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Delivery: <= 4 days. True-PDF full-copy in English will be manually translated and delivered via email. HJ 1215-2021: Water quality - Determination of phytoplankton - Filtration membrane-Microscope counting method Status: Valid
Basic dataStandard ID: HJ 1215-2021 (HJ1215-2021)Description (Translated English): Water quality - Determination of phytoplankton - Filtration membrane-Microscope counting method Sector / Industry: Environmental Protection Industry Standard Word Count Estimation: 16,136 Issuing agency(ies): Ministry of Ecology and Environment HJ 1215-2021: Water quality - Determination of phytoplankton - Filtration membrane-Microscope counting method---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order. (Water quality - Determination of phytoplankton - Membrane-microscopic counting method) National Ecological Environment Standard of the People's Republic of China Determination of Phytoplankton in Water Quality filter-microscope counting Water quality-Determination of phytoplankton- This electronic version is the official standard text, which is reviewed and typeset by the Environmental Standards Institute of the Ministry of Ecology and Environment. Published on 2021-11-29 2022-06-01 Implementation Released by the Ministry of Ecology and Environment directory Foreword...ii 1 Scope...1 2 Normative references...1 3 Terms and Definitions...1 4 Principles of the method...1 5 Reagents and materials...2 6 Instruments and equipment...2 7 Samples...3 8 Analysis steps...4 9 Result calculation and representation...6 10 Precision...6 11 Quality Assurance and Quality Control...7 12 Waste Disposal...7 Appendix A (normative appendix) Method detection limit calculation method...8 Appendix B (Informative) Whipple Eyepiece Reticle Calibration...9 Appendix C (informative appendix) Estimates of the number of phytoplankton cells in filamentous and spherical populations...12 References...13 Determination of Phytoplankton in Water Quality by Membrane-Microscopic Counting 1 Scope of applicationThis standard specifies the filter-microscopic counting method for the determination of phytoplankton in water. This standard applies to the rapid determination of phytoplankton in surface water. When the maximum sample filtration volume is 1000 ml, the detection limit of the method is 40 cells/L. The method detection limit calculation method is detailed in Appendix A.2 Normative referencesThis standard refers to the following documents or clauses thereof. For dated references, only the dated version applies to this standard. For undated references, the latest edition (including all amendments) applies to this standard. GB/T 14581 Technical Guidelines for Sampling of Water Quality in Lakes and Reservoirs HJ/T 91 Technical Specification for Surface Water and Sewage Monitoring HJ 494 Water Quality Sampling Technical Guide3 Terms and DefinitionsThe following terms and definitions apply to this standard. 3.1 phytoplankton The tiny algae that live planktonic life in water, usually phytoplankton are planktonic algae, including prokaryotic cyanobacteria and other types of eukaryotes algae. 3.2 counting unit Select the smallest unit for counting, such as a cell, under the microscope field of view. 3.3 density of phytoplankton The number of phytoplankton cells per unit volume, cells/L. 3.4 microscope counting field A defined area in the microscope field of view used to quantitatively count phytoplankton.4 Principles of the methodThe sample passes through a filter membrane with a certain pore size, and the colonies, filaments and single-celled phytoplankton are trapped on the filter membrane. Microscopic examination.5 Reagents and MaterialsUnless otherwise stated, analytical reagents that meet national standards were used in the analysis, and the experimental water was freshly prepared deionized water or distilled water. 5.1 Iodine (I2). 5.2 Potassium iodide (KI). 5.3 Formaldehyde solution. w(HCHO)=37%～40%. 5.4 Lugol's iodine solution. Weigh 60 g of potassium iodide (5.2) and dissolve it in 100 ml of water, then add 40 g of iodine (5.1), stir well to dissolve it completely, add water Dilute to 1000 ml, transfer to a brown ground-mouth glass bottle, and store at room temperature away from light. 5.5 Microscope Immersion Oil. nD 23=1.515 or nE 23=1.518, with transparent glass drop stick. 5.6 Filter membrane. mixed cellulose ester membrane, with a diameter of 25 mm and a pore size of 0.45 μm to 3 μm. Note. Each batch of filter membranes should be tested for hydration and oil transparency, and can only be used after meeting the requirements of hydration and transparency. Place 1 unused filter in the In a beaker of water, if the filter membrane is completely soaked with water, the filter membrane can be hydrated. Take 1 unused filter membrane and place it on a glass slide with microscope immersion oil. If the filter membrane is completely soaked by the immersion oil, the filter membrane can be transparent.6 Instruments and equipment6.1 No. 25 plankton net. the diameter of the mesh is 0.064 mm, the net is conical, the net mouth is sleeved on the copper ring, and the bottom end of the net has a water outlet switch piston. 