Algae quality control method based on micron particles

Abstract

The invention discloses an algae quality control method based on micron particles, which comprises the following steps: sample collection and protection: selecting representative sampling points, and collecting plankton samples by using a clean sampling device; recording sampling time, site and environment conditions; collecting the plankton through a centrifugation or filtration method, and adding an accurately prepared protective agent and fixing agent solution, so that the plankton is stored for a long time at low temperature; sample pretreatment and labeling: carrying out concentration, filtration or centrifugal pretreatment on the collected sample to remove impurities and enrich target organisms; fixing the sample by using a fixing agent to keep the biological form stable; the method has the beneficial effects that novel nano biological particles are introduced as a standard sample, and an on-site labeling recovery technology is combined, so that the problem of insufficient quality control in an existing plankton detection method is effectively solved, a scientific, systematic and effective quality control means is provided for monitoring algae and other planktons, and the method is suitable for popularization and application. Wide application prospects and popularization values are realized.

Description

Technical Field

[0001] The invention belongs to the technical field of quality control, and in particular relates to an algae quality control method based on micron particles. Background Art

[0002] Most of the existing detection methods for plankton, such as phytoplankton and zooplankton, are classic methods commonly used in aquatic biological surveys in the 1950s and 1960s. The methods are backward and have poor timeliness. In particular, there is no complete quality assurance system. In addition, the monitoring levels in various places are uneven, and there is a lack of standard samples. Therefore, the accuracy, precision, completeness, comparability and representativeness of biological monitoring data are poor, and the quality assurance of monitoring data is lacking. Taking phytoplankton as an example, quality control is mainly carried out by accurately measuring the volume, minimizing transfer, and repeating counting. Therefore, there is a lack of reasonable, effective and targeted quality control methods and systems. Summary of the invention

[0003] The purpose of the present invention is to provide an algae quality control method based on micron particles. By introducing new nanobiological particles as standard samples and combining on-site spike recovery technology, the quality of the entire process of algae sample collection, transportation, pretreatment and observation is controlled to ensure the accuracy and reliability of monitoring data.

[0004] To achieve the above object, the present invention provides the following technical solution: a method for algae quality control based on micron particles, comprising the following steps:

[0005] Sample collection and protection: Select representative sampling points and use clean sampling equipment to collect plankton samples; record the sampling time, location and environmental conditions; collect plankton by centrifugation or filtration, and add precisely configured protective agent and fixative solution to preserve the plankton for a long time at low temperature;

[0006] Sample pretreatment and spike addition: Concentrate, filter or centrifuge the collected samples to remove impurities and enrich the target organisms; use fixatives to fix the samples to keep the biological morphology stable; design a reasonable spike addition scheme, select micron-sized nano-biological particles that are similar in morphology to the target organisms and easy to identify as standard samples for spike addition to ensure that they are evenly distributed in the sample;

[0007] Morphological identification and determination: Use a high-resolution microscope to observe the unspiked and spiked samples and record the morphological characteristics of the organisms; perform morphological identification based on the principles of biological taxonomy to distinguish different species; record the recovery data of the spiked samples for subsequent analysis;

[0008] Recovery calculation and evaluation: Calculate the recovery rate and make corrections based on the spike recovery calculation formula; analyze the recovery data and evaluate the accuracy and reliability of the morphological identification method;

[0009] Quality control and quality assurance: Regularly conduct blank experiments and parallel sample measurements to evaluate the blank value and precision of the method; regularly calibrate and maintain analytical instruments to ensure their accuracy and stability; by adding standards before transportation, before sedimentation, before absorption, and before observation, analyze the losses at each stage to achieve quality control of the entire process.

[0010] As a preferred technical solution of the present invention, the protective agent is a phosphate buffer, and the preparation method of the phosphate buffer is as follows:

[0011] Weigh potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride and potassium chloride, and place them in a reaction kettle;

[0012] Add distilled water into the reactor;

[0013] Use a pH meter to adjust the pH to 7.2-7.4;

[0014] Make up to the required volume;

[0015] Filter sterilize.

[0016] As a preferred technical solution of the present invention, the fixative solution includes formaldehyde solution and Lugol's solution.

