CN114184757B - Method for measuring denitrification rate of suspended matters in water body - Google Patents
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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; determination of the solubility N in the samples obtained 2 Concentration, N for different incubation times 2 Performing 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 injection 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 injection mass spectrometry, and performs N culture for different culture times 2 The denitrification rate was calculated by linear regression of concentration and time.
Description
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, 15 N 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 collection 15 N 2 Blowing 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 of 2 Direct quantitation method and N 2/ The Ar method can directly quantify the denitrification rate of the system, but cannot measure the denitrification rate of the water suspended matter. N is a radical of hydrogen 2 The direct quantitative method utilizes gas flow culture technique, and comprises sealing sample in airtight container, flushing with inert gas, replacing air with He or Ar gas, and measuring N generated in denitrification process in closed system by gas chromatography 2 . This method has yet to be optimized due to the great difficulties in the containment of the culture system, the accuracy of gas collection and the absence of contamination.
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: placing the pre-cultured column sample into a water simulation device for culturing, timing after the beginning of culturing and sampling, and sampling every 2 hours;
s4: measurement of solubility N in sample obtained in S3 2 Concentration, N for different incubation times 2 Performing 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 collecting the column sample includes vertically driving the culture column into the sediment of the water body to be detected by using a non-disturbance sediment sampler, collecting an undisturbed sediment column sample with a surface layer of 0-10cm, ensuring that the sediment keeps the original structure and level, and keeping the water on the column sample fully overflowed
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 the step S2, the pre-culture is to immerse the column sample in a container filled with the column sample in-situ covering water for 4 to 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 sample 2 Rate of change of concentration in μmol N 2 -N·L -1 ·h -1 。
Further, the denitrification rate D in the column sample e Comprises the following steps:
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 in step S5, the denitrification rate D of the suspended matter per unit mass is determined in step S5 ss Comprises the following steps:
Preferably, the first and second electrodes are formed of a metal,
wherein D is e1 Is the denitrification rate in the first column sample, D e2 As the denitrification rate in the second column, D e3 Is the denitrification rate in the third column, D e4 Is the denitrification rate in the fourth column; m is a unit of 1 Represents the amount of suspended matter in the first column sample, m 2 Represents the mass of suspended matter in the second column, m 3 Represents the mass of the suspension in the third column, m 4 The amount of suspended matter in the fourth column is shown.
In step S5, the sediment denitrification rate D Mud Comprises the following steps:
wherein H is the height of the water in the column sample, and H is the total height of the water 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). The collection device is used for collecting in-situ water and sediments, the in-situ water and the sediments are placed in the water simulation device for simulation culture, and the membrane sample introduction mass spectrometry is used for directly measuring the soluble nitrogen in the water sample, wherein the measuring precision can reach 0.03%. By different incubation times N 2 The denitrification rate is calculated by linear regression of the concentration and the time, and the denitrification product N of the suspended matters in the water body under the sealed condition is realized 2 The 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 introduce 15 N and the like without using inhibitors, has important significance for the research of natural environment and reduces the costMeanwhile, the method 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 in 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 is to be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. 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 and may be, for example, fixedly connected, detachably connected, or integrally connected; 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.
Furthermore, 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 figure 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 covering water in the simulation culture barrel, the acquisition device comprises a culture column 5 with two open ends and one end provided with a sealing cover and the other end provided with a sealing rubber plug, one end of the acquisition device is fixed on the sealing cover, the other end faces a magnetic stirring mechanism extending in the culture column 5 and runs through the water inlet pipe 7 and the water outlet pipe 6 which are arranged on the sealing cover side by side, the outlet of the water outlet pipe 6 is connected with a sample injector of a membrane sample injection 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 part 2, a plurality of baffles 3 which radially face towards the outer wall surface of the installation column are arranged along the circumferential interval on the inner wall surface of the simulation culture barrel, the baffles 3 divide the inside the simulation culture barrel into a plurality of non-communicated compartments, and are randomly arranged at the bottom of the fixed positions, and the outer wall of the simulation culture barrel is provided with each compartment and provided with a water outlet 4.
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 culture columns 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 and D, and each column sample collects three parallel samples. The sediment is ensured to keep the original structure and layer, and the water on the column sample must be overflowed. At the same time, 25L of in-situ water is taken out by a clean plastic bucket and is brought back 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 simulated 2 And (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 abscissa 2 The 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 water 2 The rate of change of concentration multiplied by the volume of the overlying water V =1.10 (L), divided by the internal diameter area of the column S =0.005 (m) 2 ) To obtain the denitrification rate D e (μmol N 2 ·m -2 ·h -1 )
The suspension concentrations and denitrification rates after averaging three replicates of the four columns are shown in table 1:
TABLE 1 sample Denitrification Rate and suspended solids 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 body ss (μmol N 2 ·m -2 ·h -1 ·mg -1 ):
Height h of upper water in the culture column:
the denitrification rate D of the sediment in the water body can be obtained Mud (μmol N 2 m -2 ·h -1 ):
Example 2:
this example measured the denitrification rate of a normally mature aquaculture water, 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 solids content
| Sample(s) | A | B |
| Denitrification Rate (. Mu.mol.N) 2 ·m -2 ·h -1 ) | 32.87 | 48.84 |
| Mass of suspended matter (mg/L) | 11 | 14 |
The volume V of the water to be coated is 1.10L, and the suspended substance masses of A and B are 12.1g and 15.4 respectively.
