U.S. patent number 3,585,863 [Application Number 04/814,324] was granted by the patent office on 1971-06-22 for method and device for introducing samples into a chromatographic column.
This patent grant is currently assigned to Ceskoslovenska akademie ved. Invention is credited to Jiri Hrdina.
United States Patent |
3,585,863 |
Hrdina |
June 22, 1971 |
METHOD AND DEVICE FOR INTRODUCING SAMPLES INTO A CHROMATOGRAPHIC
COLUMN
Abstract
A method and apparatus for carrying out liquid chromatography. A
plurality of cartridges are transported sequentially into fluid
communication with the column. Each cartridge contains a solid
porous sorption material retained between perforated plates.
Immediately before moving into the location where the cartridge is
in communication with the column, the cartridge is clamped in a
fluid circuit and deaeration of the sorption material is carried
out by pumping a liquid through the cartridge. After deaeration of
the porous material, eluent liquid is pumped through the cartridge
and into the column and the danger of the inclusion of air bubbles
in the stream is eliminated.
Inventors: |
Hrdina; Jiri (Prague,
CS) |
Assignee: |
Ceskoslovenska akademie ved
(Progue, CS)
|
Family
ID: |
5363947 |
Appl.
No.: |
04/814,324 |
Filed: |
April 8, 1969 |
Foreign Application Priority Data
|
|
|
|
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Apr 9, 1968 [CS] |
|
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PV2629/68 |
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Current U.S.
Class: |
73/864.85;
73/61.56 |
Current CPC
Class: |
G01N
30/20 (20130101); G01N 1/405 (20130101); G01N
30/14 (20130101); G01N 2030/8881 (20130101); G01N
30/24 (20130101); G01N 2030/201 (20130101) |
Current International
Class: |
G01N
30/14 (20060101); G01N 30/00 (20060101); G01N
30/20 (20060101); G01N 1/34 (20060101); G01N
30/24 (20060101); G01N 30/88 (20060101); G01n
001/00 () |
Field of
Search: |
;73/23,23.1,53,61.11C,421 ;23/230,253 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
Primary Examiner: Swisher; S. Clement
Claims
I claim:
1. A method for introducing liquid samples into a chromatrographic
column in the absence of air bubbles comprising: adsorbing a
quantity of a sample liquid on a porous material within a
cartridge, connecting said cartridge in a first fluid circuit,
pumping a first liquid stream through said cartridge, subsequently
disconnecting said cartridge from said first circuit and connecting
said cartridge with a second fluid circuit, and pumping a second
liquid stream through said cartridge and into said column, whereby
said first liquid stream deaerates said cartridge.
2. The method according to claim 1 wherein said porous material is
substantially identical with porous material contained in said
column, said sample is substantially insoluble in said first
liquid, whereby the first liquid deaerates the cartridge without
disturbing the bond between the sample and said porous
material.
3. A method for introducing liquid samples into a chromatographic
column in the absence of air bubbles comprising: adsorbing a
quantity of a sample liquid on a porous material within a
cartridge, supporting a plurality of said cartridges for sequential
movement through a first position and through a second position,
pumping a first liquid stream through said cartridge at said first
position, and subsequently pumping a second liquid stream through
said cartridge at said second position and into said column,
whereby said first liquid stream deaerates said cartridge.
4. The method according to claim 3 wherein said material in said
cartridge is substantially identical with material contained in
said column, and said first liquid is substantially identical with
said second liquid.
5. Apparatus for introducing samples into a chromatographic column
comprising: a plurality of cartridges, means mounting said
cartridges for sequential movement between a deaerating position
and a sample introduction porition, first means for pumping liquid
through said cartridge while in said deaerating position to
displace air from the interior of said cartridge, and second means
for pumping eluent liquid through said cartridge while in said
sample introduction position, said eluent pumping means including a
conduit communicating between said cartridge and said
chromatographic column while said cartridge is in said sample
introducing position.
6. The apparatus according to claim 5 wherein said first liquid
pumping means and said eluent pumping means are operable
simultaneously for deaerating one of said cartridges while a
preceding cartridge in said sample introducing position is
receiving elution liquid and a sample is being carried from said
preceding cartridge into said column.
