EP0781417A1 - Method for analysis and device for carrying out the method - Google Patents
Method for analysis and device for carrying out the methodInfo
- Publication number
- EP0781417A1 EP0781417A1 EP95932284A EP95932284A EP0781417A1 EP 0781417 A1 EP0781417 A1 EP 0781417A1 EP 95932284 A EP95932284 A EP 95932284A EP 95932284 A EP95932284 A EP 95932284A EP 0781417 A1 EP0781417 A1 EP 0781417A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- sample
- tube
- reagent
- supply tube
- detector
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
- G01N35/1004—Cleaning sample transfer devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L13/00—Cleaning or rinsing apparatus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
- B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
Definitions
- the present invention relates to a method of chemically analyzing liquid substances with the aid of at least one throughflow sensor or detector, hereinafter referred to as "a detector", and at least one reagent, wherein a sample volume of the liquid substance to be analyzed is introduced into a flexible tube through a supply tube and the sample and a volume of reagent are mixed and caused to react with one another, and then transported through the flexible tube to the detector.
- a detector throughflow sensor or detector
- reagent a sample volume of the liquid substance to be analyzed is introduced into a flexible tube through a supply tube and the sample and a volume of reagent are mixed and caused to react with one another, and then transported through the flexible tube to the detector.
- the invention also relates to analysis apparatus designed to carry out the method, and to a supply tube which is adapted particularly for use with such appara- tus.
- the analysis method is preferably a batch-wise method and is mainly intended for use in clinical applications, such as in connection with intensive care, surgery, transplantations and other clinical applications where continuous monitoring of the chemical-physiological values of patients is required.
- the method is particularly suited for use when only a relatively few samples are to be analyzed, for instance in the analysis of samples taken from one or a few bed-ridden patients in a hospital ward.
- batch-analyzers Many different types of analysis apparatus are known to the art. In principle, these apparatus can be divided into two main groups: continuous analyzers and batch-wise working analyzers (hereinafter referred to as batch-analyzers) .
- the various samples are drawn by suction from their respective containers into a tube in which the samples are moved in mutually one and the same direction throughout the entire analysis procedure.
- the samples form a continuous flow to which reagent is introduced at predetermined points, either batch-wise or continuously.
- the flow of liquid finally reaches the measuring cell in which the liquid is assayed.
- All liquid transports in continuous analyzers are regulated with the aid of pumps, generally rotating displacement pumps, which also effect the outfeed of the samples.
- continuous analyzers can be divided between those in which the various samples in the tube are mutually separated, segmented, with the aid of air bubbles for instance, and those in which the samples in the tube constitute delimited liquid plugs in essentially laminar liquid carrier flows, which may possibly contain a reagent.
- Flow Injection Analysis or the FIA principle.
- One problem with continuously working analyzers is that the reagent or reagents is/are introduced through separate supply tubes, valves, pumps, etc., which requires a complicated apparatus construction.
- Another problem associated with continuous analysis methods is, of course, the risk of contamination between the samples.
- a continuous analysis method and system are known from US-A- 4,853,336, wherein successive liquid segments containing related components in a liquid packet in which an analysis mixture is established for instance, are mutually separated by at least one immiscible segment, for instance air. These liquid segments are mixed within the apparatus at a later point in time, somewhere in a transport line, such as to achieve delayed on-line mixing of the various components. Mixing of the various components is achieved by either eliminating, fragmenting or physically removing the immisci- ble segment at selected sites along the line. In order to prevent the samples contaminating one another, an immiscible liquid is used to cover the inner surfaces of the apparatus and the outer surface of the probe.
- the batch analyzer containers generally function as measuring cells in themselves, for instance cuvettes in different types of photometers, there ⁇ with obviating the need to transport part of the sample- reagent mixture to a separate measuring analyzer cell.
- One drawback with known batch processes is that relatively large quantities of sample and reagent are required. The amount of sample available must be at least sufficient to produce in practice a sample-reagent mixture in which the ratio between sample and reagent quantities is well adjusted. It is very difficult to attain acceptable precision with regard to this ratio when small quantities of sample are used.
- the sample-reagent mixture is contained in a more or less open container from the time of being mixed to the actual time of assaying the sample, the mixture is more sensitive to evaporation in the case of smaller volumes.
