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. 2015 Sep-Oct;77(5):586-91.
doi: 10.4103/0250-474x.169038.

Development and Validation of Reversed-Phase High Performance Liquid Chromatographic Method for Hydroxychloroquine Sulphate

A Singh  1 , Roopkishora  1 , C L Singh  1 , R Gupta  1 , S Kumar  1 , M Kumar  2
Affiliations

Development and Validation of Reversed-Phase High Performance Liquid Chromatographic Method for Hydroxychloroquine Sulphate

A Singh et al. Indian J Pharm Sci. 2015 Sep-Oct.

Abstract

In the present work new, simple reversed-phase high performance liquid chromatographic method was developed and validated for the determination of hydroxychloroquine sulphate in blood plasma. Chloroquine sulphate was used as an internal standard. The chromatographic separation was achieved with octadecyl silane Hypersil C18 column (250×6 mm, 5 μm) using water and organic (acetonitrile:methanol: 50:50, v/v) mobile phase in 75:25 v/v ratio, with sodium 1-pentanesulfonate and phosphoric acid. This organic phase was maintained at pH 3.0 by orthophosphoric acid. The flow rate of 2.0 ml/min(.) with detection at 343 nm was used in the analysis. The calibration curve of standard hydroxychloroquine sulphate was linear in range 0.1-20.0 μg/ml. The method was validated with respected to linearity, range, precision, accuracy, specificity and robustness studies according to ICH guidelines. The method was found to be accurate and robust to analyze the hydroxychloroquine sulphate in plasma samples.

Keywords: Hydroxychloroquine sulphate; ICH guidelines; antimalarial agent; chloroquine sulphate; rheumatoid arthritis.

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Figures

Fig. 1
Fig. 1
Structures of analytes. (a) Hydroxychloroquine sulphate (C18H26ClN3O.H2SO4), (b) chloroquine (C18H26ClN3).
Fig. 2
Fig. 2
Chromatograms of analytes. Chromatogram of hydroxychloroquine sulphate and chloroquine.
Fig. 3
Fig. 3
Calibration curve of hydroxychloroquine sulphate. Calibration curve of hydroxychloroquine sulphate, by peak area ratio of drug and internal standard.
Fig. 4
Fig. 4
Chromatogram of blank plasma.
See this image and copyright information in PMC

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