MassChrom® Steroids in Serum/Plasma with 96-SPE-Well-Plate - LC-MS/MS

MassChrom® Steroids in Serum/Plasma with 96-SPE-Well-Plate - LC-MS/MS

   
  • 11-Deoxycortisol
  • 17-OH-progesterone
  • Aldosterone
  • Androstenedione
  • Corticosterone
  • Cortisol
  • Cortisone
  • Dehydroepiandrosterone (DHEA)
  • Dehydroepiandrosterone sulfate (DHEA-S)
  • Dihydrotestosterone (DHT)
  • Estradiol
  • Progesterone
  • Testosterone

Order No.: 72000/96/RUO, 72000/480/RUO
96 / 480 Tests


13 Steroids – 2 panels – 1 analytical column
With 96 SPE Well Plate

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Description

The MassChrom® Steroid reagent kit allows the quantitative determination of 13 steroid hormones in serum/plasma by LC-MS/MS. Sample preparation is the same for both steroid panels using an optimised and efficient SPE sample clean up procedure in 96 SPE well plates. The use of stable isotope-labelled internal standards for every single analyte ensures reproducible and reliable quantitation of the steroids. The 6PLUS1® multilevel calibrators offer additional precision, and the method also does not require a high-end LC-MS/MS system to be carried out.

Chromatogram

Technical Data

Method of Analysis

LC-MS/MS

Parameter

11-Deoxycortisol, 17-OH-progesterone, Aldosterone, Androstenedione, Corticosterone, Cortisol, Cortisone, Dehydroepiandrosterone (DHEA), Dehydroepiandrosterone sulfate (DHEA-S), Dihydrotestosterone (DHT), Estradiol, Progesterone, Testosterone

Analysis Time

Steroidpanel 1: 10.5 min Steroidpanel 2: 12.5 min

Gradient

binary

Column temperature

32 °C

Flow rate

0.4-0.8 ml/min

Specimen

Serum/Plasma

Sample Preparation

Equilibration

  • Equilibrate Steroid 96 SPE plate with 0.8 ml
  • Equilibration Reagent 1 into each well.
  • Centrifuge 1 min at 200 x g.
  • Repeat with 0.8 ml Equilibration Reagent 2.

 

SPE clean up procedure

  • Pipette 500 μl research sample/calibrator/MassCheck® control, 50 μl Internal Standard Mix and 450 μl Extraction Buffer into each well of the Steroid 96 SPE plate.
  • Vortex for 2 min at 600 rpm.
  • Add 1 ml Wash Buffer into each well, centrifuge for 1 min at 200 x g and repeat.
  • Centrifuge for 2 min at 3000 x g to dryness, discard effluent.
  • Place plate onto the steroid collection plate.
  • Add 500 μl Elution Buffer and centrifuge for 1 min at 200 x g.

 

Clean-up and injection

  • Place steroid collection plate under nitrogen at 45°C and evaporate the eluates.
  • Add 100 μl Reconstitution Buffer into each well.
  • Shake collection plate for 2 min at 900 rpm.
  • Centrifuge 5 min at 3000 x g.
  • Seal collection plate with adhesive seal, transfer to autosampler.
  • Inject 10–40 μl eluate into the LC-MS/MS system.

 

Sample Stability

Stability of research samples depends on the specific steroid. Further information can be obtained from the instruction manual.

Injection Volume

10-40 µl

Ionisation

Steroid Panel 1: ESI negative and positive (ESI switch), Steroid Panel 2: ESI positive

MS/MS-Mode

Steroid Panel 1: MRM Steroid Panel 2: MRM

Please note

The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.

Kit Components

Components available separately

Mobile Phase A


Order no.: 72001/RUO

Mobile Phase B


Order no.: 72002/RUO

Elution Buffer


Order no.: 72003/RUO

Internal Standard Mix


Order no.: 72004/RUO

Extraction Buffer


Order no.: 72005/RUO

Reconstitution Buffer


Order no.: 72006/RUO

Equilibration Reagent 1


Order no.: 72007/RUO

Equilibration Reagent 2


Order no.: 72008/RUO

Rinsing Solution


Order no.: 72009/RUO

Wash Buffer


Order no.: 72010/RUO

Steroid Collection Plates


Order no.: 72056/RUO

Steroid 96 SPE Well Plate


Order no.: 72057/RUO

Calibrators

Controls