Determination of Harmaline and Harmine in Whole Blood Using High-performance Liquid Chromatography-tandem Mass Spectrometry
Peganum harmala is a plant widely distributed in the arid areas of northwest China,the alkaloid harmaline and harmine contained in which shows toxic effects in human such as acute central neurological symptoms,cardiovascular effects and death.In order to meet the qualitative and quantitative detection requirements for cases of Peganum harmala poisoning,a high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was developed for the simultaneous detection of harmaline and harmine in human whole blood.The blood samples were deproteinized with acetonitrile,separated on an analytical column(Agilent SB-C18 100 mm×2.1 mm×1.8 μm)with 0.1%formic acid and acetonitrile as the mobile phase by gradient elution at a flow rate of 0.4 mL/min,determined by electrospray ionization(ESI+)and multiple reaction monitoring(MRM)modes after diluted and filtrated through membrane,and quantified by external standard calibration method.The result showed that there was good linear relationship for harmaline and harmine in human whole blood over the range of 10 to 1 000 ng/mL(R2>0.999).The limit of detection(LOD)and quantitation(LOQ)of harmaline and harmine in human whole blood were 3 and 10 ng/mL,separately.At 10,100,and 500 ng/mL,the recoveries of harmaline and harmine showed a range of 84.9%to 97.0%and the matrix effects of different human whole blood were less than±25%,with the precisions of intra-day and inter-day ranged from 3.6%to 9.8%and 7.3%to 12.8%respectively,and the accuracy ranged from 2.4%to 9.5%,which meets the general rules for forensic toxicology in China.The method was successfully applied to a real case of abnormal death caused by improper use of Peganum harmala,where the concentration of harmaline and harmine in the blood of the deceased were detected to be 1.2 μg/mL and 0.2 μg/mL respectively.In conclusion,the results of this study could be applied to forensic science on the simultaneous detection of harmaline and harmine in human whole blood with the advantages of simplicity and accuracy and provide reliable technical support for the subsequent investigation of practical cases.
toxicological analysisharmalineharminehigh-performance liguid chromatography-tandem mass spectrometry(HPLC-MS/MS)whole blood
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维普
