MassChrom® Steroids in Serum/Plasma with Sample Clean Up Columns - LC-MS/MS

MassChrom® Steroids in Serum/Plasma with Sample Clean Up Columns - LC-MS/MS

Order no.: 72000/C/RUO
96 Tests
  • 11-Deoxycortisol
  • 17-OH-progesterone
  • Aldosterone
  • Androstenedione
  • Corticosterone
  • Cortisol
  • Cortisone
  • Dehydroepiandrosterone (DHEA)
  • Dehydroepiandrosterone sulfate (DHEA-S)
  • Dihydrotestosterone (DHT)
  • Estradiol
  • Progesterone
  • Testosterone

13 Steroids – 2 panels – 1 analytical column
With Sample Clean Up Columns

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 procedure with sample clean up columns. 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

Number of Tests

96

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

  • Put Sample Clean Up Columns into tubes for sample preparation.
  • Pipette 0.8 ml Equilibration Reagent 1 into each Sample Clean Up Column.
  • Centrifuge 1 min at 200 x g, check for complete run through, discard effluent.
  • Repeat with 0.8 ml Equilibration Reagent 2.

Sample preparation with Sample Clean Up Columns

  • Pipette 500 μl of the well homogenised research sample/calibrator/MassCheck® control into each Sample Clean Up Column.
  • Add 50 μl Internal Standard Mix and 450 μl Extraction Buffer and homogenise (vortex slightly for 15 seconds).
  • Centrifuge 1 min at 200 x g, check for complete run through, discard effluent.
  • Add 2 x 1 ml Wash Buffer and centrifuge 1 min at 200 x g respectively.
  • Centrifuge 2 min at 3000 x g to dryness, discard effluent.
  • Put labelled autosampler vials with conical bottom into tubes for sample preparation, then put Sample Clean Up Columns on top.
  • Add 500 μl Elution Buffer and centrifuge 1 min at 200 x g.

Concentration and injection

  • Evaporate eluates in the labelled autosampler vials under nitrogen or compressed air at 45°C to dryness.
  • Reconstitute with 100 μl Reconstitution Buffer in the labelled autosampler vials, then close using crimping pliers and vortex for 15 sec.
  • Centrifuge 5 min at 15000 x g.
  • Transfer to autosampler.
  • Inject 10–40 μl of 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

Autosampler vials


Order no.: 72054/RUO

Sample Clean Up Columns


Order no.: 72055/RUO

Calibrators

Controls