ordering information proven steroid profiling by mass...

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C M Y CM MY CY CMY K

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D I A G N O S T I C S B Y H P L C & L C-M S/MSChromSystemS

Proven Steroid Profilingby Mass Spectrometry

13 steroids –1sample prep –1analytical column

MassChrom® Steroids in Serum/Plasma

Management system certifiedaccording to DIN EN ISO 9001,DIN EN ISO 13485, ISO 13485 CMDR

Chromsystems Instruments & Chemicals GmbH, Am Haag 12, 82166 Gräfelfing/GermanyPhone: +49 89 18930-0, [email protected], www.chromsystems.comPMA72000 EN-V1 07-2015

w w w . c h r o m s y s t e m s . c o m

Ordering Information

These products are not available in all countries. Please contact yourlocal Chromsystems sales representative for availability in your region.

Order no. Product

72000/96 MassChrom® Steroids by LC-MS/MS in serum/plasmaKit for 96 tests


72000/C MassChrom® Steroids by LC-MS/MS in serum/plasmawith Sample Clean Up ColumnsKit for 96 tests

Components also available separately:

72001 Mobile Phase A, 1000 ml

72002 Mobile Phase B, 1000 ml

72003 Elution Buffer, 50 ml

72004 Internal Standard Mix, 5 ml

72005 Extraction Buffer, 45 ml

72006 Reconstitution Buffer, 10 ml

72007 Equilibration Reagent 1, 80 ml


72009 Rinsing Solution, 1000 ml

72010 Wash Buffer, 200 ml

Components available only for 72000/96 and 72000/480:

72056 Steroid Collection Plates, 3 pcs.

72057 Steroid 96 SPE Well Plate, 1 pc.

72059 Pierceable adhesive seals for 96 SPE Well Plates, 3 pcs.

Components available only for 72000/C:

72054 Autosampler vials with conical bottom, 100 pcs.

72055 Sample Clean Up Columns, 96 pcs.

Order no. Product

Analytical Column:

72100 Analytical Column Steroids, equilibrated,with test chromatogram, 1 pc.

Multilevel Calibrators and Accessories:

72038 6PLUS1® Multilevel Serum Calibrator SetMassChrom® Steroid Panel 1 (lyoph.), 7 x 3 ml


72019 Tuning Mix MassChrom® Steroid Panel 1, Analytes andInternal Standards, 1 ml


72088 System Check Solution MassChrom® Steroid Panel 1, 1 ml


Accessories only for 72000/96 and 72000/480:

72058 Waste plates, 3 pcs.

Accessories only for 72000/C:

72053 Tubes for sample preparation, 50 pcs.

J0406 Crimp Caps, 11 mm, with rubber/PTFE septa, 100 pcs.

J0509 Adapter for autosampler vials with conical bottom, 50 pcs.

Chromsystems Controls (lyoph.), 5 x 3 ml:

0341 MassCheck® Steroid Panel 1 Serum Control, Level I

0342 MassCheck® Steroid Panel 1 Serum Control, Level II

0343 MassCheck® Steroid Panel 1 Serum Control, Level III





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D I A G N O S T I C S B Y H P L C & L C-M S/MSChromSystemSw w w . c h r o m s y s t e m s . c o m

Sample Preparation(72000/96 and 72000/480)

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

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, discard effluent.� Repeat with 0.8 ml Equilibration Reagent 2.

Sample Preparation with Sample Clean UpColumns� Pipette 500 μl of the well homogenised sample/

calibrator/MassCheck® control into each SampleClean Up Column.

� Add 50 μl Internal Standard Mix and 450 μl Extraction Buffer and homogenise (vortex slightly for15 seconds).

� Centrifuge 1 min at 200 x g, 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 SampleClean Up Columns on top.

� Add 500 μl Elution Buffer and centrifuge for 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 crimpingpliers and vortex for 15 s.

� Centrifuge 5 min at 15000 x g.� Transfer to autosampler.� Inject 10–40 μl of eluate into the LC-MS/MS system.

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.

Sample Preparation with 96 SPE Well Plates� Pipette 500 μl sample/calibrator/MassCheck® control,

50 μl Internal Standard Mix and 450 μl ExtractionBuffer 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 2 min at 3000 x g.� Place plate onto the steroid collection plate.� Add 500 μl Elution Buffer and centrifuge for 1 min

at 200 x g.

Concentration and Injection� Place steroid collection plate under nitrogen at 45°C

and evaporate the eluates.� Add 100 μl Reconstitution Buffer into each well.� Vortex collection plate for 2 min at 900 rpm.� Centrifuge 5 min at 3000 x g.� Seal collection plate and transfer to autosampler.� Inject 10–40 μl into the LC-MS/MS system.

