CN109115756A - A kind of method of small system Fast Continuous Determination sea water solubility ammonium salt - Google Patents
A kind of method of small system Fast Continuous Determination sea water solubility ammonium salt Download PDFInfo
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Abstract
The invention discloses a kind of methods that small system quickly measures water body ammonium salt.The present invention devises the reaction system of 1ml, pass through each solution usage in optimization 1ml reaction system, and the reaction time, so that the stability of system is good, reduction rate is high, operation of the present invention is simple, it is time-consuming less, need sample size few etc., solve conventional water body nitrate measuring method using the problems such as sample size is more, time-consuming, process is cumbersome, be it is a kind of it is simple at sea, accurately, the method for the micro nitrate of the real time measure water body.
Description
Technical field:
The invention belongs to analytical geochemistry fields, and in particular to a kind of small system quickly measures the side of water body propylhomoserin salt
Method.
Background technique:
Ammonia nitrogen (NH4 +- N) one of main indicator as natural water nutrition condition, Water quality, pollution journey can be evaluated
The quality of degree, self-cleaning situation, is the key element in natural water during nitrogen biogeochemical cycle[1–2].To water body
The Accurate Determining of ammonium salt is the biogeochemical cycle process for studying form, distribution characteristics and the various nitrogen forms conversion of nitrogen
Important foundation.The measuring method of ammonium salt has spectrophotometry, fluorescence method and nutritive salt automatic analysis technology etc. at present[7–8].Because glimmering
Light method reaction system needs to heat and generate product and is not sufficiently stable, and is subject to certain restrictions the application of the method；Nutritive salt
Though automatic analysis technology can high-volume automatic analysis of samples, be related to instrument price valuableness, be difficult to promote in most laboratory general
And；Spectrophotometry because its measurement is simple, accurate and the features such as that is not required to expensive instrument due to be widely adopted.Spectrophotometry is main
Including sodium hypobromite oxidizing process[9]And indigo spectrophotometry[10]Two kinds, but ammonia-nitrogen sensitivity it is low, it is cumbersome and
There are matrix effects[11],And sodium hypobromite oxidizing process the advantages that poisonous reagent, simplicity, high oxygenation efficiency are not necessarily to it and by many real
Test room use.But need to clean a large amount of glass instruments in the experimentation of " standard of marine survey " " sodium hypobromite oxidizing process surveys ammonium salt "
Device measures sample absorbance using spectrophotometer one by one, has the characteristics that time-consuming and heavy workload, and waste liquid amount is big, easily
Cause secondary pollution.On the other hand, Wu Zhuozhi thinks inside glass there are organonitrogen impurities, encounters hypobromite or NaOH solution
Ammonia can be generated, the measurement of severe jamming ammonium salt is easy to appear phenomena such as poor repeatability, blank value are high and linear poor, and uses modeling
Material products can reduce blank value, improve linear relationship[12]。
This method generally uses 25ml reaction system, reacts 10min, finally uses spectrophotometric determination.However, carrying out
It needs to carry out the work such as cleaning, the drying of wide mouth glass bottle before experiment, process is cumbersome, time-consuming, and glass apparatus aboard ship operates
It is inconvenient.Meanwhile when carrying out sample measurement every time, the sample size used is more, even if not carrying out on-site measurement aboard ship, also needs to spend
Take certain manpower and material resources and sample is taken back into laboratory, moreover sample should not save too long.
Due to above-mentioned many influence factors, urgent need develops a kind of simple aboard ship, accurate, micro nitric acid of the real time measure water body
The method of salt.
