Determination Of Phytotoxins In Biological Samples By QuEChERS-HPLC-MS/MS

There are many kinds of poisonous plants and phytotoxins all over the world.The cases and accidents caused by plant toxins often occur.In such cases,the detection of phytotoxins in blood,urine,tissues and other biological samples is the key to the diagnosis of poisoning or the identification of related cases.Therefore,it is of great significance to establish a rapid and sensitive detection method for common phytotoxins in biological samples.In this paper,based on the improved QuEChERS（quick,easy,cheap,effective,rugged,safe）pre-treatment method,combined with high performance liquid chromatography-mass spectrometry（HPLC-MS / MS）,24 common phytotoxins in blood,liver and urine were analyzed.These plant toxins are Cytisine,Physostigmine,Hyoscyamine,Scopolamine,Anisodamine,Koumine,Gelsemine,Gelsenicine,Adynerine,Oleandrin,Bullatine A,Colchicine,Hypaconitine,Neoaconitine,Aconitine,Yunaconitine,Alpha-chaconine,Solanine,Wilfortrine,Wilforgine,Wilforine,Wtlfordine,Strychnine and Brucine.The results are as follows:1.A HPLC-MS / MS method for simultaneous injection and analysis of 24 phytotoxins was established.MRM scanning,ion pair qualitative analysis and standard curve quantitative analysis were used.2.In this paper,we established a pre-treatment method for three kinds of common biological samples（blood,urine,liver）,which can deal with the above three biological samples,extract and analyze the above 24 kinds of phytotoxins.In this paper,for blood samples,2 mL acetonitrile was used as extractant,70 mg NaCl was used as salting out agent,20 mg PSA and 30 mg C₁₈mixed powder were used as purification agent;for liver samples,2 mL methanol was used as extractant,60 mg PSA,50 mg C₁₈and 15 mg GCB mixed powder were used as purification agent;and for urine samples,50 mg PSA and 50 mg C₁₈mixed powder were used for purification.The improved QuEChERS method can effectively remove the interference of complex matrix in biological samplesr,reduce the matrix effect,and improve the recovery rate and sensitivity of the detection.3.The methods of QuEChERS-HPLC-MS / MS analysis of 24 phytotoxins in blood,urine and liver were confirmed.The LOD of 24 phytotoxins in the blood was 0.03⁰.3 μg·L^-1,the LOQ was 0.2¹.0 μg·L^-1,the intra-and inter-day RSD was 0.5⁶.9%,the recovery rate was 80.4% 118.0% in the three levels,the LOD of 24 phytotoxins in the liver was 0.1⁰.3 μg·L^-1,the LOQ was 0.2¹.0 μg·L^-1,intra-and inter-day RSD was 0.6⁶.5%,the recoveries were 82.9% ¹19.8% in the three levels;LOD of 24 phytotoxins in urine was 0.03⁰.3 μg·L^-1,LOQ was 0.1¹.0 μg·L^-1,intra-and inter-day RSD was 0.1⁶.0%,and the recoveries were 80.3% 121.3% in the three levels.4.Finally,the QuEChERS-HPLC-MS / MS detection method established in this paper was applied to practical cases,which proved that this method is convenient,fast,with good detection limit,and can be used in forensic toxicology.