[0017] As a preferred technical solution of the present invention, the method for preparing Lugol's solution is as follows:

[0018] Weigh potassium iodide and dissolve it in distilled water;

[0019] Slowly add iodine while stirring until the solution no longer darkens;

[0020] Adjust the ratio of iodine and potassium iodide as needed to obtain the desired concentration of Lugol's solution.

[0021] As a preferred technical solution of the present invention, the sampling device includes a plankton net and a water sampler.

[0022] As a preferred technical solution of the present invention, the high-resolution microscope includes an optical microscope and an electron microscope.

[0023] As a preferred technical solution of the present invention, the morphological characteristics of the organism include cell shape, size, color, and structure.

[0024] As a preferred technical solution of the present invention, the parallel sample determination implementation method is as follows:

[0025] Select representative samples and accurately weigh or measure the specified amount;

[0026] Under the same experimental conditions, the same batch of samples was measured repeatedly;

[0027] Record the results of each measurement and calculate the mean value and standard deviation parameters.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] By introducing new nano-bioparticles as standard samples and combining them with on-site spike recovery technology, the problem of insufficient quality control in existing plankton detection methods has been effectively solved, providing a scientific, systematic and effective quality control method for the monitoring of algae and other plankton, which has broad application prospects and promotion value. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The figure is a flow chart of the quality control method of the present invention. DETAILED DESCRIPTION

[0031] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0032] Example 1

[0033] See also Figure 1 , which is the first embodiment of the present invention, provides an algae quality control method based on micron particles, comprising the following steps:

[0034] Sample collection and protection: Select representative sampling points and use clean sampling equipment to collect plankton samples; record the sampling time, location and environmental conditions; collect plankton by centrifugation or filtration, and add precisely configured protective agent and fixative solution to preserve the plankton for a long time at low temperature;

[0035] Sample pretreatment and spike addition: Concentrate, filter or centrifuge the collected samples to remove impurities and enrich the target organisms; use fixatives to fix the samples to keep the biological morphology stable; design a reasonable spike addition scheme, select micron-sized nano-biological particles that are similar in morphology to the target organisms and easy to identify as standard samples for spike addition to ensure that they are evenly distributed in the sample;

[0036] Morphological identification and determination: Use a high-resolution microscope to observe the unspiked and spiked samples and record the morphological characteristics of the organisms; perform morphological identification based on the principles of biological taxonomy to distinguish different species; record the recovery data of the spiked samples for subsequent analysis;

[0037] Recovery calculation and evaluation: Calculate the recovery rate and make corrections based on the spike recovery calculation formula; analyze the recovery data and evaluate the accuracy and reliability of the morphological identification method;

[0038]

[0039] Spiked sample measurement value: refers to the result obtained by adding a known amount of standard substance to the original sample and then measuring it according to the same experimental steps;

[0040] Sample measurement value: refers to the measurement result of the original sample (not spiked);

[0041] Spiked amount: refers to the amount of standard substance added, usually expressed in concentration or mass;

[0042] Quality control and quality assurance: Regularly conduct blank experiments and parallel sample measurements to evaluate the blank value and precision of the method; regularly calibrate and maintain analytical instruments to ensure their accuracy and stability; by adding standards before transportation, before sedimentation, before absorption, and before observation, analyze the losses at each stage to achieve quality control of the entire process.

[0043] In this embodiment, preferably, the protective agent is a phosphate buffer, and the preparation method of the phosphate buffer is as follows:

[0044] Weigh potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride and potassium chloride, and place them in a reaction kettle;

[0045] Add distilled water into the reactor;

[0046] Use a pH meter to adjust the pH to 7.2;

[0047] Make up to the required volume;

[0048] Filter sterilize.

[0049] In this embodiment, preferably, the fixative solution is a formaldehyde solution.

[0050] In this embodiment, preferably, the sampling device is a plankton net.

[0051] In this embodiment, preferably, the high-resolution microscope is an optical microscope.

[0052] In this embodiment, preferably, the morphological characteristics of the organism include cell shape, size, color, and structure.

