title: Development and validation of a UPLC-MS/MS method for the determination of 7-hydroxymitragynine, a μ-opioid agonist, in rat plasma and its application to a pharmacokinetic study.
abstract: A simple, sensitive and specific ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated to determine the concentrations of 7-hydroxymitragynine in rat plasma. Following a single-step liquid-liquid extraction of plasma samples using chloroform, 7-hydroxymitragynine and the internal standard (tryptoline) were separated on an Acquity UPLC(TM) BEH C18 (1.7 µm, 2.1 × 50 mm) column using an isocratic elution at a flow rate of 0.2 mL/min. The mobile phase consisted of 0.1% acetic acid in water and 0.1% acetic acid in acetonitrile (10:90, v/v). The run time was 2.5 min. The analysis was carried out under the multiple reaction-monitoring mode using positive electrospray ionization. Protonated ions [M + H](+) and their respective product ions were monitored at the following transitions: 415 → 190 for 7-hydroxymitragynine and 173 → 144 for the internal standard. The calibration curve was linear over the range of 10-4000 ng/mL (r(2) = 0.999) with a lower limit of quantification of 10 ng/mL. The extraction recoveries ranged from 62.0 to 67.3% at concentrations of 20, 600 and 3200 ng/mL). Intra- and inter-day assay precisions (relative standard deviation) were <15% and the accuracy was within 96.5-104.0%. This validated method was successfully applied to quantify 7-hydroxymitragynine in rat plasma following intravenous administration.
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, The University of Mississippi, University, MS, 38677, USA.
authors.forename: Pradeep K
authors.initials: PK
authors.lastname: Vuppala
authors.forename: Seshulatha
authors.initials: S
authors.lastname: Jamalapuram
authors.forename: Edward B
authors.initials: EB
authors.lastname: Furr
authors.forename: Christopher R
authors.initials: CR
authors.lastname: McCurdy
authors.forename: Bonnie A
authors.initials: BA
authors.lastname: Avery
mesh_terms: D000701:Analgesics, Opioid / Q000819:agonists; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D016014:Linear Models; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D046948:Secologanin Tryptamine Alkaloids / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000701:Analgesics, Opioid; D046948:Secologanin Tryptamine Alkaloids; C482678:7-hydroxymitragynine
keywords: 7-hydroxymitragynine; mass spectrometry; method validation; mitragyna speciosa; pharmacokinetics; rats
references.citation: J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Apr 1;891-892:1-6
references.pmid: 22406103
references.citation: J Med Chem. 2002 Apr 25;45(9):1949-56
references.pmid: 11960505
references.citation: Life Sci. 2004 Mar 12;74(17):2143-55
references.pmid: 14969718
references.citation: J Anal Toxicol. 2012 Nov-Dec;36(9):616-25
references.pmid: 23024321
references.citation: Clin Toxicol (Phila). 2008 Feb;46(2):146-52
references.pmid: 18259963
references.citation: Life Sci. 2005 Nov 19;78(1):2-7
references.pmid: 16169018
references.citation: Talanta. 2006 Jan 15;68(3):908-18
references.pmid: 18970409
references.citation: Neuropharmacology. 2008 Aug;55(2):154-65
references.pmid: 18550129
references.citation: Curr Top Med Chem. 2011;11(9):1165-75
references.pmid: 21050173
references.citation: Life Sci. 1997;60(12):933-42
references.pmid: 9061050
references.citation: Br J Pharmacol. 2006 Jan;147 Suppl 1:S153-62
references.pmid: 16402099
references.citation: Planta Med. 2005 Mar;71(3):231-6
references.pmid: 15770543
references.citation: Eur J Pharmacol. 2006 Nov 7;549(1-3):63-70
references.pmid: 16978601
references.citation: Nat Prod Commun. 2009 Jul;4(7):907-10
references.pmid: 19731590
references.citation: Planta Med. 1994 Dec;60(6):580-1
references.pmid: 17236085
references.citation: Clin Biochem Rev. 2009 Feb;30(1):19-34
references.pmid: 19224008
references.citation: J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Aug 15;855(2):255-61
references.pmid: 17574935
references.citation: J Fluoresc. 2012 Nov;22(6):1425-9
references.pmid: 22875639
references.citation: Biomed Chromatogr. 2013 Aug;27(8):1034-40
references.pmid: 23558564