Full-automatic chlorophyll extraction and analysis device and extraction method

Abstract

The invention belongs to the technical field of chlorophyll extraction methods, and particularly relates to a full-automatic chlorophyll extraction and analysis device and an extraction method. The full-automatic chlorophyll extraction and analysis device comprises an extraction device and an analysis device, the extraction device and the analysis device are installed in the full-automatic chlorophyll extraction device, and the extraction device is connected with the analysis device; the extraction device comprises a condenser, a freezing module, a heating module and a compressor; the condenser is sequentially connected with the freezing module and the compressor to form a loop, and the heating module wraps the lower portion of the freezing module. The extraction device is used for extracting chlorophyll, has the advantages of simplicity and convenience in operation, accurate control of parameters such as temperature and illumination, good extraction effect, small personal error, short extraction period, high extraction efficiency and the like, and does not cause degradation of chlorophyll in the extraction process.

Description

Technical Field

[0001] The invention belongs to the technical field of chlorophyll extraction methods, and specifically relates to a fully automatic chlorophyll extraction and analysis device and an extraction method. Background Art

[0002] Chlorophyll is an important photosynthetic pigment in the process of photosynthesis. Algae are the main primary producers in water bodies, providing energy sources for the entire aquatic ecosystem. Chlorophyll a is the central pigment of photosynthetic reactions in most algae. By measuring the chlorophyll content in water bodies, the primary productivity of water bodies can be understood.

[0003] Chlorophyll a is one of the important indicators in the evaluation of lake eutrophication. Therefore, accurate, efficient and rapid determination of chlorophyll a content in water is the basis for rational evaluation of the nutrient status of water. External factors that affect the extraction of chlorophyll a mainly include temperature, light, human error, etc. The influence of temperature on the extraction of chloroplast pigments mainly includes: the influence of room temperature on the activity of chlorophyll a degrading enzymes and the destruction of chloroplast pigments by high temperature. Chlorophyll a will degrade at high temperature. Isolated chlorophyll a is very easy to be oxidatively degraded under strong light, and the degree of degradation varies with the duration of light, light quality and type of pigment.

[0004] At present, the extraction methods of chlorophyll a mainly include: manual grinding method, machine grinding bead grinding method, ultrasonic cell disruption method, etc.

[0005] The manual grinding method includes: placing the sample in a mortar, adding 3ml-4ml of 90% acetone solution, and grinding into a paste. The manual grinding method is cumbersome to operate, and the inability to control temperature, light and other conditions during the operation will result in poor extraction effect and large human errors.

[0006] The machine bead grinding method includes: placing the sample in a centrifuge tube with a scale plug, adding zirconium oxide grinding beads of different sizes after using 90% acetone solution, and placing the centrifuge tube in a grinder. Due to high-speed vibration grinding, the friction temperature of the grinding beads is too high, which damages the chlorophyll. Moreover, after grinding, the sample must be manually extracted and placed in a glass test tube, and the controllability of light is poor. The machine bead grinding method has the disadvantages of cumbersome extraction process, low efficiency, and large error in the transfer process. The grinding method will cause the grinding temperature to be too high due to high-speed vibration, which will cause damage to some chlorophylls. The light cannot be controlled, resulting in poor extraction effect and failure to completely extract chlorophyll a.

[0007] The ultrasonic cell disruption method has the disadvantages of long cycle, poor effect, uncontrollable temperature which may cause chlorophyll degradation, and incomplete extraction.

[0008] The patent application with application number: 201410162725.0 discloses a method for extracting chlorophyll, the steps of which include: taking green plant stems or leaves, boiling them in boiling water, drying them in the shade, and crushing them; adding a triton X-114 aqueous solution with a volume concentration of 1-8%, ultrasonic extraction, taking the supernatant, adding NaCl, stirring evenly, standing, separating, removing the lower aqueous phase, taking the upper layer and adding methanol, filtering with an ultrafiltration membrane, and concentrating the filtrate to a viscous state; adding anhydrous ethanol to the concentrate, shaking, filtering, and concentrating the filtrate to obtain an extract. The chlorophyll content is determined by an ultraviolet spectrophotometer. This method has the advantages of low extraction temperature, simple process equipment, and high product yield. It has the advantages of economy, safety, efficiency, simplicity, time saving, and environmental protection. This method does not destroy the original properties of chlorophyll during the extraction process, and maintains the Mg content in chlorophyll. 2+ The presence of chlorophyll gives it the original bright green color. However, it has the disadvantages of cumbersome manual operation, poor extraction effect and large human error. Summary of the invention

