中文摘要：

目的：研究丹参酚酸中丹参素（tanshinol，TSN）、原儿茶醛（protocatechualdehyde，PCA）、丹酚酸A（salvianolic acid A，SAA）、丹酚酸B（salvianolic acid B，SAB）及丹酚酸D（salvianolic acid D，SAD）的电喷雾串联质谱（ESI-MS／MS）检测特性，为开展复方丹参滴丸药代研究，建立灵敏可靠的定量分析血浆样品中丹参酚酸浓度的方法。方法：研究丹参酚酸的电离模式及效率、二级质谱的碎片离子及打碎效率；建立有效可靠的从血浆样品中提取被测丹参酚酸物质的前处理方法；优化色谱条件、减少分析干扰、促进质谱检测；围绕准确性、精密度和提取回收率等进行方法学验证。结果：电喷雾（ESI）负离子模式能有效实现丹参酚酸被测化合物的电离。以下离子对m／z197→135、137→108、493→295、717→519、417→197分别为被测化合物偈N、PCA、SAA、SAB、SAD提供最佳的选择离子对监测（SRM）检测条件。为取得可靠的分析结果，建立了两个分析方法：其一用于测定血浆样品中的TSN；其二用于测定血浆样品中的PCA、SAA、SAB和SAD。两个分析方法均以HCI酸化血样后用乙酸乙酯提取的方法处理血浆样品，其中，TSN的提取回收率在57．7％～61．6％之间；PCA、SAA、SAB和SAD的提取回收率在38．0％～58．9％之间。偈N的线性范围为2．7～2000．0ng／mL，相关系数r为0．999，最低检测限（LLOQ）为2．7ng／mL；PCA、SAA、SAB及SAD的线性范围为1．4～1000．0ng／mL或4．1～1000．0ng/mL，相关系数r〉0．99，LLOQ分别为1．4ng／mL（PCA）和4．1ng/mL（SAA、SAB和SAD）。两个分析方法的日内准确性和精密度分别为90％～112％和2．5％～14．5％；日间准确性和精密度分别为91％～110％和0．7％～9．8％。结论：成功建立了能够灵敏可靠地测定血浆样品中5种丹参酚酸化合物浓度的分析方法，此方法可用于复方丹参滴丸的药代

英文摘要：

AIM： To develop sensetive and specific LC-MS/MS-based methods for measurement of plasma tanshinol （TSN）, protocatechualdehyde （PCA）, and salvianolic acid A, B, and D （SAA, SAB, and SAD） in supporting the pharmacokinetic evaluation of Cardiotonic pills, a standardized danshen-containing cardiovascular phytomedicine. METHODS： Electrospray ionization （ESI） patterns and efficiency of the analytes, along with their fragmentation, were investigated to achieve sensitive ESI-MS/MS detection. Sample preparation was also studied to effectively recover the analytes from plasma. Meanwhile, chromatographic conditions were optimized to avoid matrix component interference on analyte ESI-detection. The newly developed analytical methods were evaluated with respect to accuracy, precison, selectivity, sensitivity, and stability. RESULTS： The negative ion ESI gave a high ionization efficiency for the analytes. The precursor-to-product ion pairs m/z 197→135, 136→108, 493→295, 717→519, and 417→197 were used for sensitive and specific SRM of TSN, PCA, SAA, SAB, and SAD, repectively. For the best measurement of the analytes, two methods were developed, i.e., one for determination of plasma TSN and the other one for simultanious determination of plasma PCA, SAA, SAB, and SAD. The sample preparation for the two quantification methoids involved acidifying the plasma sample with HCl before EtO-Ac-based liquid-liquid extraction; the extraction efficiency was 57.7% -61.6% for TSN from plasma or 38.0% -58.9% for PCA,SAA,SAB, and SAD in parallel from plasma. Matrix-matched standard curves of the peak area of a given analytes versus the nominal plasma concentration were linear for concentrations of TSN between 2.7 ng/mL and 2 000,0 ng/mL, with a correlation coefficient 0,999 and a lower limit of quantification 2.7 ng/mL, while linear dynamic ranges for PCA, SAA, SAB, and SAD were 1.37 or 4.1 - 1000.0 ng/mL, with correlation coefficients 〉 0.99 and lower limits of quantification 1.4 ng/mL for PCA and 4.1 ng/mL