Simple, Sensitive and Rapid Quantification of Teriparatide in Human Plasma by LCMS-8060

Significance of the Topic

Quantification of therapeutic peptides such as teriparatide in human plasma is essential for pharmacokinetic studies and dose optimization in osteoporosis treatment. Teriparatide exhibits rapid absorption and elimination at low picogram levels, creating challenges in sensitivity, selectivity, and reproducibility. Developing robust bioanalytical methods ensures accurate monitoring of peptide levels, supports regulatory compliance, and advances peptide drug development.

Objectives and Article Overview

This work describes the development and validation of a sensitive, rapid LC–MS/MS assay for teriparatide quantification in human plasma using the Shimadzu LCMS-8060 triple quadrupole mass spectrometer coupled with Nexera X2 UHPLC. The main goals were to achieve a low limit of quantification (LLOQ) of 5 pg/mL, optimize sample preparation via single-step solid-phase extraction (SPE), and demonstrate method performance across regulatory validation parameters.

Methodology and Instrumentation

The analytical workflow combined protein precipitation-free SPE with LC–MS/MS detection.

• Sample Preparation: Plasma was extracted under basic conditions using a single-step SPE procedure with conditioning (methanol, water), dual washes, elution, nitrogen evaporation, and reconstitution in 100 µL solution.
• Chromatography: Separation on a Shim-pack Velox C18 column (100 × 2.1 mm, 2.7 µm) at 0.3 mL/min with a gradient of 0.1% formic acid in water and acetonitrile; run time 10 minutes, retention time 5.74 min.
• Mass Spectrometry: Shimadzu LCMS-8060 with heated ESI and UF-Qarray ion guide. Positive MRM mode monitoring the 687.25→787.45 m/z transition with optimized collision energy (−20.2 V). Interface temperature 300 °C, heat block 300 °C, DL 250 °C.

Main Results and Discussion

• Linearity: Calibration from 5 to 300 pg/mL with r² > 0.98; signal-to-noise >10 at LLOQ.
• Precision and Accuracy: Intra- and inter-day accuracy ranged 93–112% of nominal with RSD ≤ 15% at LLOQ and ≤ 7% at higher concentrations.
• Recovery: Mean recovery 80.4% (RSD 7.7%) across LQC, MQC, HQC levels, demonstrating reproducibility.
• Matrix Effects: Matrix factor ~1.03 (CV ≤ 9%), indicating minimal ion suppression or enhancement.
• Carryover: No detectable carryover observed after highest calibrator injections.

Benefits and Practical Applications

• High sensitivity and selectivity suitable for pharmacokinetic and bioequivalence studies.
• Simple, single-step SPE increases throughput and reduces sample handling.
• Validated method meets US FDA bioanalytical guidelines, ready for routine clinical and research applications.
• Optimized instrumentation and ready-to-use protocols facilitate rapid implementation in analytical laboratories.

Future Trends and Potential Applications

Advances in high-resolution mass spectrometry and microflow liquid chromatography may further improve sensitivity and reduce sample consumption. Integration with automated sample preparation platforms can enhance throughput. The method can be adapted for other peptide hormones and biomarkers, supporting personalized medicine and drug development.

Conclusion

The validated LCMS-8060 method provides a robust, sensitive, and streamlined approach for quantifying teriparatide in human plasma at picogram levels. The combination of single-step SPE and optimized UHPLC–MS/MS parameters ensures reproducibility, regulatory compliance, and suitability for pharmacokinetic studies.

References

1. DrugBank. Teriparatide. Available at https://go.drugbank.com/drugs/DB06285 (Accessed March 8, 2022)
2. RxList. Fortéo (Teriparatide). Available at https://www.rxlist.com/forteo-drug.htm (Accessed March 8, 2022)