title: Validated high-performance liquid chromatographic method utilizing solid-phase extraction for the simultaneous determination of naringenin and hesperetin in human plasma.
abstract: Naringenin and hesperetin, the aglycones of the flavanone glucosides naringin and hesperidin occur naturally in citrus fruits. They exert a variety of pharmacological effects such as antioxidant, blood lipid-lowering, anticarcinogenic and inhibit selected cytochrome P-450 enzymes resulting in drug interactions. A specific, sensitive, precise, and accurate solid-phase extraction high-performance liquid chromatographic (HPLC) assay for the simultaneous determination of naringenin and hesperetin in human plasma was developed and validated. After addition of 7-ethoxycoumarin as internal standard, plasma samples were incubated with beta-glucuronidase/sulphatase, and the analytes were isolated from plasma by solid-phase extraction using C(18) cartridges and separated on a C(8) reversed phase column with methanol/water/acetic acid (40:58:2, v/v/v) as the eluent at 45 degrees C. The method was linear in the 10-300 ng/ml concentration range for both naringenin and hesperetin (r>0.999). Recovery for naringenin, hesperetin and internal standard was greater than 76.7%. Intra- and inter-day precision for naringenin ranged from 1.4 to 4.2% and from 1.9 to 5.2%, respectively, and for hesperetin ranged from 1.3 to 4.1% and from 1.7 to 5.1%, respectively. Accuracy was better than 91.5 and 91.3% for naringenin and hesperetin, respectively.
authors.affiliation: Department of Pharmacy, School of Health Sciences, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
authors.forename: Feras Imad
authors.initials: FI
authors.lastname: Kanaze
authors.forename: Eugene
authors.initials: E
authors.lastname: Kokkalou
authors.forename: Manolis
authors.initials: M
authors.lastname: Georgarakis
authors.forename: Ioannis
authors.initials: I
authors.lastname: Niopas
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002957:Citrus / Q000737:chemistry; D044950:Flavanones / Q000097:blood*; D005638:Fruit / Q000737:chemistry; D006569:Hesperidin / Q000097:blood*; D006801:Humans; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D044950:Flavanones; D006569:Hesperidin; C005273:naringenin; C013015:hesperetin