Method for detecting free amino acid in biogas slurry

A free amino acid and detection method technology, applied in the field of comprehensive development of biogas slurry, can solve the problems of unreasonable and safe treatment and disposal, and achieve the effect of avoiding selectivity, strong operability and simple steps

Inactive Publication Date: 2016-05-04
TONGJI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • Method for detecting free amino acid in biogas slurry
  • Method for detecting free amino acid in biogas slurry
  • Method for detecting free amino acid in biogas slurry

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Embodiment 1

[0034] The biogas slurry discharged from the 6L sludge anaerobic digestion small test device with a solid content of 5% and 20% and the residual sludge fed into it were used as experimental materials. Pass all the biogas slurry above through a 0.22μm filter membrane to remove particulate matter, adjust the pH to 10.2 with 1M sodium hydroxide, take 5ml of the above biogas slurry in a 50ml rotary evaporating eggplant-shaped bottle, and vacuum rotary evaporation at 40 degrees Celsius to dryness To remove high-concentration ammonia nitrogen and volatile biogenic amines, redissolve with 5ml of 0.1M hydrochloric acid, and vortex for 30s to mix. Take 200 μL of the above-mentioned re-dissolution solution in a 0.5ML3KMliipore new ultrafiltration centrifuge tube, centrifuge at 13000g until all the liquid passes through, vortex mix and place at 4°C for use.

[0035] The above sample was automatically injected by Agilent HPLC1260 autosampler as follows: 0.5 μL mixed with 0.5 μL of OPA and...

Embodiment 2

[0048] The biogas slurry discharged from the anaerobic digestion test device with a solid content of 10% was used as the experimental material. Biogas slurry samples were filtered through a 0.22 μm membrane filter. Filtered biogas slurry with 1mol L -1 NaOH solution to adjust the pH to 10.2, evaporate to dryness with 0.1mol L -1 HCL was re-dissolved; the sample used in the strong cationic solid-phase extraction (SCX-SPE) experiment was filtered and passed through 1mol·L -1 HCL adjusted the pH to 2.2 for use.

[0049] (1): SCX solid phase extraction cartridge experiment;

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Abstract

The invention relates to a method which comprises the following steps: removing high-concentration ammonia nitrogen and volatile biogenic amine in a way of rotary evaporation after biogas slurry is regulated to be alkaline; removing macromolecular disruptors by applying a 3K Millipore ultra-filtration membrane; performing pre-column derivatization on amino acids by using OPA and FMOC derivatization reagents, so that the amino acid has fluorescence emission characteristics to improve the detection sensitivity and the separation and selection characteristics; then performing qualitative and quantitative analysis on 24 kinds of amino acids through a reverse-high-performance liquid chromatography. Compared with a conventional method, the method avoids the selectivity of an SCX-SPE (Strong Cation-Exchange Solid-Phase Extraction Column) for the amino acids caused by different isoelectric points of the amino acids and overcomes the defects of low recovery rate and poor reproducibility during sample determination. The method provided by the invention is simple in step and strong in operability and provides a technical basis for the study on a protein degradation mechanism in the process of anaerobic digestion and a scientific basis for biogas slurry and biogas residue agricultural resource development and utilization.

Description

technical field [0001] The invention relates to the technical field of comprehensive development of biogas slurry, relates to the technology required for pretreatment of samples, and a developed detection method adapted to the one-time analysis of 24 kinds of free amino acids in biogas slurry by modern detection instruments. Background technique [0002] At present, the amount of surplus sludge is increasing rapidly with the rapid development of urban sewage treatment capacity. As of 2013, the annual production of excess sludge in my country has reached 6.25 million tons (on a dry basis). Most of them have not been properly and safely handled and disposed of. The residual sludge is rich in nutrients such as nitrogen, phosphorus, and potassium, and the organic matter content can reach 50%-60%. Anaerobic digestion, a technology that achieves stabilization of sludge while generating biogas as an energy source, can be a suitable option for sludge treatment and disposal. In th...

Claims

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Application Information

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IPC IPC(8): G01N30/89G01N30/06G01N30/14
CPCG01N30/89G01N30/06G01N30/14G01N2030/146
Inventor 李宁刘文静李建华
Owner TONGJI UNIV
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