Medroxyprogesterone Acetate and Progesterone Measurement in Human Serum: Assessments of Contraceptive Efficacy

Abstract

Medroxyprogesterone (MPA) is widely used as a contraceptive or for hormone replacement therapy after menopause. The use of MPA, specifically in adolescent girls, is linked to excess weight gain and decreases in bone mineral density. Metabolism and clearance of MPA is highly variable making this agent a candidate for individualized dosing strategies which could possibly decrease unwanted side effects. MPA has been measured in animal tissues and fluids for many years as a contaminant resulting from the use of growth hormones in food animals.MPA has been measured by radioimmunoassay, gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS/MS). LC-MS/MS techniques have proven to be both more specific than radioimmunoassay and required much simpler sample preparation techniques than GC-MS. Here we describe an LC-MS/MS method specifically designed for monitoring MPA and progesterone levels in human serum. The technique in this report involves minimal sample preparation and yields results with precision and accuracy suitable for clinical use

Cervical and systemic concentrations of long acting hormonal contraceptive (LARC) progestins depend on delivery method: Implications for the study of HIV transmission

“…Cervical secretions from visit 1, visit 2 (all contraceptive progestin treatments), and visit 3 (Mirena only), were eluted using 100% methanol and secretion volumes quantified utilizing previously published protocols [37]. Progesterone and progestin levels were quantified using liquid chromatography-mass spectrometry/mass-spectrometry (LC-MS/MS) methodologies [38, 39].…”

Simultaneous quantitation of multiple contraceptive hormones in human serum by LC–MS/MS

“…In fact, because of the improvements in sensitivity and specificity, MS assays are frequently referred to as the “gold standard” of serum steroid measurement [2]. Despite this, there are only a few methods for determining multiple progestins in a single analysis [4–6] with many devoted to a single progestin and few including simultaneous analysis of estrogens [5,7–9]. Many methods require large sample volumes to achieve adequate sensitivity [5–8] while those that use smaller sample volumes are often less sensitive [4].…”

Rapid Quantification of Medroxyprogesterone Acetate (MPA) in Human Plasma by LC-MS/MS

“…Previously published methods for MPA are laborious and involve analyte extraction through iterative rounds of liquid-liquid extractions, pooling of eluents, and high volume requirements, typically in the 1.0 -3.0 mL range (15)(16)(17). Further, analytical run times typically range from 10 to 14 min, which is unrealistic for high-throughput testing required for supporting larger clinical trials (15)(16)(17)(18)(19). Through the optimization of sample extraction and chromatographic separation parameters, a method was developed that requires 600 μL plasma and has an analytical run time of 4 min with a C18 column with a 5.0-μm bore size.…”

“…The majority of methods for MPA quantification via LC-MS/MS require large sample volumes, typically in excess of 1 mL; extensive sample preparation; and analytical run times in excess of 10 min per sample (15)(16)(17)(18)(19). Efforts have been made recently to reduce sample volumes and improve sample preparation; however, issues such as long analytical run times and assay sensitivity were not overcome (19).…”

“…Various methods have described the quantification of MPA in serum or plasma, and the majority of approaches exploit LC-MS/MS techniques (12)(13)(14)(15)(16)(17)(18)(19)(20). The majority of methods for MPA quantification via LC-MS/MS require large sample volumes, typically in excess of 1 mL; extensive sample preparation; and analytical run times in excess of 10 min per sample (15)(16)(17)(18)(19).…”