Prerequisites
To be able to carry out the procedure as described, you need the following privileges. Privileges are configured in the Control Panel.
Project Management > Edit content of project
Sequence > Create and modify sequence
Sequence > Edit method override parameters
Blanks
Add three blanks at the beginning of the sequence to allow equilibration of column and detector (UV).
In the Volume column, enter 25 µL as an injection volume.
Calibration standards
Add four calibration standards (one per calibration level). For each calibration standard:
In the Level column, enter the correct calibration level number.
In the Volume column, enter the injection volumes 10, 20, 25, and 30 µL.
For the third calibration standard (the one with 25 µL), enter QC in the Label column. This calibration standard is at the same time a quality control.
Quality Control
For each Quality Control Sample:
In the Label column, enter QC.
In the Volume column, enter 25 µL as an injection volume.
Samples
In the Label column, enter labels as follows:
S1 and H1 for the first sample standard and harsh
S2 and H2 for the second sample standard and harsh
etc.
In the Volume column, enter 25 µL as an injection volume.
In the Sample amount column:
In case of Drug substance assay or Drug product gravimetric assay, enter the weight in mg that you used for your sample preparation.
In case of Drug product volumetric assay, enter the volume in mL that you used for your sample preparation.
Add the following information in the Sample custom parameters tab of the window below the sequence table:
Parameter |
Description |
---|---|
Lot number | Lot number for the test samples |
Moisture result #1 (%) Moisture result #2 (%) | Results from a Karl Fischer Titration (determination of water content). |
Density | Density in mg/mL) for drug product to be used for label claim |
Preparation factor | Use Volumetric or Gravimetric |
Sample prep. volume | Final volume of the prepared sample in mL |
Assay low limit | Drug substance concentration lower limit in mg/mL |
Assay high limit | Drug substance concentration upper limit in mg/mL |
Label claim | Label claim for the drug product assay |
Note that these parameters can also be set in Data Analysis when preparing the result set.
The following table shows an example sequence table.
Sample name |
Label | Injection Volume | Sample Type | Level | Acquisition method |
---|---|---|---|---|---|
Water Blank | 25 µL | Blank | Standard | ||
Water Blank | 25 µL | Blank | Standard | ||
Water Blank | 25 µL | Blank | Standard | ||
WSS 10 µL | 10 µL | Cal. Std. | 1 |
Standard | |
WSS 20 µL | 20 µL | Cal. Std. | 2 | Standard | |
WSS 25 µL |
QC | 25 µL | Cal. Std. |
3 | Standard |
WSS 30 µL | 30 µL | Cal. Std. | 4 |
Standard | |
QC Standard |
QC | 25 µL | QC Check | Standard | |
Sample A S1 | S1 | 25 µL | Sample |
Standard | |
Water Blank | 25 µL | Blank | Harsh | ||
Sample A H1 | H1 | 25 µL | Sample |
Harsh | |
Water Blank | 25 µL | Blank | Harsh | ||
QC Standard | QC | 25 µL | QC Check |
Standard |
If you need to add injections that do not match the criteria in the table, for example conditioning injections or post-run washes, use the sample type Spike. Samples of type Spike will be excluded from the oligo analysis. |