[Highly sensitive determination of three kinds of amanitins in urine and plasma by ultra performance liquid chromatography-triple quadrupole mass spectrometry coupled with immunoaffinity column clean-up].

Cases of toxic mushroom poisoning occur frequently in China every year. In particular, mushrooms containing amanitins can cause acute liver damage, with high mortality rates. The symptoms of acute liver damage are experienced 9-72 h after consumption of the mushrooms. At this time, the concentration of amanitins in blood and urine is too low to be detected even by the highly sensitive ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS), thus rendering clinical diagnosis and treatment difficult. To this end, a method was developed for the determination of α-amanitin, ß-amanitin and γ-amanitin in urine and plasma by UPLC-MS/MS. Urine and plasma samples were extracted and cleaned up by using an immunoaffinity column. A sample of 2.00 mL urine or 1.00 mL of plasma was diluted with 8.00 mL of phosphate buffer solution (PBS) and then loaded onto the immunoaffinity column at a flow rate of 0.5-1.0 mL/min. After washing the column with 10 mL of PBS and 13 mL of water successively, the bound amanitins were eluted with 3.00 mL of methanol-acetone (1∶1, v/v). The eluent was dried under nitrogen at 55 ℃. The residue was dissolved in 100 µL of 10% (v/v) methanol aqueous solution. The amanitins in urine were concentrated 20 times, while those in plasma were concentrated 10 times. Chromatographic separation was performed on a Kinetex Biphenyl column (100 mm × 2.1 mm, 1.7 µm) with gradient elution using methanol and 0.005% (v/v) formic acid aqueous solution as mobile phases. The three amanitins were detected by negative electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode and quantified by the solvent standard curve external standard method. Method validation was performed as recommended by the European Drug Administration (EMEA). Four levels of quality control (QC) samples were prepared, which covered the calibration curve range, viz., the limit of quantification (LOQ), within three times the LOQ (low QC), medium QC, and at 85% of the upper calibration curve range (high QC), and used to test the accuracy, precision, matrix effect, extraction recovery, and stability. The calibration curves for the three amanitins showed good linear relationships in the range of 0.1-200 ng/mL, and the correlation coefficients (r) were greater than 0.999. The matrix effects and extraction efficiencies of the three amanitins in urine and plasma were 92%-108% and 90%-103%, respectively, and the coefficients of variation were less than 13%. The accuracies of the three amanitins in urine were within -9.4%-8.0%. The repeatability and intermediate accuracies were 3.0%-14% and 3.5%-18%, respectively. When the sampling volume was 2.00 mL, the limits of detection of the three amanitins in urine were 0.002 ng/mL. The accuracies of the three amanitins in plasma were within -13%-8.0%. The repeatability and intermediate accuracies were 3.9%-9.7% and 5.5%-12%, respectively. When the sampling volume was 1.00 mL, the limits of detection of the three amanitins in plasma were 0.004 ng/mL. The developed method is simple, sensitive, and accurate. During toxic mushroom poisoning detection, 0.0067 ng/mL of α-amanitin and 0.0059 ng/mL of ß-amanitin were detected in the urine of poisoned patients 138 h after ingesting poisonous mushrooms. This method has successfully solved the problem of detecting ultra-trace levels of amanitins in the urine and plasma of poisoned patients. It has important practical significance for the early diagnosis, early treatment, and mortality reduction of suspected poisoning patients. This method can also provide reliable technical support for future research on the toxicological effects and in vivo metabolism of these toxins.

[Simultaneous determination of paraquat and diquat in plasma and urine by ion chromatography-triple quadrupole mass spectrometry].

