Set up a Sequence

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

  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

  4. For each Quality Control Sample:

    • In the Label column, enter QC.

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

  5. 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.