Use of multiresponse statistical techniques to optimize the separation of diosmin, hesperidin, diosmetin and hesperitin in different pharmaceutical preparations by high performance liquid chromatography with UV-DAD.
A new method for the separation and determination of four flavonoids: hesperidin (HES), diosmin (DIO), hesperitin (HTIN), and diosmetin (DTIN) in pure form and pharmaceutical formulations has been developed by using high performance liquid chromatography (HPLC) with UV-DAD detection. Multivariate statistics (2kfull factorial and Box Behnken Designs) has been used for the multiresponse optimization of the chromatographic separation, which was completed in 22min, and carried out on a symmetry® C18 column (250×3mm; 5µm) as stationary phase. Separation was conducted by gradient elution mode using a mixture of methanol, acetonitrile and water pH: 2.5 (CH3COOH), as mobile phase. Analytes were separated setting the column at 22°C, with a flow rate of 0.58mLmin-1and detected at 285nm. Under the optimized conditions, the flavonoids showed retention times of: 8.62, 11.53, 18.55 and 19.94min for HES, DIO, HTIN and DTIN, respectively. Limits of detection and quantification were <0.0156µgmL-1and <0.100µgmL-1, respectively. Linearity was achieved with good correlation coefficients values (r2=0.999; n=5). Intra-day and inter-day precision were found to be less than 3.44% (n=7). Finally, the proposed method was successfully applied to determine the target flavonoids in pharmaceutical preparations with satisfactory recoveries (between 95.2% and 107.9%), demonstrating that should also find application in the quality control, as well as in the pharmacokinetic studies of these drugs.
Departament of Chemistry, University of the Balearic Islands, Cra. Valldemossa km 7.5, 07122 Palma de Mallorca, Spain; Departament of Chemistry, University of Aleppo, University Square, Aleppo, Syria.
Mohamad Subhi
MS
Sammani
Sciware Systems, S.L., Spin-off of The University of the Balearic Islands, C/Pi 37, 07193 Bunyola, Spain. Electronic address: sabrina.clavijo@uib.es.
Sabrina
S
Clavijo
Departament of Chemistry, University of the Balearic Islands, Cra. Valldemossa km 7.5, 07122 Palma de Mallorca, Spain.
Lindomar
L
Portugal
Departament of Chemistry, University of the Balearic Islands, Cra. Valldemossa km 7.5, 07122 Palma de Mallorca, Spain.
Ruth
R
Suárez
Departament of Chemistry, University of Aleppo, University Square, Aleppo, Syria.
Hassan
H
Seddik
Departament of Chemistry, University of the Balearic Islands, Cra. Valldemossa km 7.5, 07122 Palma de Mallorca, Spain; Sciware Systems, S.L., Spin-off of The University of the Balearic Islands, C/Pi 37, 07193 Bunyola, Spain.
Víctor
V
Cerdà
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003627:Data Interpretation, Statistical; D004145:Diosmin / Q000032:analysis / Q000302:isolation & purification*; D005419:Flavonoids / Q000032:analysis / Q000302:isolation & purification*; D006569:Hesperidin / Q000032:analysis / Q000302:isolation & purification*; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D052617:Solid Phase Microextraction / Q000379:methods*; D014466:Ultraviolet Rays
D005419:Flavonoids; D004364:Pharmaceutical Preparations; D004145:Diosmin; D006569:Hesperidin; C013015:hesperetin; C039602:diosmetin
Diosmetin; Diosmin; Flavonoids; HPLC; Hesperidin; Hesperitin; Multiresponse statistical techniques