CN106908395A - The assay method of planktonic algae chlorophyll in water - Google Patents
The assay method of planktonic algae chlorophyll in water Download PDFInfo
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- CN106908395A CN106908395A CN201510967101.0A CN201510967101A CN106908395A CN 106908395 A CN106908395 A CN 106908395A CN 201510967101 A CN201510967101 A CN 201510967101A CN 106908395 A CN106908395 A CN 106908395A
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- chlorophyll
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- ethyl alcohol
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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Abstract
The invention discloses a kind of assay method of planktonic algae chlorophyll a in water, the method obtains the planktonic algae in water with acetate fiber membrane filtration, the chlorophyll a in alginite is extracted with soaked in absolute ethyl alcohol, by obtaining supernatant after centrifugation, reference is made with absolute ethyl alcohol, with visible spectrophotometer at 630,647,664,750nm mensuration absorbance, calculate the content of algae chlorophyll a in water sample.Method of the present invention operation sequence is simple, and safe and reliable, as a result favorable reproducibility, practical.
Description
Technical field
The invention belongs to a kind of algae chlorophyll measuring method, and in particular to using the method for algae chlorophyll a in water by Spectrophotometry body, be suitable for the measure of various surface water.
Background technology
Chlorophyll is the important photosynthetic pigments in photosynthesis of plant.By determining phytoplankton chlorophyll, the situation of primary productivity in water body can be grasped.In environmental monitoring, water body Determination of Chlorophyll a contents can well reflect the biomass of planktonic algae in water body, thus be taken as one of index of body eutrophication.
The method for determining planktonic algae chlorophyll a in water has 3 classes, i.e. AAS, fluorescence method and high performance liquid chromatography.Wherein instrument needed for AAS is simple, easy to operate, with low cost, is the current main method for using.
The U.S.《Water and effluent monitoring analysis method》The standard method that (Standard Methods for the Examination of Water and Wastewater) is proposed is to use glass fiber filter to filter water sample, plus 90% acetone grinding, more than 2h is extracted under dark condition, spectrophotometric determination absorbance, then the content for calculating chlorophyll a are used after centrifugation.The shortcoming of the method is that glass fiber filter price is of a relatively high, and volatile organic matter acetone has certain harm to human body, and grinding is time-consuming more long, is easily lost during transfer extract solution, and need Cord blood extract solution.
What State Environmental Protection Administration of China formulated《Water and effluent monitoring analysis method》With spectrophotometric determination and Chlorophyll-a Content is calculated after the method that (fourth edition) is used is 6~8h of filter membrane low temperature drying that algae is retained with acetate fiber membrane filtration water sample, plus 90% acetone milling and extracting, after centrifugation.The shortcoming of the method is that acetone has harmfulness to human body, and grinding is time-consuming more long, is easily lost during transfer extract solution.
(patent publication No. CN1631117A) such as Xi'an University of Architecture and Technology Huang court of a feudal ruler woodss describes a kind of safer ethanol extraction method, and the method uses 90% ethanol as extract solution, is improved the security of test process.But the weak point of the method mainly has at 2 points：First is to use 0.8 μm of filter membrane, and aperture is bigger than normal, and having part algal picoplankton cannot retain.Second, the method needs also exist for grinding, is easily lost during transfer extract solution, time and effort consuming.
The content of the invention
The present invention seeks to by the improvement to original algae chlorophyll measuring method, there is provided the assay method that a kind of equipment is simple, easy to operate, safe and harmless, result accuracy is high.
The purpose of the present invention is realized by solution below：
Using the planktonic algae in acetate fiber membrane filtration water, the acetate fiber filter membrane for having retained algae is directly soaked with absolute ethyl alcohol, it is placed at room temperature at dark after extracting 6~24h and takes out centrifugation, supernatant mensuration absorbance under 630,647,664,750nm wavelength is obtained, and calculates the content of water body Determination of Chlorophyll a.
