KR100794434B1 - Automatic Chlorophyll Analyzer and Analytical Method - Google Patents

Abstract

The present invention relates to an automatic chlorophyll analysis device and analysis method, and in particular a flow path through which the fluid flow; A flow path change valve disposed on the flow path, the flow path change valve having a plurality of inlet ports and selectively communicating any one of the inlet and outlet ports with the flow passage; A filter installed on the flow path, separating the particulate matter contained in the liquid sample, and filtering the extract with chlorophyll; A fluorescence detector provided on the flow path and measuring fluorescence of the extract liquid filtered by the filter; A syringe for inhaling or discharging the fluid on the flow path; A particulate material contained in the liquid sample using the chlorophyll automatic analyzer, which is installed on the flow path and connected to the syringe to selectively change the suction and discharge directions of the fluid. After concentration, chlorophyll was extracted with an organic solvent to measure the concentration automatically.

Chlorophyll, automatic analysis, filtration

Description

Chlorophyll Analyzer and Analytical Method

1 is a block diagram showing an automatic chlorophyll analysis device according to an embodiment of the present invention,

2A and 2B are cross-sectional views illustrating a cartridge including a filter of an automatic chlorophyll analyzer according to an exemplary embodiment of the present invention.

3a and 3b is a block diagram showing a syringe of the chlorophyll automatic analysis device according to an embodiment of the present invention,

Figure 4 is a block diagram showing an automatic chlorophyll analysis device including a high performance liquid chromatography (HPLC) according to another embodiment of the present invention,

5 is an assembly view showing an chlorophyll automated analysis device manufactured according to another preferred embodiment of the present invention,

6 is a result of measuring the fluorescence detector signal calculated by the concentration of the chlorophyll automatic analysis device according to a preferred embodiment of the present invention.

** Brief description of symbols for the main parts of the drawing **

10 Euro 11-14 Access port

20: flow path change valve 30: filter

31: cartridge 32: ultrasonic generator

33: front panel 34: rear panel

35: filter O-ring 36: cartridge ring

37: cartridge lower penetration portion 38: cartridge upper penetration portion

40: fluorescence detector 50: pressure gauge

60: syringe 61: plunger

62: plunger moving means 63: light source

64: light detection sensor 70: 4-way valve

75: sample filtrate storage bag 80: sample

90: organic solvent 100: HPLC injection valve

101: HPLC pump 102: HPLC solvent

103: HPLC column 104: HPLC detector

The present invention relates to a chlorophyll analysis device, and more particularly to a chemical analysis device for automatically analyzing the chlorophyll component contained in the liquid sample. More specifically, the present invention relates to a chlorophyll automatic analysis device and an analysis method for measuring the fluorescence by filtering the sample in a filter and then extracted with an organic solvent.

Chlorophyll is a pigment necessary for phytoplankton in water to be photosynthesized. In physiological and oceanography, chlorophyll has been widely used as an indicator of the biomass present in chlorophyll. Chlorophyll concentrations are primarily used to assess water quality and the production of phytoplankton. In particular, the ocean has been used to estimate chlorophyll concentrations using ocean colors taken from satellites and to use them in fishery forecasts and global climate change studies.

The most widely used method for the analysis of chlorophyll concentration in water is to filter liquid samples, filter out particulate matter on filter paper, extract filter paper with 90% acetone, measure the absorbance of the extract, and use the chlorophyll absorbance coefficient. Chlorophyll a, b, c was the method to calculate the concentration. After filtration / extraction, the fluorescence of the extract was measured instead of the absorbance, and the sensitivity was improved by more than 50 times compared to the conventional method.

As another method, a method of measuring in vivo fluorescence by directly irradiating light to a liquid sample containing live plankton without filtering the sample has been proposed, and a simple biofluorescence sensor using the same has been developed. However, the biofluorescence measuring device has a problem that the absolute value is not reliable because the amount of fluorescence varies depending on the type of phytoplankton, growth stage, physiological condition, depth, adaptation state, and measurement time.

The reason why the automatic component analysis for measuring the absorbance or fluorescence after the filtration / extraction of the sample has not been developed is because it was not possible to implement a pretreatment device for automatically filtering the liquid sample and extracting it with a solvent. Chlorophyll analysis requires a sample pretreatment process, such as filtration, extraction, and centrifugation, which cannot be automated by conventional methods.

