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«“√ “√°“√·æ∑¬ Journal of Thai Traditional & Alternative Medicine
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ªï∑’Ë 10 ©∫—∫∑’Ë 2 情¿“§¡- ‘ßÀ“§¡ 2555 Vol. 10 No. 2 May-August 2012
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Optimal Detection of Aristolochic Acid I

in Krai-krue by High Performance thin

Layer Chromatography

†
Rattana Phadungrakwittaya*, Pravit Akarasereenont ,
Natchagorn Lumlerdkij*, Sirikul Chotewuttakorn*,
†
Sirinapa Chaisri , Tawee Laohapand †

Abstract
Aristolochic acids (AA) are toxic substances found in Krai-kruea (Aristolochia sp.). They are
nitrophenanthrene carboxylic acids which contain aristolochic acid I (AA-I) and aristolochic
acid II (AA-II) as major components. The detection of AA is needed in order to avoid using AA-
containing herbal raw materials. In this study, high performance thin layer chromatography
(HPTLC) was optimized for AA-I screening in Krai-kruea. Three mobile phase systems were
tried with HPTLC to obtain the best separation and reverse-phase thin layer chromatography
(RP-TLC) was used to confirm the identification. For HPTLC, a mixture of chloroform, metha-
nol, and acetic acid (65:20:1, v/v) was the most optimal mobile phase system with the lower limit
of detection (LOD) of 8 ng, while a mixture of acetonitrile, methanol, and water (3:0.5:1, v/v) was
the most suitable one for RP-TLC with LOD of 15 ng. According to the system, the Rf values of
AA-I in Krai-kruea samples were 0.55 and 0.48 with HPTLC and RP-TLC, respectively. In sum-
mary, these HPTLC and RP-TLC systems can be used to detect AA-I in Krai-kruea single herb
and HPTLC has greater sensitivity than RP-TLC.
Key words: aristolochic acid, Krai-kruea, HPTLC, RP-TLC, herbal formulas

Introduction (AA-I) and aristolochic acid II (AA-II) are major

Aristolochic acids (AA) are nitrophenan- components. AA and their derivatives have
threne carboxylic acids found in plants of been isolated and determined by several spec-
1-3
Aristolochia sp. in which aristolochic acid I tral and chemical methods. In 2009, the U.S.
Food and Drug Administration (USFDA) is-
* Department of Pharmacology, Faculty of Medicine Siriraj sued a strong warning to consumers to avoid
Hospital, Mahidol University, Bangkok 10700, Thailand
† botanical products that contain AA because
Center of Applied Thai Traditional Medicine, Faculty of 4
the toxicities of AA had been reported. In a
Medicine Siriraj Hospital, Mahidol University, Bangkok
10700, Thailand short-term toxicity study, rats treated orally
93