Hello, Due to their low volatility and excretion predominantly as sulfate or glucuronide conjugates, urinary steroids require hydrolysis and derivatization following extraction and prior to GC-MS analysis, limiting automation and increasing extraction complexity. Nonetheless, extremely useful procedures have developed to measure up to 40 steroids in a single GC-MS analysis.

LC-MS and LC-MS/MS are commonly used for bile acid analysis in biological fluids without derivatization. Disadvantages of LC-based methods include relatively high carry-over on LC columns and low peak capacity.

GC is a robust, simple and inexpensive lab tool for volatile or semi-volatile compound analysis. The only drawback of GC versus HPLC for bile acid analysis is polar functional groups (carboxyl and hydroxyl) need to be derivatized into GC-detectable forms. Trimethylsilylating reagents, such as N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) and N-methyl-N(trimethylsilyl)trifluoroacetamide (MSTFA), are commonly used for derivatization of both hydroxyl and carboxyl groups.

Hope this info will help you.

Good luck!

Please look at the following below links which may help you in your analysis:

https://uu.diva-portal.org/smash/get/diva2:1050511/FULLTEXT01.pdf

https://academic.oup.com/chromsci/article/50/4/349/386062

Thanks