1. DHEAS-d62. DHEAS3. Estradiol-d54. Estradiol5. Testosterone-d36. Testosterone7. DHEA-d58. DHEA9. Androstenedione-d7

10. Androstenedione11. 17-OH-Progesterone-d812. 17-OH-Progesterone13. Dihydrotestosterone-d314. Dihydrotestosterone15. Progesterone-d916. Progesterone

1 2 3

4

5

6

7 8

9

10

1112

13

14

15 16

Time, min3.0 3.5 4.54.0 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0

SpecificationsSteroid Panel 2

Linearity:covering referencerange of each steroid

Limit of quantification:10–10000 ng/l

Intraassay: CV = 1–10 %Interassay: CV = 2–17 %Analysis time: 12.5 min

1. Aldosterone-d42. Aldosterone3. Cortisol-d44. Cortisol5. Cortisone-d86. Cortisone7. Corticosterone-d88. Corticosterone9. 11-Deoxycortisol-d510. 11-Deoxycortisol

912

3

Negative ESI Positive ESI

5

6

7 8 10

2.6 3.0 3.6 4.24.0 5.0 5.6 6.0 6.4 7.0 8.0 8.4 9.02.8 3.2 3.4 3.8 4.4 4.6 4.8 5.2 5.4 5.8 6.2 6.6 6.8 7.2 7.4 7.6 8.2 8.67.8 8.8 9.2 9.4 9.6

Time, min

4

SpecificationsSteroid Panel 1

Linearity:covering referencerange of each steroid

Limit of quantification:10–200 ng/l

Intraassay: CV = 1–7 %Interassay: CV = 4–9 %Analysis time: 10.5 min

Steroid Panel 2

Steroid Panel 1

One single analytical column and onesample preparation for all parameters

Clinically most relevant steroids covered

No high-end LC-MS/MS system required

Efficient workflow with 96 SPE well platesor also with sample clean up columns

MassChrom® Kit for Steroid Profilingby Tandem Mass SpectrometryAs steroid hormones serve different functions, they are consequently involved in a variety of diseases. Inborn or acquireddisorders of steroid hormone metabolism like Cushing’s syndrome, adrenal tumours, hyperaldosteronism, and many more,highlight the importance of steroid analysis in clinical diagnostics.

Chromsystems has developed a complete solution aimed at enabling customers to perform steroid analysis efficiently withoutthe need of high-end instrumentation. The new kits MassChrom® Steroids in serum/plasma allow the determination of 13 steroidsby LC-MS/MS with one column and sample preparation procedure. The parameters that can be determined with the kitinclude the clinically most important steroids. Sample preparation is the same for both steroid panels, using an optimisedand efficient clean up procedure in 96 SPE well plates or with sample clean up columns. The use of stable isotope-labelledinternal standards for every single analyte ensures reproducible and reliable quantification of the steroids.

The MassChrom® Steroid kit is available with the analytical column for all parameters together with 6PLUS1® Multilevel CalibratorSets and MassCheck® Controls.

Sample Preparation(72000/C)


Probedruck


®

D I A G N O S T I C S B Y H P L C & L C-M S/MSChromSystemS

Proven Steroid Profilingby Mass Spectrometry

13 steroids –1sample prep –1analytical column

MassChrom® Steroids in Serum/Plasma

Management system certifiedaccording to DIN EN ISO 9001,DIN EN ISO 13485, ISO 13485 CMDR

Chromsystems Instruments & Chemicals GmbH, Am Haag 12, 82166 Gräfelfing/GermanyPhone: +49 89 18930-0, [email protected], www.chromsystems.comPMA72000 EN-V1 07-2015

w w w . c h r o m s y s t e m s . c o m

Ordering Information

These products are not available in all countries. Please contact yourlocal Chromsystems sales representative for availability in your region.

Order no. Product



72000/C MassChrom® Steroids by LC-MS/MS in serum/plasmawith Sample Clean Up ColumnsKit for 96 tests

Components also available separately:

72001 Mobile Phase A, 1000 ml

72002 Mobile Phase B, 1000 ml

72003 Elution Buffer, 50 ml

72004 Internal Standard Mix, 5 ml

72005 Extraction Buffer, 45 ml

72006 Reconstitution Buffer, 10 ml



72009 Rinsing Solution, 1000 ml

72010 Wash Buffer, 200 ml

Components available only for 72000/96 and 72000/480:

72056 Steroid Collection Plates, 3 pcs.

72057 Steroid 96 SPE Well Plate, 1 pc.

72059 Pierceable adhesive seals for 96 SPE Well Plates, 3 pcs.

Components available only for 72000/C:

72054 Autosampler vials with conical bottom, 100 pcs.

72055 Sample Clean Up Columns, 96 pcs.

Order no. Product

Analytical Column:

72100 Analytical Column Steroids, equilibrated,with test chromatogram, 1 pc.

Multilevel Calibrators and Accessories:







Accessories only for 72000/96 and 72000/480:

72058 Waste plates, 3 pcs.

Accessories only for 72000/C:

72053 Tubes for sample preparation, 50 pcs.

J0406 Crimp Caps, 11 mm, with rubber/PTFE septa, 100 pcs.

J0509 Adapter for autosampler vials with conical bottom, 50 pcs.

Chromsystems Controls (lyoph.), 5 x 3 ml:







ordering information proven steroid profiling by mass...

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