- the container itself forms a measuring cell, i.e. a cuvette
- the problem is ie worse because cuvettes must be relatively large and thus require relatively large volumes of sample- reagent mixture in order to cover the beam path in the photometer and therewith prevent stray light reaching the detector of the analyzer, this problem being well known in this particular technical field.
- Concentration determinations performed with the aid of batch- analyzers are normally either end-point analyses or kinetic assays.
- End-point analyses involve recording the value at which the sample has ceased to react with the reagent, i.e. when said value is constant in a sufficient number of mutually sequential analyses.
- the measured value is related directly to the original sample concentration.
- a kinetic assay can then be carried out, i.e. a procedure in which the values obtained with a plurality of mutually sequential analyses are recorded and the derivative of the time-dependent curve described by said values calculated.
- the original con- centration of the sample can then be calculated from the derivative and the initial concentration in the sample volume.
- Batch-analyzers that include cuvettes are often designed to be placed in carousel-like devices by means of which the individual cuvettes are moved into the beam path of the detector.
- the devices operate in accordance with process sequences, analysis cycles (generally of a 25-30-second duration), in a number of steps, including among others:
- A is the total analysis time in relation to the number of samples
- M is the time taken for one analysis cycle
- m is the number of analysis cycles for each sample
- p is the number of samples.
- An overlying object of the present invention is to provide a method of analysis which enables the construction of simpler analysis apparatus in comparison with known tech- niques.
- a particular object of the present invention is to provide a method of analysis which enables very small sample quanti ⁇ ties to be analyzed without risk of contamination between the sample quantities.
- Another particular object of the invention is to provide a method of analysis which when working with a relatively small number of samples will enable each sample to be analyzed in a much shorter time than that possible when practicing known techniques.
- Still another object of the invention is to provide analyti ⁇ cal apparatus suitable for use in clinical applications, and primarily apparatus which can be set-up and used in the proximity of patients to enable their chemical-physiological values to be monitored continuously.
- the aforesaid overlying objects of the invention are achieved with the aid of an analysis method of the kind defined in the introduction in which the sample volume and the reagent volume are supplied to the analytical apparatus one after the other through an opening in the supply tube, and in which the sample quantity and the reagent quantity are mixed together in the supply tube, and optionally also in the flexible sample tube, through the medium of an air segment which precedes the sample and reagent volumes.
- the sample and reagent volumes are thus introduced immediately after one another in the absence of an intermediate air segment.
- sample and reagent volumes will constitute a coherent package between two air segments.
- sample and reagent can be mixed together satisfactorily in this simple fashion.
- One explanation may be that eddies are generated in liquid at the liquid-air segment interface and move outwards and then downwards along the full length of the liquid plug and therewith cause the contents of the full plug (i.e. the combination of sample and reagent) to mix together.
- this mixing phenomenon is independent of whether the liquid flow is turbulent or laminar.
- the mixing effect can be optimized by selecting suitable values for parameters that are relevant with regard to liquid flow in tubes or pipes, for instance such parameters as flow rate, hydraulic diameter, roughness of the tube surface, individual resistance, viscosity of the liquid flow, etc.) .
- the method according to the present invention is also effective in causing sample and reagent to mix together surprisingly quickly; the sample and reagent are mixed together in the time taken for them to move from the supply tube to the detector, which means a time consumption of up to about two seconds, generally about one second.
- the analytical apparatus used may be given a simpler construction, since only one supply tube is required for the supply of both sample and reagent. Naturally, this will present to the person skilled in this art the possibility of making other obvious constructional simplifications.
- the method is a batch method.
- This provides a method which enables very small volumes of sample to be analyzed without risk of contamination between the volumes.
- the risk of contamination is reduced because the method is of the batch kind, the possibility of working with small sample quantities is obtained with the aid of the present invention by virtue of the fact that sample and reagent are mixed precisely as they are supplied to the analytical apparatus.
- This enables small sample and reagent volumes to be taken-up with great precision and without risk of the mixing ratio being impaired by evaporation, because the sample and reagent volumes are mixed immediately prior to being assayed and are also protected within a tube or a flexible tube from the point at which they are mixed to the location of the cell.