In the chlorophyll analysis method, the filtration process is a process of separating and concentrating phytoplankton, which is a particulate matter, in water. Vacuum filtration or pressurized pump is used for sample filtration, and phytoplankton is filtered out of glass fiber filter paper using filtration device. In order to automate the process of filtering the sample, it is necessary to use a robot or several filters to pre-mount the filters and to filter them sequentially.The number of samples is limited and the size of the equipment is very large. Could not automate It is also possible to use filter paper rolled up in a roll, but it is difficult to extract the filtered portion by this method.

Sample filtration is usually based on 1 liter.However, if the concentration of chlorophyll is high, an appropriate amount of sample should be filtered to prevent the filter paper from clogging.If the concentration of chlorophyll is low, the amount of chlorophyll should be increased in consideration of the detection limit of the detector. The error can be reduced. As filter paper is blocked by particulate matter, the experimenter should adjust it to an appropriate amount in consideration of filtration speed and search. If the amount of phytoplankton present in the sample is too low and the chlorophyll concentration is judged to be low, it is necessary to increase the amount of filtration above 1 liter to filter a large amount of sample. This adjustment process can be adjusted by the experimenter in manual measurement, but in order to automate, the biofluorescence should be measured in advance so that the chlorophyll concentration in the sample can be approximated and the filtration amount adjusted accordingly.

Then, the solvent is extracted by putting the filtered glass fiber filter paper into acetone, and in order to increase the extraction efficiency, all of the filter paper is ground, or ultrasonic irradiation is used to destroy phytoplankton cells. After chlorophyll is extracted with acetone, the extracted solution must be centrifuged to remove plankton particles or crushed filter paper before measuring fluorescence or absorption. However, the process of centrifugation to remove particles was complicated and difficult to automate.

Therefore, the present invention was invented to solve the above problems, and enables sample pretreatment such as filtration and extraction necessary for chlorophyll analysis, minimizes the size of the device, and facilitates fully unattended automatic analysis in the field. The purpose is to provide an automatic analysis device that can be performed easily.

The present invention for achieving the above object is composed of a flow path change valve, a filter, a filter cartridge, an ultrasonic generator, a fluorescence detector, a pressure gauge, a syringe for inhaling or discharging a fluid, a valve attached to the syringe, It is intended to provide an automatic chlorophyll analysis device that can concentrate the particulate matter contained in the sample in the filter and then automatically measure the concentration by extracting chlorophyll with an organic solvent.

The present invention provides a flow path through which a fluid flows; A flow path change valve disposed on the flow path, the flow path change valve having a plurality of inlet ports and selectively communicating any one of the inlet and outlet ports with the flow passage; A filter installed on the flow path, separating the particulate matter contained in the liquid sample, and filtering the extract with chlorophyll; A fluorescence detector provided on the flow path and measuring fluorescence of the extract liquid filtered by the filter; A syringe for inhaling or discharging the fluid on the flow path; And a four-way valve installed on the flow path and connected to the syringe to selectively change the suction and discharge directions of the fluid.

Here, the chlorophyll automatic analysis device may further include an ultrasonic generator capable of applying ultrasonic waves to the filter, and may further include a cartridge for supporting the filter so that the fluid passes only through the filter. Preferably the cartridge consists of a removable front plate and a back plate, it is more suitable to include a filter between the front plate and the back plate. More preferably, an ultrasonic generator capable of applying an ultrasonic wave to the filter is attached to the rear plate of the cartridge.

Another embodiment of such an automatic chlorophyll analyzer further comprises an organic solvent bag having an inlet and port for communicating with the flow path change valve, wherein the extract with chlorophyll is characterized in that the particulate matter separated from the liquid sample is added to the organic solvent. Chlorophyll automatic analysis device, characterized in that extracted by.

Another embodiment of the invention is that the filter is a metal or plastic material with pores of 0.4 to 0.5 microns, and the filter is 100 to 300 microns thick with pores of 0.4 to 0.5 microns. The filter surface layer and the 1000 to 2000 micron thick support layer with pores of 5 to 50 microns can be integrated filters.

To this end, the syringe of the automatic chlorophyll analyzer according to the present invention may be provided with a predetermined means for automatically inhaling or discharging the fluid in the flow path while automatically pushing up or retracting the plunger of the syringe, the syringe It is also possible to further include a light source and a light detection sensor that can detect the intake or discharge of the fluid inside. Here, the light source and the light detection sensor is preferably attached to the syringe.

