1Timolol |
---|
Compound Name Timolol |
CID 33624 |
Molecular Formula C13H24N4O3S |
Molecular Weight 316 |
No. Hydrogen Bond Acceptors 7 |
No. Hydrogen Bond Donors 2 |
1Timolol |
---|
Compound Name Timolol |
CID 33624 |
Molecular Formula C13H24N4O3S |
Molecular Weight 316 |
No. Hydrogen Bond Acceptors 7 |
No. Hydrogen Bond Donors 2 |
Column: | ||||||||||||||||||||||||||||
Brand Name: Kinetex | ||||||||||||||||||||||||||||
Part No: | ||||||||||||||||||||||||||||
Phase Name: Biphenyl | ||||||||||||||||||||||||||||
Elution Type: Gradient | ||||||||||||||||||||||||||||
Mobile Phase: A: 0.1% formic acid in water B: 0.1% formic acid in acetonitrile | ||||||||||||||||||||||||||||
Gradient Profile:
| ||||||||||||||||||||||||||||
Flow Rate: 0.5 mL/min | ||||||||||||||||||||||||||||
Temperature: 25°C | ||||||||||||||||||||||||||||
Detection: Tandem Mass Spec (MS-MS) (22°C) | ||||||||||||||||||||||||||||
Sample Note: Strata X-C Elution Plate Protocol
Condition
200 µL methanol
Equilibrate
200uL water
Load
750 µL diluted serum
(375 µL plasma diluted 1:1 with 4% phosphoric acid in water)
Wash1
200 µL 2% formic acid in water
Wash 2
200 µL methanol
Elute
25 µL 5% ammonium hydroxide in acetonitrile:methanol(60:40)
Inject
1 µL
Note: Spiked conc. of analyte was 1 ng/mL serum |
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 |