- the detector, the flexible tube and the supply tube are washed clean with washing solution after the assaying process, said washing solution being supplied to the detector through a washing tube and thereafter through the detector, whereafter the washing solution is flushed through and out of the supply tube via said flexible tube.
- the inventive method is quicker than conventional batch methods that operate in parallel with many samples simultaneously, when a few samples require analysis over a longer analysis period.
- an inventive batch method is much better suited for such analytical processes when only a relatively few samples are to be analyzed and a quick result of the analyses is required, e.g. when analyzing samples taken from one or a few bed-ridden patients in a hospital ward.
- the present invention also relates to apparatus for carrying out the inventive method, said apparatus including at least one detector, a detector-connected flexible tube, a supply tube having a first free end and a second end connected to said flexible tube, further a washing tube connected to the detector, and at least one pump for pumping liquid substance and liquid reagent into and out of the detector respectively and for pumping washing solution through the washing tube, the detector, the sampletubeand the supply tube, said supply tube having an opening through which sample and reagent can be supplied sequentially one after the other.
- the supply tube is preferably configured for mixing of sample and reagent therein, and optionally also in the following sample tube.
- the supply tube has at least two throughflow areas of mutually different size, wherein the smaller throughflow area merges with the larger throughflow area via an abrupt increase in area in a direction from said first end and towards said second end, and wherein the abrupt increase in area is intended to assist in generating eddies or turbulence in the flow in the supply tube and possibly also in the sample tube, at least in a direction from the first end of the supply tube and towards its second end downstream of the abrupt increase in area.
- the apparatus is a batch aj ratus and is provided with a metering pump for pumping liquid substance and reagent to the detector, and a washing pump which pumps washing solution from a washing solution container into the detector through the washing tube, and from there out through the supply tube via said tube, wherein the metering pump is an oscillating displacement pump, e.g. a plunger pump or injection pump, and is connected to the washing tube at a location between the detector and the washing pump, and wherein the washing pump is a rotary displacement pump, e.g. a peristaltic pump.
- This embodiment provides a batch analyzing apparatus of particu ⁇ larly uncomplicated design in comparison with conventional apparatus.
- the particular combination of pumps eliminates at one stroke a large part of the requirement of valves and like devices with which known techniques are encumbered. This is described in more detail below.
- the apparatus is also suitable for use in clinical applications, particularly as it can be set-up and used in the proximity of patients for the purpose of monitoring continuously their chemical/physio- logical values, this being achieved by virtue of the appara ⁇ tus working with relatively few steps with each sample and therewith generating relatively little noise.
- the apparatus includes a sample loop which is connected either to the flexible sample tube or to the washing hose, said sample loop being provided with valves for activating and deactivating the sample loop so as to alternate the flow in the analysis apparatus and the flow in an external analysis apparatus, which may be a chromatographic device, e.g. an HPLC-apparatus.
- an external analysis apparatus which may be a chromatographic device, e.g. an HPLC-apparatus.
- Fig. 1 illustrates schematically one embodiment of an inventive apparatus
- Fig. 2 is a cross-sectional view of part of the supply tube of the apparatus shown in Fig. 1;
- Fig. 3 illustrates a method of washing the supply tube.
- the apparatus illustrated in Fig. 1 is used by first pumping washing solution from a container 90 for washing solution through a flexible washing tube 40 with the aid of a washing pump 60, in the illustrated case a peristaltic pump.
- the washing tube of the illustrated apparatus is made of PVC plastic.
- the washing solution is pumped through the washing tube 40, a photometric detector 10, and a sample tube 20 and out through an outlet orifice 70 of the supply tube 30 at a flow rate of about 50 ⁇ l/s.
- the sample tube 20 is made of PEEK plastic, because PEEK has a relatively low hydrophobicity which counteracts the adhesion of large air bubbles to the sample tube.
- other plastics having similar properties may be used as an alternative.
- the outer surface of the supply tube 30 is cleaned during the washing process with the aid of the washing vessel 130 (described in more detail with reference to Fig. 3) .
- the supply tube 30 is lifted from the washing vessel with the aid of a device 170 intended herefor and the apparatus is then ready for renewed analysis of samples.
- the device 170 moves the supply tube vertically and laterally, in this case in the plane of the drawing, and moves a stand for test tubes 180 in a direction perpendicular to the plane of the drawing.
