CN113075345B - Method for analyzing and determining whether aerobic high-temperature compost is decomposed stably - Google Patents
Method for analyzing and determining whether aerobic high-temperature compost is decomposed stably Download PDFInfo
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- CN113075345B CN113075345B CN202110345955.0A CN202110345955A CN113075345B CN 113075345 B CN113075345 B CN 113075345B CN 202110345955 A CN202110345955 A CN 202110345955A CN 113075345 B CN113075345 B CN 113075345B
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
Abstract
The invention discloses a method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting a high performance liquid chromatography, which comprises the steps of extracting organic matters in a sample of biomass raw material aerobic high-temperature compost at any sampling time by using an organic extracting agent to obtain an extracting solution, analyzing organic matter components in the extracting solution by using the high performance liquid chromatography to obtain an HPLC spectrogram of the organic matters in the compost, and determining the content proportion of the organic matters with different polarities according to the area percentage of peak areas at different peak emergence times in the spectrogram; and (3) analyzing and comparing HPLC spectrograms of organic matters in the compost at any sampling time, and determining that the tested compost is decomposed stably when the content proportion of the organic matters with different polarities reaches a certain value. Compared with the conventional method for judging whether the compost is rotten and stable by monitoring the temperature, the seed germination index and the like in the prior art, the method provided by the invention is a more accurate and convenient method for judging whether the compost is rotten and stable in nature.
Description
Technical Field
The invention relates to a method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting a high performance liquid chromatography, belonging to the technical field of livestock and poultry manure resource utilization.
Background
In recent years, with the increasing national economy and the continuous improvement of the living standard of residents, the market demand of livestock and poultry is increased, which promotes the development of livestock and poultry breeding to a certain extent. The livestock and poultry breeding mode is gradually changed from decentralized feeding to regionalization, mechanization and intensification, and the expansion speed of the livestock and poultry breeding mode is gradually increased year by year. Meanwhile, although the livestock and poultry breeding method meets the market and economic requirements, the amount of the livestock and poultry excrement discharged in the process is very large, and the livestock and poultry excrement also has unpleasant pungent odor. Relevant investigations have shown that the total amount of livestock and poultry manure discharged nationwide is as high as 19.00 million tons in 2010, of which about 2.27 million tons are serious pollution sources. If the fertilizer is applied to farmlands, the average manure pollution amount per hectare of farmlands can reach 1.86 tons. If no strong policy is provided for pollution control of livestock and poultry manure, the total pollution amount is further expanded and is predicted to increase to 2.98 hundred million tons in 2020. The national first pollution source census bulletin shows that the chemical oxygen demand discharged by livestock and poultry breeding in 2010 reaches 1268.26 ten thousand tons, and accounts for 96 percent of the total amount discharged by agricultural sources; the total nitrogen and phosphorus emissions were 102.48 and 16.04 million tons, accounting for 38 and 56% of the total agricultural source emissions, respectively. Currently, the relevant data of the Ministry of agriculture shows that the livestock and poultry waste production amount is about 38 hundred million tons every year in China. A large amount of livestock and poultry manure is discharged into the environment without professional environment-friendly treatment, so that heavy burden is brought to ecological environment and social environment and even serious threat is brought to human health.
Relevant researches show that composting treatment is a convenient treatment mode for agricultural waste reduction, harmlessness and recycling. Under the condition of manual control, animal and plant remains and excrement containing fertilizer components are added with soil and mineral substances and then are mixed and stacked, and under the action of microorganisms at high temperature and high humidity, organic macromolecular substances are decomposed into micromolecular substances which can be absorbed and utilized by crops, so that the process is a mineralization, humus and harmless process of organic matters. The decomposed compost can not only improve the yield and the quality of crops, but also increase the organic matter content of soil and improve the physical and chemical properties of the soil. However, after the non-decomposed compost is applied to soil, a large amount of oxygen in the root soil is consumed due to the strong decomposition of organic matters, and toxic substances such as organic acids and the like are generated to inhibit the growth of crops. At present, much research is carried out on the maturity at home and abroad, and the temperature, the germination index of seeds, the pH value, the C/N ratio and the like are common maturity judgment indexes. However, most studies show that most of the indicators can only be used as reference indicators for compost stability and maturity rather than as independent indicators. Therefore, whether the compost sample is decomposed stably can be judged more accurately and conveniently, and the method has great significance for practical production application and scientific research.
