目的 建立一測(cè)多評(píng)法測(cè)定丹酚酸A原料藥中特定雜質(zhì)迷迭香酸、紫草酸、丹酚酸B和丹酚酸C。方法 采用Agilent Zorbax Eclipse XDB-C₁₈色譜柱（250 mm×4.6 mm，5 μm），流動(dòng)相0.2%磷酸–乙腈，梯度洗脫，檢測(cè)波長(zhǎng)286 nm，體積流量1.0 mL/min，柱溫25℃，樣品室溫度4℃，進(jìn)樣量20 μL。以丹酚酸A為內(nèi)標(biāo)，建立丹酚酸A原料藥中特定雜質(zhì)（紫草酸、迷迭香酸、丹酚酸B、丹酚酸C）的相對(duì)校正因子；分別采用一測(cè)多評(píng)法和外標(biāo)法測(cè)定丹酚酸A原料藥中特定雜質(zhì)，并比較一測(cè)多評(píng)法計(jì)算值與外標(biāo)法實(shí)測(cè)值的差異性。結(jié)果 丹酚酸A、丹酚酸B、丹酚酸C、迷迭香酸、紫草酸均在0.2～20 μg/mL與峰面積呈良好線性關(guān)系。在線性濃度范圍內(nèi)，迷迭香酸、紫草酸、丹酚酸B、丹酚酸C與丹酚酸A間的相對(duì)校正因子分別為1.52、2.09、2.33、1.06。一測(cè)多評(píng)法計(jì)算值與外標(biāo)法實(shí)測(cè)值無(wú)顯著性差異。結(jié)論 一測(cè)多評(píng)法可用于丹酚酸A原料中迷迭香酸、紫草酸、丹酚酸B、丹酚酸C的測(cè)定。

Objective To establish quantitative analysis of multi-components with single marker (QAMS) for determination of specified impurities, such as rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C in salvianolic acid A crude drug. Methods HPLC method was carried out on Agilent Zorbax Eclipse XDB-C₁₈ column(250 mm×4.6 mm, 5 μm). The mobile phase consisted of 0.2% phosphoric acid and acetonitrile solution with gradient elution. The detection wavelengths were 286 nm. The flow rate was 1.0 mL/min with injection volume of 20 μL. The column temperature and sample room temperature was set at 25℃ and 4℃, respectively. Salvianolic acid A was used as the internal reference substance to establish determination of specified impurities (rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C) in salvianolic acid A crude drug. The relative correlation factors (RCFs) of rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C were calculated and established. The contents of specified related substance were determined by the external standard method (ESM) and QAMS method, respectively. The QAMS method was validated through comparison of the results obtained by the two different methods. Results The linear ranges of rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C were all 0.2-20 μg/mL. The values of RCFs of rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C were 1.52, 2.09, 2.33, and 1.06, respectively. The two methods had no significant difference in content results. Conclusion QAMS method can be used for the determination of specified impurities (lithospermic acid, rosmarinic acid, salvianolic acid B and salvianolic acid C) in salvianolic acid A crude drug.

北京市國(guó)家重大研發(fā)計(jì)劃匹配項(xiàng)目（Z161100002616024）