Measurement of serum estrogen and estrogen metabolites in pre- and postmenopausal women with osteoarthritis using high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry

Abstract

Although 17β-estradiol (E2) deficiency has been linked to the development of osteoarthritis (OA) in middle-aged women, there are few studies relating other estrogens and estrogen metabolites (EMs) to this condition. We developed a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) method to measure the levels of six EMs (i.e., estrone, E2, estriol, 2-hydroxyestrone, 2-hydroxyestradiol, and 16a-hydroxyestrone) in healthy pre- and postmenopausal women and women with OA. This method had a precision ranging from 1.1 to 3.1% and a detection limit ranging from 10 to 15 pg. Compared to healthy women, serum-free E2 was lower in the luteal and postmenopausal phases in women with OA, and total serum E2 was lower in postmenopausal women with OA. Moreover, compared to healthy women, total serum 2-hydroxyestradiol was higher in postmenopausal women with OA and total serum 2-hydroxyestrone was lower in both the luteal and follicular phases in women with OA. In conclusion, our HPLC-ESI-MS/MS method allowed the measurement of multiple biochemical targets in a single assay, and, given its increased cost-effectiveness, simplicity, and speed relative to previous methods, this method is suitable for clinical studies.

Derivatization enhances analysis of estrogens and their bioactive metabolites in human plasma by liquid chromatography tandem mass spectrometry

“…The LOD and LLOQs for estrogen MPPZ derivatives ( Table 2 ) measured in standard solutions are comparable to previous methods for the analysis of E1 and E2 [ 16 , 21 , [32] , [33] , [34] ]. The volume of plasma (0.5 mL) extracted by SPE for detection of metabolites demonstrated an improvement over recent methods for oncology studies; one study by Yang et al., 2008 used 2 mL of patient serum using a longer extraction protocol where samples were incubated overnight prior to liquid extraction of estrogens [ 35 ].…”

Current strategies for quantification of estrogens in clinical research

“…For the applications discussed here, reversed phase chromatography is almost exclusively used, using LC columns with hydrophobic stationary phase in conjunction with a polar mobile phase. C18 columns [49,[55], [56], [57],61,63,64,69,70,72,78] with their enhanced retention capabilities and robust, consistent manufacturing quality are preferred. Chemical alterations to bonded stationary phases, again exploiting interactions with the aromatic ring, can improve distinction of isomeric structures, typically required for the estrogen metabolites.…”

Are estrogen-related drugs new alternatives for the management of osteoarthritis?

“…In-vivo estrogen treatment has shown inconclusive results in OA. Gao et al [ 21 ] found that serum-free estradiol levels and total serum estradiol levels were significantly lower in postmenopausal compared with premenopausal OA women. Afzal and Khanam [ 22 ] found that estrogen deficiency may lead to increased serum IL-6 in postmenopausal patients with OA, which has been found to promote OA progression.…”