CN114184757A - Method for measuring denitrification rate of water suspended matters - Google Patents

Method for measuring denitrification rate of water suspended matters Download PDF

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CN114184757A
CN114184757A CN202111485835.7A CN202111485835A CN114184757A CN 114184757 A CN114184757 A CN 114184757A CN 202111485835 A CN202111485835 A CN 202111485835A CN 114184757 A CN114184757 A CN 114184757A
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张鹂
夏永秋
严星
颜晓元
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Institute of Soil Science of CAS
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Abstract

The invention discloses a method for measuring the denitrification rate of suspended matters in water, which comprises the following steps: firstly, randomly selecting point positions of a water body to be detected to obtain column samples; pre-culturing the collected column sample; putting the pre-cultured column sample into a water simulation device for culturing, timing after the culture starts and sampling, and sampling every 2 hours; measuring the solubility N in the sample obtained2Concentration, N for different incubation times2Performing linear regression on the concentration and the culture time to obtain a regression linear equation; and finally, calculating the unit denitrification rate of the suspended matters in the water body and the denitrification rate of the sediments. The determination method combines a culture device and a membrane sample introduction mass spectrometry, collects in-situ water and sediments by using a collection device, places the water and sediments in a water simulation device for simulation culture, directly determines the soluble nitrogen in a water sample by using the membrane sample introduction mass spectrometry, and performs N-step culture for different culture times2The denitrification rate was calculated by linear regression of concentration and time.