[0009] Based on the above technical background, the main purpose of the present invention is to provide a fully automatic chlorophyll extraction device and extraction method to overcome the shortcomings of the prior art.

[0010] In order to achieve the above-mentioned invention object, the technical solution adopted by the present invention includes:

[0011] The first aspect of the present invention is to provide a fully automatic chlorophyll extraction and analysis device, the fully automatic chlorophyll extraction and analysis device comprises an extraction device and an analysis device, the extraction device and the analysis device are installed in the fully automatic chlorophyll extraction device, and the extraction device and the analysis device are connected;

[0012] The extraction device comprises a condenser, a freezing module, a heating module and a compressor;

[0013] The condenser, the freezing module and the compressor are connected in sequence to form a loop, and the heating module is wrapped under the freezing module.

[0014] Preferably, the extraction device further comprises a temperature sensor, and the temperature sensor is located in the freezing module.

[0015] Preferably, the extraction device also includes a capillary and a filter, which are located between the condenser and the freezing module, one end of the filter is connected to the condenser, and the other end of the filter is connected to the capillary, one end of the capillary is connected to the filter, and the other end of the capillary is connected to the freezing module 4.

[0016] Preferably, a test tube slot is provided above the freezing module for mounting test tubes containing samples;

[0017] Preferably, the extraction device further comprises a thermal cover module, and the thermal cover module is arranged above the test tube.

[0018] Preferably, the analysis device includes a cuvette turntable, which is installed in the analysis device. The cuvette turntable includes a cuvette, an emitting light and a receiving light. The cuvette is installed vertically, and multiple cuvettes are arranged in sequence to form a ring. The receiving light is installed inside the ring structure formed by the cuvette, and the emitting light is installed outside the ring structure formed by the cuvette.

[0019] Preferably, an automatic sample injector and a peristaltic pump unit are installed between the extraction device and the analysis device, and the chlorophyll extracted in the extraction device is automatically transported to the analysis device through the automatic sample injector and the peristaltic pump unit.

[0020] The second aspect of the present invention is to provide a method for extracting chlorophyll using the fully automatic chlorophyll extraction device of the first aspect of the present invention, the method comprising the following steps:

[0021] The sample is added into the fully automatic chlorophyll extraction device, cooled, the cooling program is suspended, the temperature is increased and heated, the cooling and heating programs are cycled several times, and the processed sample is passed into the cuvette turntable 11 for analysis and measurement.

[0022] Preferably, the sample is added to a fully automatic chlorophyll extraction device, and a ladder program is performed to cool the sample to -50 to -40°C, and the sample is kept at this temperature for 20 to 45 minutes. The cooling program is then paused, the sample is heated to 20 to 30°C, and the sample is kept at this temperature for 3 to 7 minutes.

[0023] Preferably, the cooling and heating program cycles are executed sequentially for 2 to 5 times.

[0024] The beneficial effects of the present invention are:

[0025] (1) The fully automatic chlorophyll extraction and analysis device of the present invention comprises a compressor, a condenser, a filter, a capillary, a freezing module, a heating module, a hot cover module, an automatic sampler, a peristaltic pump unit, a cuvette turntable, a temperature sensor and an analysis device. The fully automatic chlorophyll extraction device can realize the fully automatic extraction of chlorophyll, avoiding the tediousness of manual operation and the inability to accurately control temperature and light caused by manual operation, thereby avoiding the shortcomings of poor extraction effect and large human errors.

