Jenkins R.,PPD Inc |
Duggan J.X.,Boehringer Ingelheim Pharmaceuticals |
Aubry A.-F.,Bristol Myers Squibb |
Zeng J.,Bristol Myers Squibb |
And 11 more authors.
AAPS Journal | Year: 2015
This paper represents the consensus views of a cross-section of companies and organizations from the USA and Canada regarding the validation and application of liquid chromatography tandem mass spectrometry (LC-MS/MS) methods for bioanalysis of protein biotherapeutics in regulated studies. It was prepared under the auspices of the AAPS Bioanalytical Focus Group’s Protein LC-MS Bioanalysis Subteam and is intended to serve as a guide to drive harmonization of best practices within the bioanalytical community and provide regulators with an overview of current industry thinking on applying LC-MS/MS technology for protein bioanalysis. For simplicity, the scope was limited to the most common current approach in which the protein is indirectly quantified using LC-MS/MS measurement of one or more of its surrogate peptide(s) produced by proteolytic digestion. Within this context, we considered a range of sample preparation approaches from simple in-matrix protein denaturation and digestion to complex procedures involving affinity capture enrichment. Consideration was given to the method validation experiments normally associated with traditional LC-MS/MS and ligand-binding assays. Our collective experience, thus far, is that LC-MS/MS methods for protein bioanalysis require different development and validation considerations than those used for small molecules. The method development and validation plans need to be tailored to the particular assay format being established, taking into account a number of important factors: the intended use of the assay, the test species or study population, the characteristics of the protein biotherapeutic and its similarity to endogenous proteins, potential interferences, as well as the nature, quality, and availability of reference and internal standard materials. © 2014, American Association of Pharmaceutical Scientists.
PubMed | Tandem Labs and Genentech
Type: Journal Article | Journal: Biomedical chromatography : BMC | Year: 2015
An LC-MS/MS method for the determination of GDC-0980 (apitolisib) concentrations in dog plasma has been developed and validated for the first time to support pre-clinical drug development. Following protein precipitation with acetonitrile, the resulting samples were analyzed using reverse-phase chromatography on a Metasil AQ column. The mass analysis was performed on a triple quadruple mass spectrometer coupled with an electrospray interface in positive ionization mode. The selected reaction monitoring transitions monitored were m/z 499.3341.1 for GDC-0980 and m/z 507.3341.1 for IS. The method was validated over the calibration curve range 0.250-250ng/mL with linear regression and 1/x(2) weighting. Relative standard deviation (RSD) ranged from 0.0 to 10.9% and accuracy ranged from 93.4 to 113.6% of nominal. Stable-labeled internal standard GDC-0980-d8 was used to minimize matrix effects. This assay was used for the measurement of GDC-0980 dog plasma concentrations to determine toxicokinetic parameters after oral administration of GDC-0980 (0.03, 0.1 and 0.3mg/kg) to beagle dogs in a GLP toxicology study. Peak concentration ranged from 3.23 to 84.9ng/mL. GDC-0980 was rapidly absorbed with a mean time to peak concentration ranging from 1.3 to 2.4h. Mean area under the concentration-time curve from 0 to 24hours ranged from 54.4 to 542ngh/mL.
Hussey E.K.,Glaxosmithkline |
Kapur A.,Glaxosmithkline |
O'Connor-Semmes R.,Glaxosmithkline |
Tao W.,Glaxosmithkline |
And 4 more authors.
BMC Pharmacology and Toxicology | Year: 2013
Background: The sodium-dependent glucose co-transporter-2 (SGLT2) is expressed in absorptive epithelia of the renal tubules. Remogliflozin etabonate (RE) is the prodrug of remogliflozin, the active entity that inhibits SGLT2. An inhibitor of this pathway would enhance urinary glucose excretion (UGE), and potentially improve plasma glucose concentrations in diabetic patients. RE is intended for use for the treatment of type 2 diabetes mellitus (T2DM) as monotherapy and in combination with existing therapies. Metformin, a dimethylbiguanide, is an effective oral antihyperglycemic agent widely used for the treatment of T2DM.Methods: This was a randomized, open-label, repeat-dose, two-sequence, cross-over study in 13 subjects with T2DM. Subjects were randomized to one of two treatment sequences in which they received either metformin alone, RE alone, or both over three, 3-day treatment periods separated by two non-treatment intervals of variable duration. On the evening before each treatment period, subjects were admitted and confined to the clinical site for the duration of the 3-day treatment period. Pharmacokinetic, pharmacodynamic (urine glucose and fasting plasma glucose), and safety (adverse events, vital signs, ECG, clinical laboratory parameters including lactic acid) assessments were performed at check-in and throughout the treatment periods. Pharmacokinetic sampling occurred on Day 3 of each treatment period.Results: This study demonstrated the lack of effect of RE on steady state metformin pharmacokinetics. Metformin did not affect the AUC of RE, remogliflozin, or its active metabolite, GSK279782, although Cmax values were slightly lower for remogliflozin and its metabolite after co-administration with metformin compared with administration of RE alone. Metformin did not alter the pharmacodynamic effects (UGE) of RE. Concomitant administration of metformin and RE was well tolerated with minimal hypoglycemia, no serious adverse events, and no increase in lactic acid.Conclusions: Coadministration of metformin and RE was well tolerated in this study. The results support continued development of RE as a treatment for T2DM.Trial registration: ClinicalTrials.gov, NCT00376038. © 2013 Hussey et al.; licensee BioMed Central Ltd.
