CN100520365C - Method and device for classified detecting density of phytoplankton under water in site - Google Patents
Method and device for classified detecting density of phytoplankton under water in site Download PDFInfo
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- CN100520365C CN100520365C CNB2006100414159A CN200610041415A CN100520365C CN 100520365 C CN100520365 C CN 100520365C CN B2006100414159 A CNB2006100414159 A CN B2006100414159A CN 200610041415 A CN200610041415 A CN 200610041415A CN 100520365 C CN100520365 C CN 100520365C
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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
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- G01N21/8507—Probe photometers, i.e. with optical measuring part dipped into fluid sample
Abstract
A method for classifying underwater original position of polytoplankton by detecting concentration of polytoplankton includes setting a groove at one side of closed pressure box and a light shading cover at external side of said box; erecting LED array formed by parallel-arranged LED with center wavelength of 450nm, 525nm, 570nm 590nm and 620nm in light path at side surface of said groove; dividing polytoplankton to be five light spectrum set as green, blue, brown, red and mixture; measuring 685nm florescent intensity of five different exciting wavelength to calculate out chlorophyll concentration of each light spectrum according to known coefficient.
Description
Technical field
The invention belongs to a kind of spectral measurement method and device
Background technology
The eutrophication of water body causes the excessive propagation of water phytoplankton, and excessive phytoplankton exhausts the dissolved oxygen DO in the water body, causes hydrobiont mass mortality, water body to turn black smelly.Moreover, some phytoplankton (blue-green algae etc.) discharges multiple algae toxin in water body, and health is produced serious harm.At present, reducing the damnous effective measures of body eutrophication is that phytoplankton concentration is in time monitored, and grasps the rule of alga eruption, in time responds with in advance and adopts an effective measure.Therefore, the phytoplankton concentration monitoring has become environmental monitoring department when last important task.Existing detection means mainly contains following 3 kinds:
1. spectrophotometric method: specifically can be divided into two kinds of visible absorbance spectrophotometric method and fluorescence spectrophotometries.The visible absorbance spectrophotometric method be meant to the water sample that contains the phytoplankton cell filter, organic solvent extraction, utilize measurement of ultraviolet-visible spectrophotometer to contain the absorbance of chlorophyllous organic solvent extraction liquid again, according to the concentration of certain Equation for Calculating chlorophyll a in the certain wave strong point.Accounting equation according to chlorophyll-a concentration.This method is the most frequently used water phytoplankton concentration detection method of current environment monitoring department, can be divided into two kinds of monocolor method and three-color process.Monocolor method is a kind of method that is proposed by Lorenzen in 1967, by measuring the concentration of acidifying front and back organic solvent extraction liquid at the light absorption value calculating chlorophyll a at 664nm wavelength place, topmost feature is to have eliminated the influence of phoeophytin a to the chlorophyll a measurement result.Three-color process then is the method by propositions in 1975 such as Jeffrey, this method is measured the light absorption value at 630nm, 647nm and three wavelength places of 664nm, calculate the concentration of chlorophyll a, chlorophyll b and Chlorofucsin in view of the above, eliminated the influence of chlorophyll b and Chlorofucsin the chlorophyll a measurement result.The ultimate principle of visible absorbance metric measurement is that the organic solvent of chlorophyll a shows and the positively related absorbance of chlorophyll concentration at specific wavelength, and is irrelevant with instrument system because absorbance is a relative quantity, so measurement result need not to proofread and correct.Fluorescence spectrophotometer degree method is filtered identical with the visible absorbance spectrophotometric method with the organic solvent extraction step, the organic extract of gained chlorophyll a excites down 680nm place emitting fluorescence intensity by fluorescence spectrophotometer measurement 430nm, emitting fluorescence intensity and chlorophyll-a concentration positive correlation, measurement result need to proofread and correct with the standard model that the visible absorbance spectrophotometric method records.With respect to the visible absorbance spectrophotometric method, the maximum characteristics of fluorophotometric method are highly sensitive.No matter visible absorbance spectrophotometric method or fluorescence spectrophotometry, all need sampling to treat lab analysis, sample pre-treatments step complexity is loaded down with trivial details, take time and effort, and different sample preparation processes has appreciable impact to measurement result, directly influences different sample measurement results' comparability.
