Analysis of Aristolochic Acids from Birthwort using UPLC
Anamaria Craciun, 1st year MSc student, QSAR
West University of Timisoara, Chemistry Department,
Supervisors:
Sabina Danciu
Dr. Vasile Ostafe professor
Chemistry Department, West University of Timişoara, Romania
Aristolochic acids are extracted from the Aristolochia sp. and are a mixture of
structurally related nitrophenanthrene carboxylic acids, out of which the most
commonly known compounds are the aristolochic acids I, II, C and Ia, the 7hydroxyaristolochic acid I and the aristolic acid. In the present study we separated the
aristolochic acids I and II.
Aristolochic acids and Aristolochia sp. have been a subject of debate for about
five decades, but led to an even bigger interest in 1991, when the aristolochic acids
were found to be responsible for Chinese Herbs Nephropathy. This rapidly progressive
renal fibrosis was associated with the prolonged intake of a slimming regimen
containing of Chinese herbs, at a clinic from Belgium. The nephropathy was traced
back to Aristolochia fangchi which contained the aristolochic acids [1]. This led to a
series of studies which proved that aristolochic acids formed adducts with the DNA
[2]. Moreover, there is a hypothesis stating that the long-term ingestion of aristolochic
acids is the reason for Balkan Endemic Nephropathy [3].
Although banned from sale, products based on Aristolochia sp. are still sold on
web and advertised in various magazines. In Romania there are some newspapers that
still provide recipes for treating various diseases with extracts from birthwort plants.
This is why a rapid and sound method for quantification of aristolochic acids in
products for human consumption is required.
A variety of methods for the rapid and accurate estimation of the aristolochic acids
was published. This study is the first use of an Ultra Performance Liquid
Chromatography method for the separation of aristolochic acids from different
anatomical parts of Aristolochia clematitis. The separation of these acids was
performed in 2.2 minutes. The best solvent for the extraction of the aristolochic acids
was assessed and out of methanol, ethanol, chloroform, ethylic ether and acetone, it
turned out that the best solvent for the extraction was acetone. The extraction of the
aristolochic acids was also performed at two different temperatures, room temperature
(~ 25C) and 50C. The amount of aristolochic acids was assessed in rhizome, basal
and apical stems, mature and young leaves and flowers. The highest content of
aristolochic acids was found in flowers and young leaves.
[1] Vanherweghem J.-L., Depierreux M., Tielemans C., Abramowicz D., Dratwa M.,
Jadoul M., Richard C., Vandervelde D., Verbeelen D., Vanhaelen-Fastre R.,
Vanhaelen M., Lancet., 341, 387-391 (1993)
[2] Arlt V.M., Stiborova M., Schmeiser H.H., Mutagenesis, 17 (4) 265-277 (2002)
[3] Hranjec T., Kovac A., Kos J., Croat. Med. J., 46, 116-125 (2005)