Description

Method for measuring denitrification rate of water suspended matters
Technical Field
The invention relates to the technical field of environmental monitoring devices, in particular to a method for directly measuring the denitrification rate of suspended matters in a water body.
Background
The water body suspended matter is a hot spot area for denitrification because of being rich in nutrient substances and microorganisms.The existing denitrification rate measuring methods comprise an acetylene inhibition method, a nitrogen mass balance method, a nitrate nitrogen loss method,15N isotope labeling method, etc. These measurement methods are indirect methods and cannot measure the denitrification rate of the water suspended matter, and each has drawbacks. The acetylene inhibition method is low in cost and simple, but a sample is easy to pollute, and the problems of underestimation of denitrification loss and the like can occur. Isotopic tracing methods are typically produced during the time intervals in which oxygen is exposed to the culture device after gas collection15N2Blowing out, because oxygen is always in a saturated state, the biological activity in water and sediments is influenced, multi-step treatment is required, the tasks are various, and the method is high in cost. The nitrogen mass balance method and the nitrate nitrogen loss method are easy to cause inaccurate measuring results due to accumulation of various errors in the nitrogen conversion process. N is a radical of2Direct quantitation method and N2/The Ar method can directly quantify the denitrification rate of the system, but cannot measure the denitrification rate of the water suspension. N is a radical of2The direct quantitative method uses gas flow culture technique, seals the sample in an airtight container, washes with inert gas, replaces air with He or Ar gas, and measures N generated in the denitrification process in the closed system by gas chromatography2. The method has to be optimized because of the great difficulties in the aspects of the sealing of the culture system, the accuracy of gas collection and no pollution.
Therefore, a method for directly measuring the denitrification rate of the water suspended matters is urgently needed to be found, so that the denitrification rate of the water suspended matters can be simply, accurately and effectively measured.
Disclosure of Invention
The invention aims to solve the problems in the prior art, thereby providing a convenient, effective and direct and accurate method for measuring the denitrification rate of suspended matters in water.
Therefore, the invention adopts the following technical scheme.
The invention provides a method for measuring the denitrification rate of suspended matters in water, which is characterized by comprising the following steps of:
s1: randomly selecting the point positions of the water body to be detected to obtain column samples;
s2: pre-culturing the collected column sample;
s3: putting the pre-cultured column sample into a water simulation device for culturing, timing after the culture starts and sampling, and sampling every 2 hours;
s4: measurement of solubility N in the sample obtained in S32Concentration, N for different incubation times2Performing linear regression on the concentration and the culture time to obtain a regression linear equation;
s5: and calculating the unit denitrification rate of the suspended matters in the water body and the denitrification rate of the sediments.
Further, in step S1, the method for obtaining the column sample includes vertically driving the culture column into the sediment of the water body to be measured by using the undisturbed sediment sampler, and collecting the undisturbed sediment column sample with a surface layer of 0-10cm to ensure that the sediment keeps its original structure and level, and the water on the column sample keeps overflowing fully
The column samples comprise a first column sample, a second column sample, a third column sample and a fourth column sample, the concentration of the suspended matter in the first column sample and the third column sample is the same, the concentration of the suspended matter in the first column sample and the fourth column sample is the same, and the third column sample and the fourth column sample comprise sediments with the same composition and quality.
In step S2, the pre-culturing step is to immerse the column sample in a container filled with in-situ water for 4-6 hours, wherein the water level in the container is higher than the upper surface of the column sample by more than 6 cm.
In step S4, the regression linear equation is Y ═ cX + b, where the slope c is N in the column sample2Rate of change of concentration in μmol N2-N·L-1·h-1。
Further, the denitrification rate D in the column sampleeComprises the following steps:
Figure BDA0003397500520000021
v represents the volume of the water applied to the column sample, and S represents the inner diameter area of the column sample.
The method according to claim 6, wherein the step S5 is performed in step S5In step S5, the denitrification rate D of suspended matter per unit massssComprises the following steps:
Figure BDA0003397500520000031
or
Figure BDA0003397500520000032
Or
Figure BDA0003397500520000033
Preferably, the first and second electrodes are formed of a metal,
Figure BDA0003397500520000034
wherein D ise1Is the denitrification rate in the first column sample, De2Is the denitrification rate in the second sample, De3Is the denitrification rate in the third column, De4Is the denitrification rate in the fourth column; m is1Represents the amount of suspended matter in the first column sample, m2Represents the mass of suspended matter in the second column, m3Represents the mass of the suspension in the third column, m4Represents the amount of suspended matter in the fourth column.
In step S5, the sediment denitrification rate DMudComprises the following steps:
Figure BDA0003397500520000035
wherein H is the height of the water body in the column sample, and H is the total height of the water body and the sediments in the column sample.
The technical scheme of the invention has the following advantages:
(1) the assay method of the invention combines a culture device with Membrane Injection Mass Spectrometry (MIMS). Collecting in-situ water and sediments by a collecting device, placing the water and sediments in a water simulation device for simulation culture,the method for directly measuring the soluble nitrogen in the water sample by using the membrane sample introduction mass spectrometry has the measurement precision of 0.03 percent. By different incubation times N2The concentration and the time are subjected to linear regression to calculate the denitrification rate, so that the denitrification product N of the suspended matters in the water body under the sealed condition is realized2The direct determination of (a) is carried out,
(2) the determination method is simple, the water body simulated by the water body simulation device is more in line with the in-situ conditions, the suspended matters can be ensured to be in a suspended state, the nitrogen generated in the column is uniformly mixed, the unit denitrification rate of the suspended matters in the water body and the denitrification rate of the sediments can be obtained, and meanwhile, the influence of the sudden change of the field environment is avoided.
