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Improved nitrobacteria Diphenylamine-Griess detection method

A technology of nitrifying bacteria and detection methods, applied in the field of environmental chemistry and environmental microorganisms, can solve problems such as residues, and achieve the effects of improving reaction rate and conversion rate, and improving accuracy and timeliness.

Inactive Publication Date: 2011-06-15
LIAONING BEIFANG ENVIRONMENTAL PROTECTION +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of 5-8 drops of acetic acid during the test does not guarantee the NO 2 - fully converted to N 2 , all test tubes turned red after adding Griess reagent, indicating that there was NO 2 - remnant

Method used

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  • Improved nitrobacteria Diphenylamine-Griess detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0017] 1. Preparation of culture medium

[0018] (1) Liquid culture medium for nitrifying bacteria: 1.0g NaNO 2 , 1.0g Na 2 CO 3 , 0.25g NaH 2 PO 4 , 1.0g CaCO 3 , 0.75g K 2 HPO 4 , 0.01g MnSO 4 , 0.03g MgSO 4 4H 2 O, 1000mL distilled water, pH7.2; sterilize at 121°C for 30min.

[0019] (2) Enrichment culture: According to the formula of the culture medium, weigh the drug, prepare 1000ml culture medium in the Erlenmeyer flask, adjust the pH to 7.2, and then divide it into test tubes, 90ml, 121°C, autoclave sterilization 30min. Put 10 mL of the collected mud sample into a previously sterilized Erlenmeyer flask containing glass beads and 90 mL of culture medium, and culture it on a shaker at 15°C at 160 r / min for 10 days. The purpose is to break up the bacteria micelles, make the bacteria dispersed in the water in a single-cell state, and at the same time increase the number of nitrifying bacteria in the mud sample.

[0020] (3) Preparation of nitrifying bacteria cu...

Embodiment 2

[0024] The difference from Example 1 is:

[0025] First add 5-8 drops of hydrochloric acid to acidify the culture solution, then add 25-27 ammonium sulfamate, and react at 55-60°C until the color of the medium is colorless and there is gas at the bottom of the culture solution escape, and when the degassing stops, add a grain of ammonium sulfamate to make NO 2 - fully converted to N 2 And escape, take the culture solution on the white porcelain color comparison plate, add 3-5 drops of Griess reagent, the culture solution is colorless, finally take the culture solution on the color comparison plate, add 3-5 drops of diphenylamine, the culture solution Appears blue.

[0026] The cultivation of the nitrifying bacteria is to cultivate the nitrifying bacteria in the nitrifying bacteria culture solution for 15 days at 20° C. and 150 r / min, break up the bacteria micelles, and disperse the bacteria into a single-cell state in water, while increasing the number of nitrifying bacteri...

Embodiment 3

[0028] The difference from Example 1 is:

[0029] First add 5-8 drops of hydrochloric acid to acidify the culture solution, then add 28-30 ammonium sulfamate, and react at 60°C until the color of the medium is colorless, and gas escapes from the bottom of the culture solution , when the degassing stops, add a grain of ammonium sulfamate to make NO 2 - fully converted to N 2And escape, take the culture solution on the white porcelain color comparison plate, add 3-5 drops of Griess reagent, the culture solution is colorless, finally take the culture solution on the color comparison plate, add 3-5 drops of diphenylamine, the culture solution Appears blue.

[0030] The cultivation of the nitrifying bacteria is to cultivate the nitrifying bacteria in the nitrifying bacteria culture solution for 13 days at 30° C. and 155 r / min, break up the bacteria micelles, disperse the bacteria in the water in a single cell state, and simultaneously increase the number of nitrifying bacteria ...

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Abstract

The invention relates to the fields of environmental chemistry and environmental microbiology, in particular to an improved nitrobacteria Diphenylamine-Griess detection method. The method specifically comprises the following steps of: adding 5 to 8 drops of hydrochloric acid into culture solution in which nitrobacteria are cultured to acidize the culture solution; adding ammonium sulfamate; reacting at the temperature of between 50 and 60 DEG C until a culture medium is colorless and gas escapes from the bottom of the culture solution; when gas escape stops, adding ammonium sulfamate again to convert NO2<-> into N2 and allowing the N2 to escape; and adding 3 to 5 drops of diphenylamine into the culture solution, wherein the culture solution is blue. The NO2<-> can be removed from the culture solution by the method, so that the reaction rate and conversion efficiency of the NO2<-> in the reaction system can be improved greatly.

Description

technical field [0001] The invention relates to the fields of environmental chemistry and environmental microorganisms, in particular to an improved detection method for nitrifying bacteria Diphenylamine-Griess. Background technique [0002] Chromogenic reaction is a technique commonly used in chemistry and biology to detect chemical substances. NO in the nitrifying bacteria culture solution 3 - detection, the core content of which is the NO in the culture medium 2 - completely and completely removed. Aiming at this problem, predecessors have made many attempts, but the effect is not satisfactory. In the detection of nitric acid bacteria, the chromogenic agent diphenylamine (Diphenylamine) reacts to NO2- and NO 3 - The specificity is not enough, with NO 2 - and NO 3 -The reaction will all be blue, therefore, the remaining NO in the medium 2 - will affect the measurement results. NO 2 - The original concentration is 0.1mmol / L, when NO can not be guaranteed 2 - ...

Claims

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Application Information

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IPC IPC(8): G01N21/78C12Q1/04
Inventor 赵军郎咸明张巍李晓东匡德舜
Owner LIAONING BEIFANG ENVIRONMENTAL PROTECTION
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