Summary of the invention:
The object of the present invention is to provide it is a kind of can aboard ship it is simple, accurate, the small system of the real time measure water body ammonium salt is quick
Method, this method can be effectively solved the deficiencies of conventional Zinc Cadmium Reduction process is cumbersome, time-consuming, more using sample size, survey
Fixed water body example is not limited solely to low ammonium salt concentration, and high ammonium salt concentration is also suitable.We are to it in multiple links
It is improved: first is that using disposable round bottom spiral cover 2mL plastic centrifuge tube as reaction vessel, using preceding only clear with ultrapure water
It washes, saves the time for the glass colorimetric cylinder cleaning that conventional acid soak is crossed；Second is that replacing glass beaker to prepare with plastic beaker
Hypobromite and NaOH solution reduce the interference of glass apparatus；Third is that using microplate reader carries out spectrophotometry, can criticize
Random sample product light absorption value is measured, the flux of measurement is effectively improved.Improved small system sodium hypobromite oxidizing process effectively reduces sample
Usage amount reduces human cost, improves the precision and efficiency of measurement, achieves ideal measurement effect.In addition, compare
Other methods, this method reduce the use of glass apparatus to the greatest extent, of less demanding to the operation of instrument, survey suitable for the scene on ship
It is fixed.
The method of small system Fast Continuous Determination sea water solubility ammonium salt of the invention, comprising the following steps:
A, the formulation of standard working curve:
Moved into respectively in two groups of each six round bottoms spiral cover 2mL centrifuge tubes 0.1 μm of ol/L ammonium chloride standard solution 0,10,
20,40,60,80 μ L is diluted to 1mL with ultrapure water, mixes；100 μ L 50mg/mL sodium hypobromite oxidizing agent solutions are added, mix
It is even, react 30min；100 μ L 10g/L P-aminobenzene-sulfonamide solution are added, mix, place 5min；20 μ L1g/L1- are added how
It for two hydrochloric acid solution solution of ethylenediamine, mixes well, can be measured after placing 15min, color can stablize 4h；After colour stable,
300 μ L solution to be measured are drawn into 96 hole elisa Plates of standard flat bottom, places in microplate reader and measures each sample under 543nm wavelength
Light absorption value (As) records measured value；An=As-Ab, wherein AbFor blank absorbency, An is the light absorption value deducted after blank, with An
For ordinate, ammonium salt-nitrogen concentration C s is that abscissa draws standard working curve, and finds out standard work song with linear regression method
Line intercept a and slope b；Find out its regression beeline equation；
B, water determination:
Water body example 1ml after taking centrifugation, is added in round bottom spiral cover 2mL centrifuge tube, adds 100 μ L 50mg/mL times
Sodium bromate oxidizing agent solution mixes, and reacts 30min；100 μ L10g/L P-aminobenzene-sulfonamide solution are added, mix, place
5min；It is added how 20 μ L 1g/L 1- replace two hydrochloric acid solution of ethylenediamine, mixes well, can be measured after placing 15min, color can
Stablize 4h；After colour stable, 300 μ L solution to be measured are drawn into 96 hole elisa Plates of standard flat bottom, are placed in microplate reader in 543nm
Water body example light absorption value is measured under wavelength, which subtracts the light absorption value A that reference blank light absorption value is water sample to be measuredw；
C, ammonium salt oxygenation efficiency measures
It respectively measures the artificial seawater that 1.0ml salinity is 5.0 and the artificial seawater for being 5.0 with salinity prepares and contain 1.0,5.0 μ
Mol/L ammonium chloride solution is added separately in round bottom spiral cover 2mL centrifuge tube, then is separately added into 100 μ L 50mg/mL sodium hypobromites
Oxidizing agent solution mixes, and reacts 30min；100 μ L10g/L P-aminobenzene-sulfonamide solution are added, mix, place 5min；It is added
How 20 μ L1g/L 1- replace two hydrochloric acid solution of ethylenediamine, mix well, and can measure after placing 15min, color can stablize 4h；Face
After colour-stable, 300 μ L solution to be measured are drawn into 96 hole elisa Plates of standard flat bottom, places and surveys under 543nm wavelength in microplate reader
Determine water body example light absorption value, wherein Ab1、Ab2For the light absorption value (having deducted reagent blank) of 1.0,5.0 μm of ol/L ammonium salt solutions,WithLight absorption value respectively containing 1.0,5.0 μm of ol/L nitrite solutions (has deducted reagent sky
It is white)；
D, the calculating of ammonium salt oxygenation efficiency
Ammonium salt oxygenation efficiency R is calculated according to the following formula:
Corresponding digital code is as shown in the table in formula:
Abn--- it is the light absorption value containing 1.0 or 5.0 μm of ol/L ammonium salt solutions；
--- it is the light absorption value of 1.0 or 5.0 μm of ol/L nitrite solutions.