Disclosure of Invention
In order to solve the problems of more accurately and conveniently judging whether compost is decomposed stably and saving the cost of practical production application and scientific research, the invention provides a method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting a high performance liquid chromatography.
The method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting the high performance liquid chromatography is characterized by comprising the following steps of: extracting organic matters in the aerobic high-temperature compost of the biomass raw material by using an organic extractant to obtain an extracting solution; then analyzing the organic matter components in the extracting solution by high performance liquid chromatography to obtain HPLC spectrograms of the organic matter in the samples of the aerobic high-temperature compost at different sampling times, judging the polarity performance of the organic matter in the compost material according to the peak-out time in the spectrograms, and determining the content proportion of the organic matter with different polarities according to the area percentage of peak areas at different peak-out times in the spectrograms; analyzing and comparing HPLC spectrograms of organic matters in samples at different sampling times, and judging whether the determined compost is mature and stable or not according to the change rule of the content proportion of the organic matters with different polarities; and when the content proportion of the organic matters with different polarities reaches a certain value, determining that the measured compost is decomposed stably.
The method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting the high performance liquid chromatography is characterized in that in the step one, the biomass raw materials are livestock and poultry manure, plant straws, plant residues, agricultural product processing waste, biological fermentation filter residues, kitchen waste, rural perishable waste and the like.
The method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting the high performance liquid chromatography is characterized in that the organic extracting agent is n-butyl alcohol, and the process of extracting and extracting organic matters in a sampling sample comprises the following steps: adding n-butanol into the sample, ultrasonic extracting for 60-120 min, vacuum filtering, filtering the filtrate with 0.45 μm organic filter membrane, concentrating, re-dissolving the concentrate with mobile phase of high performance liquid chromatography, and analyzing with high performance liquid chromatography.
The method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting the high performance liquid chromatography is characterized in that the use conditions of the high performance liquid chromatograph are as follows: the chromatographic column is a C18 column (5 μm, 4.6X 250 mm), and a gradient elution procedure is adopted, methanol and water are used as mobile phases, and the flow rate is kept at 0.8 mL/min; the column temperature is 25 ℃; the sample volume is 20 mu L; PDAD ultraviolet scanning, and setting 245 nm as a detection wavelength.
Further, the steps of using a gradient elution procedure are: the volume ratio of the methanol to the water is =1:9 within 0-20 min of retention time; the volume ratio of the methanol to the water is =3:7 within the retention time of 20-40 min; and the volume ratio of the methanol to the water is =9:1 within the retention time of 40-160 min.
The method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting the high performance liquid chromatography is characterized in that the polarity types of organic matters in compost materials are divided into strong polarity, medium polarity, weak polarity and non-polarity according to the peak emergence time in the high performance liquid chromatogram, the peak emergence time is 0-20 min as a strong polarity matter peak, 21-40 min as a strong polarity matter peak, 41-60 min as a medium polarity matter peak, 61-80 min as a weak polarity matter peak and 81-160 min as a non-polarity matter peak.
The method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting the high performance liquid chromatography is characterized in that the proportion of the peak area of the strong polar substance peak in the total peak area is less than 12%, and meanwhile, the proportion of the peak area of the medium polar substance peak in the total peak area is more than 70%, and the measured compost is determined to be decomposed stably.
