CN108872528A - A kind of deposit nitrogen cycle rate determination method based on continuous process - Google Patents

A kind of deposit nitrogen cycle rate determination method based on continuous process Download PDF

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Publication number
CN108872528A
CN108872528A CN201710338794.6A CN201710338794A CN108872528A CN 108872528 A CN108872528 A CN 108872528A CN 201710338794 A CN201710338794 A CN 201710338794A CN 108872528 A CN108872528 A CN 108872528A
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deposit
nitrogen
control group
continuous process
nitrogen cycle
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俞慎
丁晶
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University of Chinese Academy of Sciences
Institute of Urban Environment of CAS
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University of Chinese Academy of Sciences
Institute of Urban Environment of CAS
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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Abstract

The deposit nitrogen cycle rate determination method based on continuous process that the invention discloses a kind of, the nitrogen cycle process applied to ecological environment field measure.Its detailed process is：The fresh deposit of 10 equal portions is weighed in corresponding 100mL serum bottle；Serum bottle headspace gas is replaced into helium using vacuum bottle washing system after sealing:Oxygen ratio is 2:1 mixed gas；Urea and deionized water are added respectively；25oC constant temperature incubation 120 hours；Head space gaseous state nitrogen content and deposit activity nitrogen content are measured respectively；According to processing group and control group various nitrogen forms content, deposit denitrification, nitrification and ammonifying process activity are calculated using respective formula.It is an advantage of the invention that：It is easy to operate, reduce traditional substep measuring method bring time cost；It is based on continuous process simultaneously, it is accurate to measure each process reaction rate of deposit nitrogen cycle, and the conspiracy relation of three continuous process is reflected, avoid the measurement of substrate for induction method from over-evaluating deposit nitrogen cycle rate.

