Studies on reaction of reduced lipoic acid with Mukaiyama reagent and its application for pharmaceutical and food analysis

ABSTRACT

A new sensitive method for the determination of lipoic acid (LA) in selected food items based on its reaction with Mukaiyama reagent (2-chloro-1-methylpyridinium iodide, CMPI) was developed. It was stated that CMPI reacts with reduced form of lipoic acid (dihydrolipoic acid, DHLA) and the stable product is produced. The spectrum of the labeled form of DHLA exhibits new band at 312 nm. Based on its spectral characteristics new spectrophotometric and UV–high-performance liquid chromatography (HPLC) methods of LA determination were elaborated. Both methods allowed determination of the analyte in the concentration range of 5 × 10⁻⁶–1 × 10⁻⁴ mol L⁻¹ with limit of detection 0.39 × 10⁻⁶ and 0.77 × 10⁻⁶ mol L⁻¹ for spectrophotometric and HPLC method, respectively. The practical usability of newly developed methods was checked by determination of lipoic acid contents in its pharmaceutical preparate Revitanerw. The proposed method was precise and accurate. The relative error of determination did not exceed ±0.067%. As chromatographic method allowed the determination of analyte in the presence of complex matrix, it was applied for assay of free fraction of α-lipoic acid in selected food items. A procedure of LA isolation from biological matrix was developed. The extraction with dichloromethane allowed quantitative recovery at 102.94 ± 4.20%. The green barley appeared to be the richest source of LA.

GRAPHICAL ABSTRACT

KEYWORDS:

• Chromatographic determination
• derivatization reaction
• food analysis
• lipoic acid
• Mukaiyama reagent
• pharmaceutical analysis
• spectrophotometric determination