Jiang H.,Bristol Myers Squibb |
Zeng J.,Bristol Myers Squibb |
Kandoussi H.,Bristol Myers Squibb |
Liu Y.,Tandem Labs |
And 5 more authors.
Journal of Chromatography A | Year: 2012
A liquid-liquid extraction (LLE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods have been developed and validated for the quantification of BMS-790052 (daclastasvir) in human plasma and urine. The samples were extracted with methyl-t-butyl ether (MTBE) before analyzing by an API 4000 mass spectrometer which was operated in a multiple reaction monitoring (MRM) mode for detection of positively charged ions of BMS-790052 and its internal standard, 13C 10-BMS-790052. The standard curves ranged from 0.050 to 50.0ng/mL for BMS-790052 in human plasma, and 1.00-1000ng/mL in human urine. The intra-assay precision (%CV), based on four levels of analytical QCs (low, geometric mean, mid and high), was within 8.6%; inter-assay precision (%CV) was within 6.7% for both plasma and urine methods, and the mean assay accuracy (%Dev) was within ±3.0% for both plasma and urine methods. The ruggedness of this accurate, precise, and selective LLE-LC-MS/MS method has been demonstrated in the successful analysis of several thousand clinical study samples. © 2012 Elsevier B.V..
Tse S.,Pfizer |
Powell K.D.,Tandem Labs |
MacLennan S.J.,BioCryst Pharmaceuticals |
Moorman A.R.,Alta Vetta Pharmaceutical Consulting LLC |
And 2 more authors.
Journal of Pain Research | Year: 2012
Purpose: This study compared the pharmacokinetic profile, and systemic and local absorption of diclofenac, following dermal patch application and oral administration in Yorkshire- Landrace pigs. Patients and methods: Twelve anesthetized, female, Yorkshire-Landrace pigs were randomized to receive either the dermal patch (FLECTOR® patch, 10 × 14 cm; Alpharma Pharmaceuticals, a subsidiary of Pfizer Inc, New York, NY) or 50 mg oral diclofenac (Voltaren®; Novartis, East Hanover, NJ). Tissue (skin area of 2 × 2 cm and underlying muscles approximately 2-3 cm in depth) and blood (10 mL) samples were collected at timed intervals up to 11.5 hours after initial patch application or oral administration. The concentrations of diclofenac in plasma, skin, and muscle samples were analyzed using validated ultra performance liquid chromatography tandem mass spectrometric methods. Results: Peak systemic exposure of diclofenac was very low by dermal application compared with oral administration (maximum concentration [Cmax] values of 3.5 vs 9640 ng/mL, respectively). Absorption of diclofenac into underlying muscles beneath the dermal patch was sustained, and followed apparently zero-order kinetics, with the skin serving as a depot with elevated concentrations of diclofenac. Concentrations of diclofenac in muscles beneath the patch application site were similar to corresponding tissues after oral administration (Cmax values of 879 and 1160 ng/mL, respectively). In contrast to the wide tissue distribution of diclofenac after oral administration, dermal patch application resulted in high concentrations of diclofenac only on the treated skin and immediate tissue underneath the patch. Low concentrations of diclofenac were observed in the skin and muscles collected from untreated areas contralateral to the site of dermal patch application. Conclusion: Dermal patch application resulted in low systemic absorption and high tissue penetration of diclofenac compared with oral administration. © 2012 Tse et al, publisher and licensee Dove Medical Press Ltd.