E, it calculates
Ammonium salt-nitrogen concentration calculates according to the following formula in water sample:
In formula:
c(NH4 +) --- ammonia salt concentration in water sample, unit are μm ol/L；
--- the average absorbance value that water sample measures；
Ab--- sample blank light absorption value；
A --- standard working curve intercept；
B --- standard working curve slope.
It is preferred that the ammonium chloride standard solution be 0.1mmol/L ammonium chloride standard solution, then respectively with ultrapure water into
Row gradient dilution makes ammonium salt-nitrogen concentration be followed successively by 0,0.5,1,2,3,4,6,8,10,12,14,16 μm of ol/L, obtains each ladder
Spend diluted ammonium salt solution.
Since each test sample used in the present invention only needs 1ml, only needed when filling sample using 2ml circle
Bottom centrifuge tube, operating process is simple, does not need to scrub wide mouth glass bottle, reduces real due to scrubbing the unclean bring of vial
Error is tested, centrifuge tube is directly discarded after measurement finishes, and reduces manpower and material resources, goes to sea and does not need to carry ultrapure water, bulb glass
The sour cylinder etc. of bottle more prevents bring glass apparatus damage of jolting on ship.
The usage amount of water body example is reduced relative to conventional 25ml system using 1ml reaction system, is solved each
Flight number CTD adopts the problem that water is inadequate, needs multiple repairing weld, ship time long, and extensive oceanographic survey sampling need to carry water body
Sample size is more, sample preservation, post-processing difficulty etc. problem, while being suitble to the measurement of small system water body nitrate, such as survey
The top different time 12ml sky is determined into background ammonium salt concentration in the small system water body of sample bottle (labco), if conventionally, facing
One serious problems: water body example is insufficient, after multiple big system sampling, it is difficult to keep the situation of original system, and adopt
With 1ml system, according to sample concentration difference, need every time sample amount be 50 μ of μ l~800 l (when sample concentration is high, only need a small amount of sample liquid,
By dilution, and sample concentration is low, then needs sample liquid amount more), background nitrate concentration can be determined.
96 hole multi-function microplate readers are used when measuring light absorption value, continuous mode becomes simpler, it is only necessary to use liquid relief
Rifle successively shifts 300 μ l solution into ELISA Plate, using artificial seawater as reference, measures in microplate reader.With conventional spectrophotometric
Meter method is compared, the cumbersome step eliminate transfer solution, cuvette surface solution is wiped with paper handkerchief, measuring sample etc. manually one by one
Suddenly, it is suitable for the real time measure batch samples on ship.
Using 50 samples can be measured in innovation and creation 1h.Time-consuming as follows: experiment consumptive material prepares (centrifuge tube) altogether
15min adds sulfanilamide (SN) hybrid reaction 5min, adds salt naphthalene hybrid reaction 10min, transfer liquid to 96 orifice plate 5min, microplate reader measurement
Take 15min.
In conclusion the present invention devises the reaction system of 1ml, by optimizing each solution usage in 1ml reaction system, with
And the reaction time so that the stability of system is good, oxygenation efficiency is high, operation of the present invention is simple, it is time-consuming less, need sample size few etc.,
Conventional water body nitrate measuring method is solved using the problems such as sample size is more, time-consuming, process is cumbersome, is a kind of simple at sea
Single, accurate, the micro nitrate of the real time measure water body method.
Detailed description of the invention:
Fig. 1 is influence of the reaction time to ammonia oxidation rate.
