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

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

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

Order No.: 72072/96, 72072/480
96 / 480 Tests


Optimised and extended method
15 Steroids – 2 panels – 1 analytical column
With 96 SPE Well Plate

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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

  • 15 steroids – 2 panels – 1 analytical column
  • One sample preparation for all parameters
  • Clinically most relevant steroids covered
  • Each parameter safeguarded by its own internal standard
  •  

    This MassChrom® Steroid kit allows the quantitative determination of 15 steroids in serum/plasma by LC-MS/MS and includes the clinically most important parameters. Sample preparation is the same for both steroid panels using an optimised and efficient SPE sample clean up procedure with 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 method does not require a high-end LC-MS/MS system to be carried out.

    Alternatively, there is also a method available using Sample Clean Up Columns (72072/C).

Chromatogram

Technical Data

Method of Analysis

LC-MS/MS

Parameter

11-Deoxycorticosterone, 11-Deoxycortisol, 17-OH-progesterone, 21-Deoxycortisol, Aldosterone, Androstenedione, Corticosterone, Cortisol, Cortisone, Dehydroepiandrosterone (DHEA), Dehydroepiandrosterone sulfate (DHEA-S), Dihydrotestosterone (DHT), Estradiol, Progesterone, Testosterone

Analysis Time

Steroid Panel 1: 10.0 min; Steroid Panel 2: 11.7 min

Limit of quantification

Steroid Panel 1: down to 14 ng/l
Steroid Panel 2: down to 5 ng/l

Linearity

Steroid Panel 1: covering reference range of each steroid

Steroid Panel 2: covering reference range of each steroid

Recovery

Steroid Panel 1: 86-115 %; Steroid Panel 2: 76-118 %

Intraassay

Steroid Panel 1: CV 1-6 %
Steroid Panel 2: CV 1-13 %

Interassay

Steroid Panel 1: CV 3-11 %
Steroid Panel 2: CV 4-13 %

Gradient

binary

Column temperature

Steroid Panel 1: 32 °C, Steroid Panel 2: 27 °C

Specimen

Serum/Plasma

Sample Preparation

  • Put 96 SPE Well Plate onto Waste Plate.
  • Pipette 0.8 ml Equilibration Reagent 1 into each well of the 96 SPE Well Plate, centrifuge and repeat with Equilibration Reagent 2.
  • Pipette 500 μl sample/calibrator/MassCheck® control, 50 μl Internal Standard Mix and 450 μl Extraction Buffer into each well, mix briefly (vortex) and centrifuge.
  • Add 2 x 0.7 ml Wash Buffer and centrifuge.
  • Place plate onto the Collection Plate.
  • Add 500 μl Elution Buffer and centrifuge.
  • Evaporate eluates in the Collection Plate (nitrogen or compressed air) to dryness.
  • Add 100 μl Reconstitution Buffer.
  • Shake Collection Plate, seal with adhesive seal and inject 5–50 μl of the eluate. 

Sample Stability

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

Injection Volume

5-50 µl

Ionisation

ESI negative and positive (ESI switch)

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.: 72011

Mobile Phase B


Order no.: 72002

Elution Buffer


Order no.: 72033

Internal Standard Mix


Order no.: 72044

Extraction Buffer


Order no.: 72005

Reconstitution Buffer


Order no.: 72006

Equilibration Reagent 1


Order no.: 72077

Equilibration Reagent 2


Order no.: 72008

Rinsing Solution


Order no.: 72009

Wash Buffer


Order no.: 72012

Calibrators

Controls

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