- about 1 ⁇ l of air is first sucked into the supply tube 30 with the aid of the metering pump 50, which in the illustrated case is a plunger pump or injection pump.
- the static peristaltic pump 60 functions therewith as a sealing valve, because it prevents liquid from passing through the washing tube 40.
- the device 170 then moves the supply tube 30 to the vicinity of and down into a test tube 180, which in the illustrated case is of the kind described in the Swedish Patent Application 9303344-7, i.e. a test tube provided with a rubber septum and a capillary tube for taking-up very small volumes of sample at the tube orifice. Samples (about 0.1-2.0 ⁇ l) are sucked into the supply tube 30 through its orifice 70, with the aid of the pump 50.
- reagent (about 15 ⁇ l) is drawn correspondingly into the supply tube 30, through its orifice 70.
- the sample and reagent plugs are thus in contact with one another as they are pumped further through the tube 20 at a flow rate of about 5-50 ⁇ l/s (which in the case of the internal diameter of the tube corresponds to a speed of about 4-40 cm/s) .
- Mixing of the sample with the reagent is begun in the supply tube 30 by virtue of air-segmenting.
- the supply tube 30 includes an abrupt change in area, as described in more detail below with reference to Fig. 2, which also possibly contributes further towards the intermixing of sample and reagent.
- the detector 10 is of the kind described in SE-B-455 134, i.e. it comprises a thin tube of transparent material in which the mixture is con ⁇ tained, and light is passed through the cylindrical wall of the tube at an acute angle to the longitudinal axis thereof and is totally reflected within the tube one or more times and thereafter led out through said cylindrical surface at an acute angle.
- the illustrated embodiment is effective in achieving a kinetic analysis of the sample-reagent mixture in less than about thirty seconds, with the aid of the detector 10 and a computer (not shown) connected thereto, whereafter the mixture is washed from the apparatus with the aid of the washing pump 60 and the analysis sequence is recommenced.
- the full sequence takes about one minute.
- the apparatus illustrated in Fig. 1 is suitable for assaying such substances as glucose, lactate, glycerol, pyruvate, urea, creatinine, alcohols, glutamate and carbon dioxide.
- Analyses can also be carried out with an external analytical appara ⁇ tus, e.g. an HPLC apparatus, for analyzing amino acids, purines, lactates, pyruvates, ascorbates, histamines, polyamines, leukotriens, free radicals, ions or selected pharmaceutical preparations, with the aid of a six-port valve 2 fitted to the washingtube40.
- the valve 2 is a two-position valve retailed under product number C6W by Valco Instruments Co., Houston, Texas, U.S.A. and functions to connect a sample loop 80 with the tube 40 when the valve 2 is set to a first position, therewith enabling the loop to be filled with sample-reagent mixture with the aid of the metering pump 50, while the sample loop 80 remains in contact with an HPLC apparatus (not shown) via the lines 4 and 6 when the valve 2 is set to a second position, so as to enable the sample- reagent mixture in the sample loop to be transferred to the HPLC apparatus.
- C6W Valco Instruments Co., Houston, Texas, U.S.A.
- the supply tube 30 of the illustrated embodiment comprises two parts, i.e. a pointed cannula 100 and a tube 110.
- the cannula 100 is of the kind described in Swedish Patent Application 9303344-7 and has an outer diameter of only 0.4 mm and is particularly suitable for taking-up very small sample volumes from so-called microdialysis test tubes, as described in said Swedish patent application.
- the cannula 100 has an inner diameter of only 0.15 mm which, in combination with the injection pump 50 enables sample volumes as small as 0.1 ⁇ l to be taken-up with great precision.
- the cannula 100 terminates in the tube 110 at an abruptly increased area or section 120 and is attached to the tube in said area.
- FIG. 3 illustrates washing of the outer surface of the supply tube 30 with washing solution contained in a known washing vessel 130.
- the washing solution exiting from the supply tube 30 fills the space 140 and flows over the space-defining wall 150, as indicated by the arrows in the Figure.
- the outer surface of the supply tube 30 is therewith washed clean.
- the washing solution then exits through the outlet 160.