Paraquat (PQ) and diquat (DQ) are widely used as non-selective contact herbicides. Several cases involving accidents, suicide, and homicide by PQ or DQ poisoning have been reported. Poising by PQ, which is mainly concentrated in the lungs, causes acute respiratory distress syndrome and leads to multiple organ toxicity. The toxic effects of DQ are similar to those of PQ but relatively less intense. The mortality rates in PQ and DQ poisoning are high. Simultaneous monitoring of the PQ and DQ concentrations in plasma and urine can provide valuable information for early clinical diagnosis and prognosis. High performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) is the main analytical method used to detect PQ and DQ in plasma and urine. As both these compounds are highly polar and water soluble, they cannot be retained effectively on a reversed-phase column with conventional mobile phases. The separation of PQ and DQ by ion-pair chromatography or hydrophilic chromatography has been reported. The use of an ion-pairing reagent helps in improving the retention capabilities of PQ and DQ. However, the sensitivity of MS detection is noticeably decreased because of ion suppression caused by the ion-pairing reagent in the mobile phase; furthermore, ion-pairing reagents may contaminate the MS system. The separation of PQ and DQ by hydrophilic chromatography is easily affected by matrix components in the sample, and their retention times are not stable. Considering PQ and DQ are bicharged cation species in solution, they are more suitable for separation by cation-exchange chromatography. A method based on ion chromatography-triple quadrupole mass spectrometry was established for the determination of PQ and DQ in plasma and urine. The plasma and urine samples were diluted with water, and then purified on a solid-phase extraction column containing a polymer-reversed phase and weak ion-exchange mixed-mode adsorbent (Oasis WCX). PQ and DQ were separated on an IonPac CS 18 analytical column (250 mm×2.0 mm, 6.0 µm) with gradient elution using a methylsulfonic acid solution electrolytically generated from an on-line eluent generation cartridge. An in-line suppressor was used to remove methylsulfonate and other anions from the eluent before the eluent entered the mass spectrometer. Between the suppressor and the ion source in MS, the addition of 3% (v/v) formic acid in acetonitrile as an organic modifier (using an auxiliary pump and a T-piece) aided desolvation in the ion source, resulted in a one-or two-fold improvement of the response, and eliminated the residual effects of the adsorption of PQ and DQ caused by ion source. The analytes were detected by triple quadrupole tandem mass spectrometry using positive electrospray ionization in the multiple reaction monitoring (MRM) mode. PQ-d8 and DQ-d4 were used as internal standards. The calibration curves for PQ and DQ showed good linear relationships in the ranges of 1.0-150 µg/L and 0.5-75 µg/L, respectively, and the correlation coefficients were > 0.999. The average matrix effects of PQ and DQ in plasma were 84.2%-89.3% and 84.7%-91.1%, while the average matrix effects of PQ and DQ in urine were 50.3%-58.4% and 51.9%-59.4%. The average recoveries of PQ and DQ in plasma were 93.5%-117% and 91.7%-112%, respectively, with relative standard deviations (RSDs) of 3.4-16.7% and 2.8%-13.2%, and that in urine were 90.0%-118% and 99.2%-116%, with relative standard deviations of 5.6%-14.9% and 2.4%-17.3% (n=6). The limits of detection of PQ and DQ in plasma and urine were 0.3 µg/L and 0.2 µg/L, respectively, with the corresponding limits of quantification being 1.0 µg/L and 0.5 µg/L. This method is sensitive and accurate, and it can be used to determine PQ and DQ for clinical diagnosis and prognosis in patients.

[Simultaneous determination of cucurbitacin B, E and I in plasma, urine, and melon and fruit vegetables by ultra performance liquid chromatography-triple quadrupole mass spectrometry using atmospheric pressure chemical ionization].

A method for the determination of cucurbitacin B (CuB), cucurbitacin I (CuI) and cucurbitacin E (CuE) in plasma, urine and melon and fruit vegetables was developed by ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). The target analytes in plasma and urine were extracted and cleaned-up by solid supported liquid-liquid extraction, while those in melon and fruit vegetables were extracted with acetonitrile and then diluted with water. CuB, CuI and CuE were separated on an XBridge BEH C18 column (100 mm×3.0 mm, 2.5 µm) with gradient elution using mobile phases of methanol and 0.025% (v/v) ammonia aqueous solution. An atmospheric pressure chemical ionization interface was used as the ion source and the analysis was performed in negative ionization multiple reaction monitoring (MRM) mode. The cucurbitacins in plasma and urine were quantified by the matrix working standard curve internal standard method, while those in melon and fruit vegetables were quantified by the solvent standard curve external standard method. Oleandrin was used as the internal standard. The average recoveries were 89.0%-113% for the three cucurbitacins in plasma and urine, with RSDs of 1.7%-12.2% (n=6). The average recoveries were 87.6%-114% for the three cucurbitacins in melon and fruit vegetables, with RSDs of 4.1%-11.1% (n=6). The limit of detection (S/N=3) of the three cucurbitacins was 0.03 µg/L in plasma and urine, and 5-10 µg/kg in melon and fruit vegetables. The method is simple, sensitive and accurate. It has been used for the determination of cucurbitacins in bitter bottle gourd and in the plasma and urine of patients poisoned by bitter bottle gourd, CuB was successfully detected.

