Drug Res (Stuttg) 2013; 63(11): 558-563
DOI: 10.1055/s-0033-1347256
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

Development and Validation of a High-performance Liquid Chromatography Coupled with Ultraviolet Detection Method for the Determination of Isocorydine in Rat Plasma and its Application in Pharmacokinetics

Y.-Q. Liu
1   Department of Pharmacy, Kunming General Hospital of Chengdu Military Region, Kunming, PR China
2   School of Clinical Medicine, Kunming medical university, Kunming, PR China
H.-L. Li
1   Department of Pharmacy, Kunming General Hospital of Chengdu Military Region, Kunming, PR China
J.-C. He
1   Department of Pharmacy, Kunming General Hospital of Chengdu Military Region, Kunming, PR China
E.-F. Feng
1   Department of Pharmacy, Kunming General Hospital of Chengdu Military Region, Kunming, PR China
G.-X. Rao
3   School of Pharmacy, Yunnan College of Traditional Chinese Medicine, Kunming, PR China
G.-L. Xu
1   Department of Pharmacy, Kunming General Hospital of Chengdu Military Region, Kunming, PR China
2   School of Clinical Medicine, Kunming medical university, Kunming, PR China
› Author Affiliations
Further Information

Publication History

received 30 November 2012

accepted 18 May 2013

Publication Date:
24 July 2013 (online)


A new HPLC–UV method has been developed, validated and applied for the determination of isocorydine (CAS 475-67-2) in rat plasma after oral or intravenous (i. v.) administration. Caffeine was used as the internal standard (IS). The analyte and IS were extracted from rat plasma by liquid–liquid extraction (LLE) with methyl tert-butyl ether and they were separated on an XTerra C18 column (250×4.6 mm, 5 µm, pH 1–12) with UV detection at 264 nm. The mobile phase consisted of methanol and 0.02 mol/L potassium dihydrogen phosphate–phosphoric acid buffer solution (pH 3.2) (30:70, v/v) at a flow rate of 1 mL/min for 8.5 min. The retention times of isocorydine and caffeine were approximately 6.5 and 5.1 min, respectively. The good linearity of the calibration curves was observed over the concentration range of 0.05–8 µg/mL (n=8, r 2≥0.9995). The lower limit of quantification (LLOQ) was 0.05 µg/mL [signal to noise ratio (S/N)≥10], and the limit of detection (LOD) was demonstrated as 0.01 µg/mL (S/N≥3). The mean extraction recovery ranged from 83.7% to 89.5% at 3 quality control (QC) concentrations. Intra-day and inter-day precision (relative standard deviation, RSD%) were within 4.7% and accuracy (relative error, RE%) ranged from −1.2% to 4.5%. The developed method was successfully applied to determination of the pharmacokinetic properties of isocorydine in rats after oral administration at a dose of 20 mg/kg and i. v. injection at 5 mg/kg.

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