Electrogenerated Chemiluminescence Derivatization Reagent, 3-Isobutyl-9,10-dimethoxy-1,3,4,6,7,11b- hexahydro-2H-pyrido[2,1-a]isoquinolin-2-ylamine, for Carboxylic Acid in High-Performance Liquid Chromatography Using Tris(2,2‘-bipyridine)ruthenium(II)

Abstract

3-Isobutyl-9,10-dimethoxy-1,3,4,6,7,11b-hexahydro-2H-pyrido[2,1-a]isoquinolin-2-ylamine (IDHPIA) was found to be a selective and highly sensitive derivatization reagent for carboxylic acid by high-performance liquid chromatography (HPLC) with electrogenerated chemiluminescence detection using tris(2,2'-bipyridine)ruthenium(II). Free fatty acids and phenylbutylic acid were used as model compounds of carboxylic acids, and the derivatization conditions were optimized with myristic acid. Under the mild reaction conditions of room temperature for 45 min in acetonitrile containing 2-bromo-1-ethylpyridinium tetrafluoroborate and 9-methyl-3,4-dihydro-2H-pyridol1,2-a]pyrimidin-2-one, all the fatty acids tested were reacted with IDHPIA to produce highly sensitive derivatives. The chemiluminescence intensity was essentially the same for all fatty acids. The derivatives obtained from 10 free fatty acids were completely separated by reversed-phase chromatography under isocratic elution conditions. The on-column detection limit (signal-to-noise ratio of 3) with proposed HPLC separation and chemiluminescence detection was 0.5 and 0.6 fmol for myristic acid and phenylbutylic acid, respectively. IDHPIA was 100-fold more sensitive than previously developed reagents (Morita, H.; Konishi, M. AnaL Chem. 2002, 74, 1584-1589). The free fatty acids in human serum were successfully determined using the present method.

Liquid chromatographic determination of acetylcholine based on pre‐column alkaline cleavage reaction and post‐column tris(2,2′‐bipyridyl)ruthenium(III) chemiluminescence detection

“…This is because chemiluminescence systems have many advantages, such as high selectivity, sensitivity, and a wide linear working range. More recently, various derivatization techniques and reagents for the ruthenium chemiluminescence detection method have been developed (17)(18)(19); however, there are no reports on these techniques being used for the analysis of quaternary ammonium compounds.…”

First observation of surface plasmon-coupled electrochemiluminescence

“…As a result, CL and ECL are widely used in medical testing [2][3][4][5]. ECL methods have been developed for a variety of applications including capillary electrophoresis [6,7], detection of ions [8], organic acids [9] and explosives [10]. The electrical origin of ECL allows its use with detector arrays for multi-analyte testing [11].…”

Chemiluminescence detection with separation techniques for bioanalytical applications

“…4), have been previously reported [62,63]. Both the derivatives obtained for ten free fatty acids could be completely separated by HPLC under isocratic conditions.…”