Simultaneous determination of morphine, codeine, 6‐acetylmorphine, cocaine and benzoylecgonine in hair by liquid chromatography/electrospray ionization tandem mass spectrometry

Abstract

A fast and sensitive liquid chromatography/triple quadrupole tandem mass spectrometry (LC/MS/MS) method was developed for the simultaneous determination of morphine, codeine, 6-acetylmorphine (6-AM), cocaine and benzoylecgonine (BE) in hair. Pulverized hair samples were extracted with methanol, and a 50 microL supernatant aliquot was injected into the LC/MS/MS system. Chromatography was performed with an XBridge phenyl column (3.5 microm particle size, 4.6 x 150 mm), and the mobile phase was composed of methanol and 10 mM ammonium acetate adjusted to pH 4.00 with 99% formic acid (95:5, v/v). A separation run with isocratic elution was completed in 10 min at a flow rate of 500 microL/min. Positive electrospray ionization and multiple reaction monitoring (MRM) with one precursor ion/product ion transition were used for the identification of each analyte. Deuterated analogues as internal standards were used for quantification and qualification. Linearity was established in the concentration range of 100-3000 pg/mg. The limits of detection were 10 pg/mg for morphine, codeine and 6-AM; and 1 pg/mg for cocaine and BE. The precision and accuracy were determined by spiking hair samples at six concentration levels. For all analytes, the relative standard deviations of intra- and inter-day precision were 0.1-6.3% and 1.5-10.6%, respectively. The accuracy ranged from 92.7 to 109.7%. The validated LC/MS/MS method was successfully applied to the analysis of 79 authentic hair samples.

On-line solid phase extraction coupled with liquid chromatography tandem mass spectrometry for the determination of codeine in human plasma

“…This method awards compensation over those previously reported [6][7][8][9][10][11][12][13]23 in terms of time per sample clean-up, improved separation speed by monolithic column and high-throughput due to fast chromatographic run time synchronized to sample extraction (3.0 min). On-line SPE-LC-MS/MS method allowed a rapid assay development and sample processing.…”

“…Inter-day accuracy ranged from 0.03 to 3.80%, and inter-day precision ranged from 4.35 to 7.25, as shown in Table 2. Although, the toxicological and forensic methods [6][7][8][9][10][11][12][13] described here are sufficiently precise and accurate for the measurement of codeine in human plasma, this current method showed excellent results, with intra and inter-day accuracy and precision less than 10% and within the ranges specified by ANVISA guideline. 17 Other automated and pharmacokinetics determination of the codeine 18-23 also failed to achieve intra-day precision less than 5% as was obtained in this study.…”

“…Most forensic application methods determined codeine with long chromatographic run time. [6][7][8][9][10][11][12][13] Therefore, mobile phase optimization is important to improve peak shape and sensitivity detection as well as codeine and tramadol chromatographic run time reduction (IS).…”

“…Several studies have demonstrated that it is still a drug of choice for treating acute and chronic pain. [1][2][3][4][5] Although, many bioanalytical methods have been developed for determination of codeine by LC-MS/MS and CG-MS, such as toxicological and forensic aplicattions, [6][7][8][9][10][11][12][13] most of these methods are based on off-line liquid-liquid or solid phase extraction (SPE). These procedures require complex steps and laborious repetitive work for sample clean-up, which may not meet the requirement for high-throughput, robustness and speed in bioanalysis and pharmacokinetic studies.…”

Improved forensic hair evidence for drugs of abuse by mass spectrometry

“…After the introduction of DESI, many ambient ionization techniques have been developed, which have been classified into seven categories by Monge et al [50] based on their desorption/ionization mechanism, although other subdivisions have also been proposed [51][52][53] :…”