Method of introducing a sample into a separation column and corresponding system
A method of introducing a sample into a separation column includes introducing the sample into a trap column, isolating the trap column from ambient atmosphere and pressurizing the trap column to a first pressure while the trap column is isolated from ambient atmosphere, providing a fluid connection between the trap column and the separation column after pressurizing the trap column to the first pressure, supplying the sample from the trap column to the separation column.
1 . A method of introducing a sample into a separation column of a liquid chromatography system, the method comprising:
introducing the sample into a trap column;
isolating the trap column from an ambient atmosphere and pressurizing the trap column to a first pressure while the trap column is isolated from the ambient atmosphere;
providing a fluid connection between the trap column and the separation column after pressurizing the trap column to the first pressure; and
supplying the sample from the trap column to the separation column, wherein the liquid chromatography system comprises an analytical pump adapted to provide a flow of pressurized fluid, wherein the method further comprises:
providing a fluid connection between the trap column and the analytical pump, wherein the fluid connection between the trap column and the analytical pump is provided after pressurizing the trap column to the first pressure.
2 . Method according to claim 1 , wherein the trap column is not fluidly connected to the analytical pump when pressurizing the trap column to the first pressure.
3 . Method according to claim 1 wherein the first pressure exceeds ambient pressure by at least 10 bar.
4 . Method according to claim 1 , wherein the liquid chromatography system comprises a sample pick up means, a seat for receiving the sample pick up means and a first distribution valve, wherein introducing the sample into the trap column comprises:
moving the sample pick up means to a sample reservoir,
sucking the sample into the pick up means,
moving the sample pick up means to the seat,
providing a fluid connection between the seat and the trap column by the first distributor valve, and
introducing the sample into the trap column.
5 . Method according to claim 1 , wherein the liquid chromatography system comprises a sample pick up means, a metering device fluidly connected to the sample pick up means, a seat for receiving the sample pick up means, a first distributor valve, wherein the first distributor valve comprises a plurality of ports and a plurality of connecting elements for changeably connecting the ports of the first distributor valve, wherein
one port is directly fluidly connected to the seat,
two ports are directly fluidly connected to the trap column,
one port is directly fluidly connected to the separation column,
one port is directly fluidly connected to the analytical pump,
wherein the method comprises
isolating the trap column from ambient atmosphere,
pressurizing the trap column to a trap column pressure exceeding ambient pressure, wherein the trap column is not fluidly connected to the separation column.
6 . Method according to claim 1 , wherein the liquid chromatography system comprises a pressure sensor, and wherein the method comprises the step of the pressure sensor sensing a pressure.
7 . Method according to claim 1 , wherein the liquid chromatography system comprises a first distributor valve, and wherein the trap column comprises a first end and a second end, the method comprising:
providing the trap column with the sample via the first end, wherein the first end is upstream to the second end;
switching the first distributor valve such that the first end is downstream of the second end, and
pushing the sample back out the first end by the analytical flow.
8 . Method according to claim 1 , wherein the liquid chromatography system comprises a first distributor valve, the method comprising pushing the sample further in the direction of a trap flow by an analytical flow such that the flow direction corresponds to the flow direction with which the trap column was supplied with the sample.
9 . Method according to claim 1 , wherein the liquid chromatography system comprises a second distributor valve, a waste and a metering device comprising a piston, the method comprising:
connecting a side of the trap column facing away from the sample to a waste via the second distributor valve,
pushing the sample with previously raised trap solvent to the trap column by means of the piston of the metering device moving forward,
pushing components which do not adhere to the trap column out to waste.
10 . Method according to claim 1 , wherein the liquid chromatography system comprises a metering device comprising a piston, a first distributor valve, a second distributor valve, a tubing configured to fluidly connect the first distributor valve and the second distributor valve and a further tubing configured to fluidly connect the first distributor valve and the metering device comprising the piston,
wherein pressurizing the trap column comprises compressing a volume in the tubing, the trap column, the metering device and the connections by means of the piston in the metering device moving forward,
wherein the volume is compressed until the first pressure is reached, to bring the sample in the trap column to the first pressure.
11 . Method according to claim 1 , wherein the liquid chromatography system comprises a metering device comprising a piston, a first distributor valve, a second distributor valve, a tubing configured to fluidly connect the first distributor valve and the second distributor valve, and further tubing configured to fluidly connect the first distributor valve and the metering device, the method comprising:
fluidly connecting the trap column to the tubing which connects the first distributor valve and the second distributor valve to provide a connection to the metering device,
reducing the pressure still present in the portion of the system fluidly connected to the trap column by means of the piston moving back.
12 . Method according to claim 1 , wherein the liquid chromatography system comprises a second distributor valve, a waste, a solvent reservoir and a metering device comprising a piston, the method comprising:
fluidly connecting the trap column to the waste,
switching the second distributor valve to the waste to dissipate residual pressure remaining in the trap column and the components fluidly connected to the trap column,
reconnecting the metering device with a solvent reservoir,
drawing up solvent, and
washing the trap column and components fluidly connected to the trap column by means of the second distributor valve.
13 . Method according to claim 1 , wherein the liquid chromatography system comprises a first distributor valve, a second distributor valve and a metering device, the method comprising:
iteratively switching the second distributor valve while the first distributor valve remains in one position,
drawing solvent into the metering device,
washing components fluidly connected to the metering device including the trap column, and
equilibrating by fluidly connecting the separation column and the analytical pump and iteratively switching the second distributor valve.
14 . Method according to claim 1 , wherein the liquid chromatography system comprises a first distributor valve, a second distributor valve, a plurality of solvent reservoirs, a tubing section and a metering device comprising a piston, a port and a further port, the method comprising:
connecting the further port of the metering device with one of the solvent reservoirs by means of the second distributor valve to allow the metering device to raise fresh trap solvent into the section of the system fluidly connected to the trap column,
connecting the tubing to one of the solvent reservoirs by means of moving the second distributor valve to open the further port and close the port of the metering device,
drawing solvent from one of the solvent reservoirs into the metering device by moving the piston back,
subsequently closing the further port and opening the port, and
supplying solvent into the tubing section by moving the piston forward to supply more solvent towards the trap column.
15 . Method according to claim 6 , wherein the liquid chromatography system further comprises a metering device comprising a piston, a first distributor valve, a second distributor valve, a tubing configured to fluidly connect the first distributor valve and the second distributor valve and a further tubing configured to fluidly connect the first distributor valve and the metering device comprising the piston, wherein pressurizing the trap column comprises compressing a volume in the tubing, the trap column, the metering device and the connections by means of the piston in the metering device moving forward, wherein the volume is compressed until the first pressure is reached, to bring the sample in the trap column to the first pressure, the first pressure being determined to have been reached by means of the pressure sensor.
16 . Method according to claim 5 , wherein the trap column pressure exceeds the ambient pressure by at least 10 bar.
17 . Method according to claim 1 , wherein the method comprises a separation method, wherein the separation method comprises separating constituents of the sample in the separation column at an analytical pressure, wherein the first pressure is at least 10% of a maximum analytical pressure.
18 . Method according to claim 1 , wherein the trap column is isolated from the ambient atmosphere by connecting the trap column to dead ends at both sides.