1a.Objectives (from AD-416)
The pilot study will examine the metabolism, safety and efficacy of increased intake of citrus liminoids by human subjects. The main study will determine their health benefits, particularly the effects on serum lipids and markers of inflammation.
1b.Approach (from AD-416)
In the pilot study 6 healthy subjects (18-65y) with LDL cholesterol greater than 130 mg/dL will be recruited and fed citrus liminoids. The metabolites of limonoids in the serum and urine samples will be determined by the use of mass spectrometry. NMR technology will be used to determine the number and sizes and of different lipoprotein fractions in plasma. Markers of inflammation in the serum and also their ex-vivo production after stimulation of blood cells with mitogens will be determined by ELISA and flowcytometry methods. The design, number of subjects, and the dose of liminoids to be used will depend upon the results obtained from the pilot study.
We needed ten subjects to complete this cross over study involving eight week supplementation with each of the placebo and limonoid containing drinks. We have completed sample collection from all ten subjects. Analysis of samples was delayed because the original contract with the Beverage Institute expired 11/30/2009, and the new contract was not established until 10/01/2010. We are currently analyzing samples and data from this study. We intend to complete all data and sample analysis and submit the annual report to the Beverage Institute before the contract expiration date of 11/30/2012. The progress of the study has been frequently discussed by phone and e-mails with the sponsor of the study (Beverage institute) and collaborators, director of human studies and director of Bioanalytical laboratory.