A simple and selective detection method for aristolochic acid in crude drugs using solid-phase extraction
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A simple and selective detection method for aristolochic acid in crude drugs using solid-phase extraction Takaomi Tagami • Akihiro Takeda • Akiko Asada • Airin Aoyama • Takahiro Doi • Masami Kawaguchi • Keiji Kajimura • Yoshiyuki Sawabe • Hirotaka Obana • Katsuhiro Yamasaki
Received: 27 October 2012 / Accepted: 21 December 2012 Ó The Japanese Society of Pharmacognosy and Springer Japan 2013
Abstract The official Japanese method for analyzing aristolochic acid I (AA-I) in Asiasarum root using conventional high-performance liquid chromatography (HPLC) is described in the Japanese Pharmacopoeia, Sixteenth Edition. Interfering peaks of AA-I sometimes appear after HPLC analysis of crude drugs. A selective analytical method is needed to determine definitively whether AA-I is present in crude drugs. In this study, we developed a selective method that combined solid-phase extraction and liquid chromatography/mass spectrometry (LC/MS) which may be useful for identifying AA-I in crude drugs and for quality control. Keywords LC–MS Aristolochic acid Crude drug Solid-phase extraction
issued safety information about crude drug products and Kampo formulations that consist of natural herbs containing AA. In particular, they called attention to four crude drugs [Akebia stem (Mokutsu), Sinomenium stem and rhizome (Boui), Saussure root (Mokkou), and Asiasarum root (Saishin)]. In Japan, Asiasarum root is examined to ensure that it does not contain AA-I; the official method of analyzing AA-I in Asiasarum root using conventional highperformance liquid chromatography (HPLC) is described in the Japanese Pharmacopoeia, Sixteenth Edition. Interfering peaks sometimes appear in the course of HPLC analysis of crude drugs. In this study, we developed a simple and selective method of analyzing AA-I by combining solid-phase extraction and liquid chromatography/ mass spectrometry (LC/MS).
Introduction Materials and methods Between 1990 and 1992, there were reports from Belgium of a severe renal disorder caused by aristolochic acid (AA), one of the main components of Aristolochia fangchi (Aristolochiaceae), which is an ingredient found in health supplements that are taken as a weight loss regimen [1]. Similar problems have been reported in Japan [2, 3], and there is concern regarding health hazards caused by AA. The Japanese Ministry of Health, Labor, and Welfare T. Tagami (&) A. Takeda A. Asada A. Aoyama T. Doi M. Kawaguchi K. Kajimura Y. Sawabe H. Obana Osaka Prefectural Institute of Public Health, 1-3-69 Nakamichi, Higashinari-ku, Osaka 537-0025, Japan e-mail: [email protected] K. Yamasaki Iwaki Meisei University, 5-5-1 Chuodai Iino, Iwaki, Fukushima 970-8551, Japan
Chemicals and reagents Aqueous ammonia solution, phosphoric acid, LC/MSgrade acetonitrile, methanol, and formic acid were purchased from Wako Pure Chemical Industries (Osaka, Japan). AA-I and LC/MS-grade ammonium formate were purchased from Sigma-Aldrich (St. Louis, MO, USA). SepPak Accell Plus QMA (QMA) cartridge (360 mg) was purchased from Wa
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