Simultaneous determination of 18 steroids in the hypothalamic pituitary gonadal axis based on UPLC-MS/MS with...

Area of Science:

• Environmental Science
• Analytical Chemistry
• Endocrinology

Background:

• Environmental contaminants can disrupt endocrine systems by altering steroid hormone levels.
• Accurate quantification of multiple steroid hormones is crucial for assessing these disruptions.
• Existing methods may require large sample volumes or struggle with specific analytes like cholesterol.

Purpose of the Study:

• To develop a simple, sensitive, and simultaneous ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method.
• To quantify 18 key steroid hormones involved in the hypothalamic-pituitary-gonadal (HPG) axis.
• To enable objective assessment of environmental load impacts on steroid hormone profiles.

Main Methods:

• Development of a UPLC-MS/MS method utilizing liquid extraction and a multimode electrospray/atmospheric pressure chemical ionization (ESCi) source.
• Optimization of mass spectrometry, liquid phase, and pretreatment steps for 18 steroid hormones, including cholesterol.
• Validation of the method for linearity (r² > 0.99), recovery (76.22-113.66%), and precision (7.52-1.14%) using minimal serum volume (100 μL).

Main Results:

• Successful simultaneous determination of 18 steroid hormones: cholesterol, aldosterone, cortisone, hydrocortisone, 21-deoxycortisol, corticosterone, 11-deoxycortisol, androstenedione, estradiol, estrone, 2-methoxyestradiol, 21-hydroxyprogesterone, 17-α hydroxyprogesterone, testosterone, dehydroepiandrosterone, progesterone, dihydrotestosterone, and pregnenolone.
• The method demonstrated high sensitivity, linearity, recovery, and precision.
• The method was successfully applied to serum samples from *Mus musculus*, *Carassius auratus*, *Rana catesbeiana Shaw*, and *Rana nigromaculata*.

Conclusions:

• The developed UPLC-MS/MS method provides an efficient and sensitive tool for simultaneous steroid hormone analysis.
• This method requires a small serum volume and can quantify challenging steroids like cholesterol.
• It is highly valuable for assessing environmental impacts on the HPG axis across diverse species.