The denitrification rate D of suspended matters in unit mass of the water body ss (μmol N 2 ·m -2 ·h -1 ·mg -1 ):
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, 25L of in-situ water was taken from a clean plastic bucket and brought back 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 simulated 2 And (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 extends 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-sized 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 supply bottle placed at a higher position, so that water flows out under the action of gravity to supply the upper overlying water lost in the culture column due to sampling. 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 force rotating piece 2 to reach about 350 rpm.
(4) Starting a motor in the water simulation device, adjusting the gear to ensure that the rotating speed of the magnetic rotating piece reaches 350rpm, and culturingAfter the nutrition system is debugged, sampling is respectively carried out for 0, 2, 4, 6 and 8 hours, the time is taken as the abscissa, and N is taken as the N 2 The 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 N 2 Correlation of concentration with time
| a | b | c | |
| Regression equation | y＝0.3394x+531.87 | Y＝0.3337x+533.77 | y＝0.335x+534.75 |
| R 2 | 0.9781 | 0.9606 | 0.9815 |
(5) The denitrification rate was calculated and N in a well-done aquaculture water was determined as shown in Table 4 2 The rate of change of concentration was 0.3337. Mu. MolN 2 -N·L -1 ·h -1 -0.3394μmolN 2 -N·L -1 ·h -1 The different spots have little spatio-temporal variability (CV = 0.89%);
TABLE 4N of a normally mature aquaculture water 2 Rate of change of concentration
Coating the culture column with N in water 2 The rate of change of concentration multiplied by the volume of overlying water V =1.10 (L), divided by the internal diameter area of the column S =0.005 (m) 2 ) To obtain the denitrification rate D e (μmol N 2 ·m -2 ·h -1 ):
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 (4)
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 sample obtained in S3 2 Concentration, N for different incubation times 2 Performing linear regression on the concentration and the culture time to obtain a regression linear equation;
s5: calculating the unit denitrification rate of suspended matters in the water body and the denitrification rate of sediments;
the column samples comprise a first column sample, a second column sample, a third column sample and a fourth column sample, the concentration of suspended matters in the first column sample and the third column sample is the same, the concentration of suspended matters in the second 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 components and mass;
the regression linear equation in step S4 is Y = cX + b, where the slope c is N in the column sample 2 Rate of change of concentration in mmol N 2 - N × L -1 × h -1 ；
Denitrification Rate D in column sample e Comprises the following steps:
v represents the volume of the water body in the column sample, and S represents the inner diameter area of the column sample;
in step S5, the unit denitrification rate D of the water body suspended solids ss Comprises the following steps:
Wherein D is e1 Is the denitrification rate in the first column sample, D e2 Is the denitrification rate in the second sample, D e3 Is the denitrification rate in the third column, D e4 Is the denitrification rate in the fourth column; m is 1 Represents the amount of suspended matter in the first column sample, m 2 Represents the mass of suspended matter in the second column, m 3 Represents the mass of suspended matter in the third column, m 4 Represents the amount of suspended matter in the fourth column;
the water body simulation device comprises a simulation culture barrel, a driving assembly and a collection device; wherein the top of the simulated culture barrel is open and is filled with the upper water, and the bottom is provided with a fixed position; the driving component is arranged on an installation column arranged in the simulation culture bucket and is provided with a motor and a magnetic force rotating part connected with the motor; collection system places below the liquid level of the water-covering on fixed position and in the submerged simulation cultivation bucket, including the uncovered and one end in both ends be equipped with sealed lid, the other end is equipped with the cultivation post of sealed plug, one end is fixed in sealed covering, the other end is towards cultivateing the magnetic stirring mechanism that extends in the post and link up side by side and set up in inlet tube and the outlet pipe of sealed lid, the export of outlet pipe is connected with the injector that the membrane advances kind mass spectrograph, the import and the supply bottle of inlet tube communicate mutually, all be equipped with the stagnant water clamp on inlet tube and the outlet pipe, magnetic stirring mechanism rotates along with the rotation of magnetic force rotation piece, simulation cultivation bucket internal face is equipped with a plurality ofly along radial orientation the circumference interval the baffle that the outer wall of erection column extends, the baffle will separate into a plurality of not communicating compartments in the simulation cultivation bucket, arbitrary the bottom of compartment is equipped with threely fixed position to be convenient for do the parallel experiment under the same environment, correspond every on the outer wall of simulation cultivation bucket be equipped with a delivery port.
2. The determination 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 measured by using a non-disturbance sediment sampler, and collecting an undisturbed sediment column sample with a surface layer of 0-10cm to ensure that the sediment keeps the original structure and level and the water on the column sample keeps overflowing.
3. The method according to claim 2, wherein the pre-incubation in step S2 is carried out by immersing the column sample in a vessel filled with in-situ water for 4 to 6 hours, the water level in the vessel being 6cm or more above the upper surface of the column sample.
4. The method according to claim 3, wherein in step S5, the denitrification rate D of the sediment Mud Comprises the following steps:
wherein H is the height of the water in the column sample, and H is the total height of the water and the sediments in the column sample.
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| N2∶Ar法直接测定淹水环境反硝化产物N2的产生速率;李晓波等;《农业环境科学学报》;20130630;第1285页右栏倒数第1-2段，第1286页图1，第1288页左栏第1段 * |
| 鄱阳湖水体悬浮物反硝化潜力模拟研究;姚晓龙等;《中国环境科学》;20150320(第03期);第853页右栏第2段 * |
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