Separation optimization of quercetin, hesperetin and chrysin in honey by micellar liquid chromatography and experimental design

Abstract

The chemometrics approach was applied for the separation optimization of flavonoid markers (quercetin, hesperetin and chrysin) in honey using micellar liquid chromatography (MLC). The investigated method combines SPE of flavonoids from honey using C(18) cartridge and their separation and quantification by micellar liquid chromatography. A two level full factorial design was carried out to evaluate the effect of four experimental factors including concentration of SDS, alkyl chain length of the alcohol used as the organic modifier (N), volume percentage of the organic modifier (V(m)) and volume percentage of acetic acid (AcOH) in mobile phase on analytes retention times. Experiments for analytes retention times modeling and optimization of separation were performed according to central composite design. Multiple linear regression method was used for the construction of the best model based on experimental retention times. Pareto optimal method was used to find suitable compatibility between resolution and analysis time of analytes in honey. The optimum mobile phase composition for separation and determination of analytes in honey were [SDS]=0.124 mol/L; 7.8% v/v ethanol and 5.0% v/v AcOH. Limits of detection and linear range of flavonoid markers were 0.0079-0.0126, 0.05-50.0 mg/L, respectively.

Micellar Liquid Chromatography from Green Analysis Perspective

“…Another interesting application of MLC is the analysis of food samples [81][82][83][84][85][86][87][88][89][90][91][92][93][94][95]. This includes the determination of several compounds in chicken muscles and liver [81][82][83]95]; bovine meat, liver, and kidney [81,82,84,95]; swine kidney [89]; eggs [81,83,93,95]; milk [82,87,93,95]; baby food [82,83,95]; honey [92]; fish sauce [91,94]; feeding staff [86,90]; olive extract samples [85]; and dietetic supplements [88].…”

“…This includes the determination of several compounds in chicken muscles and liver [81][82][83]95]; bovine meat, liver, and kidney [81,82,84,95]; swine kidney [89]; eggs [81,83,93,95]; milk [82,87,93,95]; baby food [82,83,95]; honey [92]; fish sauce [91,94]; feeding staff [86,90]; olive extract samples [85]; and dietetic supplements [88]. MLC has been shown to be a very efficient technique for food analysis, avoiding long and tedious extractions of such complex matrices while providing high percentage recoveries with high accuracy and precision.…”

Medicinal Uses of Honey

“…Honey contains vitamins [11] and phenolic compounds [18] in trace quantities. Some of the phenolic compounds in honey responsible for antioxidant effect are flavonols, flavones, flavonones, benzoic and cinnamic acids.…”

“…Honey has been found to contain significant antioxidant activity attributed to glucose oxidase, catalase, ascorbic acid, flavonoids, phenolic acids, carotenoid derivatives, organic acids, Maillard reaction products, amino acids, proteins [18,[40][41][42].…”

Optimization of Quercetin Extraction from Green Tea ( Camellia sinensis ) Using Central Composite Design, and the Pharmacological Activity of the Extract

“…For identification of the extracted quercetin, preparative TLC 12 , RP-HPLC 11,13,14,[20][21][22] , micellar liquid chromatography 23 and GC-MS 24 were used. A traditional approach, well known as the "One Variable At a Time" (OVAT), is commonly applied for investigation of the effects of extraction param- eters on the yield of the desired compounds.…”