[0026] (2) The fully automatic chlorophyll extraction and analysis device of the present invention can realize the reverse process of PCR, that is, the extraction and analysis device can realize cooling first and then heating. The extraction and analysis device can realize greater heating and cooling. The temperature span of the extraction and analysis device of the present invention can reach -50 to 120°C, with higher accuracy.

[0027] (3) The chlorophyll extraction method of the present invention uses the extraction device of the present invention to extract chlorophyll, which has the advantages of simple operation, precise control of parameters such as temperature and light, good extraction effect, small human error, short extraction cycle, high extraction efficiency, etc. The extraction process will not cause the degradation of chlorophyll, destroy other effective substances in chlorophyll, and can maintain the original properties of chlorophyll. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 A schematic diagram showing the front view of a fully automatic chlorophyll extraction and analysis device according to a preferred embodiment of the present invention; the accompanying drawings cannot include shadows, background colors, coordinates, etc.

[0029] Figure 2 A schematic diagram showing the back of a fully automatic chlorophyll extraction and analysis device according to a preferred embodiment of the present invention

[0030] Figure 3 A schematic diagram showing the internal structure of a fully automatic chlorophyll extraction and analysis device according to a preferred embodiment of the present invention;

[0031] Figure 4 A schematic structural diagram of a cuvette turntable according to a preferred embodiment of the present invention is shown.

[0032] Description of Figure Numbers

[0033] 1- Extraction device;

[0034] 2- Condenser;

[0035] 3-capillary;

[0036] 4- Freezing module;

[0037] 5- Heating module;

[0038] 6- Temperature sensor;

[0039] 7-Compressor;

[0040] 8-Peristaltic pump unit;

[0041] 9- Filter;

[0042] 10-Analytical device;

[0043] 11- cuvette turntable;

[0044] 111- cuvette;

[0045] 112-emitting light;

[0046] 113-receiving light;

[0047] 12-Heat cover module. DETAILED DESCRIPTION

[0048] The present invention will be described in detail below, and the characteristics and advantages of the present invention will become clearer and more distinct with the description.

[0049] The first aspect of the present invention is to provide a fully automatic chlorophyll extraction and analysis device, the fully automatic chlorophyll extraction and analysis device comprises an extraction device 1 and an analysis device 10, such as Figure 1 and Figure 2 As shown, the extraction device 1 and the analysis device 10 are installed in the fully automatic chlorophyll extraction and analysis device, and the extraction device 1 and the analysis device 10 are connected. The fully automatic chlorophyll extraction and analysis device is a fully enclosed device, so that the fully automatic chlorophyll extraction device of the present invention can transport the extracted chlorophyll to the analysis device for analysis and detection, and realize that the chlorophyll extraction and analysis are all carried out in a fully enclosed device, avoiding the exposure of chlorophyll to light and causing chlorophyll degradation.

[0050] An automatic sample injector and a peristaltic pump unit 8 are installed between the extraction device 1 and the analysis device 10. Figure 1 and Figure 2 As shown, the chlorophyll extracted in the extraction device is fully automatically transported to the analysis device 10 through the automatic sample injector and the peristaltic pump unit.

[0051] The automatic sample injector is used to send the sample to be analyzed and tested into the extraction device through the automatic sample injector.

[0052] The automatic sample injector is connected to a peristaltic pump unit, and the sample is controlled to enter the automatic sample injector through the peristaltic pump.

[0053] The extraction device 1 includes a condenser 2 , a freezing module 4 , a heating module 5 , a temperature sensor 6 and a compressor 7 .

[0054] The condenser 2 is connected to the refrigeration module 4 and the compressor 7 in sequence to form a closed loop. Figure 3 The heating module 5 is coated under the freezing module 4, which is used to heat the sample on the one hand, and prevent the cold air from dissipating during the refrigeration process on the other hand, and has a cold preservation effect, so that the temperature can reach -50°C or below during refrigeration.

[0055] The freezing module is used to cool the sample, the condenser is used to convert gas or vapor into liquid and transfer heat to the surrounding medium through the heat release process, and the evaporator surrounds the freezing module. The role of the evaporator in the device is to absorb heat through the evaporation of liquid refrigerant to achieve the purpose of refrigeration. The compressor is the heart of the refrigeration device.