[0053] Example 2

[0054] See also Figure 1 , which is the second embodiment of the present invention, provides an algae quality control method based on micron particles, comprising the following steps:

[0055] Sample collection and protection: Select representative sampling points and use clean sampling equipment to collect plankton samples; record the sampling time, location and environmental conditions; collect plankton by centrifugation or filtration, and add precisely configured protective agent and fixative solution to preserve the plankton for a long time at low temperature;

[0056] Sample pretreatment and spike addition: Concentrate, filter or centrifuge the collected samples to remove impurities and enrich the target organisms; use fixatives to fix the samples to keep the biological morphology stable; design a reasonable spike addition scheme, select micron-sized nano-biological particles that are similar in morphology to the target organisms and easy to identify as standard samples for spike addition to ensure that they are evenly distributed in the sample;

[0057] Morphological identification and determination: Use a high-resolution microscope to observe the unspiked and spiked samples and record the morphological characteristics of the organisms; perform morphological identification based on the principles of biological taxonomy to distinguish different species; record the recovery data of the spiked samples for subsequent analysis;

[0058] Recovery calculation and evaluation: Calculate the recovery rate and make corrections based on the spike recovery calculation formula; analyze the recovery data and evaluate the accuracy and reliability of the morphological identification method;

[0059]

[0060] Spiked sample measurement value: refers to the result obtained by adding a known amount of standard substance to the original sample and then measuring it according to the same experimental steps;

[0061] Sample measurement value: refers to the measurement result of the original sample (not spiked);

[0062] Spiked amount: refers to the amount of standard substance added, usually expressed in concentration or mass;

[0063] Quality control and quality assurance: Regularly conduct blank experiments and parallel sample measurements to evaluate the blank value and precision of the method; regularly calibrate and maintain analytical instruments to ensure their accuracy and stability; by adding standards before transportation, before sedimentation, before absorption, and before observation, analyze the losses at each stage to achieve quality control of the entire process.

[0064] In this embodiment, preferably, the protective agent is a phosphate buffer, and the preparation method of the phosphate buffer is as follows:

[0065] Weigh potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride and potassium chloride, and place them in a reaction kettle;

[0066] Add distilled water into the reactor;

[0067] Use a pH meter to adjust the pH to 7.3;

[0068] Make up to the required volume;

[0069] Filter sterilize.

[0070] In this embodiment, preferably, the fixative solution is Lugol's solution; the preparation method of Lugol's solution is as follows:

[0071] Weigh potassium iodide and dissolve it in distilled water;

[0072] Slowly add iodine while stirring until the solution no longer darkens;

[0073] Adjust the ratio of iodine and potassium iodide as needed to obtain the desired concentration of Lugol's solution.

[0074] In this embodiment, preferably, the sampling device is a water sampler.

[0075] In this embodiment, preferably, the high-resolution microscope is an electron microscope.

[0076] In this embodiment, preferably, the morphological characteristics of the organism include cell shape, size, color, and structure.

[0077] In this embodiment, preferably, blank experiments and parallel sample determinations are performed regularly to evaluate the blank value and precision of the method, which is an important means to achieve laboratory quality control. The following is a specific implementation method:

[0078] A blank experiment is a test that is performed without adding the sample to be tested, using the same operating steps, reagents and dosage as the test sample. Its purpose is to eliminate the influence of factors such as reagent background, environment and contamination during operation on the test results, thereby improving the accuracy of the test results.

[0079] Procedure

[0080] Preparation stage: ensure that all experimental equipment has been cleaned and dried, and all reagents are freshly prepared and meet purity requirements;

[0081] Operation process: Follow the sample determination steps, but do not add the sample to be tested. Pay attention to the consistency of experimental conditions, such as temperature, time, stirring speed, etc.

[0082] Data recording: record the results of the blank experiment, that is, the blank test value, which reflects the comprehensive influence of factors such as the noise of the test instrument, impurities in the reagents, the environment and contamination during the operation;

[0083] Evaluation and Improvement

[0084] Evaluate blank value: Analyze the size and discreteness of blank value. Blank value should be as low and stable as possible to reduce interference with sample measurement results.

[0085] Improvement measures: If the blank value is high or unstable, check and improve the cleanliness of experimental water, reagents, utensils, laboratory environment, and the technical level of operators;

[0086] Parallel sample measurement refers to repeated measurement of samples from the same batch to evaluate the precision of the measurement results; precision reflects the reproducibility or consistency of the measurement results;

[0087] Procedure

[0088] Sample preparation: Select representative samples and accurately weigh or measure the specified amount;

[0089] Operation process: Repeat the measurement of the same batch of samples under the same experimental conditions; pay attention to the consistency of experimental conditions, such as temperature, time, reagent dosage, etc.

