Application Of Ultra-High Performance Liquid Chromatography-Tamdem Mass Sepctrometry In Analysis Of Small Electrical Appliance And Drug Metabolism

As its high separation ability, high sensitivity, wide application scope and specificity, liquid chromatography-mass spectrometry technology (LC-MS) had become more and more important for all kinds of analysis fields. The paper was doing the two following research using LC-MS:1. Determination of toxic and harmful substances in small electrical appliances products of PoHS instructions(1) The experiment established detection of hexabromocyclododecane (HBCD) in small household electrical appliances products by heating reflux extraction-ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Comparing the ultrasonic extraction, accelerated solvent extraction, soxhlet extraction and heating reflux extraction, the optimum pretreated method for extracting three isomerides of HBCD in small household electrical appliances products was heating reflux extraction. The experiment used toluene:methanol = 10:1 as extracting solvent and heating backflow 4 hours. The extract solution was dried by N2, dissolved by the initial mobile phase, vortexed, centrifuged and filmed. After the separation by BEH C18 column, the three isomerides of HBCD were detected by mass spectrometry in the negative electrospray ionization with multiple reaction monitoring (MRM) mode. The three compounds were separated effectively in 3 min. The linear ranges of the compounds were 1.6-32.4μg·mL-1 for HBCDs with correlation coefficient greater than 0.996. The limit of detection(LOD) and the limit of quantitation(LOQ) for HBCD were 0.1 and 0.5μg·mL-1, and the recoveries was 68%-75.2%. This method was successfully used in determination the real samples.(2) The experiment established simultaneously detection of bisphenol A(BPA), tetrabromobisphenol A(TBBPA), and perfluorooctanoic acid (PFOA) in small household electrical appliances products by accelerated solvent extraction (ASE)-UPLC-MS/MS. We designed L9(34) orthogonal table for ASE, and selected the optimum extraction conditions:extracting solvent was toluene:methanol = 10:1; the pressure was 1500psi; the temperature was 100℃; heating time was 5min; static extraction time was 20min; circular 3 times; and N2 purged time was 100s; the volume for washing was 20%. The extract solution was dried by N2, dissolved by the initial mobile phase, vortexed, centrifuged and filmed. After the separation by BEH C18 column, three compounds were detected by mass spectrometry in the negative electrospray ionization with MRM mode. The three compounds were separated effectively in 3 min. The linear ranges of the compounds were 1-100ng·mL-1,0.1-10μg·mL-1,10-1000ng-mL"1 for BPA, TBBPA, PFOA with correlation coefficient greater than 0.996. The LODs for the three compounds were 0.1,10, 1ng-mL-1 and the LOQs were 0.5,50,5ng·mL-1 individually. Through the external standard method to quantitative, the recoveries for BPA, TBBPA, PFOA were 84%-92%,76%-82%,72%-74% respectively with RSD lower than 5%. The method and the result were confirmed by liquid chromatography - ion trap - time of flight mass spectrometry (LC-IT-TOF). The results were good and the method was successfully used in determination the real samples.2. Established detection of naloxone, buprenorphine and nor-buprenorphine in human serum and determination naloxone and buprenorphine in tabletA simultaneous method was successfully established and validated for the separation and determination of buprenorphine (BP), its primary metabolite, nor-buprenorphine (NBP) and a proposed co-formulate, naloxone (NLX) in human serum. The method used buprenorphine-d4 (BP-D4), nor-buprenorphine-d3 (NBP-D3), naltrexone (NTX) as internal standards (ISs). 100μL of plasma sample fortified with the ISs was cleaned up by solid-phase extraction (SPE), and was then separated on a Waters AcquityTM BEH C18 column with gradient elution using methanol and water (containing 0.2% formic) at a flow rate of 0.25mL·min-1. The mass spectrometer was used for detection and was operated in the positive electrospray ionization with MRM mode. The three compounds were effectively separated in 4.5 min. The linear ranges of the compounds were 0.1-25,0.25-25 and 0.05-25ng·mL-1 for BP, NBP and NLX, respectively, with R≥0.9948. The method had high sensitivity (the LODs were 0.02,0.1 and 0.01ng·mL-1 and the LOQs were 0.05, 0.2 and 0.025ng·mL-1 for BP, NBP and NLX, respectively) and high recoveries (≥97.6%). The result was shown to be linear and satisfactorily met current acceptance criteria for validation of bioanalytical method:intra and inter assay precisions within the required limits of≤25% RSD. The LOQs fulfilled the LOQ requirements:precision≤25% RSD, and was fully validated according to the State Food and Drug Administration (SFDA) regulations. The results demonstrated that UPLC-MS/MS with SPE was a powerful detection tool and contributed to pharmaceutical analysis in biological matrices. The experiment had also established UPLC-TUV to detect BP, NLX in tablet. All the results were satisfied.