Process and apparatus for separation by liquid chromatography

Claim 1

A large-scale separating apparatus for separating an objective component from a large amount of a mixture sample by liquid chromatography, which has a preliminary fractionation line comprising: at least one first column, in which a first packing having a large particle size has been charged; an eluent tank provided on the upstream side of the first column; a sample injecting valve provided in a passage between the first column and the eluent bath; a sample bath communicated with the sample injecting valve; a first fraction collector provided through a passage on the downstream side of the first column; a first detector provided in a passage between the first column and the first fraction collector; a liquid transfer device provided in a liquid passage; and a pre-cut valve provided in a passage between the first column and the first fraction collector for taking out main fractions containing the objective component, and a recycling line comprising: at least one second column charged with a second packing having a large particle size, which column is to be communicated through a passage with the pre-cut valve in the aforementioned preliminary fractionation line when the main fractions are taken out; a second fraction collector provided through a passage on the downstream side of the second column; a second detector provided in a passage between the second column and the second fraction collector; a recycling valve for turning eluate from the second detector to the inlet of the second column upon recycling of the eluate and causing the objective component to flow into the second fraction collector after completion of separation; and a liquid transfer device provided in a passage between the recycling valve and the inlet of the second column; and wherein the sample injecting valve, the pre-cut valve and the recycling valve are changed over by respective controlling signals, and the operation states of the respective liquid transfer devices in the preliminary fractionation line and the recycling line are controlled according to respective controlling signals, which apparatus includes first and second fractionation valves which are respectively provided on the upstream sides of the first fraction collector and the second fraction collector and are changed over according to the respective signals for taking out samples, an operation controlling means from which controlling signals are sent out at predetermined times, a peak judging means for judging the peaks in the eluate by the detection signals respectively sent from the first detector and the second detector, and a controlling means for taking out the component which sends out taking out signals according to said judgments.