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Clinical Laboratory Science (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
SE9403071 | 1994-09-14 | ||
SE9403071A SE503669C2 (sv) | 1994-09-14 | 1994-09-14 | Förfarande för analys samt anordning för genomförande av förfarandet |
PCT/SE1995/001038 WO1996008725A1 (en) | 1994-09-14 | 1995-09-14 | Method for analysis and device for carrying out the method |
Publications (1)
Publication Number | Publication Date |
---|---|
EP0781417A1 true EP0781417A1 (en) | 1997-07-02 |
Family
ID=20395234
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP95932284A Withdrawn EP0781417A1 (en) | 1994-09-14 | 1995-09-14 | Method for analysis and device for carrying out the method |
Country Status (8)
Country | Link |
---|---|
EP (1) | EP0781417A1 (sv) |
JP (1) | JPH10508687A (sv) |
CN (1) | CN1158166A (sv) |
AU (1) | AU3537295A (sv) |
FI (1) | FI971056A (sv) |
NO (1) | NO971141D0 (sv) |
SE (1) | SE503669C2 (sv) |
WO (1) | WO1996008725A1 (sv) |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6759014B2 (en) | 2001-01-26 | 2004-07-06 | Symyx Technologies, Inc. | Apparatus and methods for parallel processing of multiple reaction mixtures |
US6913934B2 (en) * | 1998-08-13 | 2005-07-05 | Symyx Technologies, Inc. | Apparatus and methods for parallel processing of multiple reaction mixtures |
JP3735666B2 (ja) * | 2001-12-27 | 2006-01-18 | 独立行政法人産業技術総合研究所 | 糖類混合物の同時分析方法及びそれに用いる分析装置 |
DE102006053096A1 (de) | 2006-11-10 | 2008-05-15 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Fluidikeinrichtung und Verfahren zu deren Betrieb |
CN106053820A (zh) * | 2016-06-29 | 2016-10-26 | 西南大学 | 微透析采样‑流动注射化学发光联用法在小分子半抗原与单克隆抗体间亲和力测定中的应用 |
BG67391B1 (bg) * | 2018-12-12 | 2021-11-15 | "Бултех 2000" Оод | Метод за промиване на работна система в анализатор на течни храни |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE3629430A1 (de) * | 1986-08-29 | 1988-03-10 | Messerschmitt Boelkow Blohm | Messanordnung zum feststellen von rissen in strukturbauteilen von luftfahrzeugen |
US5038618A (en) * | 1986-11-11 | 1991-08-13 | British Aerospace Public Limited Company | Measurement of distortion |
WO1993025866A1 (en) * | 1992-06-05 | 1993-12-23 | Monash University | Sensing patches utilising incorporated waveguide sensor |
GB9317576D0 (en) * | 1993-08-24 | 1993-10-06 | British Aerospace | Fibre optic damage detection system |
-
1994
- 1994-09-14 SE SE9403071A patent/SE503669C2/sv not_active IP Right Cessation
-
1995
- 1995-09-14 CN CN95195090A patent/CN1158166A/zh active Pending
- 1995-09-14 EP EP95932284A patent/EP0781417A1/en not_active Withdrawn
- 1995-09-14 AU AU35372/95A patent/AU3537295A/en not_active Abandoned
- 1995-09-14 JP JP8510119A patent/JPH10508687A/ja active Pending
- 1995-09-14 WO PCT/SE1995/001038 patent/WO1996008725A1/en not_active Application Discontinuation
-
1997
- 1997-03-12 NO NO971141A patent/NO971141D0/no not_active Application Discontinuation
- 1997-03-13 FI FI971056A patent/FI971056A/sv unknown
Non-Patent Citations (1)
Title |
---|
See references of WO9608725A1 * |
Also Published As
Publication number | Publication date |
---|---|
AU3537295A (en) | 1996-03-29 |
SE9403071D0 (sv) | 1994-09-14 |
FI971056A0 (sv) | 1997-03-13 |
NO971141L (no) | 1997-03-12 |
CN1158166A (zh) | 1997-08-27 |
NO971141D0 (no) | 1997-03-12 |
SE503669C2 (sv) | 1996-07-29 |
FI971056A (sv) | 1997-03-13 |
WO1996008725A1 (en) | 1996-03-21 |
JPH10508687A (ja) | 1998-08-25 |
SE9403071L (sv) | 1996-03-15 |
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