Appendix 2 How to Perform the Instrument Test

The test methods, mobile phases, and test compounds used here are just possible examples but they have proved effective in numerous cases. However, the author is pleased to get proposals for alternative procedures since there is steady progress in the development of scientific instruments. In addition, the requirements of official testing authorities have a tendency to become more strict. 

The test procedures described in this chapter are based on reversed-phase chromatography with UV detection and can be used for almost all HPLC systems, including those which are used with non-polar or more viscous mobile phases or other chromatographic methods. In such cases it is necessary to adjust the instrument parameters such as the suction speed of the autosampler or the compressibility factor of the pump. An important prerequisite is the thorough priming of the whole HPLC system with the mobile phases and flushing liquids in the case of non-miscibility, an intermediate solvent needs to be used. Do not forget the injection valve. 

All tests are performed at ambient temperature, therefore it is advantageous to prepare all the mobile phases and test solutions on the day before in order to have them ready at an identical temperature. 

Practical High-Performance liquid Chmttuuography, Fourth edition Veronika R. Meyer 2004 John Wiley Sons, Ltd ISBN 0-470-09377-3 (Hardback) 0-470-09378-1 (Paperback) 

Note Depending on the manufacturer (i.e. the packing density) and the temperature, the pressure drop can vary between 80 and 240 bar. Columns of the same manufacturer usually give the same pressure drop. 

The present operating procedure describes the procedure and the documentation for the performance qualification (PQ) of HPLC systems It can be used for both isocratic and gradient systems with UV detection and it is independent of the instrument manufacturer. The procedure includes the tests of pump, autosampler, UV detector and column oven. It can be put into practice immediately. 

All the proposed setpoint values are well proven recommended data and can be adapted to the individual requirements. 

For laboratories working in accordance to GLP or GMP it is recommended to perform the test after the installation of a new HPLC system, later at least once a year and in any case after a change of location. If a component (e.g. the pump) needs to be repaired the relevant part of the test should be performed again. 

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