Development and validation of a liquid chromatography-mass spectrometry assay for the determination of opiates and cocaine in meconium.
A procedure based on liquid chromatography-mass spectrometry (LC-MS) is described for determination of 6-monoacetylmorphine, morphine, morphine-3-glucuronide, morphine-6-glucuronide, codeine, cocaine, benzoylecgonine and cocaethylene in meconium using nalorfine as the internal standard. The analytes are initially extracted from the matrix by methanol (6-monoacetylmorphine, morphine, codeine, cocaine, benzoylecgonine and cocaethylene) or 0.01 M ammonium hydrogen carbonate buffer (morphine-3-glucuronide, morphine-6-glucuronide). Subsequently a solid-phase extraction with Bondelut Certify columns (6-monoacetylmorphine, morphine, codeine, cocaine, benzoylecgonine and cocaethylene) or ethyl solid-phase extraction columns (morphine-3-glucuronide, morphine-6-glucuronide) was applied. Chromatography was performed on a C(8) reversed-phase column using a gradient of acetic acid 1%-acetonitrile as a mobile phase. Analytes were determined in LC-MS single ion monitoring mode with atmospheric pressure ionisation-electrospray (ESI) interface. The method was validated in the range 0.005-1.00 microg/g using 1 g of meconium per assay and applied to analysis of meconium in newborns to assess fetal exposure to opiates and cocaine.
Pichini
S
S
Clinical Biochemistry Department, Istituto Superiore di Sanità, 00161 Rome, Italy. pichini@iss.it
Pacifici
R
R
Pellegrini
M
M
Marchei
E
E
Pérez-Alarcón
E
E
Puig
C
C
Vall
O
O
García-Algar
O
O
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003042:Cocaine / Q000032:analysis*; D006801:Humans; D007231:Infant, Newborn; D008470:Meconium / Q000737:chemistry*; D009294:Narcotics / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D009294:Narcotics; D003042:Cocaine