Isolation, Characterization and Quantity Determination of Aristolochic Acids, Toxic Compounds in Aristolochia bracteolata L.

Background Aristolochic Acids (AAs) are major components of plants in Aristolochia and have been found to be nephrotoxic, carcinogenic and mutagenic. Herein reported are the isolation, identification and quantity determination methods of Aristolochic Acid-I (AA-I) and Aristolochic Acid-II (AA-II) toxic compounds of Aristolochia bracteolata indigenous to Central Sudan and medicinally used in diverse biological functions including analgesic and diuretic effects, treatment of tumors, malaria and/or fevers. Methods and results AAs mixture was extracted with methanol from the defatted material of Aristolochia bracteolata whole plant at room temperature and was isolated from the aqueous methanol extract by chloroform. Moreover, Silica-gel column chromatography and Preparative Thin Layer Chromatography (PTLC) using chloroform/methanol gradient mixtures were used to isolate AAs mixtures as a yellow crystalline solid. A preliminary detection of AAs was made by Thin Layer Chromatography (silica-gel, chloroform: methanol (6:1)). The Rf value of the acids mixture was 0.43-0.46. The presence of AAs in plant sample was confirmed by High Performance Liquid Chromatography/Ultraviolet (HPLC/UV) analysis using 1% acetic acid and methanol (40:60) as mobile phase and maximum absorption wave length of 250 nm. Quantitative determination of AA-II (49.03 g/kg) and AA-I (12.98 g/kg) was also achieved by HPLC/UV. Recommendation It is recommended that the use of Aristolochia bracteolata as a medicinal plant should be extremely limited or strictly prohibited. The chromatograms obtained in this study can serve as fingerprints to identify AAs in plant samples

The safety of Homnawakod herbal formula containing <it>Aristolochia tagala</it> Cham. in Wistar rats

<p>Abstract</p> <p>Background</p> <p>A dried root of <it>Aristolochia tagala</it> Cham. (ATC) is often used in Thai traditional medicine as an antipyretic, anti-inflammatory agent, muscle relaxant, appetite-enhancing agent, and analeptic. Homnawakod, an important herbal recipe, originally contains ATC in its formula, however, some <it>Aristolochia</it> species have been reported to cause nephrotoxicity due to aristolochic acid (AA) and its derivatives, resulting in ATC removal from all formulae. Therefore, this study investigates the chemical profiles of ATC, the original (HNK+ATC) and the present Homnawakod Ayurved Siriraj Herbal Formulary™ (HNK), and investigates whether they could cause nephrotoxicity or aggravate LPS-induced organ injuries <it>in vivo</it>.</p> <p>Methods</p> <p>HPLC and LC/MS were used for chemical profile study. Male Wistar rats were randomly divided into groups in which the rats were intragastrically administered distilled water (2 groups), ATC (10 or 30 mg/kg), HNK+ATC (540 or 1,620 mg/kg), or HNK (1,590 mg/kg) for 21 days. A positive control group was administered with single dose 100 mg/kg standard AA-I intragastrically at day 1. Serum creatinine and urea were measured at baseline and at 7, 14 and 21 days of the treatment. On day 22, a model of lipopolysaccharide (LPS)-induced endotoxemia was used. One-way and two-way analyses of variance were performed and a <it>P</it> value of less than 0.05 was considered to be significant.</p> <p>Results</p> <p>The similarity of the HPLC chromatograms of HNK+ATC and HNK could suggest that the qualities of both formulae are nearly the same in terms of chemical profile. The amount of AA-I found in ATC is 0.24%w/w. All experimental groups exhibited similar levels of serum urea at baseline and 7 and 14 days of the treatment. At 21 days, rats received AA exhibited a significant increase in serum urea, whereas the others did not exhibit such toxicity. On day 22, there were no significant changes in LPS-induced renal and liver dysfunction, or LPS-induced mean arterial pressure (MAP) reduction upon administration of ATC, HNK+ATC, HNK or AA-I.</p> <p>Conclusions</p> <p>These results suggest that ATC, HNK+ATC or HNK, at the animal dose equivalent to that used in human, do not cause the acute nephrotoxicity in rats and do not aggravate LPS-induced organ injuries even further.</p