利用 LC/MS/MS 分析細辛藥材暨其製劑中馬兜鈴酸含量   全文下載

Analysis of Aristolochic Acid in Asarum (Xixin) and Its Preparations by Liquid Chromatography/Tandem Mass Spectrometry

本研究建立細辛藥材及其製劑中馬兜鈴酸之 LC/MS/MS 分析方法。利用 LC/MS/MS 作正離子電灑法（ESI+）分析，以 piromidic acid 為內部標準品，分離管柱為 C18（2.1×150 mm），移動相溶媒為 0.1% 甲酸（含 0.1% 醋酸氨）：乙?（65:35, v/v），流速為 0.3 mL/min，選定親代離子與子代離子（馬兜鈴酸 I 之配對離子為 m/z 359 及 m/z 298，馬兜鈴酸 II 之配對離子為m/z 329 及 m/z 268）進行多重反應監測（Multiple Reaction Monitoring）定量分析。馬兜鈴酸 I 與馬兜鈴酸 II 儀器檢出極限分別為 2 與 2.8 ng/mL。結果顯示細辛檢體均含馬兜鈴酸 I，馬兜鈴酸 II 則未檢出。本研究首次探討細辛不同部位中馬兜鈴酸 I 分布情形，並比較甲醇抽提物與水抽提物中馬兜鈴酸 I 之含量。市售 20 件細辛檢體，其根、根莖、葉柄、葉之 75% 甲醇抽提物中，馬兜鈴酸 I 平均含量分別為 1.23 ± 0.64、1.72 ± 1.38、7.81 ± 4.04、27.13 ± 25.31 ppm；其根、根莖、葉柄、葉之水抽提物中，馬兜鈴酸 I 平均含量分別為 0.33 ± 0.32、0.45 ± 0.50、3.98 ± 3.09、6.32 ± 9.75 ppm。結果顯示，馬兜鈴酸 I 之含量以葉最高，根最低；水抽提量約為 75% 甲醇抽提量之 25-50%。細辛濃縮散 6 件與麻黃附子細辛湯 6 件中，馬兜鈴酸 I 含量範圍分別為 0.35-2.07 ppm 與 0.21-1.00 ppm。此項研究結果可供行政部門管理細辛藥材暨其製劑之參考。"

A method for analysis of aristolochic acids (AAs: AA-I, AA-II) in Asarum (Xixin, named in Chinese) and its preparations by liquid chromatography coupled with tandem mass spectrometry has been developed. The chromatographic analysis was carried out using a C18 column (2.1×150 mm) and a mixture of 0.1% formic acid (containing 0.1% ammonium acetate) and acetonitrile (65:35, v/v) as the mobile phase at a flow rate of 0.3 mL/min. Piromidic acid was used as an internal standard. The mass detection was measured as the positive ion electrospray ionization mode. The precursor and daughter ions for quantitation were m/z 359 and 298 for AA-I, m/z 329 and 268 for AA-II. The instrument detection limit of AA-I and AA-II was 2 and 2.8 ng/mL, respectively. The result showed that Asarum contained AA-I while AA-II was not detected. AA-I distribution in different parts of Asarum was examined in detail and AA-I content in both methanol and water extracts was also compared in this study. In our twenty samples of Asarum, the average amounts of AA-I in 75% methanol extracts of roots, rhizomes, petioles and leaves were 1.23 ± 0.64, 1.72 ± 1.38, 7.81 ± 4.04 and 27.13 ± 25.31 ppm, respectively. The average amounts of AA-I in water extracts of roots, rhizomes, petioles and leaves were 0.33 ± 0.32, 0.45 ± 0.50, 3.98 ± 3.09 and 6.32 ± 9.75 ppm, respectively. The results indicated that the amounts of AA-I in roots were less than those in petioles and leaves, and the amounts of AA-I in water extracts were about 25 to 50 percent of those in 75% methanol extracts. For Asarum preparations, the amounts of AA-I in each six of Xi-xin-extract-powder and Ma-Huang-Fu-Zi-Xi-Xin-Tang ranged between 0.35-2.07 ppm and 0.21-1.00 ppm, respectively. Results of this study could be provided for the regulatory authority as references.