In addition, the present invention may further include a sample filtrate storage bag having a port that can be connected to the four-way valve to store the filtrate passed through the filter, the component of the extract after the fluorescence measurement by the fluorescence detector It is also possible to further include a high performance liquid chromatography (HPLC) device for analysis. The high performance liquid chromatography apparatus includes an HPLC injection valve that can be connected to the flow path change valve to introduce the extract, an HPLC column that can separate chlorophyll components, and an HPLC detector that can measure chlorophyll components. It is preferable.

Another preferred embodiment for achieving another object of the present invention is a flow path change valve which is installed on the flow path through which the fluid flows, and provided with a plurality of inlet ports, and selectively communicating any one of each inlet port with the flow passage. Thereby, the liquid sample is introduced into the flow path; Separating particulate matter contained in the introduced liquid sample by a filter installed on the flow path; The separated particulate matter is extracted by an organic solvent introduced by the flow path change valve, and an extract having chlorophyll is filtered from the filter; And fluorescence is measured by passing the filtered chlorophyll extract through a fluorescence detector installed on the flow path.

Here, during the step of separating the particulate material by the filter, the filtrate of the liquid sample that does not contain the particulate material is filtered through the filter, collected in the sample filtrate storage bag, and after the fluorescence is measured In addition, by filtration of the filtrate containing no particulate matter in the sample filtrate storage bag back into the filter through a fluorescence detector, it is possible to automatically chlorophyll method characterized in that it further comprises the step of washing the filter.

Hereinafter, one preferred embodiment of the present invention will be described in detail with reference to the accompanying drawings. The invention can be better understood by the following examples, which are intended for the purpose of illustration of the invention and are not intended to limit the scope of protection defined by the appended claims.

1 is a block diagram showing an automatic chlorophyll analysis device according to an embodiment of the present invention. As shown, the automatic chlorophyll analyzer according to the present invention and the flow path 10 through which the fluid flows; A flow path change valve (20) installed on the flow path (10) and having a plurality of inlet ports, for selectively communicating any one of each inlet port with the flow path; A filter (30) installed on the flow path (10), for separating particulate matter contained in the liquid sample (80), and filtering the extract having chlorophyll; A fluorescence detector (40) installed on the flow path (10) and measuring fluorescence of the extract liquid filtered by the filter (30); A syringe 60 for sucking or discharging fluid on the flow path 10; And a four-way valve 70 installed on the flow path 10 and connected to the syringe 60 to selectively change the suction and discharge directions of the fluid.

In the method for analyzing chlorophyll using the chlorophyll automatic analyzer having such a configuration, first, a method for analyzing chlorophyll is provided on a flow path 10 through which fluid flows, and includes a plurality of access ports, and any one of each access port is a flow path. By the flow path change valve 20 to selectively communicate with, the liquid sample 80 is passed through the flow path (10).

After connecting to the liquid sample 80 through the inlet port 11 of the flow path change valve 20, the inlet port 71, 72 of the four-way valve 70 connected to the syringe 60 is in communication with each other. Then, when the plunger 61 of the syringe 60 is retracted, the sample 80 can be introduced into the flow path 10 by the suction force of the syringe 60, and the sample thus introduced reaches the filter 30. You can do it.

The present invention intends to separate particulate matter contained in the liquid sample 80 using the filter 30. In other words, the particulate matter contained in the introduced liquid sample 80 is separated by the filter 30 installed on the flow path 10. Particulate matter containing chlorophyll is separated by the filter (30), and the filtrate of the liquid sample (80) which does not contain the particulate matter is filtered through the filter (30) and thus along the flow path (10). It flows to the directional valve 70, which can enter the syringe 60 or into the sample filtrate storage bag 75 depending on the port connected with the four-way valve 70.

Here, the retracting of the plunger 61 of the syringe 60 is also possible to pull the plunger 61 by hand, but by automatically pushing or retracting the plunger of the syringe to automatically suck in the fluid in the flow path or It is also possible to use a predetermined plunger moving means 62 which can be discharged.

