| 1Cortisol |
|---|
| 2Cortisol-D4 (Internal standard) |
Compound Name Hydrocortisone |
CID 5754 |
Molecular Formula C21H30O5 |
Molecular Weight 362 |
No. Hydrogen Bond Acceptors 5 |
No. Hydrogen Bond Donors 3 |
Applications
Extraction of Total Cortisol from Human Plasma Using Novum SLE & Kinetex Biphenyl
22787
Separation Mode: Reversed Phase
Reversed Phase
Biphenyl
HPLC
Anti-Inflammatory Agents
Clinical
Clinical Research
Therapeutic / Clinical
Applications
Extraction of Total Cortisol from Human Plasma Using Novum SLE & Kinetex Biphenyl
Analytes
Details
LC Conditions (App ID: 22787)
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:
| ||||||||||||||||||||||||
Flow Rate: 0.45 mL/min | ||||||||||||||||||||||||
Temperature: 25°C | ||||||||||||||||||||||||
Detection: Tandem Mass Spec (MS-MS) (22°C) | ||||||||||||||||||||||||
Detector Info: SCIEX API 5000 |
Sample Preparation Details
Description: Novum SLE, MAX 96-Well Plate, Ea |
Part Number: |
Pre Treatment Note: Extraction Procedure
Sample pre-treatment: Dilute 200 uL of human plasma with 200 ul of 50 mM sodium phosphate dibasic heptahydrate, pH unadjusted. Vortex briefly (3-5 sec).
Sample loading: Load the sample from pretreatment step abive onto the Novum plate and apply a short and gentle pulse of vacuum (~ 5" of Hg for 5-10 sec) or until the sample has completely entered the media.
Wait for 5 minutes.
Elution: Dispense 900 uL of dichloromethane onto the SLE media and allow the solvent to elute by gravity (~ 5 min elution time) and collect the eluant.
Repeat with another 900 uL of dichloromethane and collect the eluant.
Apply vacuum at 5" of Hg for 20-30 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 the dry residue in 200 uL of initial mobile phase fortified with cortisol-D4. |
Sample PreTreatment: Extraction Procedure
Sample pre-treatment: Dilute 200 uL of human plasma with 200 ul of 50 mM sodium phosphate dibasic heptahydrate, pH unadjusted. Vortex briefly (3-5 sec).
Sample loading: Load the sample from pretreatment step abive onto the Novum plate and apply a short and gentle pulse of vacuum (~ 5" of Hg for 5-10 sec) or until the sample has completely entered the media.
Wait for 5 minutes.
Elution: Dispense 900 uL of dichloromethane onto the SLE media and allow the solvent to elute by gravity (~ 5 min elution time) and collect the eluant.
Repeat with another 900 uL of dichloromethane and collect the eluant.
Apply vacuum at 5" of Hg for 20-30 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 the dry residue in 200 uL of initial mobile phase fortified with cortisol-D4. |
Evaporation: Evaporate to dryness |
1 Shake Methylene Chloride |
2 Shake Methylene Chloride |
Order Items Used in This Application
00B-4622-AN
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 2.1 mm, Ea
Qty:
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