Described acetate fiber filter membrane uses 0.45 μm of aperture.
Filter membrane is placed in 10ml tool plug centrifuge tubes, plus 10ml soaked in absolute ethyl alcohol after filtering.
The extraction conditions are that at dark, temperature conditionss are 15-27 DEG C, and extraction time is 6~24h.
The extract solution centrifugal condition is 3500rmin-1, 10min is centrifuged.
The use of light path is the glass cuvette of 1cm, absolute ethyl alcohol carries out colorimetric as reference.
Chlorophyll-a Content is calculated according to following formula.
V --- filtering volume of water sample (L)；
Absorbance under D --- different wave length；
V1--- the volume (ml) after extract solution constant volume
Assay method of the invention has the following advantages：
Without grinding, manpower can be greatlyd save, and avoid because transfer extract solution and caused by chlorophyll lose；Using absolute ethyl alcohol as extract solution, nonhazardous safe to the human body；Extract solution can be placed at room temperature, without Refrigerator store, easy to operate, reduction energy consumption；General water determination project is more, and chlorophyll can the row colorimetric estimation again after 6~24h is extracted, it is thus possible to alleviate understaffed contradiction；Extract more completely, as a result favorable reproducibility.
Specific embodiment
With reference to specific embodiment, the invention will be further described：
Take certain water body in lake some as experiment water samples, using acetate fiber filter membrane that aperture is 0.45 μm () filtering 100ml water samples, filter membrane is put into 10ml tool plug centrifuge tubes, the pure absolute ethyl alcohol 10ml of analysis is added, stopper is covered, shake is positioned over dark place afterwards for several times.Centrifuge tube is taken out after 6~24h, using medical centrifuge with 3500rmin after shaking up-1Centrifugation, take supernatant in 1cm glass cuvettes, reference is made with absolute ethyl alcohol, with grating spectrophotometer under 630,647,664,750nm wavelength mensuration absorbance, using formula noted earlier calculate Chlorophyll-a Content.Experiment set 3 it is parallel.
It is the accuracy and reappearance of checking this method, while take same water sample, is write with State Environmental Protection Administration《Water and effluent monitoring analysis method》The chlorophyll measuring method of (fourth edition) determines one group of data (the second column data in table 1) (primary condition is grinding, acetone extraction).Additionally, also measured were under not grinding condition, useEthanolThe extracted amount of different extraction times (2h, 6h, 24h, 48h), and do not used under grinding conditionAcetoneThe extracted amount of different time, for verifying the accuracy of this method.
As can be seen from Table 1, it is 91.2 μ g/L to use State Environmental Protection Administration to recommend the chlorophyll a average value that method (grinding+acetone extraction) is measured, 6~the 24h of method recommended using the present invention is extracted the Chlorophyll-a Content for measuring and is approached with the method, and extracted amount is slightly higher (6h and 24h are respectively 94.0 μ g/L and 96.1 μ g/L).Directly extracted using acetone obtaining as a result, reaching maximum in 6h chlorophyll a extracted amounts, but its relative standard deviation is larger.Additionally, the direct extract solution Determination of Chlorophyll a of acetone less stables at room temperature, it is very big that different extraction times measure numerical value change, thus does not recommend.
By the present embodiment, it was demonstrated that the method for recommendation of the present invention not only extracts algae in water, and to take chlorophyll a more complete, and favorable reproducibility.
The Different Extraction Method of table 1 measures Chlorophyll-a Content and reappearance compares (unit：μg/L)
Note：(1) boldface type is the inventive method data measured in table；(2) ethanol is extracted and uses computing formula of the present invention；(3) acetone extraction uses State Environmental Protection Administration《Water and effluent monitoring analysis method》Formula described in (fourth edition).