7. The apparatus according to claim 5 including means for clamping
said cartridges in said deaerating position and in said sample
introduction position, said clamping means including fluid passages
in fluid circuits with said first and second pumping means,
respectively and sealing means between said clamping means and said
cartridge, whereby said cartridges are effectively transported from
one position to the other and connected in the respective fluid
pumping circuits.
Description
My invention relates to a method for introducing samples into
chromatographic columns and it also relates to a device for
carrying out said method.
In one known method and device the samples to be analyzed, for
example mixtures of amino acids, are introduced before being
analyzed into individual cartridges wherein they are held by
sorption on a small amount of an ion exchanger gripped between two
porous plates. This method has the disadvantage that the samples to
be analyzed are bonded to the ion exchanger mostly in dried form,
particularly if the samples are introduced into the respective
cartridges as early as for example one month prior to the analysis.
A relatively large amount of air is contained either in microscopic
holes between ion exchanger grains or other microscopic spaces in
the cartridge and when the liquid eluent flows through the
cartridge, the air is entrained in the liquid stream which flows
into the chromatographic column. This is not too harmful in
analytical processes of medium effectivity in which one analysis
may last about two hours. But in highly effective modern methods
this disadvantage becomes damaging substantially reducing the total
efficiency of the method and device.
It is a general object of my invention to eliminate the air bubbles
from the eluent stream.
In accordance with the invention each of the cartridges, the
contents of which are to be treated in an analytical process, is
freed of an undesirable quantity of air before being put into a
position in which the sample is transposed from the cartridge into
the chromatographic column. To free the cartridge of air it is
introduced into a hydraulic circuit close to the place at which the
sample is to be transposed from the cartridge into the
chromatographic column. In this position a suitable liquid is
pumped through it either permanently or temporarily. This liquid
fills up all spaces which heretofor have been occupied by air or by
a gas.
The invention will be best understood from the following
specification to be read with the accompanying drawing in which
FIG. 1 illustrates substantially in section an embodiment of my new
device;
FIG. 2 shows a sectional view of a detail of FIG. 1;
FIG. 3 is a partial top view of a transportation disc for the
cartridges; and
FIG. 4 is a schematic view of one modified form of the pump and
conduit apparatus of FIG. 1; and
FIG. 5 is a schematic view of another modified form of the pump and
conduit apparatus of FIG. 1.
In the following specification the reference numerals 1 and 2 used
for the cartridges also indicate their working positions later
referred to; otherwise all reference numerals in all figures
identify the same or equivalent elements.
The cartridges 1, 2 are transposed stepwise by a carrier 3 such as
a transportation disc which turns step-by-step in the direction of
arrow 4 and which also liftably frees cartridges 1, 2 from the
positions shown in FIG. 1 where they are clamped between the
pressure bodies 10 and 18 and the discharge channels 13 and 19',
respectively. Each of the cartridges 1, 2 is provided with a
longitudinally extending central boring which contains a certain
amount of for example a cation exchange resin 5 held between plates
6 and 7 made from a porous material.
In this position cartridge 1 by means of sealing rings 11, 12 is
tightly clamped between the spring-loaded pressure body 10 actuated
by pressure element 9 and the mouth of channel 13 leading the
eluent into column 8 through channel 24 in the rotary valve member
14. Another channel 25 in said valve member 14 connects pipe 15
with duct 15' and guides the elution solution from pump 14' through
conduits 16,17 into the cartridge 1 from where said solution
continues through ducts 13 and 24 into column 8.
Prior to transposing a cartridge from position 2 into position 1
and to the performance of the analysis the cartridge 2 is clamped
similar to cartridge 1 between the pressure body 18 and the mouth
of duct 19' and liquid of proper qualities is pumped from container
23 by pump 22 through conduits 21, 20 into and through cartridge 2
into conduits 19',19 leading into waste. Thereby any gaseous
content is removed from cartridge 2 before it is moved into
position 1 and before analysis is performed. During such deaeration
the liquid fed by pump 22 need not be accurately dosed and pump 22
may be of any proper type such as a peristaltic pump because the
hydraulic resistances of cartridges 1, 2 or of their porous charge
5 and of the porous plates 6, 7 are very small.