Fig. 2 is influence of the oxidizer to experimental result.
Fig. 3 is influence of the color developing agent dosage to experimental result.
Fig. 4 is two methods measurement NH4 +The standard curve (a) of-N and orthogonal verifying curve (b).
Fig. 5 is small system of determination NH4 +The detection range of-N
Specific embodiment:
The following examples are further illustrations of the invention, rather than limiting the invention.
Embodiment 1:
1, laboratory apparatus and reagent
Infinite M200Pro multi-function microplate reader (Switzerland/Tecan)；Vortex-genie2 turbine mixer (the U.S.
scientific industry)。
Ammonium chloride standard solution (10mmol/L)；Nitrite (sodium nitrite) standard solution (10mmol/L)；To amino
Benzsulfamide solution (10g/L)；How 1- replaces two hydrochloric acid solution of ethylenediamine (1g/L)；Sodium hypobromite oxidant (50mg/mL).With modeling
Expect that beaker replaces glass beaker to prepare hypobromite and NaOH solution.
2, influence factor optimization experiment
2.1 in order to determine the optimum reaction condition of improved method, this research is respectively provided with oxidization time, oxidant adds
The experiment that amount, colour developing agent dose influence ammoxidation process.
2.2 at room temperature, and the ammonium chloride that compound concentration gradient is 2,4,6 and 8 μm of olL-1 uses liquid, each concentration difference
It is arranged five groups, two Duplicate Samples of every group of setting carry out oxidization time (20,30,40,50 and 60min) to the shadow of ammoxidation reaction
Experiment is rung, to determine the optimal oxidization time of sodium hypobromite oxidant.Guarantee that oxidization time is consistent, changes oxidant at room temperature
Dosage adds the 50mg/mL sodium hypobromite oxidant of 20,40,60,80,100,120 and 140 μ L, to various concentration respectively
(2、4、6、8μmol·L-1) ammonium chloride aoxidized using liquid, to study the dosage of sodium hypobromite oxidant to ammoxidation
The influence of journey, each concentration samples set seven groups, and every group two parallel.In order to study color developing agent dosage to NH4 +The influence of measurement, it is right
Concentration gradient is 2,4,6,8 μm of olL-1Ammonium chloride successively carried out using liquid P-aminobenzene-sulfonamide (sulfanilamide (SN)) solution use
Amount and 1- naphthalene replace the experimental analysis of ethylendiamine dihydrochloride (salt naphthalene) solution usage amount, determine sulfanilamide (SN) solution, salt naphthalene respectively with this
The optimum quantum of utilization of solution, experiment are all provided with two in parallel.
2.3 according to the above results, determines suitable reaction condition are as follows: 50mg/mL sodium hypobromite oxidant usage amount
100 μ L, oxidization time 30min, 100 μ L of sulfanilamide (SN) solution usage amount, 20 μ L (Fig. 1, Fig. 2, Fig. 3) of salt naphthalene solution additional amount.
3, the formulation of standard working curve:
3.1 in two groups of each six round bottoms spiral cover 2ml centrifuge tubes, and it is molten successively to move into ammonium chloride standard using pipettor respectively
0 μ l of liquid (0.1mmol/L), 5 μ l, 10 μ l, 20 μ l, 30 μ l, 40 μ l, 60 μ l, 80 μ l, 100 μ l, 120 μ l, 140 μ l, 160 μ l are used
Ultrapure water is diluted to 1ml, mixes.This standard ammonium salt solution series ammonium salt-nitrogen concentration is followed successively by 0,0.5,1,2,3,4,6,8,10,
12、14、16μmol/L。
3.2 are added 100 μ L 50mg/mL sodium hypobromite oxidizing agent solutions in the above-mentioned centrifuge tube containing ammonium chloride, mix
It is even, react 30min；100 μ L sulfanilamide (SN) solution (P-aminobenzene-sulfonamide solution 10g/L) are added, mix, place 5min；20 μ are added
L salt naphthalene solution (how 1- replaces two hydrochloric acid solution 1g/L of ethylenediamine), mixes well, and can measure after placing 15min, color can be stablized
4h。
After 3.3 colour stables, 300 μ l solution are successively shifted using liquid-transfering gun into ELISA Plate (96 hole), in Infinite
On M200Pro multi-function microplate reader (Switzerland/Tecan), reference is made with the artificial seawater of respective corresponding salinity, in 543nm wave
Strong point measures light absorption value As.An=As-Ab, wherein AbFor blank absorbency, it is vertical with An that An, which is the light absorption value deducted after blank,
Coordinate, nitrate-nitrogen concentration C s are that abscissa draws standard working curve, and finds out standard working curve with linear regression method
Intercept a and slope b.(working curve is as shown in Figure 4).
