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Enzyme-linked immunosorbent assay for TA-2005-glucuronide in human plasma
M Matsukawa1, K Takeda, H Shima
1Analytical Research Laboratory, Tanabe Seiyaku Co., Ltd., Osaka, Japan.
Journal of Pharmaceutical and Biomedical Analysis
|June 25, 1998
Summary
A new enzyme-linked immunosorbent assay (ELISA) accurately measures TA-2005-glucuronide, a key metabolite of the drug TA-2005, in human plasma. This sensitive method is valuable for pharmacokinetic studies of TA-2005.
Area of Science:
- Pharmacology and Toxicology
- Analytical Chemistry
- Biochemistry
Background:
- TA-2005 is a novel adrenergic beta-receptor agonist.
- TA-2005-glucuronide is a primary metabolite requiring sensitive quantification.
- Existing methods may lack the sensitivity or specificity for pharmacokinetic analysis.
Purpose of the Study:
- To develop and validate a sensitive enzyme-linked immunosorbent assay (ELISA) for TA-2005-glucuronide.
- To enable direct measurement of TA-2005-glucuronide in human plasma without deconjugation.
- To apply the assay for pharmacokinetic evaluation of TA-2005 in human subjects.
Main Methods:
- Competitive ELISA format using anti-TA-2005-glucuronide antiserum.
- Hapten conjugation to bovine serum albumin (BSA) and beta-D-galactosidase via N-hydroxysuccinimide ester method.
- Double antibody technique for separation; C18 cartridge pretreatment for sample cleanup.
Main Results:
- Sensitive standard curve observed in the range of 30 pg-3 ng/mL using 25 µL human plasma.
- Low inter-day (7.0-17.5%) and intra-assay (1.0-11.7%) variations with high recoveries (95.5-123.3%).
- Minimal cross-reactivity with related compounds, with effective separation from TA-2005 post-pretreatment.
Conclusions:
- The developed ELISA is a sensitive and reliable method for quantifying TA-2005-glucuronide in human plasma.
- The assay is suitable for direct measurement, simplifying sample processing.
- This method is applicable and valuable for pharmacokinetic studies of TA-2005 in humans.