[Analysis of multi-minerals in seafoods in Zhejiang Province].

OBJECTIVE: To analyze the contents of Ca, K, Na, Mg, Fe, Zn, Cu and Mn in 33 kinds of marine products, such as fish, shrimp, crab, shellfish and mollusk. METHODS: The national standard GB 5009. 268-2016 Determination of multi elements in foods was used for determination. The correlation between multi elements was statistically analyzed. RESULTS: The highest level of Ca was found in clam(510. 2 mg/100 g edible), K in mackerel(444. 4 mg/100 g edible), Na in clam(487. 6 mg/100 g edible), Mg in conch(132. 7 mg/100 g edible). The highest level of Fe and Mn was in conch(37. 35 and 2. 6 mg/100 g edible). The highest level of Zn and Cu was in oyster(15. 92 and 8. 58 mg/100 g edible). The correlation analysis showed that Mn-Ca was highly correlated in fish and shrimp(r=0. 9438 and 0. 8585, P < 0. 05), while Cu-Mg, Cu-Zn, Zn-Na were highly correlated in shellfish(r=-0. 9102, 0. 8501 and 0. 8428, P < 0. 05). CONCLUSION: There are rich mineral elements in different seafood, which can provide reference for the reasonable diet of residents.

[Simultaneous rapid determination of 84 toxic plant constituents in plasma and urine by ultra-performance liquid chromatography- triple quadrupole/linear ion trap mass spectrometry].

An ultra-performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry (UPLC-Qtrap MS) method was developed for the determination of 84 toxic plant constiuents in plasma and urine. Plasma was precipitated by acetonitrile to remove proteins and then passed through a Prime HLB SPE column to remove phospholipids, while urine was diluted with methanol. Chromatographic separation of the analytes was achieved on an Acquity BEH C18 column (100 mm×2.1 mm, 1.7 µm) by gradient elution using the mobile phase of 0.1% (v/v) formic acid and 2 mmol/L ammonium formate both in 97% (v/v) acetonitrile aqueous solution and water. Electrospray ionization mass spectrometry was carried out in the positive ion mode with multiple reaction monitoring-information dependent acquisition-enhanced product ion scan mode (MRM-IDA-EPI). The 84 analytes were quantified by the matrix working standard curve internal standard method, and a good linear relationship was observed, with correlation coefficients of ≥ 0.9911. The limits of detection (LODs) in plasma and urine were 0.01-1 µg/L and 0.03-2 µg/L, respectively. The intra- and inter-day precisions of these analytes were 0.7%-18.4% and 1.1%-18.5%, and the accuracy of all analytes ranged from 70.6% to 124.5%. This method is simple, sensitive, and accurate for the measurement of these analytes in plasma and urine for both clinical and forensic applications.

[Determination of coriatin and corianin in plasma and urine using ultra-performance liquid chromatography-triple quadrupole mass spectrometry].

An ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS) method has been developed for the determination of coriatin and corianin in plasma and urine, which are the biomarkers of poisoning caused by Coriaria sinica Maxim. Plasma and urine samples were extracted and purified using a solid supported liquid/liquid extraction method. Chromatographic separation was performed on a Cortecs C18 column (100 mm×2.1 mm, 1.6 µm) using a gradient elution of methanol and water. Coriatin and corianin were detected using negative electrospray ionization tandem mass spectrometry in multiple reaction monitoring (MRM) mode and quantified via a matrix working standard curve internal standard method; florfenicol was used as the internal standard. The assay was linear in the calibration range of 0.03-5.0 µg/L for coriatin and 0.3-50 µg/L for corianin in plasma, and 0.1-10 µg/L and 1-100 µg/L for coriatin and corianin in urine, respectively. The average recoveries were 86.2%-110% for coriatin and corianin in plasma and urine with relative standard deviations of 5.1%-14.6% (n=6). The limits of detection (S/N=3) for coriatin and corianin were 0.01 µg/L and 0.1 µg/L in plasma, and 0.03 µg/L and 0.3 µg/L in urine, respectively. The method is simple, sensitive and accurate for the determination of coriatin and corianin in plasma and urine for toxicological purposes.