4, the linear detection range analysis of small system
The linear detection range experiment of the small system of 1mL is carried out using the experiment condition after optimization, as a result as shown in Figure 5.It can
To find out, work as NH4 +- N concentration is in 0-32 μm of olL–1In range, solution concentration is in a linear relationship with absorbance, coefficient R 2
=0.9989；Work as NH4 +- N concentration is more than 32 μm of olL–1When, with NH4 +The increase of-N concentration, light absorption value increase amplitude reduction；
Work as NH4 +- N concentration is more than 80 μm of olL–1When, light absorption value has been basically unchanged, not with NH4 +The increase of-N concentration and increase.Therefore this
Test NH in determining small system rapid Continuous Determination water body4 +The detection range of-N is 0-32 μm of olL–1, compared with " ocean tune
Look into specification: the measurement of sodium hypobromite oxidizing process ammonium salt " in 0.03 μm of olL–1-8.00μmol·L–1Measurement range expands nearly 4
Times.Small system measuring method significantly improves the determination of the upper limit of ammonium nitrogen, can effectively be measured to higher ammonium nitrogen concentration water sample,
Time spent by more gradient dilutions and drug are also saved simultaneously.
According to the requirement of " environmental monitoring and analysis method directive/guide " (HJ 168-2010), 6 blank of replication calculate mark
Quasi- deviation s is 0.13 μm of olL–1.The method detection limit is calculated according to formula MDL=t (n-1,0.99) × s, as n=6, confidence
When degree is 99%, t value takes 3.37, then the detection of the small system method measurement water body ammonium salt is limited to 0.4 μm of olL -1 (Fig. 5).
5, water determination:
Water body example 1ml after taking centrifugation, is added in round bottom spiral cover 2ml centrifuge tube, and 100 50mg/mL bromines of μ L are added
Sour sodium oxidizing agent solution mixes, and reacts 30min；100 μ L sulfanilamide (SN) solution (P-aminobenzene-sulfonamide solution 10g/L) are added, mix
It is even, place 5min；20 μ L salt naphthalene solution (how 1- replaces two hydrochloric acid solution 1g/L of ethylenediamine) are added, mix well, after placing 15min
It can measure, color can stablize 4h；After colour stable, 300 μ L solution to be measured are drawn into 96 hole elisa Plates of standard flat bottom, are placed
Water body example light absorption value is measured in microplate reader under 543nm wavelength, it is water to be measured which, which subtracts reference blank light absorption value,
The light absorption value A of samplew。
6, the measurement of ammonium salt oxygenation efficiency and calculating
6.1 it is each measure the artificial seawaters that 1.0ml salinity are 5.0 and the artificial seawater for being 5.0 with salinity prepare containing 1.0,
5.0 μm of ol/L ammonium chloride solutions are added separately in round bottom spiral cover 2ml centrifuge tube, then are separately added into 100 50mg/mL bromines of μ L
Sour sodium oxidizing agent solution mixes, and reacts 30min；100 μ L sulfanilamide (SN) solution (P-aminobenzene-sulfonamide solution 10g/L) are added, mix
It is even, place 5min；20 μ L salt naphthalene solution (how 1- replaces two hydrochloric acid solution 1g/L of ethylenediamine) are added, mix well, after placing 15min
It can measure, color can stablize 4h；After colour stable, 300 μ L solution to be measured are drawn into 96 hole elisa Plates of standard flat bottom, are placed
Water body example light absorption value is measured in microplate reader under 543nm wavelength, wherein Ab1、Ab2For the suction of 1.0,5.0 μm of ol/L ammonium salt solutions
Light value (has deducted reagent blank),WithRespectively containing 1.0,5.0 μm of ol/L nitrite solutions
Light absorption value (has deducted reagent blank)；
6.2 calculate ammonium salt oxygenation efficiency R according to the following formula:
Corresponding digital code is as shown in the table in formula:
Abn--- it is the light absorption value containing 1.0 or 5.0 μm of ol/L ammonium salt solutions；
--- it is the light absorption value of 1.0 or 5.0 μm of ol/L nitrite solutions.
