Steroid Hormone Analysis with Supel™ Swift HLB DPX

Steroid hormone analysis is critical for diagnosing and monitoring diseases like Cushing’s syndrome and Addison’s disease. The Supel™ Swift HLB DPX Tips are advantageous for hormone analysis, enabling reduced sample volume, preventing evaporation, and allowing automated preparation. This method effectively analyzes nine steroids, enhancing various diagnostic applications.

Method overview

The Supel Swift HLB sorbent provides excellent selectivity and sensitivity for steroids in neutral solutions, permitting dilution with water before injection. This method uses 100 µL of serum, yielding a final injection volume of 100 µL without solvent evaporation, with recoveries ranging from 65–86 percent. The method’s LOQs are below clinically relevant values, and it can process up to 96 samples simultaneously in about 20 minutes before LC-MS/MS analysis. This automation and efficiency make the method suitable for high-throughput clinical laboratories.

The versatility of the Supel Swift HLB sorbent, due to its hydrophilic and lipophilic functional groups, allows it to bind a wide range of analytes effectively. This ensures that the method can be applied to various clinical scenarios, providing reliable and accurate results for different steroid hormones.

Experimental conditions

A Hamilton® Microlab NIMBUS96 automates sample preparation with Supel Swift DPX HLB Tips. The analysis employs an Agilent 1290 LC system with a SCIEX Triple Quad™ 6500+ mass spectrometer and an Ascentis® Express C18 column. The LC conditions include a gradient of ammonium formate in water and methanol. The sample preparation involves serum aliquoting, internal standard addition, formic acid mixing, protein dissociation, and automated extraction with wash and elution steps.

The automated liquid handler (ALH) performs a series of steps, including adding aqueous formic acid, protein dissociation, and automated extraction with wash and elution steps, ensuring thorough and efficient sample processing. The use of automated systems minimizes human error and variability, providing more consistent and reliable results.

Results & discussion

The method ensures optimal compound separation, crucial for complex biological matrices. Precision and accuracy studies revealed high reproducibility and minimal ion suppression/enhancement effects. The method shows excellent linearity and low bias, comparable to standard methods. Recoveries range from 65–86 percent, with average ionization suppression at 33 percent. The method’s LOQs are well below clinically relevant values, ensuring that even low concentrations of steroid hormones can be accurately quantified.

The precision and accuracy study demonstrated high reproducibility and minimal ion suppression/enhancement effects. The correlation graphs comparing the Supel Swift HLB DPX method with standard methods showed excellent linearity and low bias, indicating that the two methods are interchangeable. This further validates the reliability and accuracy of the Supel Swift HLB DPX Tips for clinical steroid hormone analysis.

Conclusion

Supel™ Swift HLB DPX Tips offer a reliable, fast, and programmable method for serum steroid analysis, providing the necessary clinical sensitivity and high-throughput for quick turnaround times. This method is a valuable tool for steroid quantification in clinical settings, ensuring accurate and sensitive analysis. The automated nature of the method reduces the potential for human error, improves consistency, and increases the efficiency of sample processing. For clinical laboratories, this method offers an efficient solution for the accurate determination of steroid hormones, supporting better diagnostics and patient care.

For more information, visit SigmaAldrich.com/Clinical&Forensic.