6.2 Qualitative sampling bottle. 30 ml ~ 100 ml wide-mouth polyethylene bottle. 6.3 Water collector. stainless steel or plexiglass material, cylindrical. Capacity and depth specifications to meet sampling requirements. 6.4 Quantitative sampling bottle. 1L～2L wide-mouth polyethylene bottle. 6.5 Refrigerators. 6.6 Vacuum filtration device. including funnel (diameter 25 mm, volume.200 ml), porous polyester filter membrane support plate, collection bottle, vacuum pump (relative vacuum ≤-17 kPa, adjustable). 6.7 Glass slide. 25 mm × 76 mm. Note. Slides and coverslips should be soaked in concentrated hydrochloric acid and ethanol before use. If possible, choose a slide or coverslip that can be used directly. 6.8 Cover glass. 25 mm×25 mm. 6.9 Teethless tissue forceps. 6.10 Slide drying plate. suitable for slides of the same specification. 6.11 Oven. 70 ℃±2 ℃, temperature adjustable. 6.12 Upright biological microscope or inverted biological microscope. objective lens 4×, 10×, 20×, 40×; eyepiece 10× or 15×. 6.13 Stage micrometer. also known as stage micrometer, 1 mm/100 DIV, with a division value of 0.01 mm. Note. DIV refers to equal division, 1 mm/100 DIV means that 1 mm is divided into 100 divisions. 6.14 Whipple grid. a large square grid with a side length of 7 mm to 10 mm, divided into 100 identical Square medium, 1 medium in the center is further divided into 25 identical small squares. Note. When the Whipple eyepiece reticle is not available, the grid-type eyepiece reticle of the same specification can also be used (that is, the grid type in which the middle grid in the center is not subdivided into 25 small grids) eyepiece reticle). 6.15 Counters. 6.16 Common laboratory equipment.7 samples7.1 Sample Collection 7.1.1 Qualitative samples The point layout and sampling frequency shall be implemented in accordance with the relevant regulations of GB/T 14581, HJ/T 91 and HJ 494. Collect qualitative samples using plankton net 25 (6.1). Turn off the water outlet piston switch at the bottom of the plankton net, and at the surface of the water to At a depth of 0.5 m, reciprocate in an "∞" shape at a speed of 20 cm/s to 30 cm/s, and drag slowly for about 1 to 3 minutes, until there is obvious floating in the net. When the plankton enters, lift the plankton net (6.1) out of the water surface, and the water in the net is naturally filtered out through the mesh holes, and a little water sample (5 ml~ 10 ml), move the bottom outlet into the qualitative sampling bottle (6.2), and open the bottom piston switch to collect qualitative samples. Collect layered samples When the number of plankton net 25 (6.1) is used to filter specific water layer samples, other steps are the same as the collection of surface layer samples. After qualitative sample collection is complete Clean the plankton net in time. After the samples are collected, put them in a refrigerator (6.5) for transportation in the dark. If there is a technical specification for qualitative sample collection, follow the relevant requirements of the technical specification. 7.1.2 Quantitation of samples Collect quantitative samples in accordance with the relevant provisions of GB/T 14581, HJ/T 91 and HJ 494. Use the water collector (6.3) to collect 1 L to 2 L of sample into the quantitative sampling bottle (6.4). If the water transparency is high, the number of phytoplankton When the volume is small, the sampling volume should be increased as appropriate. After the quantitative sample collection is complete, the quantitative sampling bottle (6.4) should not be full so that it can be shaken well. Note 1.Some phytoplankton (such as blue-green algae) often float on the water surface or are distributed in patches and strips, which can be sampled in this dense bloom area as a peak reference. Note 2.Quantitative sample collection should precede qualitative sample collection. Fixed time period sampling should be maintained so that results can be compared with each other. 7.2 Sample Storage 7.2.1 Qualitative samples After the qualitative sample is collected, it should be fixed by adding Lugol's iodine solution (5.4) immediately, and the dosage is 1.0% to 1.5% of the volume of the water sample. microscopic examination Samples were fixed without Lugol's iodine solution (5.4). Qualitative samples can be stored for 3 weeks at room temperature in the dark; refrigerated at 1 ℃ ~ 5 ℃ in the dark It can be stored for 12 months. In vivo samples can be stored for 36 h under refrigerated storage at 4 °C to 10 °C in the dark. 7.2.2 Quantitation of samples Immediately after the quantitative sample collection, add Lugol's iodine solution (5.4) to fix it, and the dosage is 1.0% to 1.5% of the volume of the water sample. Lugo Add