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Qualification and application of a liquid chromatography–tandem mass spectrometric method for the determination of human Aβ1-40 and Aβ1-42 peptides in transgenic mouse plasma using micro-elution solid phase extraction

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Abstract

A liquid chromatographic–tandem mass spectrometric (LC–MS/MS) method was developed and applied for the determination of human Aβ1-40 and Aβ1-42 peptides in transgenic mouse plasma to support preclinical pharmacodynamics studies. The method consisted of micro-elution solid phase extraction for sample preparation and LC–MS/MS analysis in the negative ion mode using electrospray ionization for analysis. 15N53-Aβ1-40 and 15N55-Aβ1-42 peptides were used as internal standards. A quadratic regression (weighted 1/concentrations), with an equation y = ax 2 + bx + c, was used to fit calibration curves over the concentration range of 0.500–100 ng/mL for both Aβ1-40 and Aβ1-42 peptides. For quality control samples at 6.00, 40.0 and 80.0 ng/mL from the qualification experiment, the within-run accuracy ranged from −2.69 to 0.583 % with precision values ≤8.23 % for Aβ1-40. Within-run accuracy ranged from −4.83 to 10.1 % with precision values ≤8.87 % for Aβ1-42. Samples from a pharmacodynamics study using Tg2576 transgenic mice were analyzed by this qualified LC–MS/MS method and concentrations were compared to those generated by ELISA. The two methods were shown to be comparable for Aβ1-40 quantification of samples from the Tg2576 amyloid precursor protein transgenic mouse model, but varied slightly for Aβ1-42.

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Acknowledgments

Authors thank DMPK members at Genentech, Array Biopharma and Intertek-Alta Analytical Laboratory scientists for their support on this project. Authors also specially thank Dr. Marcel Hop for his support, review and comments on this manuscript.

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Author notes

  1. Young G. Shin

    Present address: College of Pharmacy, Chungnam National University, 99 Daehak-ro, Yuseong-gu, Daejeon, 305-764, Republic of Korea

Authors and Affiliations

  1. Drug Metabolism and Pharmacokinetics, Genentech Inc., 1 DNA Way, Mail Stop 412A, South San Francisco, CA, 94080, USA

    Young G. Shin, Kimberly Scearce-Levie, Ryan J. Watts & Xingrong Liu

  2. Alta Analytical Laboratory, Intertek Pharmaceutical Services, 1100 Windfield Way, El Dorado Hills, CA, 95762, USA

    Lee Hamm, Stanley Murakami, Kenji Buirst & Michael H. Buonarati

  3. Array Biopharma Inc., 3200 Walnut Street, Boulder, CO, 80301, USA

    April Cox, Kelly Regal & Kevin W. Hunt

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Correspondence to Young G. Shin or Xingrong Liu.

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Shin, Y.G., Hamm, L., Murakami, S. et al. Qualification and application of a liquid chromatography–tandem mass spectrometric method for the determination of human Aβ1-40 and Aβ1-42 peptides in transgenic mouse plasma using micro-elution solid phase extraction. Arch. Pharm. Res. 37, 636–644 (2014). https://doi.org/10.1007/s12272-013-0215-2

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