(3) The determination method of the invention does not need to introduce15N and other labeling substances do not need to use an inhibitor, so that the method has important significance for the research of the natural environment, reduces the cost, and is simple and convenient to operate, few in steps and convenient to popularize.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a water phantom used in an embodiment of the present invention;
FIG. 2 is a schematic view of four columns used in the test in example 1 of the present invention;
FIG. 3 is a standard curve of nitrogen concentration and time in a column sample according to example 4 of the present invention.
Description of reference numerals:
1. a motor; 2. a magnetic force rotating member; 3. a baffle plate; 4. a water outlet; 5. culturing the column; 6. a water outlet pipe; 7. a water inlet pipe; 8. a water stop clip; 9. and (5) supplying to a bottle.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the accompanying drawings, and it should be understood that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience of description and simplicity of description, but do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
In addition, the technical features involved in the different embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
The water body simulation device used in the embodiment of the invention is shown in fig. 1 and comprises a simulation culture bucket, a driving assembly and a collecting device. Wherein the top of the simulation culture barrel is open and is filled with the upper water, and the bottom of the simulation culture barrel is provided with at least three fixing positions, so that parallel experiments in the same environment can be conveniently carried out. The driving assembly is arranged on a mounting structure arranged in the simulation culture bucket and is provided with a motor 1 and a magnetic force rotating piece 2 connected with the motor 1. The acquisition device is placed on a fixed position and is immersed below the liquid level of the upper water in the simulation culture barrel, and comprises a culture column 5 with two open ends, a sealing cover at one end and a sealing rubber plug at the other end, a magnetic stirring mechanism with one end fixed on the sealing cover and the other end extending towards the interior of the culture column 5, and a water inlet pipe 7 and a water outlet pipe 6 which are arranged on the sealing cover and run through the water inlet pipe 7 and the water outlet pipe 6 side by side, wherein the outlet of the water outlet pipe 6 is connected with a sample injector of a membrane sample introduction mass spectrometer, the inlet of the water inlet pipe 7 is communicated with a supply bottle 9, water stop clamps 8 are arranged on the water inlet pipe 7 and the water outlet pipe 6, the magnetic stirring mechanism rotates along with the rotation of a magnetic rotating piece 2, a plurality of baffles 3 extending towards the outer wall surface of the installation column along the circumferential direction are arranged on the inner wall surface of the simulation culture barrel at intervals, the baffles 3 divide the interior of the simulation culture barrel into a plurality of non-communicated compartments, and three fixed positions are arranged at the bottom of any compartment, and a water outlet 4 is arranged on the outer wall of the simulated culture barrel corresponding to each compartment.
Example 1
In this embodiment, the denitrification rate of a certain aquaculture water body of normal maturity is determined by the following specific steps:
(1) randomly selecting point locations in a water body to be detected for sampling, vertically pumping a culture column into sediments of a river or a pond to be detected by using a non-disturbance sediment sampler in the sampling process, and respectively collecting four groups of column samples, wherein the concentration of suspended matters in a first column sample is the same as that in a third column sample, the concentration of suspended matters in the second column sample is the same as that in a fourth column sample, the first column sample and the second column sample do not contain sediments, the third column sample and the fourth column sample contain 10cm of original sediments with the same components and mass, the four column samples are respectively represented by A, B, C, D, and each column sample collects three parallel samples. The sediment is guaranteed to keep the original structure and the original level, and water on the column sample must be overflowed. At the same time, a clean plastic bucket is used to take 25L of in-situ coated water and return the water to the laboratory for culture.
(2) After the column sample is brought back to the laboratory, the column sample is put into a water simulation device filled with in-situ upper water to be immersed for 4 hours for pre-culture, the water surface is 6cm higher than the culture column, and the cover is not covered, so that the solubility N in the culture bucket and the culture column can be simulated2And (4) uniformly mixing. In the process of cultureThe laboratory temperature needs to be adjusted to a temperature consistent with the actual water temperature at the time of field sampling.
(3) After the pre-culture is completed, the sealing cover is stretched into the upper covering water to be screwed down, a rubber sealing ring is arranged below the cover, the sealing performance of the sealing cover can be guaranteed, and a small magnet rotor is hung below the middle of the cover and used for stirring the upper covering water when the culture and sampling are started. After the cover of the culture column is screwed down, the water inlet pipe is connected with a feeding bottle arranged at a higher position, so that water flows out under the action of gravity to feed the upper covering water lost due to sampling in the culture column. The air in the water inlet pipe is discharged and then inserted into the sealing cover, the water outlet pipe is inserted, and after the air in the water outlet pipe is discharged, the water stop clamp on the water inlet pipe and the water outlet pipe is closed. The whole process ensures that no bubbles are generated in the closed culture column, if bubbles are generated, the operation is required to be carried out again according to the flow, a motor in the culture device is started, the speed of three magnetic rods connected with the lower part of the motor is adjusted by rotating gears, the rotation of the magnetic rods can drive a small-sized magnetic rotor hung under the cover of the culture column to rotate, so that the culture device can simulate the flow of river water, the suspension state of the suspension in the culture column is ensured to be consistent with the in-situ suspension state of a field water body, meanwhile, the dissolved gas generated in the culture column in the culture process can be uniformly mixed, and the determination of the denitrification rate which is more in line with the in-situ condition is facilitated. Then the motor 1 in the culture device is started, and the gear is adjusted to enable the rotating speed of the magnetic rotating piece 2 to reach about 350 rpm.
