Reverse-phase liquid chromatography with electrospray ionization/mass spectrometry for the quantification of pseudoephedrine in human plasma and application to a bioequivalence study

Jin-Ki Kim; Jun-Pil Jee; Jeong-Sook Park; Hyung Tae Kim; Chong-Kook Kim

doi:10.1055/s-0031-1296199

Arzneimittelforschung, Table of Contents

Arzneimittelforschung 2011; 61(5): 276-281
DOI: 10.1055/s-0031-1296199

Sympathomimetics

Editio Cantor Verlag Aulendorf (Germany)

Reverse-phase liquid chromatography with electrospray ionization/mass spectrometry for the quantification of pseudoephedrine in human plasma and application to a bioequivalence study

Authors

Jin-Ki Kim

¹College of Pharmacy, Seoul National University, Seoul, Republic of Korea
Jun-Pil Jee

¹College of Pharmacy, Seoul National University, Seoul, Republic of Korea
Jeong-Sook Park

²College of Pharmacy and Institute of Drug Research and Development, Chungnam National University, Daejeon, Republic of Korea
Hyung Tae Kim

³Department of Internal Medicine, Hanyang University College of Medicine, Seoul, Republic of Korea
Chong-Kook Kim

⁴College of Pharmacy, Inje University, Gimhae Gyungnam, Republic of Korea

Abstract

A sensitive and selective reverse-phase liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) method was developed and validated to quantify pseudoephedrine (CAS 90-82-4) in human plasma. Phenacetin was used as the internal standard (I.S.). Sample preparation was performed with a deproteinization step using acetonitrile. Pseudoephedrine and I.S. were successfully separated using gradient elution with 0.5% trifluoroacetic acid (TFA) in water and 0.5% TFA in methanol at a flow-rate of 0.2 mL/min. Detection was performed on a single quadrupole mass spectrometer by a selected ion monitoring (SIM) mode via electrospray ionization (ESI) source. The ESI source was set at positive ionization mode. The ion signals of m/z 166.3 and 180.2 were measured for the protonated molecular ions of pseudoephedrine and I.S., respectively. The lower limit of quantification (LLOQ) of pseudoephedrine in human plasma was 10 ng/mL and good linearity was observed in the range of concentrations 10–500 ng/mL (R² = 1). The intra-day accuracy of the drug containing plasma samples was more than 97.60% with a precision of 3.99–11.82%. The inter-day accuracy was 99.36% or more, with a precision of 7.65–18.42%.

By using this analytical method, the bioequivalence study of the pseudoephedrine preparation was performed and evaluated by statistical analysis of the log transformed mean ratios of pharmacokinetic parameters. All the results fulfilled the standard criteria of bioequivalence, being within the 80–125% range which is required by the Korea FDA, US FDA, and EMEA to conclude bioequivalence. Consequently, the developed reverse-phase LC-ESI-MS method was successfully applied to bioequivalence studies of pseudoephedrine in healthy male volunteers.

Key words

Bioequivalence - CAS 90-82-4 - Human plasma - LC-ESI-MS - Pharmacokinetics - Pseudoephedrine

Full Text

References

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