2. high performance liquid chromatography (HPLC) method: this method is based on the principle that column chromatography separates, promptly multiple different colour components such as chlorophyll a in organic extract are separated one by one according to chronological order, measured the time resolution absorption spectrum at different wave length place again with photodiode array detector by chromatography.Compare with the time resolution absorption spectrum of standard coloring matter, respectively component is carried out qualitative and quantitative analysis by appearance time (retention time) and peak area.In addition, by different types of algae being selected a kind of colour component of feature, calculate likening to of this kind algae chlorophyll a peak area and feature pigment component peaks area and be conversion factor (Conversion Factor), sample measurement gained feature pigment peak area is this kind algae proportion in sample with the ratio and the corresponding conversion factor product of chlorophyll a peak area.Therefore, the HPLC method not only can be measured multiple pigment concentration of component such as chlorophyll a, and can calculate different algal species component proportion.However, HPLC is the same with spectrophotometric method to be needed to take a sample earlier, filters simultaneously the pre-treatment process with a series of very complicateds such as organic solvent extracting in the laboratory.In addition, the HPLC method needs expensive high performance liquid chromatography and relevant pick-up unit, can't satisfy the actual detected needs of most of environmental monitoring department.
3. microscopic counting detection method: this method is that water sample to be detected is placed microscopically, the different algal species cell is distinguished differential count finally draws the concentration of different algal species in water body.This method is directly calculated algae concentration with cell quantity, and it is more direct reliable to calculate algae concentration with chlorophyll-a concentration, can carry out category measurement to different algal species concentration according to the alga cells feature simultaneously.Yet the more above-mentioned two kinds of methods of this method are measured and are more taken time and effort, and need very professional testing staff to finish simultaneously, and therefore difficulty is found broad application in environmental monitoring.
In a word, existing method all need to water sample to be measured in advance sampling carry out further lab analysis then, sense cycle is long usually, complex steps and cost are higher, had a strong impact on monitoring coverage and frequency, can't satisfy the modern environment environmental monitoring fast, in real time, the requirement of original position.
Summary of the invention
The objective of the invention is many components algae concentration category measurement algorithm based on different algal species component characteristics standard fluorescence excitation spectrum, density of phytoplankton under water in site classification and Detection method and apparatus is proposed, with the phytoplankton concentration in the different depth water body is carried out fast, in real time, the category measurement of original position, grasp the space development law of phytoplankton concentration, for large-scale alga eruption is carried out forecast work timely.
Technical scheme of the present invention is as follows:
Density of phytoplankton under water in site classification and Detection method is characterized in that:
(1), under water, the light emitting diode of concentricity emission wavelength is not luminous successively to control 5 kinds, its centre wavelength is respectively 450nm, 525nm, 570nm, 590nm and 620nm, the light that sends reduces from axle light through diaphragm, eliminate long wavelength's light source light again by the short-pass cutoff filter, after assembling, lens converge on the sample that needs to detect, the fluorescence that sample is launched is assembled through lens, by the bandpass filter of 685nm, finally enter the detected fluorescence intensity that obtains different wave length i place of photomultiplier;
(2), the setting of spectrum group:
Green group: attached pigment is made up of chlorophyll a/b, carotenoid etc.
Blue group: attached pigment is made up of phycocyanin etc.
Palm fibre group: attached pigment is made up of chlorophyll a/c etc.
Combined group: attached pigment is made up of chlorophyll a/c, phycocyanin etc.
Red group: form by phycoerythrin etc.
(3), measuring different spectrum group algae is emitting fluorescence intensity K under 450nm, 525nm, 570nm, 590nm and the 620nm time unit chlorophyll-a concentration at wavelength i, record green, blue, brown, the mixing at wavelength i place and the mean value of red 5 kinds of spectrum group algae, note is made K respectively
I (G), K
I (Be), K
I (Bn), K
I00And K
I (R), since the additivity of fluorescence intensity, the fluorescence intensity F at mixing phytoplankton solution wavelength i place
iThere is following relationship:
F
i＝(K
j(G)×G)+(K
i(Be)×Be)+(K
i(Bn)×Bn)+(K
i(M)×M)+(K
i(R)×R)
G, Be, Bn, M and R are respectively green group, blue group, brown group, combined group and red group of phytoplankton chlorophyll-a concentration, by to above-mentioned polynary once linear equation solution, can calculate the concentration of chlorophyll a that there emerged a spectrum group phytoplankton.