the iodine solution (5.4) to the quantitative sampling bottle (6.4) in advance and bring it to the site for use. Quantitative samples can be stored for 3 weeks at room temperature in the dark; It can be stored for 12 months at 1 ℃ ~ 5 ℃ in the dark. During the storage process of the sample, the oxidation degree of Lugol's iodine solution (5.4) should be checked every week. Add an appropriate amount of Lugol's iodine solution (5.4) until the color of the sample returns to tan. Note. If the sample needs to be stored for a long time, formaldehyde solution (5.3) should be added in an amount of 4% of the volume of the water sample.8 Analysis steps8.1 Specimen Preparation 8.1.1 Preflight According to the requirements of 8.1.2 ~ 8.1.5, complete the pre-inspection and mount the slides, and judge the phytoplankton density of the sample, which is convenient for the subsequent analysis of the sample. 8.1.2 Mixing the sample Before each sampling, thoroughly mix the sample by inverting at least 30 times, and the mixing action should be light. 8.1.3 Filtration volume Estimate the density of phytoplankton according to the pre-check, adjust the filtration volume of the sample, and finally make about 40 to 105 phytoplankton cells on the filter membrane. cell. Recommended sample filtration volumes for different precheck estimated density levels are shown in Table 1. 8.1.4 Sample Filtration Quickly measure the appropriate sample volume with a graduated cylinder or pipette (6.16) and add the sample to the vacuum filtration device (6.6) equipped with the filter membrane (5.6). In the funnel, let stand for 2 min to 3 min, vacuum filtration, turn off the vacuum pump when there is a 0.5 cm liquid layer in the funnel, and let the remaining liquid pass through the funnel completely. Do not drain the filter membrane. 8.1.5 Mounting Preparation After the sample has been filtered, remove the filter with toothless tissue tweezers (6.9), keeping the phytoplankton-retaining side up, and place 2 drops of On the slide (6.7) of the microscope immersion oil (5.5), drop 2 drops of microscope immersion oil (5.5) on the filter membrane with a transparent glass drop stick, and place the microscope immersion oil (5.5). Put the glass slide (6.7) into the slide drying plate (6.10), put it in the oven (6.11), and heat it at 70 ℃±2 ℃ for 2 h. Remove the slides after 2 hours Dry the plate (6.10) and observe whether the filter membrane is transparent. If it is transparent, add 2 drops of microscope immersion oil (5.5) on the filter membrane and cover with a cover glass (6.8), the preparation of the tablet is completed. If the filter membrane is not transparent after heating for 2 hours, prolong the heating time, not more than 24 hours. When covering with a coverslip (6.8), Do not disturb the filter. 8.2 Qualitative sample analysis Observe the qualitative sample under the microscope (6.12) to identify the species of phytoplankton. The dominant species has been identified, and other species should be identified at least Set to belong. For some identification references, see References. Note. In addition to qualitative samples for species identification, quantitative samples that have been counted can also be used for observation. 8.3 Quantitative sample analysis 8.3.1 Microscope calibration Calibrate the microscope (6.12) before counting and determine the area of the counting field (Whipple field or eyepiece field). Microscope Calibration Tool Includes stage micrometer (6.13) and Whipple eyepiece reticle (6.14). See Appendix B for the calibration method of the Whipple eyepiece reticle. 8.3.2 Microscope counting The recommended visual field categories and counted visual fields of samples with different density levels are shown in Table 2.Place the mount (8.1.5) on the microscope (6.12) mount On stage, microscopic counts were performed using the Whipple field or the eyepiece field of view. For sample counting at lower density levels, it is recommended to select the eyepiece field of view count. According to the size of phytoplankton cells, choose the magnification of eyepiece 10×, objective lens 20× or eyepiece 10×, objective lens 40× magnification, and record The species and quantity of phytoplankton in each field of view. ......Tips & Frequently Asked Questions:Question 1: How long will the true-PDF of HJ 1215-2021_English be delivered?Answer: Upon your order, we will start to translate HJ 1215-2021_English as soon as possible, and keep you informed of the progress. The lead time is typically 2 ~ 4 working days. The lengthier the document the longer the lead time.Question 2: Can I share the purchased PDF of HJ 1215-2021_English with my colleagues?Answer: Yes. The purchased PDF of HJ 1215-2021_English will be deemed to be sold to your employer/organization who actually pays for it, including your colleagues and your employer's intranet.Question 3: Does the price include tax/VAT?Answer: Yes. 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