[0090] Data recording: record the results of each measurement and calculate statistical parameters such as mean value and standard deviation;

[0091] Evaluation and Improvement

[0092] Evaluate precision: Based on the results of parallel sample determination, calculate indicators such as relative standard deviation (RSD) or coefficient of variation (CV) to evaluate the precision of the determination results; the smaller the RSD or CV, the higher the precision;

[0093] Improvement measures: If the precision does not meet the requirements, factors such as experimental conditions, instrument performance, and operator technical level should be checked and improved;

[0094] Comprehensive evaluation and improvement

[0095] After performing blank experiments and parallel sample measurements regularly, the blank value and precision of the method should be comprehensively evaluated; if both do not meet the requirements, the reasons should be analyzed in depth and corresponding improvement measures should be taken; at the same time, a complete laboratory quality management system should be established, personnel training and technical exchanges should be strengthened, and the overall detection level of the laboratory should be improved.

[0096] Although the embodiments of the present invention have been shown and described, as detailed above, it will be understood by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for quality control of algae based on micron particles, characterized in that: The steps include: Sample collection and protection: Select representative sampling points and use clean sampling equipment to collect plankton samples; record the sampling time, location and environmental conditions; collect plankton by centrifugation or filtration, and add precisely configured protective agent and fixative solution to preserve the plankton for a long time at low temperature; Sample pretreatment and spike addition: The collected samples are concentrated, filtered or centrifuged to remove impurities and enrich the target organisms; Use fixatives to fix samples to keep the biological morphology stable; design a reasonable spiking scheme, select micron-sized nanobiological particles that are similar to the target biological morphology and easy to identify as standard samples for spiking to ensure that they are evenly distributed in the sample; Morphological identification and determination: Use a high-resolution microscope to observe the unspiked and spiked samples and record the morphological characteristics of the organisms; perform morphological identification based on the principles of biological taxonomy to distinguish different species; record the recovery data of the spiked samples for subsequent analysis; Recovery calculation and evaluation: Calculate the recovery rate and make corrections based on the spike recovery calculation formula; analyze the recovery data and evaluate the accuracy and reliability of the morphological identification method; Spiked sample measurement value: the result obtained by adding a known amount of standard substance to the original sample and then measuring it according to the same experimental steps; Sample measurement value: the measurement result of the original sample; Spiked amount: the amount of standard substance added, expressed as concentration or mass; Quality control and quality assurance: Regularly conduct blank experiments and parallel sample measurements to evaluate the blank value and precision of the method; regularly calibrate and maintain analytical instruments to ensure their accuracy and stability; by adding standards before transportation, before sedimentation, before absorption, and before observation, analyze the losses at each stage to achieve quality control of the entire process.

2. The algae quality control method based on micron particles according to claim 1, characterized in that: The protective agent is a phosphate buffer, and the preparation method of the phosphate buffer is as follows: Weigh potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride and potassium chloride, and place them in a reaction kettle; Add distilled water into the reactor; Use a pH meter to adjust the pH to 7.2-7.4; Make up to the required volume; Filter sterilize.

3. The algae quality control method based on micron particles according to claim 1, characterized in that: The fixative solution includes formaldehyde solution and Lugol's solution.

4. The algae quality control method based on micron particles according to claim 3, characterized in that: The method for preparing the Lugol solution is as follows: Weigh potassium iodide and dissolve it in distilled water; Slowly add iodine while stirring until the solution no longer darkens; Adjust the ratio of iodine and potassium iodide as needed to obtain the desired concentration of Lugol's solution.

5. The algae quality control method based on micron particles according to claim 1, characterized in that: The sampling device comprises a plankton net and a water sampler.

6. The algae quality control method based on micron particles according to claim 1, characterized in that: The high-resolution microscope includes an optical microscope and an electron microscope.

7. The algae quality control method based on micron particles according to claim 1, characterized in that: The morphological characteristics of the organism include cell shape, size, color, and structure.

8. The algae quality control method based on micron particles according to claim 1, characterized in that: The parallel sample determination is implemented as follows: Select representative samples and accurately weigh or measure the specified amount; Under the same experimental conditions, the same batch of samples was measured repeatedly; Record the results of each measurement and calculate the mean value and standard deviation parameters.

Images