Application of mixed cloud point extraction for the analysis of six flavonoids in Apocynum venetum leaf samples by high performance liquid chromatography

• Published in: Journal of pharmaceutical and biomedical analysis 2015-03, Vol.107, p.273-279
• Main Authors: Zhou, Jun, Sun, Jiang Bing, Xu, Xin Yu, Cheng, Zhao Hui, Zeng, Ping, Wang, Feng Qiao, Zhang, Qiong
• Format: Article
• Language: English
• Subjects: Apocynum - chemistry; Apocynum venetum; Cetrimonium Compounds - chemistry; Chromatography, High Pressure Liquid - methods; CTAB; Flavonoids; Flavonoids - chemistry; Genapol X-080; Hydrogen-Ion Concentration; Kaempferols - chemistry; Mixed cloud point extraction; Plant Extracts - chemistry; Plant Leaves - chemistry; Polyethylene Glycols - chemistry; Quercetin - analogs & derivatives; Quercetin - chemistry; Rutin - chemistry; Surface-Active Agents - chemistry
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2015.01.003

Typical chromatograms of determination of six flavonoids in Apocynum venetum leaf: (A) samples with mixed cloud point extraction; (B) samples with conventional cloud point extraction. rutin (RUT); hyperoside (HYP); quercetin-3-O-sophoroside (QER); isoquercitrin (ISO); astragalin (AST) and quercetin (QUE). •A mixed cloud point extraction combined with HPLC has been developed for the determination of six flavonoids in Apocynum venetum leaf.•The surfactant Genapol X-080 and cetyl-trimethyl ammonium bromide were chosen as the mixed extracting solvents.•The method was investigated in detail. A simple, inexpensive and efficient method based on the mixed cloud point extraction (MCPE) combined with high performance liquid chromatography was developed for the simultaneous separation and determination of six flavonoids (rutin, hyperoside, quercetin-3-O-sophoroside, isoquercitrin, astragalin and quercetin) in Apocynum venetum leaf samples. The non-ionic surfactant Genapol X-080 and cetyl-trimethyl ammonium bromide (CTAB) was chosen as the mixed extracting solvent. Parameters that affect the MCPE processes, such as the content of Genapol X-080 and CTAB, pH, salt content, extraction temperature and time were investigated and optimized. Under the optimized conditions, the calibration curve for six flavonoids were all linear with the correlation coefficients greater than 0.9994. The intra-day and inter-day precision (RSD) were below 8.1% and the limits of detection (LOD) for the six flavonoids were 1.2–5.0ngmL−1 (S/N=3). The proposed method was successfully used to separate and determine the six flavonoids in A. venetum leaf samples.