Compared with the prior art, the invention has the following beneficial effects:
the essence of the composting process is the process of organic matter stabilization and humification. Along with the prolonging of the composting time, the glucide and the like are easy to degrade organic components, lignin and the like and gradually convert to macromolecular humus with high stability. The invention provides a method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting a high performance liquid chromatography, which can judge whether the aerobic high-temperature compost is decomposed stably according to the composition of organic substances in the compost. Compared with the conventional method for judging the compost maturity time by monitoring the temperature, the seed germination index and the like in the prior art, the method provided by the invention is a method for judging whether the compost is stably matured or not more accurately and conveniently in nature.
Drawings
FIG. 1 is a graph comparing the composting temperatures of the control group and the test group with the change trend of the composting time;
in fig. 1: Δ represents a control group result, o represents a test group result, and ● represents a day-average air temperature;
FIG. 2 is a graph comparing the germination percentage of plant seeds when the plant seeds are cultured with fertilizers of the control group and the test group at different composting times according to the present invention;
in fig. 2: Δ represents the control group results, and o represents the test group results;
FIG. 3 is a comparison graph of the proportion of organic substances with different polarities in fertilizers of a control group and a test group of the invention when the fertilizers at different composting times are subjected to component analysis;
in FIG. 3: panel a shows the control results and panel b shows the test results.
Detailed Description
The present invention is further illustrated by the following examples, which should not be construed as limiting the scope of the invention.
Example 1:
1) compost raw materials to be tested:
fresh pig manure is taken from a certain scale pig farm in Hangzhou city, and the water content is close to 80 percent.
2) Regulating and controlling the water content of the compost raw materials:
control group: adding wood chips into fresh pig manure to enable the mass content of the wood chips in the final mixture to be 20%;
test groups: spreading fresh pig manure in a maggot groove padded with a base material, and then uniformly scattering a certain amount of fly maggot inoculant on the surface of the manure to allow the fly maggots to freely enter a manure pile for feeding and growing; wherein the housefly larva inoculant is a mixture of wheat bran (used for hatching housefly eggs) and housefly first-instar larvae (not more than 24 hours after hatching), each gram contains about 3000 housefly first-instar larvae, and the inoculation amount of the housefly larva inoculant is 0.75 percent of the total mass of fresh pig manure. And the residue (maggot manure) left after 8 d cultivation is carried out according to the cultivation process of DB 33/T2149-2018 standard is used as a compost raw material, and the moisture content of the maggot manure is about 65 percent after the maggot manure is subjected to biological conversion.
3) Composting treatment:
composting the fertilizers obtained from the control group or the test group in the step 2) respectively, wherein the process is as follows:
the method comprises the steps of stacking the fertilizer into a strip-shaped stack body with the length of 2.0 m, the width of 1.5 m and the height of 1.0 m, controlling the ventilation condition by adopting a manual stack turning measure, turning the stack once every four days during the temperature rise period of the stack body, turning the stack once every 2-3 days during the high-temperature period, immediately turning the stack when the temperature of the stack body exceeds 65 ℃ during the high-temperature period, and controlling the temperature of the high-temperature period to be not higher than 75 ℃. And turning the stack once every 7 days after the stack starts to cool. Samples were taken at compost 0d, 2d, 13d, 19d, 22d, 39d, 52d, 55d, 61d, 70d, 72d, 87d, 92d, respectively.
4) The determination method comprises the following steps:
ambient temperature: the temperature of the air around the stack was measured with a wall thermometer;
composting temperature: during the conversion of pig manure fly maggots, inserting a mercury thermometer into the fertilizer pile body to measure the temperature at a depth of 4 cm, and randomly measuring 3 points; during aerobic high-temperature composting, 3 mercury thermometers with a length of 50 cm were inserted daily into the surface of each heap to a depth of 30 cm.
Germination index testing process: adding 20 g of sample into 200 mL of distilled water, oscillating for 60 min, filtering supernatant by using slow filter paper, and obtaining filtrate, namely compost leaching liquor, wherein the filtrate is for later use; and (3) paving one piece of filter paper with a corresponding size in a 9 cm culture dish, uniformly putting 15 pakchoi seeds, and sucking 5 by using a pipette.0 mL of compost extract was placed in a petri dish and distilled water was used as a control. Each treatment was repeated 3 times. Then placing in dark at (25 + -1) deg.C for 48 hr, and determining germination rate and root length according to formula
The germination index GI (%) was calculated.
