SOFT 2004 Meeting Abstracts - Society of Forensic Toxicologists

P12
DETERMINATION OF ATRACTYLOSIDE IN CALLILEPIS LAUREOLA USING SOLID-PHASE
EXTRACTION AND LIQUID CHROMATOGRAPHY - ATMOSPHERIC PRESSURE
IONISATION MASS SPECTROMETRY
Paul A. Steenkamp"', Nial M. Harding', Fanie R. van Heerden b .•, Ben-Erik van Wyk C
"Forensic Chemistry Laboratory, Department ofHealth, P.O. Box 1080, Johannesburg 2000, South Africa;
bDepartment of Chemistry and Biochemistry, Rand Afrikaans University, P.O. Box 524, Auckland Park
2006, South Africa; CDepartment of Botany, Rand Afrikaans University, P.O. Box 524, Auckland Park
2006, South Africa
A selective analytical method based on high-performance liquid chromatography, combined with
atmospheric pressure ionisation mass spectrometry, was developed for the detection of atractyloside. The
tuberous root of the plant Callilepis laureola is used in traditional medicine by the Zulu and Xhosa people
of South Africa and it is well known by the Zulu name impila, which means 'health'. However, C. laureola
has also been implicated in the death of numerous patients who used medication prepared from impila in
which atractyloside, the dipotassium salt of a sulfonated kaurene glycoside, was identified as the toxic
principle. This compound, together with carboxyatractyloside, was also identified as the toxic principle of
Atractylis gummifera L. (Asteraceae), a plant linked to human fatalities in Mediterranean countries since
ancient times.
The analysis was performed on an Xterra Phenyl column utilising a gradient elution profile and a mobile
phase consisting of 10 mM aqueous ammonium acetate buffer-MeOH-acetonitriIe. The developed method
showed a good linearity for the calibration curve spanning the 10 ng/mL to 1 j.ig/mL (r2 = 0.997) range.
The limit of detection and limit of quantitation were determined and found to be 100 pglmL and 10 ng/mL,
respectively. The method was successfully applied to the analysis of C. laureola tuber, herbal medicine
mixtures and viscera samples (stomach, stomach contents, liver, kidney, blood, urine, bile) for the presence
of atractyloside.
Keywords: Atractyloside, CallUepis laureola, HPLC-MS
* Corresponding author. Tel: +27-11-725-2279; fax: +27-11-725-4731. E-mail address: fcIjhb@icon.co.za
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