[Determination of monofluoroacetic acid in plasma and urine by stable isotope dilution ion chromatography-triple quadrupole mass spectrometry].

A method was developed for the determination of monofluoroacetic acid (MFA) in plasma and urine by ion chromatography-triple quadrupole mass spectrometry (IC-MS/MS). A plasma sample was extracted with 3% (v/v) perchloric acid aqueous solution, and the extract was centrifuged to remove the protein and lipids. A urine sample was acidulated with 3% (v/v) perchloric acid aqueous solution. The target analyte was extracted with methyl tert-butyl ether (MTBE) at a pH between 0.5 and 1.0. After the MTBE was removed by blowing with nitrogen, the MFA in the residues was dissolved into 0.1% (v/v) ammonia solution. The separation of MFA was carried out on a Dionex Ionpac AS 19 analytical column (250 mm×2 mm, 7.5 µm) with gradient elution using KOH solution electrolytically generated from an on-line eluent generation cartridge. Before the eluent flow entered the mass spectrometer, an in-line suppressor was used to remove potassium ions. The MFA was detected with a negative electrospray ionization source in the multiple reaction monitoring (MRM) mode, and quantified with the stable isotope internal standard method. The correlation coefficient of the linear calibration curve of MFA was greater than 0.999 at the corresponding ranges of 0.1-1000 µg/L. The average recoveries were 96.2%-120% of MFA in plasma and urine samples with relative standard deviations of 1.1%-13.1% (n=6). The limits of detection of MFA in plasma and urine samples were 0.03 µg/L and 0.1 µg/L, respectively. The method is simple, sensitive and accurate, and can be applied for the determination of MFA in plasma and urine samples.

[Rapid determination of benzo[a]pyrene in foods by ultra performance liquid chromatography-triple quadrupole mass spectrometry with atmospheric pressure chemical ionization].

A method for the determination of benzo[a]pyrene in foods was developed by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) based on isotope dilution and molecularly imprinted solid-phase extraction (MIP-SPE). The target analyte in samples was extracted with n-hexane after spiked with benzo[a]pyrene-d12, and purified using MIP-SPE to eliminate most of the coextracts. The separation of benzo[a]pyrene was carried out on an XBridge BEH C18 column with gradient elution of methanol and water. An atmospheric pressure chemical ionization (APCI) interface was used as the ion source and the analysis was performed in the multiple reaction monitoring (MRM) mode. Benzo[a]pyrene levels in the range of 0.07-50 µg/kg were measured accurately by this method, and the limit of quantification (LOQ) was 0.07 µg/kg. The average recoveries were between 86% and 104% with the relative standard deviations within 2.3%-14%. The method was sensitive and accurate, and it has been successfully applied to the measurement of benzo[a]pyrene in food samples.

[Fast determination of pericarpium papaveris illegally added in foods by TurboFlow online purification-ultra performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry].

A fast confirmation method was developed for the determination of the six markers of pericarpium papaveris, morphine, codeine, narcotine, papavarine, thebaine and protopine in foods, by TurboFlow online purification-ultra performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry (TF-UPLC-QTRAP MS). The sample was extracted with 0.10 mol/L HCl. After the procedure of removal of lipid with hexane, the extraction solution was analyzed by TF-UPLC-QTRAP MS. The main factors influencing the purification efficiency including TurboFlow column, mobile phase and elution solution were optimized. The six opium alkaloids were detected by positive electrospray ionization tandem mass spectrometry in the multiple reaction monitoring-information-dependent acquisition-enhanced product ion (MRM-IDA-EPI) mode, and quantified by solvent standard internal standard method. The limits of detection were 0.05-0.5 µg/kg and the limits of quantification were 0.2-2 µg/kg for the six opium alkaloids. The recoveries were in the range of 81.1%-98.6% with the relative standard deviations ranging from 2.9% to 15.7% (n=6). The method is sensitive and accurate, and has been successfully applied to the detection of pericarpium papaveris illegally added in foods.

[Simultaneous rapid determination of 12 microcystins and one nodularin in water by direct injection-ultra performance liquid chromatography-triple quadrupole mass spectrometry].

