Set up a Sequence

Preparations

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

1. 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.

2. 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.

3. Quality Control

   For each Quality Control Sample:

   • In the Label column, enter QC.

   • In the Volume column, enter 25 µL as an injection volume.

4. 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 in mg/mL

   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

NOTE

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.