Naser F. Al-Tannak*, Nada A. Al-Hasawi and Ladislav Novotny Pages 295 - 304 ( 10 )
Background: Flavonoids as secondary metabolites of plants fulfill various functions in cell protection. They are of a considerable scientific interest because of their potentially medical use due to their anticancer, chemoprotective, antimicrobial, antiallergic, anti-inflammatory and antiviral activities.
Objective: The study aimed to develop a new UHPLC-UV method for morin and 2 other structurally related flavonoids - naringenin and kaempferol as the structural similarity of huge numbers of flavonoids does not limit their various biological functions and activities.
Methods: Separation of morin and 2 other structurally related flavonoids - naringenin and kaempferol - was achieved by using BEH C18 (1.7 µm, 2.1 x 50 mm) analytical column (Waters® Acquity UPLC) and a mobile phase composed of 0.05%v/v Formic acid in water and acetonitrile in proportion of 77:23 v/v and pumped at a flow rate of 0.4 ml/min. Column temperature was set at 25 ºC and samples were analyzed (3 µl injection volume) at a wavelength of 340 nm. Waters® Xevo G2-S QToF coupled with Waters® Acquity UPLC system with binary Solvent Manager (I-Class) via electrospray ionization (ESI) interface was used to confirm the identity of the peaks in biological samples.
Results: A rapid and simple UHPLC-UV separation of morin, kaempferol and naringenin is documented including methods validation. The developed method was applied to measuring morin, kaempferol and naringenin in human plasma after a solid phase extraction. Additionally, stability of morin in tissue culture medium was verified. The extraction method and UHPLC-UV elution conditions described provide a practical means to analyze morin, kaempferol and naringenin in biological matrices.
Conclusion: The developed method is fast and highly sensitive. Moreover, the flavonoids used were stable in human plasma for more than 10 days.
Morin, naringenin, kaempferol, UHPLC-UV, separation, human plasma stability, UHPLC-ESI-QToF.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kuwait University, PO Box 24923 Safat, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kuwait University, PO Box 24923 Safat, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kuwait University, PO Box 24923 Safat