I don't have any other handles
Only insecure people do so.
@master.on the one and only
accept no sunstitutes.
That's not the way it works.
If you actually had any hands-on experience you'd know how time consuming it is.
1 Order the
reference standards
2 Look for a suitable testing
method either search online, ask colleagues or ask the equipment manufacturer or column supplier.
3 Whenever possible choose a
reversed-phase method, as they're usually easier.
4 See what
column type that method uses.
5 Check if your columns supplier has columns optimized for that application
i.e. there are some columns meant to make steroid analysis easier.
6 Decide on a column length.
Personally I prefer long columns first to see if the method works, then try shorter columns to reduce the time tests take.
7 Order said column
8 See what mobile phase solvents that method requires, purchase if needed.
9 Install the new column in the HPLC machine
10 Set the machine to run solvent trough it for at least 5 minutes
11 Leave the column with solvent in it at least overnight, to equalize it
12 Do some math to see if a standard dilution works or if you need serial dilutions
13 See what the solvent in the reference standard is (i.e. methanol).
14 Do some math and a "recipe" for preparing the blank, and calibration standards
15 Put a new tip in the micropipette to avoid cross contamination
16 Prepare a blank sample including some bare solvent as in the reference standard
17 Make sure the machine has enough solvents in the bottles, refill as needed
18 turn on the machine
19 allow to warm up
20 enter the method settings
21 start the test run
22 manually inject the sample for those machines without autosampler
23 check to make sure no hoses, pump or column are leaking
24 check the chart
no high peaks should be there since it's a blank sample
no peak should be 4x higher than the background "noise"
25 if 24 shows high peaks, change settings, solvents or look for another method