Myler H.,Bristol Myers Squibb |
Felix T.,Tandem Labs |
Zhu J.,WuXi AppTec |
Hruska M.,Bristol Myers Squibb |
Piccoli S.P.,Bristol Myers Squibb
Bioanalysis | Year: 2014
Background: This article presents case study data demonstrating the importance of having a thorough understanding of matrix-interference in support of global clinical trials. Methods/Results: A ligand-binding assay used in the measurement of interferon lambda in human serum was transferred from the reference laboratory to a US-based comparator laboratory and then to a China-based comparator laboratory. The method was successfully validated at each laboratory, however, during cross-validation, there were notable differences, including 30-60% difference in incurred study sample results. The differences were attributed to matrix factors included in the serum pool used to prepare standards and quality controls. Newly procured serum (n = 75 individuals) was tested for assay interference. 12% contained either pre-existing antibodies (auto-antibodies) or were identified as pharmacokinetic assay outliers. Conclusion: Prescreening of serum to exclude reactive individuals resulted in successful cross-validation and the establishment of a high integrity pharmacokinetic assay in support of global clinical trials. © 2014 Future Science Ltd.
Meng M.,Tandem Labs |
Rohde L.,Tandem Labs |
Capka V.,Tandem Labs |
Capka V.,Astra Zeneca Pharmaceuticals LP |
And 2 more authors.
Journal of Pharmaceutical and Biomedical Analysis | Year: 2010
Traditional chiral chromatographic separation method development is time consuming even for an experienced chromatographer. This paper describes the application of computer software ACD Lab ® to facilitate the development of chiral separation for the quantitation of eszopiclone using LC-MS/MS technology. Assisted by ACD/Chrom Manager and LC Simulator software, the optimal chiral chromatographic development was completed within hours. The baseline chiral separation was achieved with a total cycle time of 3min. For sample extraction method development, a Waters Oasis ® Sorbent Selection Plate containing four different sorbents was utilized. Optimal conditions were determined using a single plate under various load, wash and elution conditions. This was followed by a GLP validation which demonstrated excellent intra- and inter-day accuracy and precision for the quantitation of eszopiclone in human plasma at 1.00-100ng/mL range using LC/MS/MS technology. This method was utilized to support multiple clinic bioequivalence studies. © 2010.
Meng M.,Tandem Labs |
Wang L.,Tandem Labs |
Voelker T.,Tandem Labs |
Reuschel S.,Tandem Labs |
And 2 more authors.
Bioanalysis | Year: 2013
In order to meet the drug discovery and development needs of pharmaceutical companies, CROs are constantly challenged by the requirements for rapid LC-MS/MS method development prior to method validation and sample analysis. In order to meet this challenge, a comprehensive method development program, nicknamed 'Amoeba™, which uses a series of written protocols for standardized and efficient method development was developed. In this paper, the genesis of the Amoeba method development program is elucidated in detail and a number of case studies are presented to showcase the execution of the Amoeba method development program. Using this program, the majority of the most critical information regarding the assay can be captured. While the Amoeba program has been proven to be effective, we recognize that the development of a robust bioanalytical method for use in pharmaceutical industry also requires the careful consideration of many critical parameters and the ability to identify and resolve potential issues. The refinement of the assay relies on further evaluation of several critical factors including, but not limited to, internal standard response, matrix effects (phospholipid or nonphospholipid related) and carryover. © 2013 Future Science Ltd.
Meng M.,Tandem Labs |
Reuschel S.,Tandem Labs |
Bennett P.,Thermo Fisher Scientific
Bioanalysis | Year: 2011
In 2007, the US FDA recommended that pharmaceutical companies and CROs conduct incurred sample reanalysis (ISR) following the analysis of study samples using validated methods. Between January 2008 and Deccember 2009, over 250 separate analytes were tested for ISR in our laboratory (a bioanalytical CRO). Among these, nine analytes initially failed for ISR. While thorough investigations were conducted to identify the root cause of ISR failure for each study, these investigations were often painfully tedious and very costly, both financially and in terms of project timelines. In this paper, three representative studies are presented to showcase the detailed investigation processes, methodologies and final conclusions of the ISR investigations. Additionally, all nine ISR failures are analyzed to identify trends or common elements of the studies or methods that might help identify potential problems before they occur. Furthermore, suggestions and recommendations to minimize future ISR failures are provided. © 2011 Future Science Ltd.
PubMed | Bristol Myers Squibb and Tandem Labs
Type: | Journal: Journal of pharmaceutical and biomedical analysis | Year: 2015
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.