A novel high-throughput assay for the measurement of salivary progesterone by liquid chromatography tandem mass spectrometry
Author(s)
Schiffer, Lina
Adaway, Joanne E
Baranowski, Elizabeth S
Arlt, Wiebke
Keevil, Brian G
Type
Journal Article
Abstract
Background
Liquid chromatography tandem mass spectrometry (LC-MS/MS) enables specific and sensitive quantification of steroids with a high throughput. Saliva sampling is advantageous for multisample profiling over longer periods of time, as it is non-invasive, cheap, can be carried out at home and does not require the attendance of clinical personnel. We developed a rapid LC-MS/MS for the measurement of salivary progesterone, frequently assessed as ovulation marker in patients desiring fertility.
Methods
Samples (300 μL) were prepared by supported liquid extraction using dichloromethane and were reconstituted in 40% methanol. Chromatography was performed using a C8 column with a water/methanol gradient containing 0.1% formic acid and 2 mmol/L ammonium acetate. Quantification was performed with a Waters TQ-S mass spectrometer.
Results
Total run time was 5.5 min. The lower limit of quantification was 20 pmol/L (1.2 fmol on column). Inter- and intra-assay comparison showed coefficients of variation and bias between measured and nominal concentrations of less than 11%. Mean recovery was 91%. Interference with a large set of natural and synthetic steroids was excluded. The assay was successfully applied to measure progesterone variation during the menstrual cycle (n = 9) and diurnal variations during luteal phase (n = 7) in regularly cycling women.
Discussion
We present a novel LC-MS/MS assay for the determination of salivary progesterone with high-throughput potential. The applicability of the assay for progesterone profiling during the menstrual cycle is demonstrated.
Liquid chromatography tandem mass spectrometry (LC-MS/MS) enables specific and sensitive quantification of steroids with a high throughput. Saliva sampling is advantageous for multisample profiling over longer periods of time, as it is non-invasive, cheap, can be carried out at home and does not require the attendance of clinical personnel. We developed a rapid LC-MS/MS for the measurement of salivary progesterone, frequently assessed as ovulation marker in patients desiring fertility.
Methods
Samples (300 μL) were prepared by supported liquid extraction using dichloromethane and were reconstituted in 40% methanol. Chromatography was performed using a C8 column with a water/methanol gradient containing 0.1% formic acid and 2 mmol/L ammonium acetate. Quantification was performed with a Waters TQ-S mass spectrometer.
Results
Total run time was 5.5 min. The lower limit of quantification was 20 pmol/L (1.2 fmol on column). Inter- and intra-assay comparison showed coefficients of variation and bias between measured and nominal concentrations of less than 11%. Mean recovery was 91%. Interference with a large set of natural and synthetic steroids was excluded. The assay was successfully applied to measure progesterone variation during the menstrual cycle (n = 9) and diurnal variations during luteal phase (n = 7) in regularly cycling women.
Discussion
We present a novel LC-MS/MS assay for the determination of salivary progesterone with high-throughput potential. The applicability of the assay for progesterone profiling during the menstrual cycle is demonstrated.
Date Issued
2019-01-01
Date Acceptance
2018-06-01
Citation
Annals of Clinical Biochemistry, 2019, 56 (1), pp.64-71
ISSN
0004-5632
Publisher
SAGE Publications
Start Page
64
End Page
71
Journal / Book Title
Annals of Clinical Biochemistry
Volume
56
Issue
1
Copyright Statement
© The Author(s) 2018
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/29792048
Subjects
ADULT
COLLECTION DEVICES
DIRECT RADIOIMMUNOASSAY
ENZYME-IMMUNOASSAY
Life Sciences & Biomedicine
liquid chromatography tandem mass spectrometry
Medical Laboratory Technology
PHASE
PLASMA
PRETERM BIRTH
Progesterone
saliva
Science & Technology
STEROID-HORMONES
VALIDATION
WOMEN
Publication Status
Published
Coverage Spatial
England
Date Publish Online
2018-06-11