Organic matter polarity change: and (3) analyzing the sample in the step 3), weighing 5 g (accurately recording to 0.0001 g) of sample of the compost material, placing the sample in a 250 ml conical flask, adding 30 ml of n-butyl alcohol as an extracting agent, performing ultrasonic leaching for 80 min, performing suction filtration by a Buchner funnel, filtering the filtrate by a 0.45-micron organic system filter membrane, concentrating the filtrate at 105 ℃, re-dissolving the concentrate by using a mobile phase of a high performance liquid chromatography, and analyzing by using a high performance liquid chromatograph. The conditions of the HPLC are as follows: the chromatographic column is a C18 column (5 μm, 4.6X 250 mm), and a gradient elution procedure is adopted, methanol and water are used as mobile phases, and the flow rate is kept at 0.8 mL/min; the volume ratio of the methanol to the water is =1:9 within 0-20 min of retention time; within the retention time of 20-40 min, the volume ratio of methanol to water is =3: 7; the volume ratio of the methanol to the water is =9:1 within 40-160 min of retention time; the column temperature is 25 ℃; the sample size was 20. mu.L. PDAD ultraviolet scanning, and setting 245 nm as a detection wavelength.
5) And (3) test results:
FIG. 1 is a graph comparing the composting temperature trend with the composting time for the control group and the test group.
As can be seen from figure 1, the temperature of the pig manure compost is raised from 26 ℃ to 51 ℃ in the temperature raising period of 0-17 days; the reactor temperature is in a small-amplitude fluctuation state within 18-24 days. After 25 days, the temperature of the pile gradually becomes stable and is kept at about 50 ℃. Then entering a cooling period at about 38 days of composting. According to the national standard GB 7959-87 for composting of Chinese excrement, when the composting temperature is kept above 50-55 ℃ for 5-7 days, the excrement can be subjected to harmless treatment. Therefore, the pig manure compost reaches the rotten requirement after 38 days.
As can be seen from figure 1, the whole compost fermentation process of maggot manure can be divided into four stages of maggot treatment period, high temperature period, medium temperature period and stabilization period. The temperature of the manure pile is slowly increased in the maggot treatment stage of 0-8 days, after the maggot manure is separated and aerobic high-temperature composting is carried out, the temperature of the pile body is rapidly increased, and the temperature is kept at 55-60 ℃ within 10-20 days, so that the compost sterilization requirements are completely met.
The study of scholars shows that when the germination index of the seeds exceeds 60 percent, the compost is decomposed. FIG. 2 shows the germination index trend of Chinese cabbage seeds. It can be seen that the seeds did not germinate neither for the first 6 days of fly maggot treatment nor for the first 20 days of pig manure composting, indicating the presence of phytotoxicity. The phytotoxicity of the two composts gradually decreased and the germination index gradually increased with time. Wherein, the germination index of the maggot manure compost is more than 60 percent about 20 days, and the germination index of the pig manure compost is more than 60 percent about 40 days, so as to reach a rotten stable state.
The essence of the composting process is the process of organic matter stabilization and humification. Along with the prolonging of the composting time, the glucide and the like are easy to degrade organic components, lignin and the like and gradually convert to macromolecular humus with high stability. According to the difference of the peak appearing time in the high performance liquid chromatogram, the organic substances are divided into 5 categories of strong polarity (A), strong polarity (B), medium polarity (C), weak polarity (D) and non-polarity (E), wherein the peak appearing time of A is 0-20 min, that of B is 21-40 min, that of C is 41-60 min, that of D is 61-80 min, and that of E is 81-160 min. The partition ratio (i.e., percentage) of the amount of each substance is equal to the proportion of the area of the substance in the total area of the substance. As shown in FIG. 3, the variation of the strongly polar and moderately polar substances with the composting time is large, and the strongly polar organic substances generally show a downward trend, and the moderately polar organic substances generally show an upward trend. With the time being prolonged, the proportion of the strong polarity organic matters and the proportion of the medium polarity organic matters in the maggot manure on the 20 th day of the compost and the proportion of the pig manure on the about 40 th day of the compost both meet the requirements that the proportion of the strong polarity organic matters is less than 12 percent and the proportion of the medium polarity organic matters is more than 70 percent, which indicates that the compost treatment has reached a rotten and stable state at the moment. This conclusion was also validated against compost temperature and seed germination index analysis.
