= Emerging. More than 5 years before clinical availability. |
= Expected to be clinically available in 1 to 4 years. |
= Clinically available now. |
Topic: Endocrinology
Authors: David Marshall, Brian Keevil
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Short Abstract Mitra devices are small polymer tips that absorb a fixed amount of blood which can be dried prior to analysis. |
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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. |
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References & Acknowledgements:
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Description | Y/N | Source |
Grants | no | Mr |
Salary | yes | Waters |
Board Member | no | |
Stock | no | |
Expenses | no |
IP Royalty: no
Planning to mention or discuss specific products or technology of the company(ies) listed above: | no |