A Validated Method for Quantifying Atractyloside and Carboxyatractyloside in Blood by HPLC-HRMS/MS, a Non-Fatal Case of Intoxication with Atractylis gummifera L.

Abstract

Atractyloside (ATR) and carboxyatractyloside (CATR) are diterpene glycosides that are responsible for the toxicity of several Asteraceae plants around the world. Mediterranean gum thistle (Atractylis gummifera L.) and Zulu impila (Callilepis laureola DC.), in particular, are notoriously poisonous and the cause of many accidental deaths, some suicides and even some murders. There is no current method for measuring the two toxins in biological samples that meet the criteria of specificity required in forensic medicine. We have endeavored to fill this analytical gap. Analysis was carried out using a solid-phase extraction and a high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry detection. The method was validated in the whole blood with quantification limits of 0.17 and 0.15 µg/L for ATR and CATR, respectively. The method was applied to a non-fatal case of intoxication with A. gummifera. To the best of the authors' knowledge, this is the first time that a concentration of ATR and CATR in blood (883.1 and 119.0 µg/L, respectively) and urine (230.4 and 140.3 µg/L, respectively) is reported. ATR and CATR were quantified in A. gummifera roots by the standard method addition (3.7 and 5.4 mg/g, respectively).

The standard addition method and its validation in forensic toxicology

“…In recent years, it seems likely that scientific reports using SAM are increasing, but in many of them except our reports, the validation data for SAM are either missing, incomplete or confusingly mixed with the validations of MMCM [48][49][50][51][52][53][54][55] (see Table 1). Only one report [56] describing postmortem distribution of mepirapim and acetyl fentanyl in a cadaver that died of drug poisoning presented satisfactory validation data for SAM, which were similar to those proposed by our group [25, 28, 31-35, 37, 39].…”

Comparação dos efeitos das folhas de Cestrum axillare Vell. com as saponinas isoladas em caprinos

“…Os sinais clínicos foram similares aos encontrados neste estudo e consistiam em anorexia, apatia, tenesmo retal com discreto prolapso de reto, desidratação progressiva com retração dos globos oculares e sinais nervosos como incoordenação motora, tremores musculares e agressividade. Além disto, os achados patológicos do fígado, caracterizados pela aparência macroscópica de aspecto de noz moscada e microscopicamente a necrose hepática, observados na intoxicação por C. axillare são similares as descritas na intoxicação de plantas que possuem como princípio tóxico os carboxiatractilosídeos (Cole et al 1989), como Xanthium cavanillesii (Driemeier et al 1999), Xantium strumarium (Turgut et al 2005, Botha et al 2014 e Atractylis gummifera (Carlier et al 2014). No entanto, ainda não foi determinada a ocorrência de carboxiatractilosídeo nas folhas de C. axillare.…”

High-resolution mass spectrometry in toxicology: current status and future perspectives

“…Another application of Q-OT for biogenic toxins is the determination of atractyloside and carboxyatractyloside, glycosides of the plant Atractylis gummifera L. in whole blood for confirmation of the diagnosis of a corresponding poisoning (Carlier et al 2014). Symptoms may include epigastric pain, diarrhea and vomiting, and respiratory depression, which causes hypoxemia, leading to tissue hypoxia, spasms, stiffness, and convulsions.…”