72000-C steroids serum plasma sample clean up columns

MassChrom® Steroids in Serum/Plasma - LC-MS/MS

Order no.: 72000/C
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

Clinical relevance

Steroid hormones are structurally derived from cholesterol and play a role in a wide range of physiological processes, including carbohydrate metabolism, fat and protein metabolism as well as in stress response. They are secreted by the adrenal cortex, testes, ovaries, and during pregnancy by the placenta. They are transported through the bloodstream to the cells of various organs where they influence and regulate the organs’ physiological functions.

The determination of hormones is important for the diagnosis of a number of diseases such as adrenogenital syndrome, hyperaldosteronism, or tumours. By using LC-MS/MS, not only high sensitivity and specificity can be achieved, but also the efficient measurement of multiple analytes in a single run.

 

Product advantages

  • 13 steroids – 2 panels – 1 analytical column
  • One sample preparation for all parameters
  • Each parameter safeguarded by its own internal standard

 

The MassChrom® Steroid reagent kit allows the quantitative determination of 13 steroid hormones in serum/plasma by LC-MS/MS and includes the clinically most relevant parameters. Sample preparation is identical for the two steroid panels and is conducted by means of an efficient solid phase extraction using individual SPE columns (Sample Clean Up Columns). Optionally, there is another method available from Chromsystems that uses 96 SPE well plates. The use of stable isotopically labelled internal standards for each analyte ensures reproducible and reliable quantification of the steroids. This method also does not require a high-end LC-MS/MS system.

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

Steroid Panel 1: 10.5 min; Steroid Panel 2: 12.5 min

Limit of quantification

Steroid Panel 1: 10-200 ng/l
Steroid Panel 2: 10-10000 ng/l

 

Linearity

Steroid Panel 1: covering reference range of each steroid
Steroid Panel 2: covering reference range of each steroid

 

Intraassay

Steroid Panel 1: CV 1-7 %
Steroid Panel 2: CV 1-10 %

Interassay

Steroid Panel 1: CV 4-9 %
Steroid Panel 2: CV 2-17 %

Gradient

binary

Column temperature

32 °C

Flow rate

0.4-0.8 ml/min

Specimen

Serum/Plasma

Sample Preparation

Equilibration

  1. Put Steroid 96 SPE Well Plate onto Waste Plate or Sample Clean Up Columns into tubes for sample preparation
  2. Add 0.8 ml Equilibration Reagent 1
  3. Centrifuge 1 min at 200 x g, check for complete run through, discard effluent
  4. Repeat steps 2 and 3 with 0.8 ml Equilibration Reagent 2

Sample preparation

  1. Pipette 500 µl of the well homogenised sample/calibrator/control into Steroid 96 SPE Well Plate or Sample Clean Up Column 0.8 ml
  2. Add 50 µl Internal Standard Mix and 450 µl Extraction Buffer
  3. Shake Steroid 96 SPE Well Plate for 2 min at 600 rpm or vortex Sample Clean Up Columns slightly for 15 seconds
  4. Centrifuge 1 min at 200 x g, check for complete run through, discard effluent
  5. Add 2 x 1 ml Wash Buffer and centrifuge 1 min at 200 x g
  6. Centrifuge 2 min at 3000 x g to dryness, discard effluent
  7. Place Steroid 96 SPE Well Plate onto Steroid Collection Plate or put autosampler vials with conical bottom into tubes for sample preparation, then put Sample Clean Up Columns on top
  8. Add 500 µl Elution Buffer and centrifuge 1 min at 200 x g

Clean-up and injection

  1. Evaporate eluates in Steroid Collection Plate or autosampler vials under nitrogen or compressed air at 45­°C to dryness
  2. Reconstitute with 100 µl Reconstitution Buffer, close autosampler vials with crimp caps
  3. Shake Steroid Collection Plate  for 2 min at 900 rpm or vortex autosampler vials for 15 s
  4. Centrifuge 5 min, collection plate at 3000 x g, autosampler vials at 15000 x g
  5. Seal Steroid Collection Plate with adhesive seal, transfer to autosampler
  6. Inject 10–40 µl eluate into the LC-MS/MS system

Sample Stability

The stability depends on the specific steroid. Further information can be obtained from the instruction manual.

Injection Volume

10-40 µl

Ionisation

ESI switch negative/positive (Panel 1), ESI positive (Panel 2)

MS/MS-Mode

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

Mobile Phase B


Order no.: 72002

Elution Buffer


Order no.: 72003

Internal Standard Mix


Order no.: 72004

Extraction Buffer


Order no.: 72005

Reconstitution Buffer


Order no.: 72006

Equilibration Reagent 1


Order no.: 72007

Equilibration Reagent 2


Order no.: 72008

Rinsing Solution


Order no.: 72009

Wash Buffer


Order no.: 72010

Sample Clean Up Columns


Order no.: 72055

Calibrators

Controls

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