ABSTRACT
A reversed-phase high-performance liquid chromatographic (RP-HPLC) method was described for the determination of chlorogenic acid (CGA) in rat plasma using protocatechuic acid as internal standard (IS). CGA in plasma was extracted with acetonitrile, which also acted as deproteinization agent. Chromatographic separation was performed on a Kromasil C18 column with methanol-0.2 m acetic acid (pH 3.0, 25:75, v/v) as mobile phase at a flow-rate of 1.0 mL/min with an operating temperature of 30 degrees C and UV detection at 300 nm. The standard curve was found to be linear over the concentration ranges of 0.4-2.5 microg/mL and 2.5-40 microg/mL, and the limit of quantification (LOQ) was 0.4 microg/mL. The analytical precision and accuracy were validated by relative standard deviation (RSD) and relative error, which were in ranges 3.14-10.78% and -2.20-5.00%, respectively. The average recovery of CGA was 87.59%. The method was successfully applied to the pharmacokinetic study of CGA in Yin-Huang granules.
Subject(s)
Chlorogenic Acid/pharmacokinetics , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/administration & dosage , Animals , Chlorogenic Acid/analysis , Lonicera/chemistry , Male , Rats , Rats, Wistar , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
In the present study, a reversed phase high performance liquid chromatographic (RP-HPLC) method was established for simultaneous determination of chlorogenic acid, caffeic acid, ferulic acid, protocatechuic acid and protocatechuic aldehyde in a Chinese herbal preparation (Fufang-Pugongying-Mixture). The separation was performed on a Hypersil ODS-2 column by isocratic elution with methanol and 0.2 M acetate buffer (pH 3.6) (15 : 85, v/v) as the mobile phase at the flow-rate of 1.0 ml/min with operating temperature of 30 degrees C, and detection wavelength of 300 nm. A good linear regression relationship between peak-areas and concentrations was obtained over the range of 2-200 microg/ml for the five marker compounds mentioned above. The spike recoveries were within 96.72-104.07%. The variation coefficient (CV) values of the precision were in the range of 0.89-4.50%. Moreover the developed method has reference value for quantitative analysis of Taraxacum, Lonicera and Angelica.
