Applications
Automated Extraction of Prednisolone & Cortisone from Plasma Using Novum SLE & Kinetex 2.6u Biphenyl
22785
Separation Mode: Reversed Phase
Reversed Phase
Biphenyl
HPLC
Antineoplastic Agents
Clinical
Clinical Research
Therapeutic / Clinical

Applications

Automated Extraction of Prednisolone & Cortisone from Plasma Using Novum SLE & Kinetex 2.6u Biphenyl
Analytes
1
2
3
Compound Name
Cortisone
Details
LC Conditions (App ID: 22785)
Column:
Brand Name:
Part No:
Elution Type:
Gradient
Mobile Phase:
A: 10 mm ammonium acetate in water
B: 10 mm ammonium acetate in methanol
Gradient Profile:
Step No.Time(min)%A%B
105050
22595
33.1595
43.115050
555050
Flow Rate:
0.45 mL/min
Temperature:
25°C
Detection:
Tandem Mass Spec (MS-MS) @ 22°C
Sample Preparation Details
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.
SPE Load:
Load 150.00 µL Sample diluted in 150
SPE Elution 1:
1000 mL Ethyl acetate
SPE Evaporation:
Evaporate to dryness
SPE Reconstitute:
150 µL 10mM ammonium acetate in water:methanol(50:50)

Order Items Used in This Application

Similar Applications
search
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 3.0 mm, Ea
Reversed Phase
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 3.0 mm, Ea
Reversed Phase
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 3.0 mm, Ea
Reversed Phase
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 3.0 mm, Ea
Reversed Phase
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 3.0 mm, Ea
Reversed Phase
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 3.0 mm, Ea
Reversed Phase
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 3.0 mm, Ea
Reversed Phase
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 3.0 mm, Ea
Reversed Phase
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 2.1 mm, Ea
Reversed Phase
Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 4.6 mm, Ea
Reversed Phase