Synthesis of safflomide and its HPLC measurement in mouse plasma after oral administration

Authors: Park, Jae; Chen, Pei

Submitted to: Journal of Chromatography B
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/28/2006
Publication Date: 2/8/2007
Citation: Park, J.B., Chen, P. 2007. Synthesis of safflomide and its HPLC measurement in mouse plasma after oral administration. Journal of Chromatography B: Biomedical Applications. 852(1-2):398-402.

Interpretive Summary: N-caffeoyltryptamine (Safflomide) is a phytochemical synthesized from caffeic acid and tryptamine. N-caffeoyltryptamine and its natural analogues are found in numerous plants, including coffee, cocoa, and safflower. Recent studies indicate that the compounds may produce biological activities implicated in preventing and/or treating conditions arising from human chronic diseases, such as inflammation and atherosclerosis. However, only limited information is currently available regarding the effect of N-caffeoyltryptamine on human disease. The bio-availability of the compound is nearly unknown, making it impossible to accurately propose and/or assess its effects on humans. In this study, N-caffeoyltryptamine was first synthesized and a method developed to measure the concentrations of N-caffeoyltryptamine in biological samples. This study produced two significant outcomes: 1) the synthesis of N-caffeoyltryptamine as a standard and 2) the development of an HPLC method quantifying N-caffeoyltryptamine with excellent peak resolution, high sensitivity, and reliability. These outcomes can be utilized by researchers in nutrition, molecular biology, and medicine to quantify N-caffeoyltryptamine in biological samples, as well as its bioavailability after oral intake.

Technical Abstract: N-caffeoyltryptamine is a compound belonging to a group of phenylpropanoid amides found in plants. For this study, N-caffeoyltryptamine was chemically synthesized and a high performance liquid chromatography (HPLC) method was developed for quantifying N-caffeoyltryptamine in biological samples. The synthesis was simple, and the yield of N-caffeoyltryptamine was greater than 50%. Using synthesized N-caffeoyltryptamine as a standard, HPLC separation was performed on a Nova-Pak C18 column using an isocratic buffer, and separation was performed using a coulometric electrochemical detector with four electrode channels. The detection of N-caffeoyltryptamine yielded an excellent peak resolution at a retention time of 18 min. The lower limit of the detection was as little as 100 fmol. Using this HPLC method, the plasma concentrations of N-caffeoyltryptamine were determined in mouse blood, collected at 5, 10, 15, 20, 25, 30, and 35 min following oral administration (1 and 3 mg/30 g body weight). This HPLC method, standardized with N-caffeoyltryptamine, is the first reported method able to quantify N-caffeoyltryptamine in standard and plasma samples with a good detection limit and consistent reproducibility.