A High-throughput Method for Accurate Measurement of FT in Serum using ED Coupled with ID-UHPLC-MS/MS

[madman]

Introduction

Testosterone is the most abundant circulating androgen in both men and women. It is a biomarker for identifying androgen excess and deficiency, and to monitor their treatment. Measurement of the metabolically active form, free testosterone (FT), has been part of the clinical workup, especially in individuals with impaired sex hormone binding globulin (SHBG) levels[1]. FT is currently measured using direct commercial immunoassays, assays using equilibrium dialysis (ED), or other separation techniques. Though the ED approach is recognized as the recommended approach, it is laborious and technically challenging for routine clinical use. The CDC Clinical Standardization Program is developing a high throughput method using the ED procedure coupled with isotope dilution ultra-high-performance liquid chromatography-tandem mass spectrometry (ID-UHPLC-MS/MS) that can be used in patient care, public health, and clinical research.




Nov2/2022 (original thread)

Measurement of FT in Serum using ED Coupled with ID-UHPLC-MS/MS: in Comparison to cFTV

Reproductive Endocrinology ODP419 Measurement of Free Testosterone in Serum using Equilibrium Dialysis Coupled with ID-UHPLC-MS/ MS: in Comparison to Calculated FT Hui Zhou, Ph.D., Ashley Ribera, BS, Uliana Danilenko, Ph.D., Otoe Sugahara, BS, and Hubert Vesper, PhD Assessment of free...

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[madman]

Method

 

[madman]

Analytical Parameters

 

[madman]

Results

 

[madman]

Summary

• The CDC ED-UPLC/MS/MS FT method shows good agreement with FT measurements using reference device and has sufficient accuracy, precision, and sensitivity for use in patient care, public health, and clinical research.

• The ED conditions, such as dialysis buffer, are consistent with those used in established reference methods.

• The method employs a well-plate-based ED device that can be automated allowing for processing 150 patient samples within 24 hours.

• Consistent with previous reports, the method shows a bias to calculated FT, which might be related to limitations of the Vermeulen equation