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Beckman Coulter Biomek FX-ADMETox Quick Start Manual

Permeability analyzer

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TM
Biomek FX-ADMETox
Workstation
PAMPA Evolution96
Permeability Analyzer
Quick Start Guide
A28712-AA
July 2005
© 2005 Beckman Coulter, Inc.
4300 N. Harbor Blvd., Fullerton, CA 92834-3100
Printed in U.S.A.
Quick Start Preliminaries for Using the
PAMPA EVOLUTION96 Software and
Biomek FX Workstation
Preparing a Blank
Default file which contains the UV spectra of a Blank may be
replaced with new one.
1.
Launch PAMPA Evolution-96 Command Software on the
computer connected to the PAMPA Evolution
instrument.
2. Go to menu File | Save as and save new file as
""YYMMDD_Blank" (for example: 040701_Blank) where
the first 6 characters are the year-month-day numbers.
3. Be careful not to touch the underside of the UV plates,
place 4 new 96-well plastic UV plates stacked on top of
each other into position P 11.
4. Put High Profile, 12 Trough Reservoir filled with System
Solution into position P4.
5. Click the Start Assay button on the toolbar. The Start
Assay dialog displays. Make sure that all check-boxes
are not selected.
6. Click the Start button.
7. A dialog box displays Transfer the System Solution to
UV plate.
When the instrument finishes this step, Spectrophotometer
will start reading UV spectra of the blank plate. Wait till
Spectrophotometer finishes reading.
1.
To inspect the spectrophotometric data, click View 96
Spectra button.
2. 96 spectra view displays.
3. Inspect UV spectra of the blank in order to exclude
bacterial contamination.
4. In 96 spectra view go to menu File | Export Plate
Spectra
5. The Save Spectra dialog box displays.
6. Select Blank Plate from Plate to be Exported drop-
down list.
7. Click the Save button.
8. A special spectra file will be saved in the same folder
where data file is under the name Data file name_Blank
Plate. If appropriate care is taken of the System
Solution, one Blank may be reused for many
experiments.
9. Abort the assay and close the method.
10. Start a new PAMPA method and go to the main menu
Instrument | Settings.
11. The internal UV blank spectrum must be loaded into the
file.
12. Browse for the file Data file name_Blank Plate.spc and
click the Open button.
13. Select Use pre-measured blank.
14. Select Tell me about low -UV wells...
15. Select Auto-refine. The results of the experiment will
have the results automatically calculated.
16. Click the Ok button when done.
Run PAMPA Assay
1.
Prepare system buffer with different pH conditions
a.Prepare 1 Liter of System Solution by adding 25 mL of
System Solution Concentrate to total 1 L of distilled
water using a graduated cylinder
b.Pour equal aliquots of System Solution in 3 clean
glass bottles (about 333 mL each)
c.Measure initial pH of System Solution using pH meter
d.Adjust pH to 7.4, 6.2 and 5.0 by adding 0.5M NaOH
according to the table below.
e.Verify System Solution pH using pH meter.
.pHstart
.pHstart=
=2.6
2.8
pHtarget
NaOH
pHtarget
NaOH
(mL)
(mL)
5.0
5.5
5.0
5
6.2
8.3
6.2
7.8
7.4
11
7.4
10.5
f.Add 15 mL of the pH-adjusted System Solution in each
partition of the High Profile 12 Trough Reservoir
:
according to the table below
1
2
3
4
5
6
7
pH
pH
pH
pH
pH
pH
pH
7.4
7.4
7.4
7.4
6.2
6.2
6.2
2. Prepare GIT-Lipid.
a.Thaw 2 glass ampules of GIT lipid solution.
b.Break amples using ampule breaker.
c.Empty GIT lipid into the first column of a Low Profile
12 Trough Reservoir.
d.Transfer 18 µL of lipid into each well of the LipidPlate.
3. Prepare Acceptor System Buffer (ASB 7.4).
a.Place 50 mL of ASB 7.4 into a Low Profile Reservoir
.
.
.
.pHstart=3.0
pHtarget
NaOH
(mL)
5.0
4.5
6.2
7.3
7.4
10.2
8
9
10
11
12
pH
pH
pH
pH
pH
6.2
5.0
5.0
5.0
5.0
.

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Summary of Contents for Beckman Coulter Biomek FX-ADMETox

  • Page 1 Plate. If appropriate care is taken of the System Solution, one Blank may be reused for many d.Transfer 18 µL of lipid into each well of the LipidPlate. © 2005 Beckman Coulter, Inc. experiments. 4300 N. Harbor Blvd., Fullerton, CA 92834-3100 3.
  • Page 2 4. Prepare test compounds c.Browse to open PAMPA Template.xlt file located in the PAMPA Evolution command Software directory. a.Spectroscopic purity DMSO for test compound preparation. d.Enter each compound in the same well location that it was loaded in the StockPlate. b.Add DMSO to test compounds according to the table below: e.Physiochemical constants are optional.

This manual is also suitable for:

Pampa evolution96