Claims (6)
1. in a kind of water planktonic algae chlorophyll a assay method, it is characterized in that：Using the swim alga in acetate fiber membrane filtration water
Class, the acetate fiber filter membrane for having retained algae is directly soaked with absolute ethyl alcohol, is placed at room temperature at dark after extracting 6~24h and is taken out
Centrifugation, obtains supernatant mensuration absorbance under 630,647,664,750nm wavelength, and calculate water body Determination of Chlorophyll a's
Content.
2. in water according to claim 1 planktonic algae chlorophyll a assay method, it is characterized in that：The acetate fiber filter membrane
Using 0.45 μm of aperture.
3. in water according to claim 1 planktonic algae chlorophyll a assay method, it is characterized in that：Filter membrane is placed in after filtering
10ml tool plug centrifuge tubes, plus 10ml soaked in absolute ethyl alcohol.
4. in water according to claim 1 planktonic algae chlorophyll a assay method, it is characterized in that：The extraction conditions are black
Dark place, temperature conditionss are 15-27 DEG C, and extraction time is 6~24h.
5. in water according to claim 1 planktonic algae chlorophyll a assay method, it is characterized in that：The extract solution is centrifuged bar
Part is 3500rmin-1, 10min is centrifuged.
6. in water according to claim 1 planktonic algae chlorophyll a assay method, it is characterized in that：The use of light path is 1cm's
Glass cuvette, absolute ethyl alcohol carries out colorimetric as reference.
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| CN201510967101.0A CN106908395A (en) | 2015-12-22 | 2015-12-22 | The assay method of planktonic algae chlorophyll in water |
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| CN201510967101.0A CN106908395A (en) | 2015-12-22 | 2015-12-22 | The assay method of planktonic algae chlorophyll in water |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108827891A (en) * | 2018-06-21 | 2018-11-16 | 上海海事大学 | Ballast water for ship microalgae cell biology amount detection systems and method |
| CN109852664A (en) * | 2018-12-14 | 2019-06-07 | 浙江易测环境科技有限公司 | A method of for detecting single algae bio amount in wawter bloom or red tide water |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1631117A (en) * | 2004-12-30 | 2005-06-29 | 西安建筑科技大学 | Hydrobiontic algae chlorophyll measuring method |
| CN1793846A (en) * | 2005-12-20 | 2006-06-28 | 沈阳建筑大学 | Method for monitoring alpha chlorophyll |
| CN1963466A (en) * | 2006-11-20 | 2007-05-16 | 南开大学 | Testing method for chlorophyll of phytoplankton in water |
| CN101858857A (en) * | 2010-05-14 | 2010-10-13 | 同济大学 | Analyzing and measuring method for chlorophyll a of phytoplankton in eutrophic water |
-
2015
- 2015-12-22 CN CN201510967101.0A patent/CN106908395A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1631117A (en) * | 2004-12-30 | 2005-06-29 | 西安建筑科技大学 | Hydrobiontic algae chlorophyll measuring method |
| CN1793846A (en) * | 2005-12-20 | 2006-06-28 | 沈阳建筑大学 | Method for monitoring alpha chlorophyll |
| CN1963466A (en) * | 2006-11-20 | 2007-05-16 | 南开大学 | Testing method for chlorophyll of phytoplankton in water |
| CN101858857A (en) * | 2010-05-14 | 2010-10-13 | 同济大学 | Analyzing and measuring method for chlorophyll a of phytoplankton in eutrophic water |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108827891A (en) * | 2018-06-21 | 2018-11-16 | 上海海事大学 | Ballast water for ship microalgae cell biology amount detection systems and method |
| CN109852664A (en) * | 2018-12-14 | 2019-06-07 | 浙江易测环境科技有限公司 | A method of for detecting single algae bio amount in wawter bloom or red tide water |
| CN109852664B (en) * | 2018-12-14 | 2024-05-17 | 浙江易测环境科技有限公司 | Method for detecting single algae biomass in water body of water bloom or red tide |
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Application publication date: 20170630 |