Waters Acquity/Quattro Premier, Maintenance and Service Procedures

Page: 1/2 OPERATIONAL GUIDELINES Version no: 4 MS laboratory Date: 17.10.2008 Compiled by: JPS M402 Accepted by: HK

M402 Waters Acquity/Quattro Premier, maintenance and service procedures

The instrumentation is comprised of two separate units: 1) Acquity UPLC unit for ultra- performance liquid chromatography (consists of Solvent Manager, Sample Manager, PDA detector, and column oven) and 2) Quattro Premier tandem quadrupole mass spectrometer.

Acquity

Maintenance procedures

At the beginning of the day, solvent lines, wash syringes, sample syringe, and sample needle are flushed with the automatic priming functions as instructed in the manual. When changing the solvent bottles, five wash cycles are needed so that no air remains in the solvent lines or wash syringes. The presence of air is checked after the priming procedures. The instrument is visually checked for leakages when the solvent flow is led through a column.

Note: If there is no solvent in the seal wash line or if the line contains so much air that the automatic priming function does not expel it, the line has to be filled manually using syringe filled with the solvent. After this the priming is performed.

When starting the solvent flow, it is increased in increments of 0.1 ml/min (UPLC columns) or 0.2 ml/min (HPLC columns) in order to avoid sudden pressure peaks and column damage. In order to avoid precipitates, microbial growth and corrosion, buffer solutions, solvents with over 95% water, and acidic/basic solvents, need to be flushed out of the instrument after the runs have been completed.

The following basic rule applies when tightening the ferrulae of Acquity (does not apply to finger-tight screws): a new ferrula is first tightened finger-tight. After this it is tightened max. 3/4 round with a wrench. A used ferrula is first tightened finger-tight. After this it is tightened max. 1/4 round. Be careful not to overtighten! First, a moderate tightening is performed, and after that the screw may be tightened more in case a leakage is observed – the maximum being as mentioned above.

The inner and outer parts of the instrument as well as its nearby area are to be kept free from dust and dirt.

Service

The functioning of Acquity Solvent Manager is controlled daily by observing pressure stability (delta-value) and the pressure graphs on UPLC Console. If something abnormal is observed, the presence of possible leakage is checked. The union at the location of the leakage may be opened and retightened. If this does not help, the ferrula, check valve, or pump seals may need to be replaced. All the consumables/spare parts replaced are to be written down in the instrument diary S401 and in the follow-up sheet S402 (on the wall).

In order to test the condition of the valves and the seals of Acquity, Static Decay Test is performed with methanol as instructed in the manual. The test is performed whenever check valves or pump seals are replaced, or else at least once every six months. When the test has been performed, it is to be mentioned in the instrument diary S401 and in the follow-up sheet S403 (on the wall).

If there are problems in the injection of the sample, the condition of the sample needle is checked. There may also be a need to perform z- and/or x,y,zp-calibration of Sample Manager. (instructions in the manual). The calibrations are to be mentioned in the instrument diary S401 and in the follow-up sheet S403 (on the wall).

Quattro Premier

At the beginning of the day, it is made sure that no leakages are present and that the first runs look normal (baseline, sample peaks). If problems are observed, the solvent spray in the ion source of Quattro Premier is checked (if electrospray ionization is used). Inconstant spray may be a sign of problems related to Acquity or to solvent splitter, or of a blockage in/a bad union of the ESI capillary (the capillary can be removed, cleaned/replaced, tested and reattached as instructed in the manual).

A weak signal is often a sign of a dirty ion source. The walls of the ion source, cone gas nozzle, and sample cone can be cleaned (as instructed in the manual) without the evacuation of the vacuum. These cleaning procedures should always be performed if the ion source looks dirty.

If there is a reason to suspect that also the inner parts of the ion source are dirty, the vacuum needs to be evacuated first and the instrument needs to be turned off as instructed in the manual. Source T-wave between the ion source and the mass analyzer gets dirty only rarely in normal usage.

Mass calibration to m/z value 2000 (or higher) is to be performed with cesium iodide whenever the decimals of the known peaks of mass spectra start to systematically deviate from the accurate monoisotopic m/z values, or else at least once a year. The mass calibration is to be mentioned in the instrument diary S401 and in the follow-up sheet S403 (on the wall).

All the consumables/spare parts replaced are to be written down in the instrument diary S401 and in the follow-up sheet S402 (on the wall).

The seals of the prevacuum pump are to be replaced at least every 10 months.

Note! The solvent flow may NEVER be led to the ion source if the gases are not on. The voltages of the ion source and the analyzer (“operate”) are to be turned on and off simultaneously with the solvent flow. The gases have to be on before “operate” is turned on. Also, the gases may not be turned off before “operate” is turned off.1

The instrument is to be kept free from dust and dirt.

1According to new guidelines, a small, continuous gas flow is kept on all the time.