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Monitoring of the non-steroid anti-inflammatory drug indomethacin: Development of immunochemical methods for its purification and detection
Year:
2011
Authors :
Altstein, Miriam
;
.
Skalka, Nir
;
.
Volume :
400
Co-Authors:
Skalka, N., Department of Entomology, Institute of Plant Protection, Volcani Center, Bet Dagan 50250, Israel, Department of Anatomy and Anthropology, Sackler School of Medicine, Tel-Aviv University, Ramat-Aviv, Tel-Aviv 69978, Israel
Krol, A., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot 76703, Israel
Schlesinger, H., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot 76703, Israel
Altstein, M., Department of Entomology, Institute of Plant Protection, Volcani Center, Bet Dagan 50250, Israel
Facilitators :
From page:
3491
To page:
3504
(
Total pages:
14
)
Abstract:
The present research focused on the development of an immunoassay and an immunochemical sol-gel-based immunoaffinity purification (IAP) method for purification and detection of the non-steroid anti-inflammatory drug (NSAID) indomethacin (IMT). A polyclonal antibody (Ab) for IMT was generated, and two sensitive microplate assays for the detection of IMT were developed (termed OV and HRP formats), based on the enzyme-linked immunosorbent assay (ELISA) method. The limits of detection of the assays were 15±1.25 ng mL-1 (n∈=∈50) and 12±0.17 ng mL-1 (n∈=∈4) for the OVA and HRP formats, respectively. The Abs exhibited slight cross-reactivity with other NSAIDs. The Abs were also used to develop a sol-gel-based IAP method for clean-up and concentration of IMT. Several sol-gel formats with various amounts of antibodies were examined; the best and most reproducible format was at a TMOS:HCl molar ratio of 1:6 in which 120 μL of IMT Abs was entrapped. The binding capacity under these conditions was ca. 100 to 250 ng of IMT with very low non-specific binding (less than 5% of the applied amount). The sol-gel IAP method, combined with solid-phase extraction, successfully eliminated serum interference to a degree that enabled analysis of spiked serum samples by ELISA. The method was also found to be fully compatible with subsequent chemical analytical methods, such as liquid chromatography followed by mass spectrometry. The approaches developed in this study form a basis for analysis of IMT in biological samples in order to monitor their pharmacokinetic properties, and may be further used to study population exposure to IMT, and to monitor the occurrence of IMT contamination in water samples. © 2011 Springer-Verlag.
Note:
Related Files :
Blood
Body Fluids
Microarray analysis
nonsteroid antiinflammatory agent
Residue monitoring
standard
Show More
Related Content
More details
DOI :
10.1007/s00216-011-5027-y
Article number:
0
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
25777
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:17
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Scientific Publication
Monitoring of the non-steroid anti-inflammatory drug indomethacin: Development of immunochemical methods for its purification and detection
400
Skalka, N., Department of Entomology, Institute of Plant Protection, Volcani Center, Bet Dagan 50250, Israel, Department of Anatomy and Anthropology, Sackler School of Medicine, Tel-Aviv University, Ramat-Aviv, Tel-Aviv 69978, Israel
Krol, A., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot 76703, Israel
Schlesinger, H., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot 76703, Israel
Altstein, M., Department of Entomology, Institute of Plant Protection, Volcani Center, Bet Dagan 50250, Israel
Monitoring of the non-steroid anti-inflammatory drug indomethacin: Development of immunochemical methods for its purification and detection
The present research focused on the development of an immunoassay and an immunochemical sol-gel-based immunoaffinity purification (IAP) method for purification and detection of the non-steroid anti-inflammatory drug (NSAID) indomethacin (IMT). A polyclonal antibody (Ab) for IMT was generated, and two sensitive microplate assays for the detection of IMT were developed (termed OV and HRP formats), based on the enzyme-linked immunosorbent assay (ELISA) method. The limits of detection of the assays were 15±1.25 ng mL-1 (n∈=∈50) and 12±0.17 ng mL-1 (n∈=∈4) for the OVA and HRP formats, respectively. The Abs exhibited slight cross-reactivity with other NSAIDs. The Abs were also used to develop a sol-gel-based IAP method for clean-up and concentration of IMT. Several sol-gel formats with various amounts of antibodies were examined; the best and most reproducible format was at a TMOS:HCl molar ratio of 1:6 in which 120 μL of IMT Abs was entrapped. The binding capacity under these conditions was ca. 100 to 250 ng of IMT with very low non-specific binding (less than 5% of the applied amount). The sol-gel IAP method, combined with solid-phase extraction, successfully eliminated serum interference to a degree that enabled analysis of spiked serum samples by ELISA. The method was also found to be fully compatible with subsequent chemical analytical methods, such as liquid chromatography followed by mass spectrometry. The approaches developed in this study form a basis for analysis of IMT in biological samples in order to monitor their pharmacokinetic properties, and may be further used to study population exposure to IMT, and to monitor the occurrence of IMT contamination in water samples. © 2011 Springer-Verlag.
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