GEA Pharma Systems
The lipseal of the impeller shaft was considered to be a critical part in the Single Pot Processor because it is not flush with thelid and no spray balls are available in the lid for this size of machine. The intention of including this part in the cleaning validationstudy was to see whether the movement of the water by using the mixer is enough to clean this centrally-located part as well.The reason for including a chopper knife and a 100 cm² surface of the bowl as critical locations is clear: these parts are in directcontact with the product and, if residues remain there, they will almost certainly contaminate a next batch. A similar reasoning was followed for the outlet valve: although the contact between the product and the outlet valve is short,there is a considerable risk that residues that remain in the outlet valve will contaminate the next batch. Although the bowl seal has no direct contact with the product due to the design of the equipment, it was included as critical areabecause of the possibility that water could infiltrate the small gap between the two metal surfaces of bowl rim and lid rim.Finally, the T
™ opening was included to assess the possibility of water seeping through the T
™ filter and beingtrapped. At the end of the cleaning cycle samples of the final rinse water were collected to determine whether the detergent had beenremoved.
Analysis of the swab samples
After transfer of the swab in a borosilicate tube (Corning Glass Works, Corning, NY, USA), 5 ml distilled water was added. Thetube was vortex-mixed for 10 s, homogenised in an ultrasonic bath for 1 minute and 250
l of the solution was injected onto thecolumn. The chromatographic system consisted of an isocratic pump (L-7110), a variable wavelength UV-detector (L-7400) anda PC-interface (D-7000). The integration of the peak area was performed using the HPLC System Manager V3.0 (Merck-Hitachi,Darmstadt, Germany). The analytical column (250 mm x 4 mm ID) was packed with 5
m RP-C18 particles (Licrosphere, Merck,Darmstadt, Germany). Samples were manually injected using a syringe-loaded injector (Valco six-channel injector, ValcoInstruments, Houston, TX, USA) and a loop of 250 µl. The mobile phase consisting of 50% (v/v) MeOH and 50% (v/v) distilledwater was degassed before use; its flow rate was 1.0 ml/min. The UV-detector monitored the theophylline concentration at 280nm. All experiments were run at ambient temperature. Stock solutions of theophylline at a concentration of 100
g/ml wereprepared in distilled water, stored at 8°C in stoppered flasks and were used to prepare a set of six calibration standards; their concentrations varied between 0.01-100 µg/ml. The method was linear over the entire concentration range (R²=0.9991). Thewithin-day variability (n=5) was 0.28%, while the inter-day variability over the above-mentioned concentration range wasdetermined at 0.33%. The detection limit was determined at 2.1ng/ml, thus the quantification threshold is 14.7 ng/swab.
Analysis of the rinse samples
The residues of the detergent in the final rinse water were established by measuring the conductivity using a conductivity meter type LF325’ (WTW, Weilheim, Germany) in accordance with the specifications of the detergent supplier (4). The samples of thefinal rinsing water were compared with blank samples of the demineralised water used for rinsing and samples of a 1:1000dilution of the detergent solution.