Ultrasensitive sub-pg/ml determination of tiotropium bromide in human plasma by 2D-UHPLC-MS/MS: challenges and solutions

  • Bioanalysis. 2016;8(5):385-95. doi: 10.4155/bio.15.256.
Jingduan Chi  1 ,  Fumin Li  1 ,  Rand Jenkins  2
Affiliations
  • 1. PPD® Laboratories, 3230 Deming Way, Middleton, WI 53562, USA.
  • 2. PPD® Laboratories, 2240 Dabney Rd, Richmond, VA 23230, USA.
PMID: 26893186 DOI: 10.4155/bio.15.256
Abstract

Background: To adequately support pharmacokinetic evaluations of tiotropium bromide in planned clinical studies. It was desirable to measure it with a LLOQ of sub pg/ml.

Results: A sensitive bioanalytical method for the determination of tiotropium in human plasma sample was successfully developed and validated in the range of 0.2-100 pg/ml. The method was successfully applied to support two clinical studies of over 3000 samples. The overall incurred sample reanalysis passing rate was 93.7%.

Conclusion: The combination of a dual stage liquid-liquid extraction and a 2D ultra-HPLC greatly reduced matrix effects and increased assay sensitivity. When developing effective ultrasensitive assays, it is imperative to balance the aspects related to sensitivity with those that will ensure assay ruggedness.

Keywords
2D-UHPLC; dual-stage liquid–liquid extraction; inhaled therapy; tiotropium bromide; ultrasensitive bioanalysis (sub pg/ml).
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