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Forensics                                                                                                                                              107







                                                          Fig. 1  Shimadzu UHPLC-MS/MS Nexera-8040 system












                                                                 Chromatographic conditions The analytical conditions allowed the chromatographic separation of  two couples of isomers: norephedrine and norpseudoephedrine;  ephedrine and pseudoephedrine (Fig. 2). A typical chromatogram of  the 58 compounds is presented in Fig. 3.(qualifier), respectively.   B







                                                              3. Results                                                          Fig. 2  Chromatograms obtained after an injection of a 5 µL whole blood extract spiked at 200 µg/L.                                      Order of retention - A: norephedrine and norpseudoephedrine /B: ephedrine and pseudoephedrine












                            UHPLC conditions (Nexera MP system, Fig. 1)





                              Column: Restek Pinnacle DB PFPP  50ʷ2.1 mm 1.9 µm Mobile phase A: 5mM Formate ammonium with 0.1% formic acid in water B: 90% CH3OH/ 10% CH3CN (v/v) with 0.1 % formic acid  Flow rate: 0.474 mL/min Time program: B conc. 15% (0-0.16 min) - 20% (1.77 min) - 90% (2.20 min) –                        100%(4.00 min) – 15% (4.10-5.30 min) Column temperature: 50 °C MS conditions (LCMS-8040, Fig. 1) Ionization: ESI, Positive MRM mode Ion source temperatures: Desolvation line: 300°C                                       Heater Block: 500°C Gases: Nebulization: 2.5 L/min            Drying: 10 L/min MRM Transitions: 2 Transitions per compound were dynamically  scanned for 1 min except pholcodine (2 min)  Pause time: 3 msec



























                                                                                      Finally, 5 µL were injected in the UHPLC-MS/MS system. The whole
                                                              amphetamines, as well as cocaine and 4 of its metabolites) and 18
                                                                     isotopically labeled internal standards (in order to improve method
                                                                         µL of acetonitrile. After a 15 s shaking, the mixture was placed at
                                                                       precision and accuracy) at 20 µg/L in acetonitrile (20 µL), and 200
                                                                                sesquihydrate) were added and the mixture was shaken again for
                                                                                  15 s and centrifuged for 10 min at 12300 g. The upper layer was
                                                                                    diluted (1/3; v/v) with a 5 mM ammonium formate buffer (pH 3).
                                                                  To 100 µL of sample (urine, whole blood or plasma) were added
                                                                           -20°C for 10 min. Then approximately 50 mg of QuEChERS salts
                                                                isotopically labeled internal standards (designed with *) (Table1).
                                                            This method involves 40 compounds of interest (13 opiates, 22
                                                                                                     -6-monoacetylmorphine*  -Dextromethorphan  -Dihydrocodeine*  -Ethylmorphine  -Hydrocodone  -Hydromorphone  -Methylmorphine*  -Morphine*  -Naloxone*  -Naltrexone*  -Noroxycodone*  -Oxycodone*  -Pholcodine
                                            1  CHU Limoges, Department of Pharmacology and Toxicology, Unit of clinical and forensic toxicology, Limoges, France ;
                                                    *Part of this work was presented in a poster session of the 62 th  ASMS Conference, 15 - 19 June, 2014, Baltimore.
                                                                                                Opiates
                                           Sylvain DULAURENT 1 , Mikaël LEVI 2 , Jean-michel GAULIER 1 , Pierre MARQUET 1,3  and Stéphane MOREAU 2
                                              2  Shimadzu France SAS,  Le Luzard 2, Boulevard Salvador Allende, 77448 Marne la Vallée Cedex 2
                                   plasma and urine by UHPLC-MS/MS using a QuEChERS sample preparation
                                                         2. Methods and Materials  (MgSO4/NaCl/Sodium citrate dehydrate/Sodium citrate   acquisition method lasted 5.5 min.
                               Determination of opiates, amphetamines and cocaine in whole blood,


                                                                                                Amphetamines or related compounds  -2-CB  -2-CI  -4-MTA  -Ritalinic acid  -Amphetamine*  -BDB  -Ephedrine*  -MBDB  -m-CPP  -MDA*  -MDEA*  -MDMA*  -MDPV  -Mephedrone  -Metamphetamine*  -Methcathinone  -Methiopropamine  -Methylphenidate  -Norephedrine  -Norfenfluramine  -Norpseudoephedrine  -Pseudoephedrine Table 1 list of analyzed compounds with their associate internal standard (*)
























           global w430×h280                      3  Univ Limoges, Limoges, France  1. Introduction  The determination of drugs of abuse (opiates, amphetamines,  cocaine) in biological fluids is still an important issue in toxicology,  in cases of driving under the influence of drugs (DUID) as well as  in forensic toxicology. At the end of the 20th century, the  analytical methods able to determine these three groups of  narcotics were mainly based on a liquid-liquid-extraction with  derivatization followed by GC-MS. Then LC-MS/MS was proposed,  coupled with off-line sample preparation. Recently, on-line  Solid-Phase-Extraction coupled with UHPLC-MS/MS was described,  but in our hands it gave rise to significant carry-over after highly  co








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