Quantitative Bioanalytical Methods Validation and Implementation: Best Practices for Chromatographic and Ligand Binding Assays

Viswanathan, C. T.; Bansal, Surendra K.; Booth, Brian; DeStefano, Anthony J.; Rose, Mark J.; Sailstad, Jeffrey; Shah, Vinod P.; Skelly, Jerome P.; Swann, Patrick G.; Weiner, Russell

doi:10.1007/s11095-007-9291-7

Cited by 690 publications

(506 citation statements)

References 3 publications

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“…4); the mean matrix factors ranged from 88 to 101% in human plasma (CVs, 7 -9%), and from 93 to 101% in rat plasma (CVs, 6.0 -7.0%). Precision and accuracy values were within acceptable limits ( [18,21], US Food and Drug Administration. Guidance for industry.…”

Section: Validationmentioning

confidence: 77%

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Quantitation of imidazo[1,2‐a]quinoxaline derivatives in human and rat plasma using LC/ESI‐MS

Khier

Moarbess²,

Deleuze‐Masquéfa³

et al. 2009

J of Separation Science

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Original PaperQuantitation of imidazo[1,2-a]quinoxaline derivatives in human and rat plasma using LC/ESI-MS Since several years, our group developed quinoxalinic compounds. Among the synthesized compounds, in the imidazo[1,2-a]quinoxaline series, EAPB0203 has shown interesting activities both on melanoma and lymphoma. The structure of EAPB0203 has been modulated and a new compound, EAPB0503, exhibits an in vitro cytotoxic activity on melanoma cancer cell line 7 -9 times higher than EAPB0203. We validated an LC/ESI-MS method to simultaneously quantify EAPB0503 and its metabolite EAPB0603 in human and rat plasma. Chromatography was performed on a C8 Zorbax eclipse XDB column with a mobile phase consisting of acetronitrile and formate buffer gradient elution. LC-MS data were acquired in SIM mode at m/z 305, 291, and 303 for EAPB0503, EAPB0603, and the internal standard, respectively. The drug/ internal standard peak area ratios were linked via quadratic relationships to concentrations (low range: 5 -300 lg/L, high range: 100 -1000 lg/L). The method is precise (precision, f14%) and accurate (recovery, 92 -113%). Mean extraction efficiencies, A72% for each analyte, were obtained. The lower LOQs were 5 lg/L. This highly specific and sensitive method was successfully used to investigate plasma concentrations of EAPB0503 and EAPB0603 in a pharmacokinetic study carried out in rat and would also be useful in clinical trials at a later stage.Keywords: Human and rat matrices / Imidazo[1,2-a]quinoxaline / LC/ESI-MS / Lymphoma / Melanoma /

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Section: Validationmentioning

confidence: 77%

“…The matrix effect (i.e., suppression or enhancement of ionization of analytes by the presence of matrix components) was assessed by calculating the matrix factor [18]. It was defined as a ratio of the analyte peak response in the presence of matrix ions to the analyte peak response in the absence of matrix ions.…”

Section: Matrix Effect and Selectivitymentioning

confidence: 99%

Quantitation of imidazo[1,2‐a]quinoxaline derivatives in human and rat plasma using LC/ESI‐MS

Khier

Moarbess²,

Deleuze‐Masquéfa³

et al. 2009

J of Separation Science

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“…Those acceptance limits could be, for example, the limits chosen for assessing the validity of the method, or two or three times the intermediate precision SD of the method. The acceptance limits shown in Figures 3A through D are settled by allowing a degradation of maximum Ϫ30%, i.e., a minimum recovery of 70% (dashed line) (14). The maximum storage times when using the Liaison method with either Dry tubes or EDTA ones depicted in Figures 3A and B are the shortest and are estimated at 9 and 2 mo, respectively.…”

Section: Resultsmentioning

confidence: 98%

Estimation of the Stability of Parathyroid Hormone when Stored at −80°C for a Long Period

