The fields with the red border are required.
Create HLB Method
Prepare Standards
Method Protocols
Optimize HLB Method
BioAnalysis Method Builder for an Oasis HLB Generic Method
To find the right Oasis HLB SPE Product for your method
1) Select type of compound you want to isolate?
Basic compounds
Polar Basic compounds
Acidic compounds
Neutral compounds
2) Select preferred SPE format?
Cartridge
96 well Plate
3) Select use of µElution format to eliminate the evaporation step* (Y/N)
Yes
No
4) Select your usual plasma load volumes range (min-max)
?
10 - 100 µL
100 - 375 µL
375 - 500 µL
500 -1000 µL
1000 - 2000 µL
To calculate the optimized Method Development Protocol for your method
5) Input the plasma load volume you expect to use for
this new method
(µL)
6) Input the
mid-point
of the
concentration range
for
this new method
(ng/mL)?
7) Select the number of Data points (replicates) used for your Recovery Calculations
3
4
6
8
SPE Product Details
Compound Type
Oasis SPE format
Format details
Waters Part Number to Order
© Copyright Waters Corporation 2014
SPE Method Calculated for selected SPE product
Plasma (sample) volume
Pre-SPE Treatment (diluent) volume
Condition
Equilibrate
Load
Wash
Elute
Total Elution volume
SPE Concentration or Dilution Factor
Flow Rate for SPE method
* It is recommended to avoid the evaporation step for volatile and/or heat labile Compounds
*The µElution format allows sample concentration up to 15x without evaporation (when all the compound elutes in the first 25 µL)
OASIS HLB Recovery and Matrix Effects Experimental Set-up
Mid-point of the concentration range for this method
ng/mL
Number of Data points (replicates) required
STANDARDS PREPARATION
Concentrated Standard Stock Solution preparation (S1)
Standard Solution preparation (S2)
Pre-Treatment
(diluent) Preparation
© Copyright Waters Corporation 2014
PLASMA PREPARATION
Estimated total volume of bulk Plasma needed
Pre-Spike Plasma preparation (P1)
Post-Spike Plasma preparation (P2)
OASIS HLB Calculating Recovery and Matrix Effects
Pre-SPE Spiked Samples
Post-SPE Spiked Samples
Standard Solution (Diluted S2)
Load Pre-Spike Plasma (P1)
Load Post-Spike Plasma (P2)
Standard (S2) conc
(ng/mL)
Apply SPE Process
Apply SPE Process
Take Aliquot of
Elute
Elute
Standard Solution (S2)
Add Water
Add Standard Spike (S2)
Inject onto LC/MS
Inject onto LC/MS
Inject onto LC/MS
Final concentration (ng/mL)
Final concentration (ng/mL)
Final concentration (ng/mL)
Enter Average LC/MS Area Counts
Enter Average LC/MS Area Counts
Enter Average LC/MS Area Counts
BioAnalysis Results
Recovery
%
Matrix Effects
%
Matrix Factor
© Copyright Waters Corporation 2014
Oasis HLB Method Optimization
Numbers shown are for example. Overwrite them with your experimental results
Acid Elution (% MeOH in Formic Acid)
Base Elution (% MeOH in Ammonium Hydroxide)
0
10
20
30
40
50
60
70
80
90
0
10
20
30
40
50
60
70
80
90
Analyte 1
Analyte 1
Analyte 2
Analyte 2
Analyte 3
Analyte 3
% MeOH at lift-off for Acid Elutes (A1)
Enter values
% MeOH at lift-Off for Base Elutes (B1)
Enter values
% MeOH at Area Count maxima (A2)
from graphs
% MeOH at Area Count maxima (B2)
from graphs
Unable to load Flash content. The Charts Control requires Flash Player 9.0.45 or higher. You can download the latest version of Flash Player from the
Adobe Flash Player Download Center
Unable to load Flash content. The Charts Control requires Flash Player 9.0.45 or higher. You can download the latest version of Flash Player from the
Adobe Flash Player Download Center
RESULT
Wash with
Elute with
© Copyright Waters Corporation 2014
Original HLB Method
Optimized HLB Method
Condition
Condition
Equilibrate
Equilibrate
Load
Load
Wash
Wash
Elute
Elute
The browser does not support JavaScript. The calculations created using
SpreadsheetConverter
will not work. Please access the web page using another browser.
