Identification and quantitation of amphetamine, methamphetamine, MDMA, pseudoephedrine, and ephedrine in blood, plasma, and serum using gas chromatography-mass spectrometry (GC/MS).

Amphetamine, methamphetamine, MDMA, pseudoephedrine, and ephedrine are measured in blood, serum, and plasma using gas chromatography coupled to mass spectrometry (GC/MS). Following a simple liquid-liquid extraction, analytes are derivatized with heptafluorobutyric anhydride (HFBA) and 1 microL injected onto a HP-5MS 15-meter capillary column. Quantitation of each analyte is accomplished using a multi-point calibration curve and deuterated internal standards. The method provides a simple, robust, and reliable means to identify and measure these analytes.

Simultaneous determination and quantification of 12 benzodiazepines in serum or whole blood using UPLC/MS/MS.

The benzodiazepines are a large, commonly prescribed family of psychoactive drugs. We describe a method permitting the simultaneous detection and quantification of 12 benzodiazepines in serum using ultra-performance liquid chromatography (UPLC) coupled with tandem mass spectrometry (MS/MS). Analytes included alprazolam, temazepam, oxazepam, nordiazepam, clonazepam, lorazepam, diazepam, chlordiazepoxide, midazolam, flunitrazepam, 7-aminoclonazepam, and 7-aminoflunitrazepam. Sample pretreatment is simple consisting of protein precipitation using cold acetonitrile (ACN) mixed with the deuterated internal standards. Samples were capped and vortexed for 5 min to ensure maximum precipitation. Following a 5-min centrifugation period, 400 microL of the supernatant was transferred to a clean tube and evaporated down under nitrogen. Samples were reconstituted in 200 microL of a deionized water:ACN (80:20) mixture and transferred to appropriate vials for analysis. Chromatographic run time was 7.5 min, and the 12 analytes were quantified using multiple reaction monitoring (MRM) and 6-point calibration curves constructed for each analyte at concentrations covering a clinically significant range.

Identification and quantitation of cocaine, benzoylecgonine, and cocaethylene in blood, serum, and plasma using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS).

Cocaine is a widely abused stimulant. Numerous methods exist for the identification of the drug, or more commonly, one of its metabolites in urine. Urine testing is useful for most cases, but it is necessary to use other matrices in forensic situations and when subjects are anuric. We describe a novel method for the analysis of cocaine, benzoylecgonine, and cocaethylene in blood, serum, and plasma utilizing ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). Sample preparation has been minimized to a simple deproteinization step in which each specimen is mixed with an acetonitrile-internal standard mixture. The method has excellent precision across the linear range of 25-2,000 ng/mL for each analyte. With a run-time of 4 min, this method provides a significant improvement over traditional GC/MS methods.

Detection and quantification of cocaine and benzoylecgonine in meconium using solid phase extraction and UPLC/MS/MS.

The simultaneous determination and quantification of cocaine and its major metabolite, benzoylecgonine, in meconium using UPLC-MS/MS is described. Ultra-performance liquid chromatography (UPLC) is an emerging analytical technique which draws upon the principles of chromatography to run separations at higher flow rates for increased speed, while simultaneously achieving superior resolution and sensitivity. Extraction of cocaine and benzoylecgonine from the homogenized meconium matrix was achieved with a preliminary protein precipitation or protein 'crash' employing cold acetonitrile, followed by a mixed mode solid phase extraction (SPE). Following elution from the SPE cartridge, eluents were dried down under nitrogen, reconstituted in 200 microL of DI water:acetonitrile (ACN) (75:25), and injected onto the UPLC/MS/MS for analysis. The increased speed and separation efficiency afforded by UPLC, allowed for the separation and subsequent quantification of both analytes in less than 2 min. Analytes were quantified using multiple reaction monitoring (MRM) and six-point calibration curves constructed in negative blood. Limits of detection for both analytes were 3 ng/g and the lower limit of quantitation (LLOQ) was 30 ng/g.

Quantitation of morphine, codeine, hydrocodone, hydromorphone, oxycodone, oxymorphone, and 6-monoacetylmorphine (6-MAM) in urine, blood, serum, or plasma using liquid chromatography with tandem mass spectrometry detection.

Opiates and opioids currently rank among the most commonly prescribed pain medications. We describe two liquid chromatography tandem mass spectrometry (LC-MS-MS) methods for the quantification of morphine, codeine, hydrocodone, hydromorphone, oxycodone, oxymorphone, and 6-monoacetylmorphine (6-MAM). In the first, urine samples are pretreated by acidifying with sodium acetate containing appropriate deuterated internal standards and hydrolyzed with beta-glucuronidase. Samples are cooled, diluted with water, vortexed, centrifuged, and a portion is transferred to an autosampler vial for analysis. The second method allows for the measurement of the compounds in blood, serum, or plasma specimens. Analysis of these samples involves pretreatment with acetonitrile containing deuterated internal standards to deproteinize the sample, which is subsequently vortexed and centrifuged. A portion of the organic layer is transferred to a clean test tube, dried under nitrogen, and reconstituted with water for analysis. Quantitation of analytes is accomplished using a commercially available single-point calibrator (urine samples) or an in-house prepared six-point standard curve (blood samples).