(4) Starting a motor in the water simulation device, adjusting gears to enable the rotating speed of the magnetic rotating piece to reach 350rpm, sampling for 0, 2, 4, 6 and 8 hours respectively after the culture system is debugged, taking time as a horizontal coordinate and N as an abscissa2The concentration is plotted on the ordinate, and the slopes of the four column samples are obtained.
(5) Calculating the denitrification rate, and covering the culture column with N in water2The rate of change of concentration multiplied by the volume V of the water cover is 1.10(L), and divided by the area S of the inner diameter of the column is 0.005 (m)2) To obtain the denitrification rate De(μmol N2·m-2·h-1)
The suspension concentrations and denitrification rates for the four column samples, three replicates for each column sample, are averaged as shown in Table 1:
TABLE 1 sample Denitrification Rate and suspended matter content
Sample (I) A B C D
Denitrification Rate (. mu. mol N)2·m-2·h-1) 28.09 83.88 52.29 96.39
Suspended matter concentration (mg/L) 9 9 12 12
The volume V of the overlying water was 1.10L, and the suspended matter masses of A and C were 9.9g and 13.2g, respectively.
The denitrification rate D of suspended matters in unit mass of the water bodyss(μmol N2·m-2·h-1·mg-1):
Figure BDA0003397500520000081
Height h of upper water in the culture column:
Figure BDA0003397500520000082
the denitrification rate D of the sediment in the water body can be obtainedMud(μmol N2m-2·h-1):
Figure BDA0003397500520000083
Example 2:
in this example, the denitrification rate of a mature aquaculture water body is determined by the specific steps as described in example 1.
The denitrification rate and the suspended matter content of the water sample of the water body are measured as shown in the table 2.
TABLE 2 sample Denitrification Rate and suspended matter content
Sample (I) A B
Denitrification Rate (. mu. mol N)2·m-2·h-1) 32.87 48.84
Mass of suspension (mg/L) 11 14
The volume V of the overlying water was 1.10L, and the suspended matter mass of A, B was 12.1g and 15.4, respectively.
The denitrification rate D of suspended matters in unit mass of the water bodyss(μmol N2·m-2·h-1·mg-1):
Figure BDA0003397500520000084
Example 3
In this embodiment, the denitrification rate of a certain aquaculture water body of normal maturity is determined by the following specific steps:
(1) randomly selecting point positions in a water body to be detected for sampling, vertically driving a culture column into the sediment of a river or a pond to be detected by using a non-disturbance sediment sampler in the sampling process, and collecting an undisturbed sediment column sample with the surface layer of 0-10cm to ensure that the sediment keeps the original structure and level, and water on the column sample must overflow. At the same time, a clean plastic bucket is used to take 25L of in-situ coated water and return the water to the laboratory for culture.
(2) After the column sample is brought back to the laboratory, the column sample is put into a water simulation device filled with in-situ upper water to be immersed for 4 hours for pre-culture, the water surface is 6cm higher than the culture column, and the cover is not covered, so that the solubility N in the culture bucket and the culture column can be simulated2And (4) uniformly mixing. In the culture process, the laboratory temperature needs to be adjusted to be consistent with the actual water temperature in field sampling.
(3) After the pre-culture is completed, the sealing cover is stretched into the upper covering water to be screwed down, a rubber sealing ring is arranged below the cover, the sealing performance of the sealing cover can be guaranteed, and a small magnet rotor is hung below the middle of the cover and used for stirring the upper covering water when the culture and sampling are started. After the cover of the culture column is screwed down, the water inlet pipe is connected with a feeding bottle arranged at a higher position, so that water flows out under the action of gravity to feed the upper covering water lost due to sampling in the culture column. The air in the water inlet pipe is discharged and then inserted into the sealing cover, the water outlet pipe is inserted, and after the air in the water outlet pipe is discharged, the water stop clamp on the water inlet pipe and the water outlet pipe is closed. The whole process ensures that no bubbles are generated in the closed culture column, if bubbles are generated, the operation is required to be carried out again according to the flow, a motor in the culture device is started, the speed of three magnetic rods connected with the lower part of the motor is adjusted by rotating gears, the rotation of the magnetic rods can drive a small-sized magnetic rotor hung under the cover of the culture column to rotate, so that the culture device can simulate the flow of river water, the suspension state of the suspension in the culture column is ensured to be consistent with the in-situ suspension state of a field water body, meanwhile, the dissolved gas generated in the culture column in the culture process can be uniformly mixed, and the determination of the denitrification rate which is more in line with the in-situ condition is facilitated. Then the motor 1 in the culture device is started, and the gear is adjusted to enable the rotating speed of the magnetic rotating piece 2 to reach about 350 rpm.
(4) Starting a motor in the water simulation device, adjusting gears to enable the rotating speed of the magnetic rotating piece to reach 350rpm, sampling for 0, 2, 4, 6 and 8 hours respectively after the culture system is debugged, taking time as a horizontal coordinate and N as an abscissa2The concentration is the ordinate, and the slope is obtained. As shown in fig. 3, a, b, and c represent three repeated samplings, and the results are shown in table 3:
TABLE 3 solubility N2Correlation of concentration with time
a b c
Regression equation y＝0.3394x+531.87 Y＝0.3337x+533.77 y＝0.335x+534.75
R2 0.9781 0.9606 0.9815
(5) The denitrification rate was calculated and N in a water body of a normally mature aquaculture was shown in Table 42The rate of change of concentration was 0.3337. mu. molN2-N·L-1·h-1-0.3394μmolN2-N·L-1·h-1The time-space variability of different sample points is not large (CV is 0.89%);
TABLE 4N of a normally mature aquaculture water2Rate of change of concentration
Figure BDA0003397500520000101
Coating the culture column with N in water2The rate of change of concentration multiplied by the volume V of the water cover is 1.10(L), and divided by the area S of the inner diameter of the column is 0.005 (m)2) To obtain the denitrification rate De(μmol N2·m-2·h-1):
Figure BDA0003397500520000102
Figure BDA0003397500520000103
Figure BDA0003397500520000104
Figure BDA0003397500520000105
From the above, it can be seen that the denitrification rates obtained by 3 times of repeated sampling are substantially consistent, which indicates that the technical scheme of the application has high consistency.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.

