Research Use Cases for Mass Spect Products: MSG1000

The research paper titled:

A Sensitive and Cost-Effective LC–MS-MS Method for Determination of 1α,25-Dihydroxyvitamin D3 in Human Plasma

by Bruce A. Stanley, Jenny P. Dai, Allan Xu, Eric J. Battaglioli, and Robin T. Wilson, published in 2015.

- Full paper can be found here.

The research paper discusses the development of a sensitive and cost-effective method for quantifying 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3) in human plasma.

Importance of 1,25(OH)2D3: 1,25(OH)2D3 is the biologically active form of vitamin D, which plays a crucial role in maintaining bone health and regulating various biological processes, including immune function, cell proliferation, differentiation, and apoptosis.

Challenges in Quantification: Accurate quantification of 1,25(OH)2D3 is challenging due to its extremely low concentrations in human plasma, typically in the low picograms-per-milliliter range.

Previous Methods: Prior methods to increase the sensitivity of liquid chromatography–tandem mass spectrometry (LC–MS-MS) assays have used more sensitive derivatization reagents or microflow LC–MS-MS concepts, but these methods had drawbacks related to cost, availability, stability of reagents, and system complexity.

New Method: The study presents a novel LC–MS-MS method that combines immunoaffinity extraction (IAE), 4-phenyl-1,2,4-triazole-3,5-dione (PTAD) derivatization, and methylamine adduction. This method aims to improve the accuracy and sensitivity of 1,25(OH)2D3 quantification.

Immunoaffinity Extraction (IAE): IAE is used to separate 1,25(OH)2D3 from matrix interferences, resulting in higher selectivity and a cleaner sample. It reduces the need for extensive separation on an LC column, shortens the run time, and is cost-effective.

Derivatization Reagent: PTAD is chosen as the derivatization reagent due to its practicality, lower cost, and relative stability. It provides sensitive and reliable measurement of 1,25(OH)2D3 when used in combination with methylamine.

Optimization: The study discusses the optimization of various parameters, including PTAD concentration, methylamine concentration in the mobile phase, and choice of organic solvent, to maximize sensitivity and reduce run time.

Method Validation: The developed method is validated for linearity, sensitivity, accuracy, precision, selectivity, recovery, and matrix effect. It is shown to be highly accurate and precise with a low limit of quantification (LLOQ) of 5 pg/mL.

Comparison with Prior Method: The new method, using PTAD and methylamine, is compared with the previous method using a more expensive derivatization reagent. Both methods yield similar results with the same LLOQ, but the new method is more cost-effective and faster.

Conclusion: The study concludes that the combination of IAE, PTAD derivatization, and methylamine adduction offers a sensitive and cost-effective solution for the analysis of 1,25(OH)2D3 in human plasma. This method is practical, has lower costs, and can be widely adopted in LC–MS-MS laboratories.

Mass Spect Gold Human Serum Ultra-Low Vitamin D (MSG1000) product played a crucial role in the researchers' objective of developing a sensitive and cost-effective method for quantifying 1,25(OH)2D3 in human plasma. Here's how the MSG1000 was used to achieve their goal:

Calibration Standards: MSG1000 was utilized to create calibration standards for the analysis. These standards involved spiking MSG1000 with known concentrations of 1,25(OH)2D3, ranging from low to high levels.

Testing Accuracy: These calibration standards, prepared using MSG1000, were used to assess the accuracy and precision of the developed method. The researchers compared the results from analyzing the calibration standards to the known concentrations, ensuring that the method provided accurate measurements.

Method Validation: MSG1000 played a role in the method validation process. It was employed to verify the linearity, sensitivity, and precision of the method across a range of concentrations.

By using MSG1000 to create calibration standards and validate their method, the researchers ensured the accuracy and reliability of their approach for quantifying 1,25(OH)2D3 in human plasma. This was a critical step in achieving their objective of developing a practical and cost-effective method for this analysis.

References:

Stanley, B. A., Dai, J. P., Xu, A., Battaglioli, E. J., & Wilson, R. T. (2015, February 28). A sensitive and cost-effective lc–ms-ms method for determination of 1α,25-dihydroxyvitamin d3 in human plasma. Spectroscopy . Retrieved October 15, 2023, from https://www.spectroscopyonline.com/view/sensitive-and-cost-effective-lc-ms-ms-method-determination-1-25-dihydroxyvitamin-d3-human-plasma.