7, the calculating of ammonium salt measurement
Ammonium salt-nitrogen concentration calculates according to the following formula in water sample:
In formula:
c(NH4 +) --- ammonia salt concentration in water sample, unit are μm ol/L；
--- the average absorbance value that water sample measures；
Ab--- sample blank light absorption value；
A --- standard working curve intercept；
B --- standard working curve slope.
Specific measurement example:
This experiment water sample picks up from the website PRE-A of the mouth of the Zhujiang River (PRE) four (113.43 ° of E, 23.09 ° of N), PRE-B
(113.69 ° of E, 22.59 ° of N), PRE-C (113.86 ° of E, 22.36 ° of N), PRE-D (113.72 ° of E, 21.99 ° of N) superficial water and south
Sea (SCS) SCS28# (118.01 ° of E, 21.50 ° of N) level be 25m, 50m, 75m, 100m, 200m, 300m, 500m, 1000m and
The seawater sample of 1400m.After water body example acquisition, it is directly loadable into aseptic PET G material plastic bottle, a little saturation mercury chloride is added
Solution is kept in dark place under the conditions of 4 DEG C.Sample send back to behind laboratory be respectively adopted small system measuring method (see above-mentioned steps 1-7) and
Two methods of ocean specification conventional method carry out NO simultaneously2 –-N、NH4 +- N measurement.With two methods respectively simultaneously measure the mouth of the Zhujiang River and
South Sea sample.Because the mouth of the Zhujiang River ammonium nitrogen content is higher, concentration reaches as high as 500 μm of olL–1, the dilute of corresponding multiple is carried out before measuring
It releases.Measurement result is compared using orthogonal proof method, using 1mL system of determination result as abscissa, 25mL system of determination result
Linear fit (Fig. 4) is carried out for ordinate.The fitting a straight line related coefficient of two kinds of measurement results of PRE and South Sea 28# water sample point
Not Wei 0.9971,0.9917, linear relationship is good, for the feasibility of further verifying 1mL system of determination, to two kinds of sample surveys
Determine result and does F inspection and t inspection.It is examined through the F that confidence level is 95%, 1mL system and standard 25mL system of determination method are to PRE sample
The F value of product test result is 1.01, is less than F table 6.39, and illustrating the precision of two methods, there was no significant difference.It is through confidence level
95% t is examined, and Sig.=0.842 > 0.05 of two methods illustrates that there was no significant difference for 2 kinds of methods.Therefore, small using 1mL
It is feasible that system sodium hypobromite oxidizing process, which measures NH+4-N in river mouth water sample,.It is examined through the F that confidence level is 95%, 1mL system
It is 3.58 with F value of the standard 25mL system of determination method to South Sea 28# different levels seawater sample test result, is less than F table 6.39,
Illustrating the precision of two methods, there was no significant difference.It is examined through the t that confidence level is 95%, the Sig.=0.775 of two methods >
0.05, illustrate that there was no significant difference for 2 kinds of methods.Therefore, using in the small system sodium hypobromite oxidizing process measurement seawater sample of 1mL
NH4 +- N is feasible.
