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1 Acta Chromatographica

1 2004

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Development and validation of an HPLC method with post-column derivatisation to assay n-acetylcysteine in plasma

Vander Heyden Y., Mangelings D., Van Brempt J., Spapen H.

Acta Chromatographica

2004

No. 14

149-164

A quantitative reversed-phase HPLC method has been developed that enables determination of both low endogenous and high therapeutic concentrations of N-acetylcysteine (NAC) in plasma. The compound is detected fluorimetrically after derivatisation with ortho-phthalaldehyde in the presence of a primary amine. Validation of the method revealed injection and method repeata-bility were good. The linear range was adequate and the limit of quantifi-cation was between 0.4 and 0.6 žM. Recovery of N-acetylcysteine from plasma samples was also acceptable. This method was applied to plasma samples from patients with a clinical septic shock who had received very high doses of N-acetylcys-teine. Six samples were taken at different times after administration of N-acetylcysteine. The blood-concentration profiles obtained indicate the me-thod is suitable for following the evolution of NAC in plasma under these conditions and can therefore be used for pharmacokinetic profiling.