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Direct quantification of morphine glucuronides and free morphine in urine by liquid chromatography-tandem mass spectrometry

Koen Deventer (UGent) , OJ Pozo, Frans Delbeke (UGent) and Peter Van Eenoo (UGent)
(2012) FORENSIC TOXICOLOGY. 30(2). p.106-113
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Abstract
A simple and robust liquid chromatographic-tandem mass spectrometric method was developed and validated for the direct quantification of the structural isomers morphine-3-glucuronide (M3G), morphine-6-glucuronide (M6G), and morphine in urine. Method development showed that dilution of the urine samples was mandatory to obtain reproducible chromatography and that the use of deuterated internal standards was needed to compensate for ion suppression. Sample preparation consisted of a 20-fold dilution of urine into the internal standard solution. Chromatography was performed on a C-8-column using gradient conditions. The mobile phase consisted of water/methanol, both containing 0.1 % acetic acid and 1 mM ammonium acetate. Calibration curves were constructed between 0.05 and 2 mu g/ml. Validation data showed biases ranging from 1.5 to 5.2 % for M3G, -2.7 to -5.9 % for M6G, and 3.7 to 7.1 % for morphine. Imprecision never exceeded 5.2, 5.9, and 5.7 % for M3G, M6G, and morphine, respectively. The applicability of the method was checked by the analysis of 20 urine samples that were analyzed concurrently with a routine gas chromatography-mass spectrometry method. Results showed good correlation between the methods with a correlation coefficient of approximately 0.95.
Keywords
LC-MS, Doping, DIRECT-INJECTION, ELECTROSPRAY-IONIZATION, ENZYMATIC-HYDROLYSIS, PRESCRIPTION OPIOIDS, INCOMPLETE RECOVERY, LC-MS/MS, METABOLITES, VALIDATION, OPIATES, 6-GLUCURONIDE, Detection, Urine, Opiate, Morphine

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Chicago
Deventer, Koen, OJ Pozo, Frans Delbeke, and Peter Van Eenoo. 2012. “Direct Quantification of Morphine Glucuronides and Free Morphine in Urine by Liquid Chromatography-tandem Mass Spectrometry.” Forensic Toxicology 30 (2): 106–113.
APA
Deventer, Koen, Pozo, O., Delbeke, F., & Van Eenoo, P. (2012). Direct quantification of morphine glucuronides and free morphine in urine by liquid chromatography-tandem mass spectrometry. FORENSIC TOXICOLOGY, 30(2), 106–113.
Vancouver
1.
Deventer K, Pozo O, Delbeke F, Van Eenoo P. Direct quantification of morphine glucuronides and free morphine in urine by liquid chromatography-tandem mass spectrometry. FORENSIC TOXICOLOGY. 2012;30(2):106–13.
MLA
Deventer, Koen, OJ Pozo, Frans Delbeke, et al. “Direct Quantification of Morphine Glucuronides and Free Morphine in Urine by Liquid Chromatography-tandem Mass Spectrometry.” FORENSIC TOXICOLOGY 30.2 (2012): 106–113. Print.
@article{3128517,
  abstract     = {A simple and robust liquid chromatographic-tandem mass spectrometric method was developed and validated for the direct quantification of the structural isomers morphine-3-glucuronide (M3G), morphine-6-glucuronide (M6G), and morphine in urine. Method development showed that dilution of the urine samples was mandatory to obtain reproducible chromatography and that the use of deuterated internal standards was needed to compensate for ion suppression. Sample preparation consisted of a 20-fold dilution of urine into the internal standard solution. Chromatography was performed on a C-8-column using gradient conditions. The mobile phase consisted of water/methanol, both containing 0.1 \% acetic acid and 1 mM ammonium acetate. Calibration curves were constructed between 0.05 and 2 mu g/ml. Validation data showed biases ranging from 1.5 to 5.2 \% for M3G, -2.7 to -5.9 \% for M6G, and 3.7 to 7.1 \% for morphine. Imprecision never exceeded 5.2, 5.9, and 5.7 \% for M3G, M6G, and morphine, respectively. The applicability of the method was checked by the analysis of 20 urine samples that were analyzed concurrently with a routine gas chromatography-mass spectrometry method. Results showed good correlation between the methods with a correlation coefficient of approximately 0.95.},
  author       = {Deventer, Koen and Pozo, OJ and Delbeke, Frans and Van Eenoo, Peter},
  issn         = {1860-8965},
  journal      = {FORENSIC TOXICOLOGY},
  language     = {eng},
  number       = {2},
  pages        = {106--113},
  title        = {Direct quantification of morphine glucuronides and free morphine in urine by liquid chromatography-tandem mass spectrometry},
  url          = {http://dx.doi.org/10.1007/s11419-012-0135-8},
  volume       = {30},
  year         = {2012},
}

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