In this way, when the syringe 60 of the automatic chlorophyll analyzer according to the present invention is retracted, a fluid, that is, a liquid sample is introduced into the flow path 10, and phytoplankton or particulate matter including chlorophyll contained in the sample 80 is provided. Silver is separated from the filter 30, the filtrate does not include this is introduced into the syringe (60). When the plunger 61 of the syringe 60 is fully retracted, the inlet port 72 of the four-way valve 70 connected to the syringe 60 communicates with the other inlet port 73, and the plunger 61 is advanced. The solution contained in the syringe may be discharged to the outside of the flow path 10 through the access port 73.

In addition, as described above, the solution contained in the syringe 60, that is, the filtrate of the sample not containing chlorophyll may be stored using a separate sample filtrate storage bag 75. Here, the sample filtrate storage bag 75 is capable of storing the filtrate passing through the filter, it is characterized in that it has a port that can be connected to the inlet port 73 of the four-way valve (70). That is, after lowering the plunger 61 of the syringe 60 to the end, the other inlet port 72 of the four-way valve 70 connected to the syringe 60 is replaced with another inlet port 73 (sample filtrate storage bag). And an entrance port connected to the (75), the plunger 61 is moved upward to store the solution contained in the syringe (60) in the sample filtrate storage bag (75).

Filtration of the chlorophyll-containing liquid sample 80 may be continued by repeating the above-described procedure, and the volume of the sample to be filtered may be varied by adjusting the number of operations of the plunger 61. When the capacity of the sample filtrate storage bag 75 is exceeded, the inlet 72 of the valve 70 connected to the syringe 60 communicates with the other inlet 74, and the filtrate is transferred to the outside of the flow path 10. It can also be discharged.

Filtration of the liquid sample for the analysis of chlorophyll should first go through the step of separating the particulate matter containing chlorophyll by the filter 30 in this way, this separation step is to filter the filter 30 according to the present invention More preferably.

The filter 30 is preferably a metal or plastic material with a fine hole of 0.4 to 0.5 microns. Further, in order to minimize the clogging of the filter, it is more preferable that the filter uses an asymmetrical form, and the filtration surface layer and the size of the pores having a thickness of 100 to 300 microns, preferably 200 microns, having a pore of 0.4 to 0.5 microns are several. Filters that are much larger than tens (5 to 50) microns and more than 1000 to 2000 microns (1 mm to 2 mm) thick with integrated support layers can be used to speed up the filtration, which later uses filtrate You can also easily backwash.

Since the filter 30 has a constant area, the amount of filtering the particulate matter contained in the sample 80 is limited. Therefore, by adjusting the volume of the sample to be filtered according to the amount of particulate matter contained in the sample 80, the filter 30 is filtered within the appropriate sensitivity that can be measured by the fluorescence detector 40 while preventing the filter 30 from clogging. It is necessary.

The chlorophyll automatic analysis device should increase the measurement sensitivity by intelligently increasing the filtration volume of the sample when the chlorophyll concentration in the sample 80 is low, and when the filter 30 is clogged due to the high concentration of particulate matter in the sample 80. In addition, the sample 80 filter volume should be intelligently reduced to prevent failure. In the present invention, a method for intelligently and automatically adjusting the filtration volume of the sample 80 can be used in various ways, (a) to measure the biofluorescence and turbidity of the sample to be measured by mounting a separate sensor in advance Method, (b) a method of measuring the pressure in the flow path 10 during the filtration process, (c) a method of mounting a light source and a sensor for monitoring whether the sample 80 filtrate is inhaled in the syringe (60).

First, (a) a method of equipping a separate sensor for preliminarily measuring the biofluorescence and turbidity of the sample 80 is a method of automatically adjusting the filtration volume by grasping the state of the sample in advance. It can be equipped with an additional sensor.

Second, (b) a method for measuring the pressure in the flow path 10 during the filtration process is by lowering the plunger 61 of the syringe 60, thereby filtering the sample to the pressure inside the flow path 10 to the pressure gauge 50 By measuring, it is possible to find out when the filter 30 begins to become clogged. When the filter 30 starts to be clogged, even if the plunger 61 is lowered, the sample is not filtered, and thus the pressure inside the flow path 10 is reduced. By monitoring the signal of the pressure gauge, it is determined whether the filter 30 is clogged in the filtration process. And intelligently stop filtration.

