Steroid Hormone Analysis in Serum Using XTR tips with Supel™ Swift HLB

Automation Compatibility: Hamilton Robotics

Routine hormone analysis is necessary for establishing and monitoring patient diseases. For example, the continuous monitoring of cortisol levels can help diagnose a patient with Cushing disease (high cortisol) or Addison disease (low cortisol). A robust method for steroid hormone determination in serum is imperative in diagnostics and treatment. XTR tips with 3 mg of Supel™ Swift HLB sorbent chemistry were used in Hamilton format. This allowed for reduced sample volume, sample evaporation mitigation, and offered a fully automated approach. In this method, a total of 9 steroids (cortisone, cortisol, 11-deoxycortisol, androstenedione, testosterone, dehydroepiandrosterone, 5-dihydrotestosterone, 17-hydroxyprogesterone, and progesterone) were analyzed as a panel to cover a variety of testing applications and disease diagnostics.

Method
Condition20% MeOH/ H2O
Aspirate/ Dispense x2
BindAspirate/ Dispense Diluted Sample x5
Wash 1100% H2O
Aspirate/ Dispense x3
Wash 220% MeOH
Aspirate/ Dispense x3
Elute75 µL Elution (50/50 MeOH/ACN)
Aspirate/ Dispense x3
+Final Dilution with 25 µL water

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Supel™ Swift HLB from Millipore Sigma is a proprietary and patent pending copolymer having both hydrophilic and lipophilic functional groups. The sorbent’s dual polarity nature makes it ideal for extracting a broad range of compounds from aqueous matrices, and is an appropriate choice in applications for food & environmental samples as well as biological samples such as urine, serum and plasma. The hydrophilic and lipophilic balance (HLB) property of the polymer material enables the retention of a wide range of compounds having a wide range of polarities and Log P values.

You can order XTR tips with Supel™ Swift HLB Online via Sigma Aldrich

Sigma Aldrich carries XTR tips with this propriety sorbent in Universal (manual) format and Hamilton Robotics format. Standard sorbent chemistries amounts are available in 3, 5, 10, and 20 mg for 300 µL or 1 mL tips.