Determination of 11-nor-Delta(9)-tetrahydrocannabinol-9-carboxylic acid inurine using high-performance liquid chromatography and electrospray ionization mass spectrometry
T. Breindahl et K. Andreasen, Determination of 11-nor-Delta(9)-tetrahydrocannabinol-9-carboxylic acid inurine using high-performance liquid chromatography and electrospray ionization mass spectrometry, J CHROMAT B, 732(1), 1999, pp. 155-164
High-performance liquid chromatography with electrospray ionization mass sp
ectrometry was used to determine 11-nor-Delta(9)-tetrahydrocannabinol-9-car
boxylic acid (THC-COOH) in urine. After basic hydrolysis of conjugates, the
compound was extracted using SPEC-PLUS-3ML-C-18 solid-phase extraction col
umns. A deuterium labelled internal standard (d(3)-THCCOOH) was added prior
to hydrolysis. Separation was performed on a reversed-phase Zorbax Eclipse
XDB-C-8 analytical column (150x3.0 mm LD.) using a gradient program from 6
0 to 80% acetonitrile (4 mM formic acid) at a flow-rate of 0.5 ml/min. The
compounds were detected by single ion monitoring of m/z 345 and m/z 348 for
the protonated molecules [THC-COOH+H](+) and [d(3)-THC-COOH+H](+), respect
ively. The precision and accuracy were tested on spiked urine samples in th
e range 2.5-125 ng/ml. The mean recovery was 95% (n=58), coefficients of va
riations were 2.2-4.3% and the limit of detection 2 ng/ml. Diagnostic quali
fying ions of THC-COOH (m/z 327 and m/z 299) and d(3)-THC-COOH (m/z 330) we
re generated using up-front collision-induced dissociation. The relative io
n intensities in clinical samples (n=21) were within +/-20% deviation compa
red with standards. Using this tolerance and the presence of the ions m/z 3
27 and m/z 299 at the correct retention times as the acceptance criteria fo
r identification of THC-COOH positive samples, the limit of detection was 1
5 ng/ml. The LC-MS method complies with the current recommendations on drug
s of abuse testing, in which mass spectrometric detection is emphasized. (C
) 1999 Elsevier Science B.V. All rights reserved.