Claims (8)

1. A method for measuring the denitrification rate of suspended matters in water is characterized by comprising the following steps:
s1: randomly selecting the point positions of the water body to be detected to obtain column samples;
s2: pre-culturing the collected column sample;
s3: putting the pre-cultured column sample into a water simulation device for culturing, timing after the culture starts and sampling, and sampling every 2 hours;
s4: measurement of solubility N in the sample obtained in S32Concentration, N for different incubation times2Performing linear regression on the concentration and the culture time to obtain a regression linear equation;
s5: and calculating the unit denitrification rate of the suspended matters in the water body and the denitrification rate of the sediments.
2. The method according to claim 1, wherein in step S1, the method for collecting the column sample comprises vertically driving the culture column into the sediment of the water body to be tested by using a non-disturbance sediment sampler, and collecting the original sediment column sample with a surface layer of 0-10cm to ensure that the sediment keeps its original structure and level and the water on the column sample keeps overflowing.
3. The assay of claim 2, wherein the column sample comprises a first column sample, a second column sample, a third column sample, and a fourth column sample, wherein the first column sample and the third column sample have the same concentration of the suspension, wherein the second column sample and the fourth column sample have the same concentration of the suspension, and wherein the third column sample and the fourth column sample comprise the same composition and mass of the sediment.
4. The method according to claim 3, wherein the pre-incubation is performed in step S2 by immersing the column sample in a container filled with in-situ water for 4-6 hours, the water level in the container being 6cm or more above the upper surface of the column sample.
5. The method according to claim 4, wherein the regression linear equation in step S4 is Y ═ cX + b, where the slope c is N in the column sample2Rate of change of concentration in μmol N2-N·L-1·h-1。
6. The method according to claim 5, wherein the denitrification rate D in the column sampleeComprises the following steps:
Figure FDA0003397500510000021
v represents the volume of water in the column sample, and S represents the inner diameter area of the column sample.
7. The method according to claim 6, wherein in step S5, the denitrification rate D per unit mass of the suspension isssComprises the following steps:
Figure FDA0003397500510000022
or
Figure FDA0003397500510000023
Or
Figure FDA0003397500510000024
Preferably, the first and second electrodes are formed of a metal,
Figure FDA0003397500510000025
wherein D ise1Is the denitrification rate in the first column sample, De2Is the denitrification rate in the second sample, De3Is the denitrification rate in the third column, De4Is the denitrification rate in the fourth column; m is1Represents the amount of suspended matter in the first column sample, m2Represents the mass of suspended matter in the second column, m3Represents the mass of the suspension in the third column, m4Represents the amount of suspended matter in the fourth column.
8. The method according to claim 7, wherein in step S5, the denitrification rate D of the sedimentMudComprises the following steps:
Figure FDA0003397500510000026
wherein H is the height of the water body in the column sample, and H is the total height of the water body and the sediments in the column sample.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115504634A (en) * 2022-11-03 2022-12-23 江苏南大华兴环保科技股份公司 Preposed denitrification graded biological denitrification reaction system and device

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
姚晓龙等: "鄱阳湖水体悬浮物反硝化潜力模拟研究", 《中国环境科学》 *
李晓波等: "N2∶Ar法直接测定淹水环境反硝化产物N2的产生速率", 《农业环境科学学报》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115504634A (en) * 2022-11-03 2022-12-23 江苏南大华兴环保科技股份公司 Preposed denitrification graded biological denitrification reaction system and device

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