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Figure 3.3. Microwave assisted extraction plant as described by Lubenova [86] where 1) solvent container, 2) extractor, 3) placed in a chamber’s wall, 4) microwave source-magnetron, 5) circulation contour, 6) filter, 7) temperature sensor, 8) pump, 9) drain
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valve, 10) circulation valve, 11) pH meter, 12) inner perforated wall container, 13) extract collector, 14) control unit for temperature.
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