Cavalier

Hubert

Krzesinski

et al. 2009

Clinical Journal of the American Society of Nephrology

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Background and objectives: Stability of parathyroid hormone (PTH) at ؊80°C for long storage periods has never been studied. This can be of importance for the conclusions of studies where blood banks have been constituted. The study's aim was to evaluate stability of PTH when stored as serum or plasma EDTA samples at ؊80°C.Design, setting, participants & measurements: Samples were collected from 16 chronic hemodialysis patients using EDTA and gel-separator tubes. Plasma and serum were aliquoted; one aliquot was assayed with Elecsys and Liaison methods to determine the "baseline" values and another aliquot after 1, 3, 6, and 12 mo. The factors "method," "tubes," "subjects," and "time" were included in a mixed linear model to evaluate their effects on measured PTH values. The prediction interval methodology was used to assess where a future result could be obtained with a defined probability.Results: With the Liaison method, the maximum storage times with either dry or EDTA tubes were estimated to be 9 and 2 mo, respectively. With the Elecsys method, samples could be stored at least 2 yr with acceptable level of degradation.Conclusion: PTH stability at ؊80°C is not infinite. Maximum storage time and acceptance limits (30%) were defined, showing that with one method, samples should be stored for not more than 2 mo, whereas the other could be stored for up to 2 yr. With any PTH assay, the maximum storage time should be evaluated to ascertain that samples will keep their initial reactive profile after prolonged storage periods.

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“…Because deuterium-labeled analogs of compounds risk having a small amount of signal associated with the non-deuterium-labeled drug, we chose to minimize the concentrations of the internal standards. The standards used in this study had an isotopic purity of 90.95% D 8 Table I summarizes the data collected during the method validation experiments, which followed FDA guidance (35). Values were deemed acceptable based on % RSD and % error of ≤15% for all QC points except LOQ, where ≤20% was allowed (35).…”

Section: Methods Developmentmentioning

confidence: 99%

“…The standards used in this study had an isotopic purity of 90.95% D 8 Table I summarizes the data collected during the method validation experiments, which followed FDA guidance (35). Values were deemed acceptable based on % RSD and % error of ≤15% for all QC points except LOQ, where ≤20% was allowed (35). All calibration curves contained at least 5 points in the range of 5-100 ng/mL in brain tissue homogenate, and achieved R 2 > 0.99 for methylone, mephedrone, and MDPV using deuterium-labeled analogs as internal standards.…”

Section: Methods Developmentmentioning

confidence: 99%

Quantification of Synthetic Cathinones in Rat Brain Using HILIC–ESI-MS/MS

et al. 2016

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The abuse of synthetic cathinones, formerly marketed as "bath salts", has emerged over the last decade. Three common drugs in this class include 3,4-methylenedioxypyrovalerone (MDPV), 4-methylmethcathinone (mephedrone), and 3,4-methylenedioxymethcathinone (methylone). An LC-MS/MS method has been developed and validated for the simultaneous quantification of MDPV, mephedrone, and methylone in brain tissue. Briefly, MDPV, mephedrone, methylone, and their deuterium-labeled analogs were subjected to solid phase extraction (SPE) and separated using an HILIC Silica Column. The HPLC was coupled to a Shimadzu IT-TOF (ion trap-time of flight) system with the electrospray source running in positive mode (+ESI). The method was validated for precision, accuracy, and extraction efficiency. All inter-day and intra-day % RSD (percent relative standard deviation) and % error values were less than 15% and extraction efficiency exceeded 80%. These conditions allowed for limits of detection of 1ng/mL for MDPV, and 5 ng/mL for both mephedrone and methylone. The limits of quantification were determined to be 5ng/mL for MDPV and 10 ng/mL for mephedrone and methylone. The method was utilized to evaluate the pharmacokinetics of these drugs in adult male rats following administration of a drug cocktail including MDPV, mephedrone, and methylone. All three compounds reached peak concentrations in the brain within 15 min. Although methylone and mephedrone were administered at the same dose, the peak concentration (C max ) of mephedrone in the brain was significantly higher than that for methylone, as was the area under the curve (AUC). In summary, this quick and sensitive method for measuring synthetic cathinones may be used for future pharmacokinetic investigations of these drugs in target tissue.

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Quantitative Bioanalytical Methods Validation and Implementation: Best Practices for Chromatographic and Ligand Binding Assays

Cited by 690 publications

References 3 publications

Quantitation of imidazo[1,2‐a]quinoxaline derivatives in human and rat plasma using LC/ESI‐MS

Quantitation of imidazo[1,2‐a]quinoxaline derivatives in human and rat plasma using LC/ESI‐MS

Estimation of the Stability of Parathyroid Hormone when Stored at −80°C for a Long Period

Quantification of Synthetic Cathinones in Rat Brain Using HILIC–ESI-MS/MS

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