MSACL 2025 Abstract

LC-MS/MS Method Development and Validation for Quantification of Steroids in Serum

Younus Mohammad and Berit Jensen
Canterbury Health Laboratories, Christchurch, New Zealand

Younus Mohammad, PhD (Presenter) Canterbury Health Laboratories

Presenter Bio: I have completed a PhD in Pharmaceutical Sciences from the University of Otago, Dunedin, New Zeland

I have completed my Bachelors and Masters of Pharmacy from Jawaharlal Nehru Technological University, Hyderbad, India.

I have worked in pharmaceutical research laboratories for more than ten years after completing my under graduation. I have joined the Canterbury Health Laboratories as a Scientific Officer in March 2024.

Relevant Financial Disclosures (within past 24 months, reported on Mar 17, 2025) No relevant financial relationship(s) to disclose. Conflict mitigation NOT REQUIRED.

INTRODUCTION:
An accurate, precise and rapid measurement of steroids is crucial in diagnosis, monitoring and providing personalised treatment for many disorders, such as Cushing’s syndrome, Addison’s disease and congenital adrenal hyperplasia. Compared to traditional immunoassays LC-MS/MS provide more specificity, sensitivity and accuracy in measuring very low levels of steroids.

OBJECTIVE:
The aim of present work was to develop a simple LC-MS/MS method for the simultaneous quantification of eight steroid hormones: aldosterone, cortisol, 11-deoxycortisol, 21-deoxycortisol, androstenedione, 17-hydroxypregnenolone, testosterone, 17-hydroxyprogesterone, and dihydrotestosterone (DHT).

METHODS:
Supported liquid extraction was used for sample preparation. Chromatographic separation was achieved on a Cortecs premier C18 column, and 0.05 mM ammonium fluoride in water (+ 2% v/v methanol) and 0.05 mM ammonium fluoride in methanol (+2% v/v water) were employed as mobiles phase solvents. The analytes were detected on Water TQ-XS system, aldosterone was assessed in negative mode and all other analytes were assessed in positive mode. Chrome system calibrators and quality control samples (panel 1 and panel 2) were used. The method was validated for selectivity, sensitivity, accuracy, precision, recovery studies, effect of matrix and dilution integrity. The method also used quality assurance samples for external comparison and was compared against an external laboratory (Pathology Queensland, Australia).

RESULTS:
The analytical run was less than 10 mins. The validation results were found to be within acceptable limits, accuracy <15% compared to reference samples and % CV <10%. Passing-Bablok regression analysis for the method comparison study was within acceptable limits.

CONCLUSIONS:
The method can be a useful tool in determination of steroids in both clinical and scientific laboratory research.