In the figures 1, 2 and 3, the composting time of the test group is divided into two stages, the first stage corresponds to 0-8 days of the maggot bioconversion process in the step 2) of the example 1, and the second stage corresponds to the composting process in the step 3) of the example 1.
The statements in this specification merely set forth a list of implementations of the inventive concept and the scope of the present invention should not be construed as limited to the particular forms set forth in the examples.
Claims (2)
1. A method for analyzing and determining whether aerobic high-temperature compost is decomposed stably by adopting a high performance liquid chromatography is characterized by comprising the following steps: extracting organic matters in the aerobic high-temperature compost of the biomass raw material by using an organic extractant to obtain an extracting solution; then analyzing the organic matter components in the extracting solution by high performance liquid chromatography to obtain HPLC spectrograms of the organic matter in the samples of the aerobic high-temperature compost at different sampling times, judging the polarity performance of the organic matter in the compost material according to the peak-out time in the spectrograms, and determining the content proportion of the organic matter with different polarities according to the area percentage of peak areas at different peak-out times in the spectrograms; analyzing and comparing HPLC spectrograms of organic matters in the compost at different sampling time, and judging whether the measured compost is mature and stable or not according to the change rule of the content proportion of the organic matters with different polarities; when the content proportion of the organic matters with different polarities reaches a certain value, the measured compost is determined to be stably decomposed;
the organic extractant is n-butanol, and the process of extracting the organic matters in the compost sample comprises the following steps: adding n-butanol into sample, ultrasonic extracting for 60-120 min, vacuum filtering, filtering the filtrate with 0.45 μm organic filter membrane, concentrating, re-dissolving the concentrate with mobile phase of high performance liquid chromatography, and analyzing with high performance liquid chromatograph;
the operating conditions of the high performance liquid chromatograph are as follows: the chromatographic column is a C18 column with the diameter of 5 μm and the diameter of 4.6 multiplied by 250 mm, a gradient elution program is adopted, methanol and water are used as mobile phases, and the column temperature is 25 ℃; the sample volume is 20 mu L; PDAD ultraviolet scanning, and setting 245 nm as a detection wavelength;
the steps of the gradient elution program are as follows: the flow rate of the mobile phase was maintained at 0.8 mL/min; the volume ratio of the methanol to the water is =1:9 within 0-20 min of retention time; the volume ratio of the methanol to the water is =3:7 within the retention time of 20-40 min; the volume ratio of the methanol to the water is =9:1 within 40-160 min of retention time;
dividing the polarity types of organic matters in the compost materials into strong polarity, medium polarity, weak polarity and non-polarity according to the peak-off time in the high-performance liquid chromatogram, wherein the peak-off time is 0-20 min as a strong-polarity matter peak, 21-40 min as a strong-polarity matter peak, 41-60 min as a medium-polarity matter peak, 61-80 min as a weak-polarity matter peak, and 81-160 min as a non-polarity matter peak;
the proportion of the peak appearance area of the strong polar substance peak in the total peak appearance area is less than 12%, and the proportion of the peak appearance area of the medium polar substance peak in the total peak appearance area is more than 70%, and the detected compost is determined to be thoroughly decomposed and stable.
2. The method for determining whether aerobic high-temperature compost is decomposed stably by high performance liquid chromatography analysis as claimed in claim 1, wherein the biomass raw material is at least one of livestock and poultry manure, plant straw, plant residue, agricultural product processing waste, biological fermentation residue, kitchen waste, rural perishable waste.
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