Third, another method of monitoring whether the filter is clogged is to (d) equip the syringe 60 with a light source and sensor to monitor whether the sample 80 filtrate is being aspirated. Figure 3a and 3b is a block diagram showing a syringe of the chlorophyll automatic analysis device according to an embodiment of the present invention. As shown in FIG. 3A, the syringe 60 according to the present invention includes a light source 63 and a light detection sensor 64 capable of detecting that the fluid is sucked or discharged into the syringe 60. The light source 63 and the light detection sensor 64 are preferably attached to the syringe 60.

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As shown in FIG. 3A showing the filtration filter clogging automatic detection sensor mounted on the chlorophyll autoanalyzer according to the present invention, the filtration filter clogging automatic detection sensor mounted outside the glass syringe 60 has two semi-circular attachments that can be detached. It consists of (65, 66), one side is equipped with a light source 63 for irradiating light, and the other side is equipped with a light detection sensor 64 for detecting light. The two semi-circular attachments 65, 66 are mounted by screw 67 to the outside of the syringe at a suitable height.

When the light emitting unit light source 63 for irradiating light and the light detection sensor 64 for detecting light emitted from the glass syringe 60 are mounted, the filtrate flows into the syringe when the plunger 61 is lowered. It is possible to determine whether the filter is clogged by directly measuring whether the filtrate and whether the filtrate is not introduced into the syringe (60). When the filtrate is contained in the syringe 60, as shown in FIG. 3B, the light irradiated from the light emitter light source 63 does not reach the detector photodetection sensor 64 due to refraction, but the filter 30 is blocked. If the filtrate does not enter well, the light is detected by the light detection sensor 64, it is possible to monitor the clogging of the filter 30 by using this. Changes in the time zone in which the signal from the sensor increases can also be used to determine the degree of blockage, such as whether the filter is clogging or the filter is completely blocked.

As such, after the particulate matter contained in the introduced liquid sample 80 is separated by the filter 30 installed on the flow path 10, the separated particulate material is separated from the flow path change valve ( The extract is extracted by the organic solvent introduced by 20) and the chlorophyll extract is filtered through the filter 30. That is, the filter 30 according to the present invention functions not only to separate the particulate matter contained in the liquid sample 80 but also to filter the extract having chlorophyll.

That is, after filtration of the sample 80 is completed, the plunger 61 is raised to discharge the filtrate inside the syringe 60, and then the organic solvent 90 enters the inlet port 11 of the flow path change valve 20. The organic solvent 90 flows into the flow path 10 by moving to the inlet port 12 connected to the reagent bottle, and retracting and inhaling the plunger 61 of the syringe 60. The particulate matter separated by the filter 30 is extracted using the organic solvent to destroy the cells in the particulate matter and to obtain an extract containing chlorophyll. To this end, the present invention can use an ultrasonic generator capable of applying ultrasonic waves to the filter 30, by operating the ultrasonic generator to apply ultrasonic waves to the filter 30 to destroy the cells in the separated particulate matter, Allow solvent extraction to occur within time.

Here, the chlorophyll automatic analysis device according to the present invention may further include a cartridge 31 for supporting the filter 30 so that the fluid passes only through the filter 30, the cartridge 31 is shown in FIG. As shown in 2a, the front plate 33 and the rear plate 34 that can be separated, and the filter 30 is characterized in that between the front plate 33 and the rear plate (34).

2A and 2B are cross-sectional views showing the cartridge 31 including the filter 30 of the chlorophyll automatic analysis device according to an embodiment of the present invention. The cartridge 31 is composed of a removable front plate 33 and a rear plate 34, and the filter 30, the filter O-ring 35, and the cartridge ring 36 can be placed therein. . The two O-rings 35 installed at the front and rear of the upper and lower ends of the filter 30 are for ensuring the watertightness of the gap so that the sample 80 necessarily passes through the filter surface of the filter, and the cartridge installed in the middle of the cartridge 31. O-ring 36 is intended to prevent bubbles from leaking or leaking when pressure or empty pressure occurs inside the filtration cartridge 31 as the plunger 61 moves up and down. The O-rings 35 and 36 may use a chemically stable material even in contact with an organic solvent, or may use a rubber O-ring coated with Teflon.

And, it is preferable that the ultrasonic generator 32 is attached to the back plate 34 of the cartridge 31. As shown in FIG. 2B, the sample 80 is introduced through the hole 37 in the lower portion of the filtration cartridge 31 to pass through the filter 30 so that no air bubbles remain in the filtration cartridge 31. After that is to be discharged through the lower hole 38. After mounting the filter 30 and the O-rings 35 and 36 of the filtration cartridge, the front plate 33 and the rear plate 34 are brought into close contact with the screw 39.

