Quantification of testosterone, androstenedione and 17-hydroxyprogesterone in whole blood collected using Mitra microsampling devices

• Published in: Annals of clinical biochemistry 2020-09, Vol.57 (5), p.351-359
• Main Authors: Marshall, David J, Adaway, Joanne E, Hawley, James M, Keevil, Brian G
• Format: Article
• Language: English
• Subjects: 17-alpha-Hydroxyprogesterone - blood; Androstenedione - blood; Blood Specimen Collection - instrumentation; Blood Specimen Collection - methods; Healthy Volunteers; Humans; Testosterone - blood
• ISSN: 0004-5632; 1758-1001
• DOI: 10.1177/0004563220937735

Background Measurement of testosterone (T), androstenedione (A4) and 17-hydroxyprogesterone (17OHP) usually requires a venous serum sample which may have implications for sample stability or collection. Objective A liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay was developed for samples collected using Mitra devices. Analytical validation was completed, and sample comparisons were undertaken to assess Mitra versus venous samples. Method Sample was combined with deionized water and internal standard. After mixing, MTBE was added for extraction. The supernatant was transferred to a deep-well plate and dried prior to re-constitution. A HSS T3 column and Waters TQS Micro was used, the detected quantifier transitions were T m/z 289.2 > 96.95, A4 287.2 > 96.95 and 17OHP 331.25 > 96.95. Results Mean recovery was 102% for T, 98% for A4 and 97% for 17OHP. Lower limit of quantification was 1 nmol/L for T/A4 and 4 nmol/L for 17OHP. T was linear up to 41.6 nmol/L, A4 41.9 nmol/L and 17OHP 72.6 nmol/L. Ion suppression was