title: Multiplexed LC-MS/MS method for the simultaneous quantitation of three novel hepatitis C antivirals, daclatasvir, asunaprevir, and beclabuvir in human plasma.
abstract: Dual or triple combination regimens of novel hepatitis C direct-acting antivirals (DAA, daclatasvir, asunaprevir, or beclabuvir) provide high sustained virological response rates and reduced frequency of resistance compared to clinical monotherapy. To support pharmacokinetic (PK) assessments in clinical studies, a multiplexed liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantitation of daclatasvir, asunaprevir, beclabuvir (BMS-791325) and its active metabolite (BMS-794712) in human plasma was developed and validated. Human plasma samples were extracted with methyl-t-butyl ether followed by an LC-MS/MS analysis, which was conducted in a multiple reaction monitoring (MRM) mode. The lower limits of quantitation (LLOQ) were 1 ng/mL for daclatasvir, asunaprevir, and BMS-794712, and 2 ng/mL for beclabuvir. Intra-run precision (≤4.5% CV), inter-run precision (≤2.9% CV), and accuracy (±5.3% deviation) based on different concentration levels (low, geometric mean, mid and high) of the quality control samples (QCs) provided evidence of the methods accuracy and precision. Selectivity and matrix effect on LC-MS/MS detection, stability in plasma, and potential interference of coadministered drugs (ribavirin and interferon) were all evaluated and the results were acceptable. Method reproducibility was demonstrated by the reanalysis of a portion of study samples. The cross-validation results for QCs demonstrated the equivalency between this method and two single-analyte methods which were previously validated for quantitation of daclatasvir in human plasma. This approach of using a multiplexed LC-MS/MS method for the simultaneous quantitation of three DAAs is time- and cost-effective, and can maintain good data quality in sample analysis.
authors: Analytical and Bioanalytical Development, Bristol-Myers Squibb Co., Princeton, NJ 08540, United States. Electronic address: hao.jiang@bms.com.
authors: Hao
authors: H
authors: Jiang
authors: Analytical and Bioanalytical Development, Bristol-Myers Squibb Co., Princeton, NJ 08540, United States.
authors: Hamza
authors: H
authors: Kandoussi
authors: Analytical and Bioanalytical Development, Bristol-Myers Squibb Co., Princeton, NJ 08540, United States. Electronic address: jianing.zeng@bms.com.
authors: Jianing
authors: J
authors: Zeng
authors: Analytical and Bioanalytical Development, Bristol-Myers Squibb Co., Princeton, NJ 08540, United States.
authors: Jian
authors: J
authors: Wang
authors: Tandem Labs, West Trenton, NJ, United States.
authors: Roger
authors: R
authors: Demers
authors: Clinical Pharmacology and Pharmacometrics, Bristol-Myers Squibb Co., Princeton, NJ, United States.
authors: Timothy
authors: T
authors: Eley
authors: Clinical Pharmacology and Pharmacometrics, Bristol-Myers Squibb Co., Princeton, NJ, United States.
authors: Bing
authors: B
authors: He
authors: Discovery Chemistry Synthesis, Bristol-Myers Squibb Co., Wallingford, CT, United States.
authors: Richard
authors: R
authors: Burrell
authors: Discovery Chemistry Synthesis, Bristol-Myers Squibb Co., Wallingford, CT, United States.
authors: John
authors: J
authors: Easter
authors: Analytical and Bioanalytical Development, Bristol-Myers Squibb Co., Princeton, NJ 08540, United States.
authors: Pathanjali
authors: P
authors: Kadiyala
authors: Analytical and Bioanalytical Development, Bristol-Myers Squibb Co., Princeton, NJ 08540, United States.
authors: Janice
authors: J
authors: Pursley
authors: Tandem Labs, West Trenton, NJ, United States.
authors: Laura
authors: L
authors: Cojocaru
authors: Tandem Labs, West Trenton, NJ, United States.
authors: Chanda
authors: C
authors: Baker
authors: Tandem Labs, West Trenton, NJ, United States.
authors: John
authors: J
authors: Ryan
authors: Analytical and Bioanalytical Development, Bristol-Myers Squibb Co., Princeton, NJ 08540, United States.
authors: Anne-Françoise
authors: AF
authors: Aubry
authors: Analytical and Bioanalytical Development, Bristol-Myers Squibb Co., Princeton, NJ 08540, United States.
authors: Mark E
authors: ME
authors: Arnold
mesh_terms: D000998:Antiviral Agents / Q000097:blood / Q000737:chemistry* / Q000494:pharmacology; D001552:Benzazepines / Q000097:blood / Q000737:chemistry*; D002219:Carbamates; D002853:Chromatography, Liquid / Q000379:methods; D016174:Hepacivirus / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000097:blood* / Q000737:chemistry; D007211:Indoles / Q000097:blood / Q000737:chemistry*; D007372:Interferons / Q000097:blood / Q000737:chemistry; D007546:Isoquinolines / Q000097:blood / Q000737:chemistry*; D010949:Plasma / Q000737:chemistry*; D011759:Pyrrolidines; D015203:Reproducibility of Results; D012254:Ribavirin / Q000097:blood / Q000737:chemistry; D013449:Sulfonamides / Q000097:blood / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods; D014633:Valine / Q000031:analogs & derivatives
chemical_list: C587012:8-cyclohexyl-N-((dimethylamino)sulfonyl)-1,1a,2,12b-tetrahydro-11-methoxy-1a-((3-methyl-3,8-diazabicyclo(3.2.1)oct-8-yl)carbonyl)cycloprop(d)indolo(2,1-a)(2)benzazepine-5-carboxamide; D000998:Antiviral Agents; D001552:Benzazepines; D002219:Carbamates; D007093:Imidazoles; D007211:Indoles; D007546:Isoquinolines; D011759:Pyrrolidines; D013449:Sulfonamides; D012254:Ribavirin; D007372:Interferons; D014633:Valine; C549273:daclatasvir; C571889:asunaprevir
keywords: Asunaprevir; Beclabuvir (BMS-791325) and its metabolite; Daclatasvir; Human plasma; LC-MS/MS; Pharmacokinetics