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Determination of serum amantadine by liquid chromatography-tandem mass spectrometry.
Clin Chim Acta. 2005 Sep; 359(1-2):125-31.CC

Abstract

BACKGROUND

Amantadine (1-adamantylamine) is used for treatment of influenza, hepatitis C, parkinsonism, and multiple sclerosis. Current amantadine analysis by HPLC or gas chromatography (GC) requires a laborious sample pretreatment with extraction and/or derivatization steps. We established an LC-MS/MS method without protein precipitation, centrifugation, extraction and derivatization steps.

MATERIAL AND METHODS

50 microl sample+50 microl of 0.4 mg/l 1-(1-adamantyl)pyridinium bromide as internal standard+1000 microl water (96-well plate). Of this 25 microl+500 microl water (96-well plate; final serum dilution 1:462). LC-MS/MS: Surveyor MS pump, Autosampler, triple-quadrupole TSQ Quantum mass spectrometer (Thermo Electron). Autosampling: 2 microl of each sample. Chromatography: isocratic water/acetonitrile (60/40 v/v) with 5 g/l formic acid, flow rate 0.2 ml/min, run time 3 min, Phenomenex Luna C8(2) (100 x 2.0 mm (i.d.); 3-microm bead size) column. Mass spectrometry: electrospray atmospheric pressure ionization, positive ion and selective reaction monitoring mode, ion transitions m/z 152.0-->135.1 (at 22 eV amantadine) and 214.1-->135.1 (at 26 eV internal standard).

RESULTS

Calibration curves were constructed with spiked serum samples (amantadine 50-1000 microg/l, r>0.99). No carry over (5000 microg/l). No ion suppression with retention times similar to those of amantadine (1.8 min) and the internal standard (2.1 min). Detection limit 20 mg/l, linearity 20-5000 mg/l, intra-assay/inter-assay CV<6%/<8%, recovery 99-101%. Method comparison: LC-MS/MS=1.23 x GC-45 (Passing-Bablok regression). No significant bias between GC and LC-MS/MS (Bland-Altman plot).

CONCLUSION

We consider the sample pretreatment without deproteination, derivatization and centrifugation steps and the specificity of the tandem mass spectrometry as the most important points of our amantadine analysis method.

Authors+Show Affiliations

Arndt T
Bioscientia GmbH, Konrad-Adenauer-Strasse 17, D-55218 Ingelheim, Germany. torsten.arndt@bioscientia.de
Guessregen B
No affiliation info available
Hohl A
No affiliation info available
Reis J
No affiliation info available

MeSH

AmantadineCalibrationChromatography, High Pressure LiquidDrug MonitoringHumansReference StandardsReproducibility of ResultsSpectrometry, Mass, Electrospray Ionization

Pub Type(s)

Journal Article
Validation Study

Language

eng

PubMed ID

15913590

Citation

Arndt, Torsten, et al. "Determination of Serum Amantadine By Liquid Chromatography-tandem Mass Spectrometry." Clinica Chimica Acta; International Journal of Clinical Chemistry, vol. 359, no. 1-2, 2005, pp. 125-31.
Arndt T, Guessregen B, Hohl A, et al. Determination of serum amantadine by liquid chromatography-tandem mass spectrometry. Clin Chim Acta. 2005;359(1-2):125-31.
Arndt, T., Guessregen, B., Hohl, A., & Reis, J. (2005). Determination of serum amantadine by liquid chromatography-tandem mass spectrometry. Clinica Chimica Acta; International Journal of Clinical Chemistry, 359(1-2), 125-31.
Arndt T, et al. Determination of Serum Amantadine By Liquid Chromatography-tandem Mass Spectrometry. Clin Chim Acta. 2005;359(1-2):125-31. PubMed PMID: 15913590.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Determination of serum amantadine by liquid chromatography-tandem mass spectrometry. AU - Arndt,Torsten, AU - Guessregen,Brunhilde, AU - Hohl,Axel, AU - Reis,Janine, PY - 2005/01/31/received PY - 2005/03/17/revised PY - 2005/03/17/accepted PY - 2005/5/26/pubmed PY - 2005/11/8/medline PY - 2005/5/26/entrez SP - 125 EP - 31 JF - Clinica chimica acta; international journal of clinical chemistry JO - Clin Chim Acta VL - 359 IS - 1-2 N2 - BACKGROUND: Amantadine (1-adamantylamine) is used for treatment of influenza, hepatitis C, parkinsonism, and multiple sclerosis. Current amantadine analysis by HPLC or gas chromatography (GC) requires a laborious sample pretreatment with extraction and/or derivatization steps. We established an LC-MS/MS method without protein precipitation, centrifugation, extraction and derivatization steps. MATERIAL AND METHODS: 50 microl sample+50 microl of 0.4 mg/l 1-(1-adamantyl)pyridinium bromide as internal standard+1000 microl water (96-well plate). Of this 25 microl+500 microl water (96-well plate; final serum dilution 1:462). LC-MS/MS: Surveyor MS pump, Autosampler, triple-quadrupole TSQ Quantum mass spectrometer (Thermo Electron). Autosampling: 2 microl of each sample. Chromatography: isocratic water/acetonitrile (60/40 v/v) with 5 g/l formic acid, flow rate 0.2 ml/min, run time 3 min, Phenomenex Luna C8(2) (100 x 2.0 mm (i.d.); 3-microm bead size) column. Mass spectrometry: electrospray atmospheric pressure ionization, positive ion and selective reaction monitoring mode, ion transitions m/z 152.0-->135.1 (at 22 eV amantadine) and 214.1-->135.1 (at 26 eV internal standard). RESULTS: Calibration curves were constructed with spiked serum samples (amantadine 50-1000 microg/l, r>0.99). No carry over (5000 microg/l). No ion suppression with retention times similar to those of amantadine (1.8 min) and the internal standard (2.1 min). Detection limit 20 mg/l, linearity 20-5000 mg/l, intra-assay/inter-assay CV<6%/<8%, recovery 99-101%. Method comparison: LC-MS/MS=1.23 x GC-45 (Passing-Bablok regression). No significant bias between GC and LC-MS/MS (Bland-Altman plot). CONCLUSION: We consider the sample pretreatment without deproteination, derivatization and centrifugation steps and the specificity of the tandem mass spectrometry as the most important points of our amantadine analysis method. SN - 0009-8981 UR - https://www.unboundmedicine.com/medline/citation/15913590/Determination_of_serum_amantadine_by_liquid_chromatography_tandem_mass_spectrometry_ DB - PRIME DP - Unbound Medicine ER -
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