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Protein Precipitation Method for Determination of Zileuton in Human Plasma by LC-MS/MS

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Abstract

Purpose

The main aim of this study was to establish a rapid and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the determination of Zileuton in human plasma.

Methods

Zileuton was extracted from human plasma samples by protein precipitation with acidified acetonitrile. Following protein precipitation (PPT), analyte separation was done using Phenomenex Kinetex™ XB-C18 (50 × 2.1 mm, 1.7 μm) column using isocratic elution with a mobile phase of 0.1% formic acid (50%) and 0.1% formic acid:acetonitrile 2:3 V:V (50%) at a flow rate of 0.4 mL/min and an injection volume of 5 μL. The detection was performed on a triple quadrupole mass spectrometer by multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z 237.0 → 161.0, 241.0 → 165.0 for Zileuton and Zileuton-D4 in positive electrospray ionization mode, correspondingly. The method was validated over a concentration range of 20.0–8000.0 ng/mL on SCIEX Triple Quad 4500 MS System. Total run time was 2.1 min.

Results

The method was fully validated. Precision and accuracy results were as follows: the overall %Accuracy for intra batch LLOQ QC was within 93.7–109.5%, for the rest of QCs 98.8–111.6%, and the highest %CV for LLOQ QC was 10.1%, and for the rest of QCs 5.3%.

The variety of tests for matrix effects, extraction efficiency, selectivity, linearity, sensitivity, recovery, and stability were also performed.

Conclusions

A rapid, simple, and specific LC-ESI-MS method for determination of Zileuton in human plasma has been described. The method has several advantages compared to the previously reported methods, including shorter analysis time, better sensitivity, simple sample preparation, wider range of concentrations, and use of isotopic internal standard.

The developed and validated method can be successfully applied for the bioequivalence/pharmacokinetic studies of Zileuton.

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References

  1. Bell RL, Young PR, Albert D, Lanni C, Summers JB, Brooks DW, et al. The discovery and development of zileuton: an orally active 5-lipoxygenase inhibitor. Int J Immunopharmacol. 1992;14:505–10.

    Article  CAS  Google Scholar 

  2. Granneman GR, Braeckman RA, Erdman KA. Determination of a new 5-lipoxygenase inhibitor, Zileuton, and its inactive N-dehydroxylated metabolite in plasma by high performance liquid chromatography. Clin Pharmacokinet. 1995;29(Suppl 2):1–8.

    Article  CAS  Google Scholar 

  3. Israel E, Dermarkarian R, Rosenberg M, Sperling R, Taylor G, Rubin P, et al. The effects of a 5-lipoxygenase inhibitor on asthma induced by cold, dry air. New Eng J Med. 1990;323:1740–4.

    Article  CAS  Google Scholar 

  4. Awni WM, Braeckman RA, Granneman GR, Witt G, Dubé LM. Pharmacokinetics and pharmacodynamics of zileuton after oral administration of single and multiple dose regimens of zileuton 600 mg in healthy volunteers. Clin Pharmacokinet. 1995;29(Suppl 2):22–33.

    Article  CAS  Google Scholar 

  5. Zui Lin Change, Analytical profiles of drug substances and excipients, Vol 25, North Chicago, IL, 1998.

  6. The United States Pharmacopeia. The United States Pharmacopoeial Convention, Inc., Rock Ville, MD, USA, 3891–3893.

  7. Martindale. The complete drug reference. 34th ed. London: The Pharmaceutical Press; 2005. p. 807.

    Google Scholar 

  8. The Merck index, an encyclopedia of chemicals, drugs and biologicals, 14th Edition, Merck Research Laboratories, a division of Merck and Co. Inc., NJ, USA-2006, 10124.

  9. Pian P, Labovitz E, Hoffman K, Clavijo CF, Lynn RR, Galinkin JL, et al. Quantification of the 5-lipoxygenase inhibitor zileuton in human plasma using high performance liquid chromatography-tandem mass spectrometry. J Chromatogr B. 2013;937:79–83.

    Article  CAS  Google Scholar 

  10. Prakash K, Adiki SK, Kalakuntla RR. Development and validation of liquid chromatography-mass spectrometry method for the determination of zileuton in human plasma. Sci Pharm. 2014;82:57–583.

    Article  Google Scholar 

  11. Awni WM, Wong S, Chu S, Patterson K, Hansen R, Machinist JM, et al. Pharmacokinetics of Zileuton and its metabolites in patients with renal impairment. J Clin Pharmacol. 1997;37:395–404.

    Article  CAS  Google Scholar 

  12. FDA guidance on “Bioanalytical method validation”. 2001.

  13. EMA “Guideline on the investigation of bioequivalence”. Doc. Ref.: CPMP/EWP/QWP/1401/98 Rev. 1/Corr

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Authors and Affiliations

  1. DarmanTest Laboratories LLC, 6/1 Abelyan Street, 0038, Yerevan, Armenia

    Yeghig Armoudjian, Astghik Mikayelyan, Hasmik Zakaryan, Armine Aleksanyan & Harutyun Alaverdyan

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  1. Yeghig Armoudjian
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  2. Astghik Mikayelyan
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  3. Hasmik Zakaryan
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  4. Armine Aleksanyan
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  5. Harutyun Alaverdyan
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Correspondence to Astghik Mikayelyan.

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Armoudjian, Y., Mikayelyan, A., Zakaryan, H. et al. Protein Precipitation Method for Determination of Zileuton in Human Plasma by LC-MS/MS. J Pharm Innov 15, 581–590 (2020). https://doi.org/10.1007/s12247-019-09403-6

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  • DOI: https://doi.org/10.1007/s12247-019-09403-6

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