Highly Sensitive and Selective Method for Estimation of Formoterol at Sub-pg/mL in Human Plasma Using Shimadzu LCMS-8060NX

Significance of the Topic

Formoterol is a long acting beta2 agonist with rapid onset used in asthma and COPD management. Its low bioavailability and need for accurate quantification in plasma at sub pg mL levels drive the development of sensitive analytical methods.

Objectives and Study Overview

The study aimed to develop and validate a highly sensitive LC MS MS method using Shimadzu LCMS 8060NX and Nexera X2 for quantifying formoterol in human plasma at concentrations down to 0.2 pg mL. The method was evaluated for selectivity, linearity, precision, accuracy, recovery, matrix effects and carryover.

Methodology and Instrumentation

• Sample preparation by solid phase extraction with 200 µL extraction buffer in K2 EDTA plasma
• SPE cartridges conditioned with methanol and water, washed and eluted with 10 percent acetonitrile
• Chromatography on Shim pack Velox C18 column with isocratic mobile phase 0.1 percent formic acid in 5 mM ammonium acetate and acetonitrile at 0.3 mL per minute
• Detection on Shimadzu LCMS 8060NX with ESI source in MRM mode

Main Results and Discussion

• Method displayed excellent selectivity across six plasma lots with no interference at the retention time of formoterol
• Calibration curve linear from 0.2 to 100 pg mL with correlation coefficient above 0.99
• Intra and inter day precision and accuracy within acceptance criteria percent CV under 12 at LLOQ and under 6 at higher levels
• Extraction recovery ranged from 58 to 65 percent with overall precision below 8 percent RSD
• Matrix factor close to unity at LQC and HQC levels indicating minimal matrix effects
• No carryover observed after high concentration injections

Benefits and Practical Applications

The method supports high throughput quantification of formoterol in pharmacokinetic and clinical studies, requiring minimal plasma volume and providing fast runtime. It is suitable for drug development and therapeutic monitoring in respiratory and cardiovascular research.

Future Trends and Applications

The approach may be extended to multiplexed analysis of beta agonists and biomarker panels. Advances in ionization and microflow LC MS could further reduce sample volume and enhance sensitivity. Integration with automated sample preparation and high resolution MS holds potential for broader clinical application and metabolite profiling.

Conclusion

A robust, rapid and ultra sensitive LC MS MS method has been established for quantifying formoterol down to 0.2 pg mL in human plasma. Validation demonstrated excellent performance in specificity, linearity, precision, accuracy, recovery and minimal matrix interference, making it a valuable analytical tool for pharmacology and clinical studies.

Used Instrumentation

• Shimadzu Nexera X2 UHPLC system
• Shimadzu LCMS 8060NX triple quadrupole mass spectrometer
• Shim pack Velox C18 column 100 × 2.1 mm, 2.7 µm

Reference

• Gaikwad AB, Atmakuri CK, Arote YG Highly Sensitive and Selective Method for Estimation of Formoterol at Sub pg mL in Human Plasma Using Shimadzu LCMS 8060NX ADC Shimadzu Analytical India Pvt Ltd 2024