1Cortisone |
---|
2Cortisol-D4 (Internal standard) |
3Prednisolone |
Compound Name Cortisone |
CID 222786 |
Molecular Formula C21H28O5 |
Molecular Weight 360 |
No. Hydrogen Bond Acceptors 5 |
No. Hydrogen Bond Donors 2 |
1Cortisone |
---|
2Cortisol-D4 (Internal standard) |
3Prednisolone |
Compound Name Cortisone |
CID 222786 |
Molecular Formula C21H28O5 |
Molecular Weight 360 |
No. Hydrogen Bond Acceptors 5 |
No. Hydrogen Bond Donors 2 |
Column: | ||||||||||||||||||||||||
Brand Name: Kinetex | ||||||||||||||||||||||||
Part No: | ||||||||||||||||||||||||
Phase Name: Biphenyl | ||||||||||||||||||||||||
Elution Type: Gradient | ||||||||||||||||||||||||
Mobile Phase: A: 10 mm ammonium acetate in water B: 10 mm ammonium acetate in methanol | ||||||||||||||||||||||||
Gradient Profile:
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Flow Rate: 0.45 mL/min | ||||||||||||||||||||||||
Temperature: 25°C | ||||||||||||||||||||||||
Detection: Tandem Mass Spec (MS-MS) (22°C) | ||||||||||||||||||||||||
Detector Info: SCIEX API 5000 |
Description: Novum SLE, MINI 96-Well Plate, 1/Pk |
Part Number: |
Pre Treatment Note: Sample pre-treatment
Dilute 150 uL of human plasma (spiked with 25 ng/mL and 125 ng/mL of cortisone and prednisolone respectively) with 150 uL of 50 mM sodium phosphate dibasic heptahydrate, pH unadjusted. Mix briefly (3-5 sec).
Sample loading:
Load the sample from pre-treatment step above onto the Novum plate and apply a short and gentle pulse of vacuum (~ 10" of Hg for 20 secs) until the sample has completely entered the media.
Wait for 5 minutes.
Elution:
Dispense 1000 uL of ethyl acetate onto the SLE media and allow the solvent to elute by gravity (~ 5 min elution time) and collect the eluant.
Apply vacuum at 5" of Hg for 45 secs to complete the extraction.
NOTE: Prolonged application of vacuum will result in elution of plasma out of the SLE media and into the final extracted solvent.
Dry down:
Evaporate the final extract to complete dryness under slow stream of N2 at 40°C
Reconstitute:
Reconstitute the dry residue in 150 uL of initial mobile phase fortified with cortisol-D4. |
Sample PreTreatment: Sample pre-treatment
Dilute 150 uL of human plasma (spiked with 25 ng/mL and 125 ng/mL of cortisone and prednisolone respectively) with 150 uL of 50 mM sodium phosphate dibasic heptahydrate, pH unadjusted. Mix briefly (3-5 sec).
Sample loading:
Load the sample from pre-treatment step above onto the Novum plate and apply a short and gentle pulse of vacuum (~ 10" of Hg for 20 secs) until the sample has completely entered the media.
Wait for 5 minutes.
Elution:
Dispense 1000 uL of ethyl acetate onto the SLE media and allow the solvent to elute by gravity (~ 5 min elution time) and collect the eluant.
Apply vacuum at 5" of Hg for 45 secs to complete the extraction.
NOTE: Prolonged application of vacuum will result in elution of plasma out of the SLE media and into the final extracted solvent.
Dry down:
Evaporate the final extract to complete dryness under slow stream of N2 at 40°C
Reconstitute:
Reconstitute the dry residue in 150 uL of initial mobile phase fortified with cortisol-D4. |
Evaporation: Evaporate to dryness |
1 Shake Ethyl acetate |