It is important that the sample bonded by sorption or otherwise to
the carrier material in the cartridge is not carried away from the
cartridge even to a small extent. This can be achieved by filling
the cartridges with a small quantity for example of the same cation
exchange resin with which the columns are also filled. The liquid
used for expelling air or gas forms for example a citrate or
another buffer which is substantially identical with elution
buffers intended not only to carry the sample into the column but
also to perform chromatography in the column. One difference is
that the pH value is so low that no untolerable quantity of the
sample will be carried away into waste when the liquid flows
through the cartridge, and the entire quantity of the sample
remains bonded to the material in the cartridge.
FIG. 2 illustrates the rotary valve member 14 turned approximately
by 60.degree. clockwise to directly connect the pipe 15 through the
channel 25 in the rotary valve member 14 with column 8; the eluent
is thus fed directly into the column 8 and the cartridges 1, 2 can
be freely manipulated as they are not introduced into the hydraulic
circuit between pump 14' and column 8.
FIG. 3 shows a conventional transportation disc 3 supporting a
plurality of cartridges 1, 2 arranged in two concentric placements.
This arrangement is advantageous for automatic introduction of
samples into more than one column.
In accordance with conventional practice, the disc 3 rotates at
predetermined time intervals to advance the cartridges from one
position to the next. Thus, the cartridges are unclamped by raising
the clamping heads 10 and 18, while raising the disc 3 to engage
the collar at the upper end of each cartridge 1 and 2. In this
manner, the cartridges are disengaged from the conduits 17 and 13;
and the conduits 21 and 19' and are free to move from the position
indicated at 2 in FIG. 1 to the position indicated at 1 in FIG. 1.
The cartridge previously in the position indicated 1 advances
toward the right, as viewed in FIG. 1 and as indicated by the arrow
4. The cartridges are thus transported step by step into and out of
engagement with the clamping heads 10 and 18. When the cartridges 1
and 2 are in the position shown in FIG. 1, the pump 22 draws liquid
from the container 23 for deaeration.
During deaeration the cartridge in position 2 is being completely
filled with liquid by pump 22 from container 23. Excess liquid
passes out of the cartridge 2 through the conduit 19' to waste.
After release of the cartridge from this position 2 and during its
transposition into position 1, that is approximately for one or two
seconds, no liquid flows therethrough because of the hydraulic
resistances of the cartridge charge 5 and of the porous plates 6,
7.
The valve 14 is in the position shown in FIG. 2 while the cartridge
is being transported from one position to the other. The pump 14'
operates continuously and the eluent liquid flows through the
conduit 15 and through the valve passage 25 to the column 8. When
the cartridge is clamped in position, as shown in FIG. 1, the valve
14 is rotated to the position shown in FIG. 1 to conduct eluent
through the passage 17, through the cartridge containing the
sample, and into the column 8. The valve 14 is then returned to the
position shown in FIG. 1 to stop the flow of eluent liquid to the
cartridge, thereby allowing the cartridge to be transported by the
disc 3. The deaerating step that is performed at FIG. 2 and the
elution step that is performed at position 1 occur substantially
simultaneously.
It would not be harmful if an air bubble would form under the
pressure body 10 when the cartridge is again gripped in position 1.
At a low rate of flow this air bubble would not be carried down by
the stream and any sudden change in the flow would be entirely
negligible; the danger of transposition of many small bubbles from
the cartridge into the column is completely eliminated even in the
form of foam. In very effective chromatographic processes for
example lasting less than one hour a transposition of a larger
amount of gas into the column would be a factor which considerably
reduces the separating power of the column.
FIG. 4 shows a modified form of the pump and apparatus at the pump
and conduit apparatus of FIG. 1. The liquid container 23a supplies
liquid to the pump 22a and a valve 24a selectively controls flow
through the conduit 21a which corresponds to the conduit 21 in FIG.
1, or with the waste conduit 19a, corresponding to the waste
conduit 19 in FIG. 1. Similarly, FIG. 5 discloses a pump 22b for
supplying liquid from a container 23b to the conduit 21b or 19b, as
controlled by the operation of the valves 26 and 27.
* * * * *