The usage amount of water body example is reduced relative to conventional 25ml system using 1ml reaction system, is solved each
Flight number CTD adopts the problem that water is inadequate, needs multiple repairing weld, ship time long, and extensive oceanographic survey sampling need to carry water body
Sample size is more, sample preservation, post-processing difficulty etc. problem, while being suitble to the measurement of small system water body nitrate, such as survey
The top different time 12ml sky is determined into background ammonium salt concentration in the small system water body of sample bottle (labco), if conventionally, facing
One serious problems: water body example is insufficient, after multiple big system sampling, it is difficult to keep the situation of original system, and adopt
With 1ml system, according to sample concentration difference, need every time sample amount be 50 μ of μ l~800 l (when sample concentration is high, only need a small amount of sample liquid,
By dilution, and sample concentration is low, then needs sample liquid amount more), background nitrate concentration can be determined.
96 hole multi-function microplate readers are used when measuring light absorption value, continuous mode becomes simpler, it is only necessary to use liquid relief
Rifle successively shifts 300 μ l solution into ELISA Plate, using artificial seawater as reference, measures in microplate reader.With conventional spectrophotometric
Meter method is compared, the cumbersome step eliminate transfer solution, cuvette surface solution is wiped with paper handkerchief, measuring sample etc. manually one by one
Suddenly, it is suitable for the real time measure batch samples on ship.
Using 50 samples can be measured in innovation and creation 1h.Time-consuming as follows: experiment consumptive material prepares (centrifuge tube) altogether
15min adds sulfanilamide (SN) hybrid reaction 5min, adds salt naphthalene hybrid reaction 10min, transfer liquid to 96 orifice plate 5min, microplate reader measurement
Take 15min.
In conclusion the present invention devises the reaction system of 1ml, by optimizing each solution usage in 1ml reaction system, with
And the reaction time so that the stability of system is good, oxygenation efficiency is high, operation of the present invention is simple, it is time-consuming less, need sample size few etc.,
Conventional water body nitrate measuring method is solved using the problems such as sample size is more, time-consuming, process is cumbersome, is a kind of simple at sea
Single, accurate, the micro nitrate of the real time measure water body method.
Claims (2)
1. a kind of method of small system Fast Continuous Determination sea water solubility ammonium salt, which comprises the following steps:
A, the formulation of standard working curve:
Moved into respectively in two groups of each six round bottoms spiral cover 2mL centrifuge tubes 0.1 μm of ol/L ammonium chloride standard solution 0,10,20,40,
60,80 μ L is diluted to 1mL with ultrapure water, mixes；100 μ L 50mg/mL sodium hypobromite oxidizing agent solutions are added, mix, reaction
30min；100 μ L 10g/L P-aminobenzene-sulfonamide solution are added, mix, place 5min；It is added how 20 μ L1g/L1- replace second two
Two hydrochloric acid solution solution of amine, mixes well, and can measure after placing 15min, color can stablize 4h；After colour stable, 300 are drawn
μ L solution to be measured is placed in microplate reader into 96 hole elisa Plates of standard flat bottom and measures each sample light absorption value under 543nm wavelength
(As), measured value is recorded；An=As-Ab, wherein AbFor blank absorbency, An is the light absorption value deducted after blank, is vertical sit with An
Mark, ammonium salt-nitrogen concentration C s are that abscissa draws standard working curve, and finds out standard working curve intercept with linear regression method
A and slope b；Find out its regression beeline equation；
B, water determination:
Water body example 1ml after taking centrifugation, is added in round bottom spiral cover 2mL centrifuge tube, adds 100 μ L 50mg/mL hypobromous acid
