate of 1.0 mL/min. Diclofenac was used as the internal standard. This procedure enabled a run time of within 35 min. In the validation of this analytical method, the RSD and RE were less than 7.0%, indicating satisfactory accuracy and repeatability. The linear relationships between the concentration of analyte in samples within a given range (ferulic acid 0.175- 3.50 μg/mL, shikonin 0.5-10.0 μg/mL, acetylshikonin 0.5-10 μg/mL, β,β-dimethylacrylshikonin 0.35-7 μg/mL, isobutylshikonin 0.4-0.8 μg/mL, deoxyshikonin 0.255-5.1 μg/mL, and isovalerylshikonin 0.25-5 μg/mL) and the corresponding peak area ratio were good indications that this analytical method could be used in quantitative analysis. The lower limit of detection of these marker substances under above conditions was 0.05 μg/mL. 本研究之目的在於開發出一種快速、簡單、靈敏的HPLC方法供紫雲膏中之指標成分之定量分析用，這些指標成分包括紫草中的紫草素、丙醯紫草素、β, β-二甲基丙烯紫草素、異丁基紫草素、去氧紫草素和異戊醯紫草素以及當歸中的阿魏酸等。紫雲膏檢品以乙腈抽提後，以逆相層析管柱（C18, ThermoQuest, 250 × 4.6 mm i.d.），於室溫下分析。沖溶條件如次，移動相，乙腈47%和pH 2.56 醋酸水溶液53%；流速，每分鐘1.0 mL進行，檢定波長，UV 280 nm；內部標準品，diclofenac。整個分析時間為35分鐘內。分析方法確認方面，在特定濃度範圍內（紫草素0.5-10μg／mL、丙醯紫草素0.5-10μg／mL、β, β-二甲基丙烯紫草素0.35 -7μg／mL、異丁基紫草素0.4-8μg／mL、去氧紫草素0.255-5.1μg／mL、異戊醯紫草素0.25-5μg／mL以及阿魏酸0.175-3.5μg／mL），其分析之相對標準差和相對誤差均小於7%；而且每個指標成分在此設定濃度範圍下其濃度與得到之相關面積比均有良好的線性關係，顯示此分析方法可應用於定量分析。這些指標成分的偵測極限至少可達0.05 μg／mL。