A rapid method was developed for the simultaneous determination of 12 microcystins (MCs) and one nodularin (NOD) in water by direct injection-ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). The water samples were first diluted with equal volume of methanol, and then filtered through polyether sulfone (PES) syringe filter. The filtrates were directly injected into the UPLC system. The separation of the analytes was carried out on an ACQUITY UPLC BEH 300 C18 column (100 mm×2.1 mm, 1.7 µm) with gradient elution using mobile phases of acetonitrile containing 0.1% (v/v) formic acid and 0.2% (v/v) formic acid aqueous solution. The 12 microcystins and one nodularin were detected by positive electrospray ionization in the multiple reaction monitoring (MRM) mode, and quantified by standard solvent external standard method. The limits of detection were 0.03-0.1 µg/L and the limits of quantification were 0.1-0.3 µg/L. The recoveries were in the range of 79.5%-123% with the relative standard deviations ranging from 1.0% to 20% (n=6). The method is simple, sensitive and accurate, and has been successfully applied to the detection of the 13 kinds of algae toxins in water.

[Determination of alpha-solanine, alpha-chaconine and solanidine in plasma and urine by ultra-performance liquid chromatography-triple quadrupole mass spectrometry].

An ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS) method was developed for the determination of alpha-solanine, alpha-chaconine and solanidine in plasma and urine. The sample was acidified with aqueous solution containing 2% (v/v if not specified) formic acid, and then cleaned-up by solid-phase extraction with a mixed-mode cation exchange (MCX) cartridge. The analysis of the glycoalkaloids was carried out on an Acquity UPLC BEH C18 column (50 mm x 2.1 mm, 1.7 microm) with gradient elution of acetonitrile (containing 0.1% formic acid) and H2O (containing 0.05% formic acid and 5.0 mmol/L ammonium acetate). The analytes were detected by positive electrospray ionization tandem mass spectrometry in MRM mode, and quantified by external matrix-matched standard calibration. The cycle time of each analysis was 5.5 min. The calibration curves were linear in the range of 0.3-100 ng/mL of the glycoalkaloids in plasma and urine. The correlation coefficients were 0.997-0.999. The limits of detection (S/N = 3) and quantitation (S/N = 10) were 0.1 ng/mL and 0.3 ng/mL. The average recoveries were 82%-112% and 96%-114% for the glycoalkaloids spiked in plasma and urine, respectively, with relative standard deviations of 4.0%-16% and 2.7%-17% (n = 6). The method is simple, accurate and sensitive to detect the glycoalkaloids in plasma and urine for both clinical and forensic purposes.

[Rapid simultaneous determination of 53 beta-lactam antibiotics and their metabolites in milk by ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry].

A rapid method for the determination of 53 beta-lactams and their metabolites residues in milk was developed by ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). The method was based on protein precipitation by adding equal quantity of acetonitrile, followed by the ultra filtration of the extracts. The analysis of the residues was achieved on an ACQUITY BEH C18 column with gradient elution using mobile phases of 0.1% formic acid in water and acetonitrile. The analytes were detected by positive electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode, and quantified by matrix-matched internal standard calibration. The cycle time of each analysis was 10 min. Under the optimum operation conditions, excellent linear dynamic range was observed from the quantification limits to 200 microg/kg with the correlation coefficients better than 0.991 1 for all the compounds. The average recoveries for all the beta-lactams and their metabolites ranged from 71% to 121% with the relative standard deviations of 1.7%-19% (n = 6). This method enabled the screening for more than 50 samples per day by one person and also showed good performance for quantitation, and allowed the determination of the beta-lactams and their metabolites residues in milk below the maximum residue levels (MRLs) according to the Bulletin No. 235 (2002) of Ministry of Agriculture, P. R. China and EU Regulation 2377/90/EC (2008 Revised).

[Determination of tetrodotoxin in seafood using graphitized carbon black clean-up with hilic ultra performance liquid chromatography-triple quadrupole mass spectrometry].