Description

A kind of deposit nitrogen cycle rate determination method based on continuous process
Technical field
The invention belongs to ecological environment research fields, and in particular to a kind of deposit nitrogen cycle rate based on continuous process Measuring method.
Background technique
Nitrogen cycle is the ecosystem processes closely related with quality of water environment, Atmospheric Chemistry and Primary Production, is weight The ecosystem function wanted is related to the production service of the ecosystem.To nitrogen cycle and its active measurement quilt of related microorganisms It is widely used in the adaptation for characterizing each ecosystem function and the ecosystem to external interference.Currently, conventional soil and deposit nitrogen Cycle rate measurement is completed with substep substrate for induction method, i.e., with urea（Or amino acid）, ammoniacal nitrogen and nitrate be substrate, Culture measures its reacting final product yield after a certain period of time（Ammoniation potentiality measure NH4 +Yield, nitrification measure NO3 -Or Person NO3 -+NO2 -Yield, denitrification measure N2Or N2+N2O yield）Or disappearance of substrate amount.Or not add substrate direct Culture measures each step reacting final product yield or the original disappearance of substrate amount of medium, i.e. surrounding medium nitrogen cycle background after a certain period of time Rate determination.Conventional method substep measures, and may over-evaluate whole deposit（Or soil）Nitrogen cycle rate, and can not embody not With the continuity of process.
Summary of the invention
Traditional nitrogen cycle rate substep measuring method there are aiming at the problem that, the technical problems to be solved by the invention are exactly A kind of deposit nitrogen cycle rate determination method based on continuous process is provided.
The technical problem to be solved is that technical solutions in this way to realize for the method for the present invention, weighs the deposition of homogeneous The fresh sample of object (being about as much as 10g dry weight) is packed into 100mL serum bottle, seals；The headspace gas for closing serum bottle is set It is changed to helium:Oxygen ratio is 2:1 mixed gas, the oxygen content of the big air-water-sediment of simulated environment；10mL urea is molten Liquid (urea nitrogen containing 1mg) and 10mL deionized water are injected respectively enters processing group and control group serum bottle；Serum bottle is in incubator In 25 DEG C utilize N in gas chromatograph for determination serum bottle headspace gas immediately after culture 120 hours2And N2O concentration；Then it is added The KCl solution of 2M vibrates 60 minutes extraction NH4 +- N and NO3 -+NO2 -- N measures its concentration with flow injection analyzer；And pass through Three formula calculate separately denitrification, nitrification and the ammoniation successive reaction rate of deposit below：
Denitrification activity (mg N kg-1dw hr-1)
=[(N2+N2O)Processing group-(N2+N2O)Control group]/T*DW
Nitrification activity (mg N kg-1dw hr-1)
={ [(N2+N2O)Processing group-(N2+N2O)Control group]+[(NO3 -+NO2 -)Processing group-(NO3 -+NO2 -)Control group]}/T*DW
Ammoniation activity (mg N kg-1dw hr-1)
={ [(N2+N2O)Processing group-(N2+N2O)Control group]+[(NO3 -+NO2 -)Processing group-(NO3 -+NO2 -)Control group]+[(NH4 +)Processing group-(NH4 +)Control group]}/T*DW
(N in formula2+N2It O is) N in headspace gas2+N2O-N content (milligram)；(NO2 -+NO3 -) and (NH4 +) it is KCl in deposit Extract corresponding form N content (milligram)；T is incubation time (hour)；DW is deposit dry weight (kilogram).
Due to using above-mentioned experimental program, the method for the present invention is had the following advantages that：It is easy to operate, reduce traditional substep Measure the demand of denitrification, nitrification and ammoniation bring time cost and deposit full-page proof amount；It is based on continuous process simultaneously, Each process reaction rate of deposit nitrogen cycle can be accurately measured, and reflects the conspiracy relation of three continuous process, is avoided single Deposit nitrogen cycle rate is over-evaluated in the measurement of process substrate for induction method.Add urea can for amino acid or15N marks organic nitrogen bottom Object replaces.This method can also be used for the measurement of pedotheque.
Detailed description of the invention
Fig. 1 is the key step flow chart of the method for the present invention.
Specific embodiment
The method of the present invention is described further with reference to the accompanying drawings and embodiments：
Deposit nitrogen cycle measuring method specific embodiment of the present invention based on continuous process as shown in Figure 1, include with Lower step：
Step 1：Weigh respectively 10 equal portions mix after the fresh sample of deposit (be about as much as 10g dry weight, mixed in sample sack, Sack mixing should not be opened) in 10 100mL serum bottles, it is sealed with silica gel plug and aluminium lid；
Step 2：Serum bottle after sealing vacuumizes immediately, and washes 3 times with helium to remove N2, it is then injected into 50ml O2, put down Weighing apparatus discharges overpressure gases in serum bottle head space, helium in headspace gas after five minutes, using syringe:Oxygen is finally than being 2:1；
Step 3：10mL urea liquid (urea nitrogen containing 1mg) and 10ml deionized water are injected respectively into processing with syringe In group and control group (wherein every group of sample there are 5 repetitions) serum bottle, all serum bottles are placed on to 25 DEG C of water bath with thermostatic control culture It is cultivated 120 hours in case；
Step 4：After culture, be equipped with immediately electron capture detector (ECD) (ECD), thermal conductivity detector (TCD) (TCD) and flame from N in the gas chromatograph for determination headspace gas of sonization detector (FID)2And N2Then O content is respectively added in each serum bottle 40mL 2M KCl solution, gyrate shaker 200rpm vibrate 60 minutes, and 1000rpm is centrifuged 10 minutes, with 0.45 μm of acetic acid fibre Film filtering supernatant is tieed up, measures NH in measurement filtered fluid using Flow Analyzer4 +- N and NO3 -+NO2 -- N concentration；
Step 5：Denitrification, nitrification and the ammoniation activity of deposit are calculated separately by following three formula：
Denitrification activity (mg N kg-1dw hr-1)
=[(N2+N2O)Processing group-(N2+N2O)Control group]/T*DW
Nitrification activity (mg N kg-1dw hr-1)
={ [(N2+N2O)Processing group-(N2+N2O)Control group]+[(NO3 -+NO2 -)Processing group-(NO3 -+NO2 -)Control group]}/T*DW
Ammoniation activity (mg N kg-1dw hr-1)
={ [(N2+N2O)Processing group-(N2+N2O)Control group]+[(NO3 -+NO2 -)Processing group-(NO3 -+NO2 -)Control group]+[(NH4 +)Processing group-(NH4 +)Control group]}/T*DW
In formula, (N2+N2It O is) N in headspace gas2+N2O-N total amount (milligram)；(NO2 -+NO3 -) and (NH4 +) it is in deposit KCl extracts corresponding form N total amount (milligram)；T is incubation time (hour)；DW is deposit dry weight (kilogram).
In order to avoid pollution of the nitrogen to gas in serum bottle in air, gas displacement process described in this method is whole It carries out under water.Finally determine that 1 mg urea nitrogen of addition is that substrate concurrently sets 120 hours and is on the basis of test of many times result Incubation time come guarantee substrate addition nitrogen quantity to the minimal disturbances of system and each process rate measurement in exogenous nitrogen it is continuously anti- It answers under state.
This research can accurately measure nitrogen cycle process by the addition of substrate urea and the measurement to various nitrogen forms Middle denitrification, nitrification and ammoniation rate.The nitrogen cycle rate of control group deposit can also be directly calculated, The as deposit background nitrogen cycle rate.Substrate can for amino acid or15The organonitrogen compound of N label.