The present invention is to obtain an extract containing chlorophyll by the organic solvent using the filter or the cartridge containing the filter, the organic solvent 90 by retracting the plunger 61 of the syringe 60 It flows into the flow path 10. At this time, the plunger 61 of the syringe 60 is preferably retracted only until the organic solvent is filled in the cartridge 31 and then stopped. As described above, the automatic chlorophyll analyzer according to the present invention may further include an organic solvent (90) bag having an access port that can communicate with the flow path change valve 20, and the extract with chlorophyll is a liquid sample (80). The particulate matter separated from) may be characterized in that it is extracted by the organic solvent (90).

When the extract having chlorophyll is filtered through the filter 30, the filtered chlorophyll extract passes through the fluorescence detector 40 installed on the flow path 10 to measure fluorescence. That is, after retracting the plunger 61 of the syringe 60 to filter the extract contained in the cartridge 31 with the filter 30 to remove particulate matter, the particulate matter is removed to extract the chlorophyll containing Bay is introduced into the fluorescence detector 40.

At this time, it is preferable to reduce the descending speed of the plunger 61 while irradiating ultrasonic waves to the filter cartridge 31 to allow sufficient mixing to occur inside the filter cartridge 31. When the extract is passed through the flow cell of the fluorescence detector 40, it is irradiated with light of 430nm to measure the fluorescence at 680nm, and converted to chlorophyll concentration. After the fluorescence measurement, the access port 12 of the flow path change valve 20 may be moved to the access port 13 connected to the discharge pipe, and the plunger 61 may be advanced to discharge the extract.

Another embodiment of the present invention is a chlorophyll automatic analysis device further comprises a high performance liquid chromatography (HPLC) device capable of component analysis of the fluorescence measurement is completed by the fluorescence detector 40. Figure 4 is a block diagram showing an automatic chlorophyll analysis device including a high performance liquid chromatography (HPLC) according to another embodiment of the present invention.

As shown here, the high performance liquid chromatography apparatus, the HPLC injection valve 100, which can be connected to the flow path change valve so that the extract can be introduced, HPLC column 103 that can separate the chlorophyll component It is preferable to include the HPLC detector 104 which can measure a chlorophyll component.

Through the HPLC injection valve 100, HPLC according to the present invention may include a pump 101 that is connected to the flow path change valve 20, and transfers the solvent (102). After the filtration extraction step, when the measurement is completed in the fluorescent fluorescence detector 40, the inlet port 12 of the flow path change valve 20 is moved to the inlet port 14 connected to the HPLC, the plunger 61 is raised The extract is injected into the HPLC sample injection valve 100. The extract is stored and injected into the loop of the valve 100 in which the solvent 102 is moving by the HPLC pump 101, separated by component in the HPLC column 103, and then measured in the HPLC detector 104. . This embodiment according to Figure 4 is characterized by automating the process of pretreatment of the sample when analyzing the components using HPLC.

Meanwhile, in the method for automatically analyzing chlorophyll according to the present invention, the filtrate of the liquid sample 80 containing no particulate matter is filtered through the filter 30 during the step of separating the particulate matter by the filter 30. After the step of collecting the sample filtrate storage bag 75 and the fluorescence is measured, the filtrate containing no particulate matter in the sample filtrate storage bag 75 is filtered through the fluorescence detector 40. By back-injection into (30), chlorophyll automatic analysis method is also possible, further comprising the step of washing the filter (30).

After discharging the extract, the filter 30 is backwashed using the filtrate contained in the syringe 60. That is, the inlet 72 of the four-way valve 70 communicates with the inlet 73 connected to the sample filtrate storage bag 75, and then the plunger 61 is lowered to collect the filtrate stored in the bag. Inhale into. After the flow path change valve 20 communicates with the inlet and outlet 13 connected to the outlet, the plunger 61 is raised to backwash the filter 30 using the filtrate. At this time, when the ultrasonic generator 32 is operated to irradiate the inside of the cartridge 31 with ultrasonic waves, the ultrasonic vibration is applied to the filter 30 to remove debris of the particulate matter remaining in the filter 30 and to perform backwashing. It can also be promoted.

