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Related Experiment Videos

Chiral capillary electrophoretic method for quantification of apomorphine.

Pham Thi Thanh Ha1, Ann Van Schepdael, Laurence Van Vaeck

  • 1Laboratory of Pharmaceutical Chemistry and Drug Analysis, Faculty of Pharmaceutical Sciences, 4 Van Evenstraat, B-3000 Leuven, Belgium.

Journal of Chromatography. A
|October 27, 2004
PubMed
Summary

A new chiral separation method for apomorphine enantiomers was developed using capillary electrophoresis and hydroxypropyl-beta-cyclodextrin. This validated method accurately quantifies apomorphine in intestinal transport studies.

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Area of Science:

  • Analytical Chemistry
  • Pharmaceutical Sciences
  • Separation Science

Background:

  • Chiral determination of apomorphine enantiomers is crucial for pharmaceutical development.
  • Existing methods may face challenges in resolving enantiomers effectively.
  • Cyclodextrins are widely used as chiral selectors in separation techniques.

Purpose of the Study:

  • To develop and validate a novel capillary electrophoresis method for chiral separation of R,S-apomorphine.
  • To optimize chiral selector type, concentration, buffer pH, and applied voltage.
  • To apply the validated method for apomorphine enantiomer determination in a Caco-2 cell model.

Main Methods:

  • Screening of seven cyclodextrins for enantioselectivity towards R,S-apomorphine.

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  • Optimization of capillary electrophoresis parameters including chiral selector concentration, buffer pH, and applied voltage.
  • Validation of the developed method for limit of detection (LOD) and limit of quantification (LOQ).

    • Main Results:

    • Hydroxypropyl-beta-cyclodextrin was selected as the optimal chiral selector, resolving apomorphine enantiomers.
    • Optimal conditions were determined as 14 mM hydroxypropyl-beta-cyclodextrin, pH 3.0, and 16 kV.
    • The validated method achieved an LOD of 0.2 microM and LOQ of 0.5 microM.

    Conclusions:

    • A robust and validated capillary electrophoresis method for chiral determination of apomorphine enantiomers was successfully established.
    • The method demonstrated suitability for analyzing apomorphine enantiomers in complex biological matrices, such as in Caco-2 cell transport studies.