Described water sample is through the light shield shading.
Density of phytoplankton under water in site classification and Detection device, it is characterized in that: on an airtight pressure-bearing casing, there is a groove its side, there is a light shield in the casing outside, what comprise installation successively in the light path of groove side is respectively 450nm by centre wavelength, 525nm, 570nm, the light emitting diode matrix that 590nm and 620nm light emitting diode are formed side by side, diaphragm, the short-pass cutoff filter, lens A, transparent window on the groove side, the light path of top, groove top comprises the transparent window on the groove end face of installing successively, lens B, the 685nm bandpass filter, photomultiplier.
The present invention is based on the characteristic standard fluorescence excitation spectrum of different algal species component, composition kind according to attached pigment is divided into 5 classes with phytoplankton, the phytoplankton that similar attached pigment is formed has similar 685nm place fluorescence excitation spectrum feature usually, has developed a kind of category measurement computing method of polycomponent algae concentration thus.
Innovative point of the present invention:
A. many kinds of algae components classification and Detection simultaneously:
System by phytoplankton being divided into some spectrum groups, finally can calculate the concentration of the phytoplankton of each spectrum group based on the sorting algorithm of many components phytoplankton concentration, and this detection for phytoplankton concentration has very important significance.Because some algae has special harm, need be to its concentration special concern, so the modern environment monitoring requires phytoplankton concentration is carried out category measurement.Yet traditional fluorescence detection can only be measured the total amount of chlorophyll a, and the result who obtains at last can only represent the total concentration of phytoplankton in the water body.This system has satisfied the requirement of modern environment monitoring to phytoplankton concentration category measurement.
B. super bright luminous two pipes are as excitation source
Traditional underwater fluorometer adopts pulse xenon lamp or laser instrument as excitation source usually, and not only cost costliness, volume are big, and cutting needs high voltage supply usually, is unfavorable for being used for portable fluorescent instrument.The present invention adopts super bright light emitting diode to replace traditional light source as excitation source.Diode is with low cost, and volume is little, and supply voltage only needs several volts, and general accumulator can satisfy power supply requirement.
C. its specific structure design can be satisfied the in site measurement needs of different depth
This system adopts the design of inside and outside double-layer structure, skin is a light shield, main rising covered external natural light, internal layer is the pressure-bearing casing, it is the core of instrument, adopt the structural design of anti-high pressure, can place in 100 meters scopes the different degree of depth to measure, this just satisfied water phytoplankton plant concentration to different depth carries out in real time, the measurement requirement of original position.
The invention effect
The density of phytoplankton under water in site photofluorometer adopts the super bright light emitting diode of a plurality of different excitation wavelengths as excitation source, adopt special double-layer structure design, have anti-pressure ability, can finish to the phytoplankton concentration of different depth in the water body classify in real time, original position, measure fast.Apparatus of the present invention can be installed on the fixing small-sized monitoring platform, and phytoplankton concentration in the water body is carried out continuous measurement, by wireless network measurement data are sent it back monitoring center, so that large-scale alga eruption is sent early warning timely.Simultaneously, this system can carry out quick real-time in-situ to the water phytoplankton concentration of diverse location in conjunction with the GPS GPS and measure as a kind of Portable Measurement Instrument.In addition, the water body of this system being put into a certain position allows it sink naturally, can measure the vertical distribution of phytoplankton concentration in the water body.
This system can be widely used in environmental monitoring department to the phytoplankton concentration monitoring, also can be applied to simultaneously the waterworks to the research to algae of the measurement of the phytoplankton plant concentration of water head site and algae research department.
Description of drawings
Fig. 1 is apparatus of the present invention structural representations.
Fig. 2 is apparatus of the present invention index paths.
Embodiment
Referring to Fig. 1, Fig. 2.
