Rapid, Sensitive and Direct Quantitation of Tiotropium at sub-pg/mL in Plasma using Shimadzu LCMS-8060

User Benefits

- Rapid, simple, and most sensitive method with LLOQ of 0.2 pg/mL - Low plasma volumes in sample extraction extends the life of mass spectrometer - Single step sample extraction method increased sample productivity

Introduction

Tiotropium is an inhaled long-acting anti-cholinergic for the maintenance treatment of COPD (chronic obstructive pulmonary disease) . It is chemically described as (1α, 2β, 4β, 5α, 7β) -7- [ (Hydroxydi-2-thienylacetyl) oxy] – 9.9 – dimethyl - 3 - oxa - 9 - azoniatricyclo [3.3.1.0] nonane bromide monohydrate. It is a synthetic, non-chiral, quarternary ammonium compound . Structure of tiotropium and tiotropium D3 bromide is provided in Fig. 1 and Fig. 2. The long duration of action with tiotropium is owing to prolonged, competitive binding to M (3) muscarinic receptors. Tiotropium is administered in the form of dry powder inhalation which results in very low systemic bioavailability. This translates into significant challenges to develop a sensitive and reproducible bioanalytical method that can reliably measure plasma levels of tiotropium at very low expected levels. The required LLOQ for most inhalation dosed medications is typically in the range of pg/mL to sub pg/mL. Several analytical methods have been developed to determine tiotropium in biological samples using HPLC with tandem mass spectrometric detection (LC–MS/MS), with the lowest reported LLOQ of 0.500 pg/ml. These methods fall short of the ideal target sensitivity required by the intended low dose studies, and this motivated us for the current study. The main aim of this work is to develop a LC-MS/MS method at sub-picogram level (LLOQ - 0.2pg/mL) to support regulatory studies.

October 30, 2025 GMT