Preparations

Step 1:	Perform any required Instrument Maintenance. A Performance Qualification is a System Test, designed to qualify an HPLC that has been serviced, and is in generally good repair. Prior to starting, ensure that lamps, seals, check valves, rotor seals and other typical maintenance items have been serviced according to the Instrument manual, and/or your relevant SOP’s.
Step 2:	The pump Flow Rate must be verified for an accuracy of ±5% or better, at a flow rate of 1.0 mL/min, with the PQ Test Column in place, with the Method Mobile Phase. If the pump has already been qualified at this flow rate, you may enter the actual flow value into the software from the calibration records. If not, use a dry 5 mL volumetric flask and calibrated stopwatch to confirm the flow accuracy at 1.0 mL/min. Note that this flow rate check is not meant to replace the typically more extensive Operational Qualification checks at several flow rates. Rather, it is to document the Suitability of the pump to perform the various Performance Qualification Test Protocols described in the kit.
Step 3:	Prepare Mobile Phase(s) 13:87 Acetonitrile:0.1% Acetic Acid (1 mL/L). One Liter is sufficient for a simple isocratic qualification. For a quaternary gradient, 2L is recommended. Mobile phase is stable for 60 days, if sealed. Flush and equlibrate HPLC using the HSQ Test Column. If a Gradient Qualification is to be performed, reserve about 500 mL of mobile phase. Spike with Gradient Visualization Solution (GVS) at the rate of 6 mL/L, according to the Operating Instructions. Use this only in the “B*” solvent reservoir.
Step 4:	Set up Methods and Sequence according to the detailed requirements of your HPLC and Data System, and following your company’s internal SOP’s regarding documentation. Injection Sequence 01 shows an example of a typical Sequence performing all the Test Protocols. Adapt this Sequence to your particular instrument as necessary. For example, if only a single injection per vial is used, then multiple vials must be prepared rather than a single vial. Except for the Primary Standard Wavelength Calibration with Holmium Oxide solution, all other tests are fully automated. Once the Sequence is designed, it should operate without user intervention.The various methods all utilize the same general instrument conditions. These are: Method Wavelength: 273 nm Injection Volume: 10 µL (except for volume linearity) Flow Rate: 1 mL/min Temperature: Ambient Note that column oven temperature is not qualified with the HSQ Kit, since most companies maintain their own specific SOP’s addressing this component. The use of a column oven is not required for the PQ Tests. The operating temperature for the PQ Checks should be within the range of 15°C-27°C (60°F-80°F).
Step 5:	After the Test Protocols have been completed, enter the results into the provided Excel™ Template. There are extensive drop-down menus and instructions to help with data entry. Note that the first page of the spreadsheet contains both Instrument data and Test Result Summaries. Note that the data are automatically listed in this section only for the test protocols actually performed.