[0056] According to a preferred embodiment of the present invention, the extraction device 1 further comprises a temperature sensor 6 , and the temperature sensor 6 is located in the freezing module 4 .

[0057] The temperature sensor is used to detect the temperature and provide real-time feedback on the heating temperature of the sample, so as to facilitate more accurate temperature monitoring.

[0058] According to a preferred embodiment of the present invention, the extraction device 1 further comprises a capillary tube 3 and a filter 9, wherein the capillary tube 3 and the filter 9 are located between the condenser 2 and the freezing module 4, wherein one end of the filter 9 is connected to the condenser 2 and the other end is connected to the capillary tube 3, and one end of the capillary tube 3 is connected to the filter 9 and the other end is connected to the freezing module 4. The condenser 2, the filter 9, the capillary tube 3, the freezing module 4 and the compressor 7 are sequentially connected to form a closed loop, such as Figure 3 shown.

[0059] According to a preferred embodiment of the present invention, a test tube slot is provided above the freezing module 4 for installing a test tube containing a sample, and the extraction device 1 further comprises a thermal cover module 12, which is provided above the test tube. The thermal cover module is used to heat the sample to be processed.

[0060] The present invention heats the sample from above and below simultaneously by means of the heating module 5 coated under the freezing module 4 and the heat cover module 12 arranged above the test tube, thereby avoiding uneven heating of the sample, making the heating of the sample more precise and improving the extraction effect of chlorophyll.

[0061] The analysis device 10 includes a cuvette turntable 11 , and the cuvette turntable 11 is installed in the analysis device 10 .

[0062] According to a preferred embodiment of the present invention, the cuvette turntable 11 includes a cuvette 111, an emission light 112, and a reception light 113. Figure 4 As shown, the cuvette 111 is installed vertically, and a plurality of cuvettes 111 are arranged in sequence to form a ring. The receiving light 113 is installed inside the ring structure formed by the cuvette, and the emitting light 112 is installed outside the ring structure formed by the cuvette.

[0063] The chlorophyll extracted by the extraction device is passed into the cuvette turntable for detection. The test tube containing chlorophyll extracted in the device works through the peristaltic pump. The automatic sampler will inject the sample into the cuvette. The analysis device will control the emission light to irradiate the cuvette, and the light receiving unit will detect the light. The chlorophyll value can be obtained through analysis. The cuvette turntable can be rotated, and the samples in the 36 cuvettes pass through the emission light and the receiving light in turn under the rotation of the cuvette turntable. The chlorophyll content can be obtained through the emission light and the receiving light detection and analysis.

[0064] The second aspect of the present invention is to provide a method for extracting chlorophyll using the fully automatic chlorophyll extraction and analysis device of the first aspect of the present invention, the method comprising the following steps:

[0065] The sample is added into the fully automatic chlorophyll extraction device, cooled, the cooling program is suspended, the temperature is increased and heated, the cooling and heating programs are cycled several times, and the processed sample is passed into the cuvette turntable 11 for analysis and measurement.

[0066] Preferably, the sample is added to the fully automatic chlorophyll extraction device, and the cooling program is performed by the freezing module 4 to cool it to -50 to -40°C, and it is kept warm at this temperature for 20 to 45 minutes, then the cooling program is paused, and the heating module 5 and the hot cover module 12 are used to heat it to 20 to 30°C, and it is kept warm at this temperature for 3 to 7 minutes, and the cooling and heating program cycles are performed in sequence for 2 to 5 times, and the treated sample is passed into the cuvette turntable 11 for analysis and measurement.

[0067] More preferably, the sample is added into a fully automatic chlorophyll extraction device, and the temperature is lowered to -45°C by a ladder program using the freezing module 4, and the sample is kept warm at this temperature for 30 minutes. Then the refrigeration program is paused, and the temperature is raised to 25°C by the heating module 5 and the hot cover module 12, and the sample is kept warm at this temperature for 5 minutes. The refrigeration and heating program is cycled 3 times, and the treated sample is passed into the cuvette turntable 11 for analysis and measurement.

