Quantitative analysis of mitragynine in human urine by high performance liquid chromatography-tandem mass spectrometry

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 877; no. 24; pp. 2499 - 2505
• Main Authors: Lu, Shijun, Tran, Buu N., Nelsen, Jamie L., Aldous, Kenneth M.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.08.2009; Elsevier
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Forensic; Fundamental and applied biological sciences. Psychology; General pharmacology; HPLC-MS/MS; Humans; Kratom; Medical sciences; Mitragynine; Pharmacology. Drug treatments; Secologanin Tryptamine Alkaloids - urine; Street Drugs - urine; Substance Abuse Detection - methods; Tandem Mass Spectrometry - methods; Urine drug test; Forensic; HPLC-MS/MS; Urine drug test; Mitragynine; Kratom; Human; Drug; Urine; Biological fluid; HPLC chromatography; Quantitative analysis
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2009.06.024

Mitragynine is the primary active alkaloid extracted from the leaves of Mitragyna speciosa Korth, a plant that originates in South-East Asia and is commonly known as kratom in Thailand. Kratom has been used for many centuries for their medicinal and psychoactive qualities, which are comparable to that of opiate-based drugs. Kratom abuse can lead to a detectable content of mitragynine residue in urine. Ultra trace amount of mitragynine in human urine was determined by a high performance liquid chromatography coupled to an electrospray tandem mass spectrometry (HPLC-ESI/MS/MS). Mitragynine was extracted by methyl t-butyl ether (MTBE) and separated on a HILIC column. The ESI/MS/MS was accomplished using a triple quadrupole mass spectrometer in positive ion detection and multiple reactions monitoring (MRM) mode. Ajmalicine, a mitragynine's structure analog was selected as internal standard (IS) for method development. Quality control (QC) performed at three levels 0.1, 1 and 5 ng/ml of mitragynine in urine gave mean recoveries of 90, 109, and 98% with average relative standard deviation of 22, 12 and 16%, respectively. The regression linearity of mitragynine calibration ranged from 0.01 to 5.0 ng/ml was achieved with correlation coefficient greater than 0.995. A detection limit of 0.02 ng/ml and high precision data within-day and between days analysis were obtained.