PROCESSING AND BIOTECHNOLOGICAL IMPROVEMENT OF FOODS TO PREVENT OBESITY RELATED AND OTHER DEGENERATIVE DISEASES
Location: Processed Foods Research
Title: Quantification of the Sulfated Cholecystokinin CCK8 in Hamster Plasma Using Immunoprecipitation-Liquid Chromatography-Mass Spectrometry/Mass Spectrometry
Research conducted cooperatively with:
| The Dow Chemical Company|
Submitted to: Analytical Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 20, 2009
Publication Date: November 1, 2009
Citation: Young, S.A., Julka, S., Bartley, G.E., Gilbert, J.R., Wendelburg, B., Hung, S., Anderson, W., Yokoyama, W.H. 2009. Quantification of the Sulfated Cholecystokinin CCK8 in Hamster Plasma Using Immunoprecipitation-Liquid Chromatography-Mass Spectrometry/Mass Spectrometry. Analytical Chemistry. 81:9120-8.
Interpretive Summary: The amino acid sequence of the satiety hormone, CCK8, from hamsters was determined by mass spectrometry. Active and inactive forms of cholecystokinin (CCK) exist and CCK8 is biologically active. To improve the accuracy of analysis, immunoprecipitation was used.
Cholecystokinin (CCK) and the different molecular forms of CCK are well established as biomarkers for satiety. CCK hormone and the different biologically active and inactive molecular forms have been shown to influence food intake associated with satiety and are predominately secreted from the gut. Despite the importance of CCK in satiety, endogenous CCK-8 has not been well characterized in Syrian Golden hamsters. In this study, we have cloned and sequenced hamster CCK and identified and characterized endogenous CCK-8 from hamster plasma. Hamster CCK-8 is comprised of 8 amino acid residues which are highly conserved amongst other species. Following accurate identification and characterization of hamster CCK-8, we have developed a highly specific and sensitive immunoprecipitation based LC-MS/MS assay for its quantification. The present assay enables quantification of active CCK-8 over a concentration range from 0.1 to 2.5 ng/mL in hamster plasma samples. This range covers both the basal and postprandial levels of CCK-8. Assay accuracy (% RE) and precision (% CV) were measured at three concentrations on each of four days and did not exceed x.x and xx.x%, respectively. Additionally, the method was shown to be applicable for determining the increased concentration of endogenous CCK-8 in hamster plasma following being gavaged high fat diet compared to a low fat diet.