= Emerging. More than 5 years before clinical availability.
= Expected to be clinically available in 1 to 4 years.
= Clinically available now.
MSACL 2018 EU : Marshall

MSACL 2018 EU Abstract

Topic: Endocrinology

Quantification of Testosterone, Androstenedione and 17-Hydroxyprogesterone Collected Using Mitra Micro Sampling Devices

David Marshall (Presenter)
Manchester University NHS Foundation Trust

Presenter Bio: I am a trainee Clinical Scientist at Wythenshawe Hospital in Manchester. I have worked with clinical mass spectrometry for the last 3 years, including routine operation, method development and troubleshooting. I am also in the process of completing a Masters in Clinical Science as part of my training

Authors: David Marshall, Brian Keevil
Manchester University NHS Foundation Trust

Short Abstract

Mitra devices are small polymer tips that absorb a fixed amount of blood which can be dried prior to analysis.

A simple liquid:liquid extraction was used following reconstitution of the mitra device in distilled water. Separation was carried out using a Waters Acquity HSS T3 column which resolved the peaks well.

We have developed an assay for the simultaneous quantification of testosterone, androstenedione and 17-hydroxyprogesterone using a single mitra tip.

Long Abstract

Introduction

Measurement of testosterone, androstenedione and 17-hydroxyprogesterone (17-OHP) usually requires a venous serum sample. A highly sensitive and specific assay was developed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) after extraction from a volumetric absorptive microsampling (VAM) device combined with a simple liquid-liquid extraction clean up procedure.

Methods

Pre-mixed calibrator/sample (10 µL) was combined with deionised water and mixed internal standard, after mixing MTBE was added and mixed by inversion for 1 minute. The supernatant was transferred to a clean 2 mL deep-well plate, dried down and re-constituted with 100 µL 50:50 MEOH/water. Separation was then performed on an HSS T3 2.1 x 50mm column (Waters, Manchester, UK), flow rate of 0.5 ml/min using a gradient of aqueous and organic mobile phases. We detected multiple reaction monitoring transitions for androstenedione were m/z 287.2>96.95, testosterone 289.2>96.95 and 17OHP 331.25>96.95 using a Waters TQD in electrospray-positive mode.

Results

Androstenedione and d7-androstenedione eluted at 2.40 minutes, testosterone and [13C]-testosterone eluted at 2.64 minutes, 17-OHP and [13C]-17-OHP eluted at 2.82 minutes. Mean recovery was 86% for androstenedione, 102% for testosterone and 97% for 17OHP, lower limit of quantification was 1 nmol/L for androstenedione, 1 nmol/L for testosterone and 4 nmol/L for 17-OHP. Androstenedione was linear up to 41.9 nmol/L, testosterone was linear up to 41.6 nmol/L and 17-OHP was linear up to 72.6 nmol/L. Ion suppression was within acceptability criteria for all the analytes measured.

Conclusions & Discussion

We have developed a rapid assay for LC-MS/MS measurement of VAMS for androstenedione, testosterone and 17-OHP in a routine clinical laboratory. It is simple, reproducible and easy to perform.


References & Acknowledgements:


Financial Disclosure

DescriptionY/NSource
GrantsnoMr
SalaryyesWaters
Board Memberno
Stockno
Expensesno

IP Royalty: no

Planning to mention or discuss specific products or technology of the company(ies) listed above:

no