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Liu X.,China Pharmaceutical University | Zhi H.,Beijing Zhongyan Tongrentang Chinese Medicine R and D Co. | Du F.,Beijing Zhongyan Tongrentang Chinese Medicine R and D Co. | Ye Z.,Chinese Institute of Materia Medica | And 4 more authors.
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences | Year: 2010

A sensitive and reproducible HPLC method for quantitative determination of puerarin (PUE) in rat plasma was developed and validated using 4-hydroxybenzaldehyde as an internal standard. The separation of PUE was performed on a CAPCELL PAK C18 column by gradient elution with 0.2% aqueous phosphoric acid and acetonitrile as the mobile phase. The method was validated and found to be linear in the range of 80-12,000. ng/mL. The limit of quantification was 80. ng/mL based on 100 μL of plasma. The variations for intra- and inter-day precision were less than 8.3%, and the accuracy values were between 98% and 105.2%. The extraction recoveries were more than 85%. The method was successfully applied in the comparative study of pharmacokinetics of PEGylated puerarin (PEG-PUE) versus PUE in rats. Compared with PUE, PEG-PUE showed a 5.2-fold increase in half-life of PUE and a 4.7-fold increase in mean residence time. In addition, this method was also successfully applied to determine the low plasma concentration of PUE regenerated from PEG-PUE in vitro. © 2010 Elsevier B.V. Source


Liu X.,China Pharmaceutical University | Yu B.,China Pharmaceutical University | Wang N.,Beijing Zhongyan Tongrentang Chinese Medicine R and D Co. | Zhang B.,Beijing Zhongyan Tongrentang Chinese Medicine R and D Co. | And 4 more authors.
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences | Year: 2010

The aim of this study was to develop a validated specific stability-indicating HPLC method for the quantitative determination of PEGylated puerarin (PEG-PUE) in aqueous solutions. The method was validated by subjecting PEG-PUE to forced degradation under stress conditions of acid, alkali, water hydrolysis, and oxidation. Both PEG-PUE and puerarin (PUE) were simultaneously determined and separated on CAPCELL PAK C18 column by gradient elution with 0.2% aqueous phosphoric acid and acetonitrile as the mobile phase. The flow rate was 1.0mLmin-1 and detection wavelength was set at 250nm. Both calibration curves showed good linear regression (r≥0.9998) within test ranges. The LOD and LOQ of PEG-PUE were determined to be 3 and 9μgmL-1 respectively. Degradation of PEG-PUE followed pseudo-first-order kinetics with t1/2 of 59min at pH 9.0 and 17.79h at pH 7.4. However, at pH 5.0 and 2.0, there was no significant degradation of PEG-PUE over time. In conclusion, the method was observed to have the necessary specificity, precision, and accuracy, and to be suitable for quantity monitoring the degradation process of PEG-PUE during stability studies. The degradation studies may give insight into useful information for formulation development of PEG-PUE. © 2010 Elsevier B.V. Source

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