Density of phytoplankton under water in site classification and Detection device, constitute by three formants, 1., the light source activation unit be respectively:, comprise super bright LED array and relevant optical module thereof, 2., input and control module, 3., data processing and memory cell comprise photomultiplier and microcontroller,, comprise the externally connected portable formula computer that contains data collecting card and related software.Its concrete structure is mounted on the airtight pressure-bearing casing 9, and there is a groove 10 its side, and there is a light shield 11 in casing 9 outsides, is open between casing and the light shield 11, and seawater can enter groove, to be used for measurement.All there is transparent window groove 10 end faces and side, and optical element is installed in the casing 9 of close groove 10 end faces and side.
The core of total system is the design of optical texture, as shown in Figure 2,1, be light-emitting diodes optical arrays excitation source, by 5 kinds not the super bright light emitting diode of concentricity emission wavelength form, its centre wavelength is respectively 450nm, 525nm, 570nm, 590nm and 620nm, and concrete parameter sees Table 1; 2, be diaphragm, main effect is to reduce from the influence of axle light to measuring; 3, be the short-pass cutoff filter, it sees through scope is 400nm-620nm, and 650nm～900nm ends, and effect is to eliminate the influence of light source long wavelength light to measurement result; 4, be lens A, the light of light emitting diode is converged concentrate on the sample; 5, sample; 6, be lens B, the fluorescence of dispersing that sample is launched converges; 7, be the bandpass filter of 685nm; 8, be photomultiplier, detect 685nm and go out fluorescence intensity.The course of work of whole optical path system is that microprocessor controls light-emitting diodes light is luminous successively, the light that sends reduces from axle light through diaphragm 2, eliminate long wavelength's light source light by short-pass cutoff filter 3 again, after assembling, lens A converges on the test sample 5, the fluorescence that sample 5 is launched is assembled through lens B, by the bandpass filter 7 of 685nm, it is detected finally to enter photomultiplier 8, and sample 5 is the seawater in the groove 10.
The concrete parameter of table 1. stimulated luminescence diode
Claims (1)
1, density of phytoplankton under water in site classification and Detection device, it is characterized in that: on an airtight pressure-bearing casing, there is a groove its side, there is a light shield in the casing outside, what comprise installation successively in the light path of groove side is respectively 450nm by centre wavelength, 525nm, 570nm, the light emitting diode matrix that 590nm and 620nm light emitting diode are formed side by side, diaphragm, the short-pass cutoff filter, lens A, transparent window on the groove side, the light path of top, groove top comprises the transparent window on the groove end face of installing successively, lens B, the 685nm bandpass filter, photomultiplier.
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Cited By (3)
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| CN101957320A (en) * | 2010-08-11 | 2011-01-26 | 中国科学院安徽光学精密机械研究所 | Portable optical mechanical system of underwater in-situ luminoscope |
| CN102279174A (en) * | 2011-07-15 | 2011-12-14 | 中国科学院苏州纳米技术与纳米仿生研究所 | Alga identification and measurement sensor and method |
| CN103439303A (en) * | 2013-08-09 | 2013-12-11 | 中国长江三峡集团公司 | Alga anabiosis and vertical migration monitoring device and monitoring method for early warning of algal bloom |
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| FR2916848B1 (en) * | 2007-06-01 | 2010-06-04 | Force A | METHOD AND SYSTEM FOR CHARACTERIZING PIGMENT BIOLOGICAL TISSUE |
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| CN101957320A (en) * | 2010-08-11 | 2011-01-26 | 中国科学院安徽光学精密机械研究所 | Portable optical mechanical system of underwater in-situ luminoscope |
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| CN102279174A (en) * | 2011-07-15 | 2011-12-14 | 中国科学院苏州纳米技术与纳米仿生研究所 | Alga identification and measurement sensor and method |
| CN102279174B (en) * | 2011-07-15 | 2015-04-01 | 中国科学院苏州纳米技术与纳米仿生研究所 | Alga identification and measurement sensor and method |
| CN103439303A (en) * | 2013-08-09 | 2013-12-11 | 中国长江三峡集团公司 | Alga anabiosis and vertical migration monitoring device and monitoring method for early warning of algal bloom |
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