OBJECTIVE: To develop a rapid hilic ultra performance liquid chromatography (UPLC)-mass spectrum (MS)/MS method for determination of tetrodotoxin in seafood. METHODS: The sample of muscle and liver of puffer fish and nassarius were extracted with aqueous solution containing 0.2% (V/V) acetic acid (the extract of liver must be purified through HLB cartridge), and then cleanup of extract was accomplished by solid-phase extraction with a graphitized carbon black cartridge. The analysis of tetrodotoxin was carried out on a chromatographic column (Acquity UPLC BEH Amide, 100 mm×2.1 mm×1.7 µm) with gradient elution of 95% (V/V) acetonitrile-H2O both containing 0.1% (V/V) formic acid and 2.0 mmol/L ammonium formate, and detected by positive electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode, and quantified by matrix-match standard solution. RESULTS: The calibration curves were linear in the range of 30 - 10 000, 50 - 10 000 and 30 - 10 000 µg/kg of tetradotoxin in muscle and liver of puffer fish and in muscle of nassarius, respectively. The correlation coefficients were within 0.9963 - 0.9990. The limits of detection were 10, 20 and 10 µg/kg, and that of quantitation were 30, 50 and 30 µg/kg for muscle and liver of puffer fish and muscle of nassarius, respectively. The average recoveries were 81.5% - 93.1%, 82.3% - 106.0% and 83.5% - 95.2% for tetrodotoxin spiked in muscle and liver of puffer fish and in muscle of nassarius, respectively, with relative standard deviation (RSD) of 2.3% - 11%, 4.3% - 14.0% and 3.5% - 13.0% (n = 6). CONCLUSION: The method was simple, accurate and sensitive, and could be successfully applied to the measurement of tetrodotoxin in puffer fish and nassarius.

[Isolation and identification of shellfish toxins from contaminated blue mussel (Mytilus edulis) from the East China Sea].

OBJECTIVE: To investigate the contamination of shellfish poisoning of mussels, the poisonous constituents in that were isolated and identified. METHODS: The mussel tissue homogenate was extracted by acetone, and then the acetone extract was partitioned between diethyl ether and water. The ether extract was fractionated by column chromatography over silica gel and further isolated by semi-preparative RP-HPLC, monitored by ultra performance liquid chromatography coupled with triple quadrupole mass spectrometry. RESULTS: Four poisonous constituents were isolated. Two of them were elucidated as pectenotoxin-2 seco acid and 7-epi-pectenotoxin-2 seco acid, respectively, on the basis of mass spectral data and compared with the production of enzymatic hydrolysis of PTX-2, and others were identified as okadaic acid and dinophysistoxin-1 by UPLC-MS/MS analysis compared with standard substances. CONCLUSION: OA, DTX-1, 7-epi-PTX-2sa and PTX-2sa had been isolated from the mussel, respectively. The concentrations of free OA, DTX-1 and total OA in which were surpassed the maximum permitted levels in EU. OA and DTX-1 were confirmed to be the main toxins responsible for this DSP outbreak.

[Rapid simultaneous determination of 42 psychoactive drugs and their metabolites in human plasma and urine by ultra performance liquid chromatography-tandem mass spectrometry].

A rapid method for the simultaneous determination of 42 psychoactive drugs and their metabolites (barbitals, benzodiazepines, tricyclic antidepressants, phenothiazines, etc. ) in human plasma and urine was developed using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). After a simple protein precipitation step, the analysis of the drugs and the metabolites was achieved on an Acquity UPLC BEH C18 column by using the gradient elution with ammonium acetate and methanol-acetonitrile (1: 1, v/v) as the mobile phases, and detected by electrospray ionization-tandem mass spectrometry in the multiple reaction monitoring (MRM) mode operated simultaneously in both positive and negative modes by rapid switching, and quantified by matrix-match standard solution. The average recoveries were 60.2% - 125% and 64.5% - 126% for the drugs in plasma and urine except those of perphenazine, thioridazine and chloropromazine in plasma were 37.6% - 57.5%, 36.3% - 48.3%, 52.4% -67.4%, respectively; trazodone and diazepam in urine were 100% -142% and 108% - 177%, respectively. The relative standard deviations (RSDs) of all the drugs in plasma and urine were within 0.8% - 26% and 2.6%18% (n = 6), respectively. The detection limits of the 4 barbitals ranged from 20 to 100 mg/L and those of the other drugs ranged from 0.05 to 2.0 mg/L. The method is simple, selective and sensitive to detect drugs for both clinical and forensic purposes.