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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Liu, Liling Cui, Zhiyi Deng, Yuzhong Dean, Brian Hop, Cornelis E. C. A. Liang, Xiaorong |
| Description | Country affiliation: United States Author Affiliation: Liu L ( Drug Metabolism and Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, United States.); Cui Z ( Drug Metabolism and Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, United States.); Deng Y ( Drug Metabolism and Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, United States.); Dean B ( Drug Metabolism and Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, United States.); Hop CE ( Drug Metabolism and Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, United States.); Liang X ( Drug Metabolism and Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, United States. Electronic address: Liang.Xiaorong@gene.com.) |
| Abstract | A high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for the quantitative determination of NAD(+) in human whole blood using a surrogate analyte approach was developed and validated. Human whole blood was acidified using 0.5N perchloric acid at a ratio of 1:3 (v:v, blood:perchloric acid) during sample collection. 25µL of acidified blood was extracted using a protein precipitation method and the resulting extracts were analyzed using reverse-phase chromatography and positive electrospray ionization mass spectrometry. (13)C5-NAD(+) was used as the surrogate analyte for authentic analyte, NAD(+). The standard curve ranging from 0.250 to 25.0µg/mL in acidified human blood for (13)C5-NAD(+) was fitted to a 1/x(2) weighted linear regression model. The LC-MS/MS response between surrogate analyte and authentic analyte at the same concentration was obtained before and after the batch run. This response factor was not applied when determining the NAD(+) concentration from the (13)C5-NAD(+) standard curve since the percent difference was less than 5%. The precision and accuracy of the LC-MS/MS assay based on the five analytical QC levels were well within the acceptance criteria from both FDA and EMA guidance for bioanalytical method validation. Average extraction recovery of (13)C5-NAD(+) was 94.6% across the curve range. Matrix factor was 0.99 for both high and low QC indicating minimal ion suppression or enhancement. The validated assay was used to measure the baseline level of NAD(+) in 29 male and 21 female human subjects. This assay was also used to study the circadian effect of endogenous level of NAD(+) in 10 human subjects. |
| File Format | HTM / HTML |
| ISSN | 15700232 |
| Volume Number | 1011 |
| e-ISSN | 1873376X |
| Journal | Journal of Chromatography B |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2016-02-01 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Analytical Chemistry Chromatography, Liquid Methods Nad Blood Tandem Mass Spectrometry Adolescent Adult Aged Circadian Rhythm Female Humans Linear Models Male Middle Aged Reproducibility Of Results Sensitivity And Specificity Spectrometry, Mass, Electrospray Ionization Young Adult Journal Article |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Medicine Analytical Chemistry Clinical Biochemistry Biochemistry |
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