Transl Clin Pharmacol.  2014 Jun;22(1):35-42.

Development of a LC-MS/MS for Quantification of Venlafaxine in Human Plasma and Application to Bioequivalence Study in healthy Korean Subjects

• Kang HK
• Kang MA
• Kim HJ
• Park YS
• Kim SH
• Kang JS

Abstract

A simple, rapid and selective liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) is developed and validated for quantification of venlafaxine in human plasma with simple liquid-liquid extraction step consisted of extraction with ether and dichloromethane for 10 min and mixing with 1 M sodium acetate in human plasma using fluoxetine as an internal standard (IS). The analyte are separated using an isocratic mobile phase consisted of acetonitrile and 5 mM ammonium formate (4/3, v/v) on a isocratic YMC hydrosphere C18 (2.0x50.0 mm, 3.0 microm) column and analyzed by MS/MS in the multiple reaction monitoring (MRM) mode using the transitions of respective [M+H](+) ions, m/z 278.2-->260.3 and m/z 310.1-->148.1 for quantification of venlafaxine and IS, respectively. The standard calibration curves showed good linearity within the range of 1.0-200.0 ng/mL (r2=0.9986, 1/chi2 weighting). The lower limit of quantification (LLOQ) was 1.0 ng/mL. The retention times of venlafaxine and IS were 0.6 min and 0.7 min that means the potential for the high-throughput potential of the proposed method. In addition, no significant metabolic compounds were found to interfere with the analysis. Acceptable precision and accuracy were obtained for the concentrations over the standard curve range. The validated method was successfully applied to bioequivalence study after 75-mg of venlafaxine sustained-release (SR) capsule in 24 healthy Korean subjects.

Keyword

Venlafaxine; LC-MS/MS; Bioequivalence study; Pharmacokinetics

MeSH Terms

Ammonium Compounds
Calibration
Chromatography, Liquid
Ether
Fluoxetine
Humans
Ions
Liquid-Liquid Extraction
Methylene Chloride
Pharmacokinetics
Plasma*
Sodium Acetate
Tandem Mass Spectrometry
Therapeutic Equivalency*
Venlafaxine Hydrochloride
Ether
Fluoxetine
Ions
Methylene Chloride
Sodium Acetate

Figure

• 

  Figure 1. Chemical structures of (A) venlafaxine [(RS)-1-[2-dimethylamino-1-(4-methoxyphenyl)-ethyl]-cyclohexanol, MW=277.402 g/ mol, C17H27NO2] and (B) fluoxetine [IS, (RS)-N-methyl-3-phenyl-3-[4-(trifluoromethyl) phenoxy] propan-1-amine, MW=309.33 g/mol, C17H18F3NO] (from ChemSpider free chemical data base).

• 

  Figure 2. Chromatograms of (A) double blank plasma without venlafaxine and IS, (B) with IS (10.0 μg/mL), (C) with venlafaxine (LLOQ, 1.0 ng/mL) and IS (10.0 μg/mL), and (D) subject's plasma taken 10.0 h after a single oral dose of 75-mg venlafaxine capsule spiked with IS (10.0 μg/mL).

• 

  Figure 3. Full-scan mass spectra of precursor (A, C) and product ions (B, D) of venlafaxine (m/z 278.2→260.4) and fluoxetine (m/z 310.2→148.1), respectively.

• 

  Figure 4. Mean plasma concentrations versus time plots after a single oral dose of 75-mg venlafaxine SR capsule to the 24 healthy male subjects (○: reference, ●: test) (n=24, Mean±SD).

Reference

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