Validation of HPLC-UV Assay of Caffeic Acid in Emulsions.
An accurate, sensitive, precise and rapid reversed-phase high-performance liquid chromatographic method was successfully developed and validated for the determination of caffeic acid (CA) in emulsions. The best separation was achieved on a 250 × 4.6 mm, 5.0 µm particle size RP18 XDB Waters column using ethanol and purified water (40:60 v/v) adjusted to pH 2.5 with acetic acid as the mobile phase at a flow rate of 0.7 mL/min. Ultraviolet detection was performed at 325 nm at ambient column temperature (25°C). The method was linear over the concentration range of 10-60 µg/mL (r(2) = 0.9999) with limits of detection and quantification of 1.44 and 4.38 µg/mL, respectively. CA was subjected to oxidation, acid, base and neutral degradation, as well as photolysis and heat as stress conditions. There were no interfering peaks at or near the retention time of CA. The method was applied to the determination of CA in standard and pharmaceutical products with excellent recoveries. The method is applicable in the quality control of CA.
Department of Drugs and Medicines, Faculty of Pharmaceutical Sciences of Araraquara, Universidade Estadual Paulista "Júlio de Mesquita Filho"-UNESP, Araraquara, São Paulo, Brazil.
Caroline Magnani
CM
Spagnol
Department of Drugs and Medicines, Faculty of Pharmaceutical Sciences of Araraquara, Universidade Estadual Paulista "Júlio de Mesquita Filho"-UNESP, Araraquara, São Paulo, Brazil.
Vera Lucia Borges
VL
Isaac
Department of Drugs and Medicines, Faculty of Pharmaceutical Sciences of Araraquara, Universidade Estadual Paulista "Júlio de Mesquita Filho"-UNESP, Araraquara, São Paulo, Brazil correama@fcfar.unesp.br.
Marcos Antonio
MA
Corrêa
Department of Drugs and Medicines, Faculty of Pharmaceutical Sciences of Araraquara, Universidade Estadual Paulista "Júlio de Mesquita Filho"-UNESP, Araraquara, São Paulo, Brazil.
Hérida Regina Nunes
HR
Salgado
D019342:Acetic Acid; D002109:Caffeic Acids / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000592:standards*; D056148:Chromatography, Reverse-Phase / Q000592:standards*; D004355:Drug Stability; D004655:Emulsions; D000431:Ethanol; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057230:Limit of Detection; D010084:Oxidation-Reduction; D010782:Photolysis; D010936:Plant Extracts / Q000737:chemistry*; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet / Q000592:standards*; D013607:Tablets; D014867:Water
D002109:Caffeic Acids; D004655:Emulsions; D010936:Plant Extracts; D012997:Solvents; D013607:Tablets; D014867:Water; D000431:Ethanol; D019342:Acetic Acid; C040048:caffeic acid