[Determination of morphine, codeine and 6-monoacetylmorphine in saliva of substance-abuse patients using HPLC/MS methods].

Abstract

BACKGROUND/AIM:

Saliva represents an alternative specimen for substances abuse determination in toxicology. Hence, the aim of this study was to optimize a method for saliva specimen preparation for heroin metabolites, morphine and 6-monoacetylmorphine (6-mam), and codeine determination by liquid chromatography-mass spectrometry (LC/MS), and to apply this method on saliva samples taken from the patients.

METHODS:

Saliva specimen was prepared using liqiud/liquid extraction of morphine, codeine and 6-mam by mixture of chloroform and isopropanol (9 : 1; v/v). Extracts were analysed by HPLC/MS technique: separation column Waters Spherisorb 5 microm, ODS2, 4.6 x 100 mm; mobile phase: ammonium acetate : acetonitile (80 : 20; v/v), mobile phase flow rate 0.3 mL/min; mass detection range: 100-400 m/z. Regression and correlation analyses were performed with the probalility level of 0.05. Concentrations of morphine, codeine and 6-mam were determined in saliva samples of the patients with "opiates" in urine identified by the test strips.

RESULTS:

Calibration for each analysed substance was done in the concentration range from 0.1 to 1 mg/L and the coefficient of correlation was R2 > 0.99. We obtained following calibration curves: y = 385531x + 14584; y = 398036x + 31542; and y = 524162x - 27105, for morphine, codeine and 6-mam, respectively. Recovery for morphine and codeine determination was 99%, while for 6-mam it was 94%. Limits of detection and quantification of a proposed method were 0.01 mg/L and 0.05 mg/L, respectively. Concentration of morphine in the saliva of the heroin users ranged between 0.54 and 5.82 mg/L, concentration of codeine between 0.05 and 5.33, and 6-mam between 0.01 and 0.68 mg/L. A statistically significant correlation between codeine and 6-mam concentrations was obtained.

CONCLUSION:

A proposed HPLC/MS method for morphine, codeine and 6-mam determination in saliva is accurate, simple, cheap and suitable for routine analysis and monitoring of heroin abuse.

• Authors: Vesna Milovanović, Biljana Cirić, Jasna Milenković, Vesna Kilibarda, Marijana Curcić, Slavica Vucinić, Biljana Antonijević
• Journal: Vojnosanitetski pregled (Vojnosanit Pregl) Vol. 69 Issue 2 Pg. 141-6 (Feb 2012) ISSN: 0042-8450 [Print] Serbia
• PMID: 22500367 (Publication Type: Journal Article)

Chemical References

• Morphine Derivatives
• Morphine
• 6-O-monoacetylmorphine
• Codeine

Topics

• Chromatography, High Pressure Liquid (methods)
• Codeine (analysis)
• Humans
• Mass Spectrometry (methods)
• Morphine (analysis)
• Morphine Derivatives (analysis)
• Saliva (chemistry)
• Substance Abuse Detection