[0068] The present invention uses a compressor cascade refrigeration to ensure that the temperature reaches -50 to -40°C within a few minutes after power is turned on. The temperature is repeatedly raised to -45°C and 25°C under multi-sensor monitoring, so that the ice crystals in the chlorophyll in the sample melt, which increases the liquid concentration in the cells and causes the cells to rupture. After multiple cycles, the sample in the water body can completely release the chlorophyll. By adopting the fully automatic chlorophyll extraction device described in the first aspect of the present invention, the error of human operation is avoided in the entire extraction process, and the sample is extracted for chlorophyll under completely light-proof conditions, and finally enters the analysis device of the extraction device at one time through the sampling device to obtain the result. This extraction method will not cause the degradation of chlorophyll, the chlorophyll extraction is more complete, the measurement is more accurate, the error caused by human operation is avoided, the operation is simple, and the extraction cycle is short.

[0069] Example

[0070] The present invention is further described below by means of specific examples. These embodiments are only intended to illustrate the present invention but are not intended to limit the scope of the present invention.

[0071] Example 1

[0072] The fully automatic chlorophyll extraction and analysis device comprises an extraction device 1 and an analysis device 10. Figure 1 and Figure 2 As shown, the extraction device 1 and the analysis device 10 are installed in the fully automatic chlorophyll extraction device, the extraction device 1 and the analysis device 10 are connected, and the fully automatic chlorophyll extraction device is a fully enclosed device. An automatic sampler and a peristaltic pump unit 8 are installed between the extraction device 1 and the analysis device 10.

[0073] The extraction device 1 includes a condenser 2, a freezing module 4, a heating module 5, a temperature sensor 6, a compressor 7, a capillary tube 3, a filter 9 and a thermal cover module 12. The condenser 2, the filter 9, the capillary tube 3, the freezing module 4 and the compressor 7 are sequentially connected to form a closed loop. The temperature sensor 6 is located in the freezing module 4, the heating module 5 is wrapped under the freezing module 4, and a test tube slot is provided above the freezing module 4 for installing a test tube containing a sample. The extraction device 1 also includes a thermal cover module 12, and the thermal cover module 12 is provided above the test tube. Figure 3 shown.

[0074] The analysis device 10 includes a cuvette turntable 11, which is installed in the analysis device 10. The cuvette turntable 11 includes a cuvette 111, an emission light 112, and a receiving light 113. Figure 4 As shown, the cuvette 111 is installed vertically, and multiple cuvettes 111 are arranged in sequence to form a ring, the receiving light 113 is installed inside the ring structure formed by the cuvette, and the emitting light 112 is installed outside the ring structure formed by the cuvette. Figure 4 As shown in the figure, the samples in the 36 cuvettes are tested in turn as the cuvette turntable rotates, and the chlorophyll content can be obtained through analysis.

[0075] Example 2

[0076] A method for extracting chlorophyll using the fully automatic chlorophyll extraction device described in Example 1, the method comprising the following steps:

[0077] The sample is added into the fully automatic chlorophyll extraction device, and the temperature is lowered to -45°C by the freezing module 4 in a ladder program, and kept at this temperature for 30 minutes. Then the refrigeration program is paused, and the temperature is raised to 25°C by the heating module 5 and the hot cover module 12, and kept at this temperature for 5 minutes. The refrigeration and heating program is cycled 3 times, and the processed sample (extracted chlorophyll) is passed into the cuvette turntable 11 for analysis and measurement.

[0078] Example 3

[0079] A method for extracting chlorophyll using the fully automatic chlorophyll extraction device described in Example 1, the method comprising the following steps:

[0080] The sample is added into the fully automatic chlorophyll extraction device, and the temperature is lowered to -50°C by the refrigeration module 4 in a ladder program, and kept at this temperature for 20 minutes. Then the refrigeration program is paused, and the temperature is raised to 20°C by the heating module 5 and the hot cover module 12, and kept at this temperature for 7 minutes. The refrigeration and heating program cycles are performed twice in sequence, and the processed sample (extracted chlorophyll) is passed into the cuvette turntable 11 for analysis and measurement.

