Determination of α-amanitin and β-amanitin in human biological fluids by high-performance liquid chromatography

Jehl, F.; Gallion, C.; Birckel, P.; Jaeger, Albert; Flesch, F.; Minck, R

doi:10.1016/0003-2697(85)90474-9

Cited by 37 publications

(13 citation statements)

References 19 publications

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“…The detection limit for both amanitins was about 0.5 ng/ml. In previous reports, the detection limits were 3 ng/ml [7] or 1 ng/ml for urine and 0.1 ng/ml for plasma [8] by RIA, 10 ng/ml by LC-UV [10,11], 2 ng/ml for plasma by LC-ECD [13], and 1 ng/ ml [19] or 2.5 ng/ml [20] for urine samples by CE-DAD; however, these methods cannot identify the amanitins in biological fl uids. An LC-MS method for determining α-amanitin and β-amanitin in urine after immunoaffi nity extraction was reported; their detection limit for the compounds was 2.5 ng/ml [15].…”

Section: Quantitation Of Both Amanitins In Human Plasma and Validatiomentioning

confidence: 99%

“…For analysis of amatoxins, radioimmunoassay (RIA) [7][8][9], liquid chromatography (LC)-ultraviolet detection (UV) [10,11], LC-electrochemical detection (ECD) [12,13], LC-mass spectrometry (MS) (-MS) [14][15][16][17][18], and capillary electrophoresis (CE) with diode array detection Abstract A number of reports are available in the literature that describe liquid chromatography-mass spectrometry (LC-MS) and LC-tandem mass spectrometry (LC-MS-MS) analysis of amanitins, very toxic mushroom toxins, in biological samples. However, the extractive pretreatment methods and LC separation column materials vary remarkably according to the different reports.…”

Section: Introductionmentioning

confidence: 99%

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Simple analysis of α-amanitin and β-amanitin in human plasma by liquid chromatography-mass spectrometry

et al. 2010

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m/z 919-921 and m/z 920-922, respectively, gave symmetrical peaks and good separation of both amanitin peaks. Using an external calibration method, linearity, detection limits, recovery rates, and precision were tested; they were all satisfactory. To our knowledge, the present method gives the simplest LC-MS analysis for amanitins among those so far reported. We recommend the method for use in actual forensic and clinical toxicological analysis of amanitins in biological samples.

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Section: Quantitation Of Both Amanitins In Human Plasma and Validatiomentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Simple analysis of α-amanitin and β-amanitin in human plasma by liquid chromatography-mass spectrometry

et al. 2010

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“…Currently, there are several high-performance liquid chromatography (HPLC)-ultraviolet (UV) [11][12][13][14][15][16][17], liquid chromatography (LC)-mass spectrometry (MS) [18][19][20] and ultra performance liquid chromatography (UPLC)-MS/MS [21] methods to determine ␣-amanitin in mushrooms, liver and biological fluids (Table 1). HPLC-MS methods for ␣-amanitin detection are accurate, sensitive, and specific; nevertheless HPLC-MS is an expensive technique and is not available in many laboratories.…”

Section: Introductionmentioning

confidence: 99%

Quantification of alpha-amanitin in biological samples by HPLC using simultaneous UV- diode array and electrochemical detection

Garcia

Costa

Baptista

et al. 2015

Journal of Chromatography B

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α-Amanitin is a natural bicyclic octapeptide, from the family of amatoxins, present in the deadly mushroom species Amanita phalloides. The toxicological and clinical interests raised by this toxin, require highly sensitive, accurate and reproducible quantification methods for pharmacokinetic studies. In the present work, a high-performance liquid chromatographic (HPLC) method with in-line connected diode-array (DAD) and electrochemical (EC) detection was developed and validated to quantify α-amanitin in biological samples (namely liver and kidney). Sample pre-treatment consisted of a simple and unique deproteinization step with 5% perchloric acid followed by centrifugation at 16,000×g, 4°C, for 20min. The high recovery found for α-amanitin (≥96.8%) makes this procedure suitable for extracting α-amanitin from liver and kidney homogenates. The resulting supernatant was collected and injected into the HPLC. Mobile phase was composed by 20% methanol in 50mM citric acid, and 0.46mM octanessulfonic acid, adjusted to pH 5.5. The chromatographic runs took less than 22min and no significant endogenous interferences were observed at the α-amanitin retention time. Calibration curves were linear with regression coefficients higher than 0.994. The overall inter- and intra-assay precision did not exceed 15.3%. The present method has low interferences with simple and fast processing steps, being a suitable procedure to support in vivo toxicokinetic studies involving α-amanitin. In fact, the validated method was successfully applied to quantify α-amanitin in biological samples following intraperitoneal α-amanitin administration to rats. Moreover, human plasma was also used as matrix and the purposed method was adequate for detection of α-amanitin in that matrix. The results clearly indicate that the proposed method is suitable to investigate the pharmacokinetic and tissue distribution of α-amanitin. Additionally, the method will be very useful in the development of novel and potent antidotes against amatoxins poisoning and to improve the knowledge of α-amanitin toxicity.

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“…Detection can be carried out with a UV-spectrometer [1,3,[16][17][18] or by coupling to a mass spectrometer [5]. Besides also radioimmunoassays were used [4].…”

Section: Introductionmentioning

confidence: 99%

Identification of toxic oligopeptides in Amanita fungi employing capillary electrophoresis‐electrospray ionization‐mass spectrometry with positive and negative ion detection

2008

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The identification of toxic oligopeptides employing CE-ESI-MS is presented. The analytes studied ama- and phallotoxins are of significant forensic interest because over 90% of the lethal cases of fungus poisoning in man are caused by species of Amanita which contain these toxins. A CE method was developed to separate the toxins alpha-, beta- and gamma-amanitin, phalloidin and phallacidin. Their fragmentation patterns in MS(n) experiments were investigated in the positive and in the negative ion mode, also the influence of the sheath liquid mixture of the used interface on the S/N. Method validation included the determination of the LOD and the repeatability of the migration time and peak area for both detection modes. With the optimized method LODs of 13-79 ng/mL (17-87 nmol/L) were reached. The CE-MS procedure was successfully applied to the identification of ama- and phallotoxins in extracts of air-dried mushroom samples.

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Determination of α-amanitin and β-amanitin in human biological fluids by high-performance liquid chromatography

Cited by 37 publications

References 19 publications

Simple analysis of α-amanitin and β-amanitin in human plasma by liquid chromatography-mass spectrometry

Simple analysis of α-amanitin and β-amanitin in human plasma by liquid chromatography-mass spectrometry

Quantification of alpha-amanitin in biological samples by HPLC using simultaneous UV- diode array and electrochemical detection

Identification of toxic oligopeptides in Amanita fungi employing capillary electrophoresis‐electrospray ionization‐mass spectrometry with positive and negative ion detection

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