In addition, Figure 5 is an assembly view showing an automatic chlorophyll analysis device manufactured according to another preferred embodiment of the present invention, the present invention as shown herein flow path change valve 20, filter 30, fluorescent The detector 40, the syringe 60, the four-way valve 70, and the HPLC injection valve 100 may be implemented as an analytical device representing the outside.

6 is a result of measuring the fluorescence detector signal calculated by the concentration of the chlorophyll automatic analysis device according to a preferred embodiment of the present invention. As shown, the output signal of the detector increases as the concentration of the sample increases, and the chlorophyll concentration can be calculated using the area or height of the peak. For quantitative analysis of chlorophyll concentration, a calibration curve is prepared using chlorophyll a standard solution, and the sample is analyzed and converted to chlorophyll concentration.

As described above, according to the present invention, chlorophyll automatic analysis capable of sample pretreatment such as filtration and extraction necessary for chlorophyll analysis, minimizing the size of the device, and smoothly performing full unattended automatic analysis in the field can be performed. There is an effect that can provide a device.

Claims (16)

delete delete A flow path through which the fluid flows; A flow path change valve disposed on the flow path, the flow path change valve having a plurality of inlet ports and selectively communicating any one of the inlet and outlet ports with the flow passage; A filter installed on the flow path, separating the particulate matter contained in the liquid sample, and filtering the extract with chlorophyll; A fluorescence detector provided on the flow path and measuring fluorescence of the extract liquid filtered by the filter; A syringe for inhaling or discharging the fluid on the flow path; And a four-way valve installed on the flow path and connected to the syringe to selectively change the suction and discharge directions of the fluid. Chlorophyll automatic analysis device further comprises a cartridge for supporting the filter so that the fluid passes through only the filter. The chlorophyll automatic analysis device according to claim 3, wherein the cartridge is formed of a detachable front plate and a back plate, and the filter is included between the front plate and the back plate. The apparatus of claim 4, wherein an ultrasonic generator capable of applying an ultrasonic wave to a filter included between the front plate and the back plate is attached to the back plate of the cartridge. delete The apparatus of claim 3, wherein the filter is a metal or plastic material having a pore size of 0.4 to 0.5 microns. 4. The filter of claim 3, wherein the filter is a filter in which a 100 to 300 micron thick filtration surface layer having a pore size of 0.4 to 0.5 microns and a 1000 to 2000 micron thick support layer having a pore size of 5 to 50 microns are integrated. Chlorophyll automatic analyzer characterized in. delete According to claim 3, Chlorophyll automatic analysis device further comprises a light source and a light detection sensor for detecting the intake or discharge of the fluid into the syringe. The chlorophyll automatic analysis device according to claim 10, wherein the light source and the light detection sensor are attached to a syringe. 4. The chlorophyll automated analysis device according to claim 3, further comprising a sample filtrate storage bag having a port that can be connected to the four-way valve to store the filtrate passed through the filter. 4. The chlorophyll automated analysis device according to claim 3, further comprising a high performance liquid chromatography (HPLC) device capable of component analysis of the fluorescence measured extract by the fluorescence detector. The method of claim 13, wherein the high performance liquid chromatography apparatus, HPLC injection valve which can be connected to the flow path change valve to allow the extract to flow in, HPLC column capable of separating chlorophyll components, Chlorophyll automated analyzer comprising an HPLC detector capable of measuring the chlorophyll component. delete A liquid sample is introduced into the flow path by a flow path change valve installed on the flow path through which the fluid flows, and having a plurality of inlet ports, and selectively communicating any one of each inlet port with the flow path; Separating particulate matter contained in the introduced liquid sample by a filter installed on the flow path; The separated particulate matter is extracted by an organic solvent introduced by the flow path change valve, and an extract having chlorophyll is filtered from the filter; The filtered chlorophyll extract is passed through a fluorescence detector installed on the flow path comprising the step of measuring the fluorescence, During the step of separating the particulate matter by the filter, the filtrate of the liquid sample containing no particulate matter is filtered through the filter and collected in the sample filtrate storage bag, After the fluorescence is measured, the filtrate containing no particulate matter in the sample filtrate storage bag is back-injected into the filter through a fluorescence detector, thereby further comprising the step of washing the filter. Chlorophyll automated analysis method.

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