Sodium oxidizing agent solution mixes, and reacts 30min；100 μ L10g/L P-aminobenzene-sulfonamide solution are added, mix, place 5min；Add
Enter how 20 μ L 1g/L 1- replace two hydrochloric acid solution of ethylenediamine, mix well, can be measured after placing 15min, color can stablize 4h；
After colour stable, 300 μ L solution to be measured are drawn into 96 hole elisa Plates of standard flat bottom, are placed in microplate reader under 543nm wavelength
Water body example light absorption value is measured, which subtracts the light absorption value A that reference blank light absorption value is water sample to be measuredw；
C, ammonium salt oxygenation efficiency measures
It respectively measures the artificial seawater that 1.0ml salinity is 5.0 and the artificial seawater for being 5.0 with salinity prepares and contain 1.0,5.0 μm of ol/
L ammonium chloride solution is added separately in round bottom spiral cover 2mL centrifuge tube, then is separately added into the oxidation of 100 μ L 50mg/mL sodium hypobromites
Agent solution mixes, and reacts 30min；100 μ L10g/L P-aminobenzene-sulfonamide solution are added, mix, place 5min；20 μ are added
How L1g/L 1- replaces two hydrochloric acid solution of ethylenediamine, mixes well, and can measure after placing 15min, color can stablize 4h；Color is steady
After fixed, 300 μ L solution to be measured are drawn into 96 hole elisa Plates of standard flat bottom, places in microplate reader and measures water under 543nm wavelength
Body sample light absorption value, wherein Ab1、Ab2For the light absorption value (having deducted reagent blank) of 1.0,5.0 μm of ol/L ammonium salt solutions,WithLight absorption value respectively containing 1.0,5.0 μm of ol/L nitrite solutions (has deducted reagent sky
It is white)；
D, the calculating of ammonium salt oxygenation efficiency
Ammonium salt oxygenation efficiency R is calculated according to the following formula:
Corresponding digital code is as shown in the table in formula:
Abn--- it is the light absorption value containing 1.0 or 5.0 μm of ol/L ammonium salt solutions；
--- it is the light absorption value of 1.0 or 5.0 μm of ol/L nitrite solutions.
E, it calculates
Ammonium salt-nitrogen concentration calculates according to the following formula in water sample:
In formula:
c(NH4 +) --- ammonia salt concentration in water sample, unit are μm ol/L；
--- the average absorbance value that water sample measures；
Ab--- sample blank light absorption value；
A --- standard working curve intercept；
B --- standard working curve slope.
2. the method according to claim 1, wherein the ammonium chloride standard solution is 0.1mmol/L chlorination
Then ammonium standard solution carries out gradient dilution with ultrapure water respectively, so that ammonium salt-nitrogen concentration is followed successively by 0,0.5,1,2,3,4,6,
8,10,12,14,16 μm of ol/L, obtain the ammonium salt solution of each gradient dilution.
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| CN109827916A (en) * | 2019-03-13 | 2019-05-31 | 国家海洋局东海环境监测中心(国家海洋局东海海洋工程勘察设计研究所) | It is a kind of that the water quality nitrate detection method without salt effect is restored based on zinc cadmium |
| CN111855597A (en) * | 2020-07-27 | 2020-10-30 | 温氏食品集团股份有限公司 | Method for rapidly detecting total phosphorus in feed |
| CN113324930A (en) * | 2021-05-25 | 2021-08-31 | 广东石油化工学院 | Method for continuously and rapidly measuring nitrite nitrogen in fresh water by using small system |
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| CN109827916A (en) * | 2019-03-13 | 2019-05-31 | 国家海洋局东海环境监测中心(国家海洋局东海海洋工程勘察设计研究所) | It is a kind of that the water quality nitrate detection method without salt effect is restored based on zinc cadmium |
| CN111855597A (en) * | 2020-07-27 | 2020-10-30 | 温氏食品集团股份有限公司 | Method for rapidly detecting total phosphorus in feed |
| CN113324930A (en) * | 2021-05-25 | 2021-08-31 | 广东石油化工学院 | Method for continuously and rapidly measuring nitrite nitrogen in fresh water by using small system |
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