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Simultaneous determination of quercetin, kaempferol, and isorhamnetin in phytopharma‐ceuticals of Hippophae rhamnoides L. by high‐performance liquid chromatography with chemiluminescence detection
Author(s) -
Zhang Qunlin,
Cui Hua
Publication year - 2005
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200500055
Subject(s) - isorhamnetin , kaempferol , chemiluminescence , chromatography , chemistry , quercetin , high performance liquid chromatography , flavones , organic chemistry , antioxidant
A novel method based on reversed‐phase high‐performance liquid chromatography with chemiluminescence detection has been developed for the simultaneous determination of three flavonols including quercetin, kaempferol, and isorhamnetin. The procedure was based on the chemiluminescent enhancement by flavonols of the cerium(IV)‐rhodamine 6G system in sulfuric acid medium. The effects of several parameters on the HPLC resolution and CL emission were studied systematically. Good separation was achieved with isocratic elution using a mixture of methanol and aqueous 1.0% acetic acid (37 : 63, v/v ) within 25 min. Under optimized conditions, the linear working range covers 3 orders of magnitude with relative standard deviations below 4.5% for 11 replicate injected flavonol samples, and detection limits ( S/N = 3) were 1.6×10 –8 , 3.5×10 –9 , and 6.5×10 –9 g mL –1 for quercetin, kaempferol, and isorhamnetin, respectively. The chemiluminescence reaction was compatible with the mobile phase of high‐performance liquid chromatography. The proposed method has been successfully applied to the determination of three active flavonols in phytopharmaceuticals of Hippophae rhamnoides L. After a simple extraction procedure, the repeatability and recovery were satisfactory.