[0081] Example 4

[0082] A method for extracting chlorophyll using the fully automatic chlorophyll extraction device described in Example 1, the method comprising the following steps:

[0083] The sample is added into the fully automatic chlorophyll extraction device, and the temperature is lowered to -40°C by the refrigeration module 4 in a ladder program, and kept at this temperature for 45 minutes. Then the refrigeration program is paused, and the temperature is raised to 30°C by the heating module 5 and the hot cover module 12, and kept at this temperature for 3 minutes. The refrigeration and heating programs are cycled 5 times in sequence, and the processed sample (extracted chlorophyll) is passed into the cuvette turntable 11 for analysis and measurement.

[0084] The present invention has been described in detail above in conjunction with specific implementations and exemplary examples, but these descriptions cannot be understood as limiting the present invention. Those skilled in the art understand that, without departing from the spirit and scope of the present invention, a variety of equivalent substitutions, modifications or improvements may be made to the technical solution of the present invention and its implementation methods, all of which fall within the scope of the present invention. The scope of protection of the present invention shall be subject to the attached claims.

Claims

1. A fully automatic chlorophyll extraction and analysis device, characterized in that: The fully automatic chlorophyll extraction device comprises an extraction device and an analysis device, the extraction device and the analysis device are installed in the fully automatic chlorophyll extraction device, and the extraction device and the analysis device are connected; The extraction device comprises a condenser, a freezing module, a heating module and a compressor; The condenser, the freezing module and the compressor are connected in sequence to form a loop, and the heating module is wrapped under the freezing module.

2. The fully automatic chlorophyll extraction and analysis device according to claim 1, characterized in that: The extraction device further comprises a temperature sensor, which is located in the freezing module.

3. The fully automatic chlorophyll extraction and analysis device according to claim 1, characterized in that: The extraction device also includes a capillary tube and a filter, which are located between the condenser and the freezing module. One end of the filter is connected to the condenser, and the other end of the filter is connected to the capillary tube. One end of the capillary tube is connected to the filter, and the other end of the capillary tube is connected to the freezing module.

4. The fully automatic chlorophyll extraction and analysis device according to claim 1, characterized in that: A test tube slot is provided above the freezing module for installing test tubes containing samples; The extraction device further comprises a thermal cover module, and the thermal cover module is arranged above the test tube.

5. The fully automatic chlorophyll extraction and analysis device according to claim 1, characterized in that: The analysis device includes a cuvette turntable, which is installed in the analysis device. The cuvette turntable includes a cuvette, an emitting light and a receiving light. The cuvette is installed vertically, and multiple cuvettes are arranged in sequence to form a ring. The receiving light is installed inside the ring structure formed by the cuvette, and the emitting light is installed outside the ring structure formed by the cuvette.

6. The fully automatic chlorophyll extraction and analysis device according to claim 1, characterized in that: An automatic sample injector and a peristaltic pump unit are installed between the extraction device and the analysis device, and the chlorophyll extracted from the extraction device is fully automatically transported to the analysis device through the automatic sample injector and the peristaltic pump unit.

7. A method for extracting chlorophyll using the fully automatic chlorophyll extraction and analysis device according to any one of claims 1 to 6, characterized in that: The method comprises: The sample is added to the fully automatic chlorophyll extraction device, cooled, the cooling program is paused, the temperature is increased and heated, the cooling and heating program is cycled several times, and the processed sample is passed into the cuvette turntable for analysis and measurement.

8. A method according to claim 7, characterized in that Add the sample to the fully automatic chlorophyll extraction device, execute the ladder program to cool down to -50 ~ -40 ℃, keep it at this temperature for 20 ~ 45 minutes, then pause the refrigeration program, heat it to 20 ~ 30 ℃, and keep it at this temperature for 3 ~ 7 minutes.

9. The method according to claim 7, characterized in that: Execute the cooling and heating program cycles 2 to 5 times in sequence.

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