Claims (5)

1. a kind of deposit nitrogen cycle rate determination method based on continuous process, it is characterised in that：Its detailed process is：
Step 1：The fresh deposit of 10 equal portions is weighed in 100mL serum bottle according to deposition water content of matter, is sealed；
Step 2：Serum bottle headspace gas is replaced into helium using vacuum bottle washing system:Oxygen ratio is 2:1 mixed gas；
Step 3：1mg urea nitrogen is added with syringe respectively（Processing group）And deionized water（Control group）, 25oC constant temperature incubation 120 Hour；
Step 4：It is utilized respectively living in gas-chromatography and flow injection analyzer measurement head space gaseous state nitrogen content and deposit mud Property nitrogen（Ammonium state and nitrate nitrogen）Content；
Step 5：According to processing group and control group various nitrogen forms content, using respective formula calculate separately deposit denitrification, Nitrification and ammoniation rate.
2. a kind of deposit nitrogen cycle rate determination method based on continuous process according to claim 1, feature exist In：Headspace gas is replaced into helium in above-mentioned steps 2:Oxygen ratio is 2:1 mixed gas can effectively simulate normal atmosphere pressure The oxygen content of big air-water-sediment.
3. a kind of deposit nitrogen cycle rate determination method based on continuous process according to claim 1, feature exist In：It is 1mg urea nitrogen that substrate is added in above-mentioned steps 3, it is ensured that addition substrate nitrogen content is smaller to the disturbance of whole system.
4. a kind of deposit nitrogen cycle rate determination method based on continuous process according to claim 1, feature exist In：The constant temperature incubation time sets 120 hours in above-mentioned steps 3, it is ensured that each process of nitrogen cycle has carried out continuously additional nitrogen source Reaction.
5. a kind of deposit nitrogen cycle rate determination method based on continuous process according to claim 1, feature exist In：The denitrification of deposit, nitrification and ammoniation activity calculation formula are respectively in above-mentioned steps 5：
Denitrification activity (mg N kg-1dw hr-1)
=[(N2+N2O)Processing group-(N2+N2O)Control group]/T*DW
Nitrification activity (mg N kg-1dw hr-1)
={ [(N2+N2O)Processing group-(N2+N2O)Control group]+[(NO3 -+NO2 -)Processing group-(NO3 -+NO2 -)Control group]}/T*DW
Ammoniation activity (mg N kg-1dw hr-1)
={ [(N2+N2O)Processing group-(N2+N2O)Control group]+[(NO3 -+NO2 -)Processing group-(NO3 -+NO2 -)Control group]+[(NH4 +)Processing group-(NH4 +)Control group]}/T*DW
(N in formula2+N2It O is) N in headspace gas2+N2O-N amount (milligram)；(NO2 -+NO3 -) and (NH4 +) it is KCl in culture deposit Extract corresponding form N amount (milligram)；T is incubation time (hour)；DW is deposit dry weight (kilogram).
CN201710338794.6A 2017-05-15 2017-05-15 A kind of deposit nitrogen cycle rate determination method based on continuous process Pending CN108872528A (en)

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CN109828041A (en) * 2019-01-25 2019-05-31 北京建工环境修复股份有限公司 The measuring method of volatile organic matter total amount in a kind of underground water
CN114002361A (en) * 2021-11-02 2022-02-01 中国科学院南京土壤研究所 Dry land soil background N2Method for measuring discharge rate
CN114814184A (en) * 2022-05-10 2022-07-29 中国科学院城市环境研究所 Method for measuring degradation rate of degradable plastic based on carbon 13 isotope method

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109828041A (en) * 2019-01-25 2019-05-31 北京建工环境修复股份有限公司 The measuring method of volatile organic matter total amount in a kind of underground water
CN114002361A (en) * 2021-11-02 2022-02-01 中国科学院南京土壤研究所 Dry land soil background N2Method for measuring discharge rate
CN114814184A (en) * 2022-05-10 2022-07-29 中国科学院城市环境研究所 Method for measuring degradation rate of degradable plastic based on carbon 13 isotope method

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Application publication date: 20181123