Method Development 

Accurate steroid analysis is far more complex than it appears. It requires not only advanced instrumentation, but extensive method development, validation, and control over multiple variables in the analytical process.  Our methods have been built through significant time, experimentation, and technical refinement.

Accuracy

• We utilize certified reference standards alongside isotopically labeled internal standards to ensure precise compound identification and accurate quantification.

• All internal standard additions are performed using positive displacement pipettes, minimizing volumetric error and ensuring consistent accuracy

Extensive Method Development

We have systematically developed and optimized our methods by evaluating:

• Multiple chromatographic columns, including variations in stationary phase and column length

• Mobile phase compositions and gradient ramping strategies

• Temperature conditions affecting compound separation

• Resolution capabilities on high-resolution Orbitrap instrumentation

Advanced Mass Spectrometry Techniques

To maximize both sensitivity and specificity, we have tested and implemented multiple acquisition strategies, including:

• Parallel Reaction Monitoring (PRM) for highly specific targeted analysis

• Targeted SIM (tSIM) for improved detection of low-abundance compounds

• Resolution and scan parameter optimization for confident compound identification

Why This Matters 

This level of development allows us to:

• Accurately distinguish between closely related compounds

• Detect low-level impurities and contaminants

• Provide consistent and reproducible results