title: A Validated Method for Quantification of Dolutegravir Using Ultra Performance Liquid Chromatography Coupled With UV Detection.
abstract: Dolutegravir (DTG) is an integrase strand transfer inhibitor, which is a newly approved antiretroviral drug used for the treatment of HIV-infected naive and experienced individuals. Many aspects of DTG pharmacology remain to be studied. Our aim was to develop and fully validate a robust analytical method for the quantification of DTG in plasma using liquid chromatography coupled with UV detection.
  
  
  
  A simple and rapid protein precipitation method was used for analyte extraction from 100 µL plasma. The separation was achieved on a C8 reverse-phase analytical column using a gradient elution with 50 mmol/L formic acid and 50 mmol/L ammonium acetate in water (mobile phase A), and 100% acetonitrile (mobile phase B) and at a flow rate of 0.3 mL/min and a total run time of 10 minutes. The detector wavelength was set at 258 nm.
  
  
  
  The linearity of the calibration curve (r > 0.9999, n = 6) was validated over a concentration range of 0.25-10 mcg/mL. Intra-assay variability ranged from 3.3% to 6.1% and inter-assay variability ranged from 4.5% to 5.7%. The overall accuracy ranged from 90.7% to 97.7% for the 3 different concentrations of quality control samples. Recovery efficiency of extraction ranged from 94.3%-100%. This method is highly selective with no interferences from commonly concomitant antiretroviral drugs or endogenous metabolites.
  
  
  
  The described method is simple, robust, selective, accurate, precise, and cost-effective. Thus, this assay can be readily transferred and implemented in clinical settings and used for pharmacokinetic studies and therapeutic drug monitoring programs.
authors: *Jefferiss Trust Laboratories, Faculty of Medicine, Imperial College London, St Mary's Hospital Campus; †Department of Molecular and Clinical Pharmacology, University of Liverpool; and ‡St Stephen's Centre, Chelsea and Westminster Foundation Trust, London, United Kingdom.
authors: Xinzhu
authors: X
authors: Wang
authors: Sujan Dilly
authors: SD
authors: Penchala
authors: Alieu
authors: A
authors: Amara
authors: Laura
authors: L
authors: Else
authors: Myra
authors: M
authors: McClure
authors: Marta
authors: M
authors: Boffito
mesh_terms: D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods*; D003362:Cost-Benefit Analysis; D016903:Drug Monitoring / Q000191:economics / Q000379:methods*; D019428:HIV Integrase Inhibitors / Q000493:pharmacokinetics*; D006575:Heterocyclic Compounds, 3-Ring / Q000493:pharmacokinetics*; D006801:Humans; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet / Q000191:economics / Q000379:methods; D000099226:Dolutegravir
chemical_list: D019428:HIV Integrase Inhibitors; D006575:Heterocyclic Compounds, 3-Ring; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D000099226:Dolutegravir