CN117517517A - Method for detecting long-chain fatty acid in anaerobic fermentation liquid - Google Patents
Method for detecting long-chain fatty acid in anaerobic fermentation liquid Download PDFInfo
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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
- G01N30/02—Column chromatography
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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
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
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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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- G01N30/28—Control of physical parameters of the fluid carrier
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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
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
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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
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
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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
- G01N30/02—Column chromatography
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- G01N30/30—Control of physical parameters of the fluid carrier of temperature
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Abstract
The invention relates to the technical field of analysis and detection, in particular to a method for detecting long-chain fatty acid in anaerobic fermentation liquid. Comprising the following steps: the anaerobic fermentation liquid is subjected to freeze vacuum drying and dehydration treatment to obtain a sample; mixing the sample with an extractant, extracting, and separating a liquid phase; the volume ratio of the extractant is 20-30: 1 and concentrated hydrochloric acid, and then detecting the liquid phase by gas chromatography-mass spectrometry. The invention establishes a gas chromatography-mass spectrometry detection method for long-chain fatty acid in anaerobic fermentation liquid. The detection method is simple and quick, quantitative and qualitative is accurate, and the detection requirement of long-chain fatty acid in anaerobic fermentation liquid can be met.
Description
Technical Field
The invention relates to the technical field of analysis and detection, in particular to a method for detecting long-chain fatty acid in anaerobic fermentation liquid.
Background
Along with the improvement of economy and material living standard, the proportion of the kitchen waste in the urban household waste is increased year by year, a large amount of kitchen waste is not recycled, not only occupies land resources, but also causes great risks for physical and mental health of residents due to putrefaction, malodor emission and pathogenic bacteria breeding of the kitchen waste. The traditional treatment methods such as crushing and direct discharge, sanitary landfill, incineration and the like have the defects of high cost, non-energy conservation and the like in different degrees. Anaerobic fermentation technology is widely used in the field of kitchen waste recycling due to the advantages of mature technology, high added value of products and the like. The organic waste is used as common biomass energy and can be converted into clean energy, namely methane, through an anaerobic digestion technology. The main components of the organic waste are carbohydrate, protein and grease, and the methane production potential of the grease is far higher than that of the carbohydrate and the protein, so the high-grease waste is a biomass energy source with development potential. The grease waste has huge output and wide sources, and if the high-efficiency anaerobic digestion treatment of the grease waste can be realized, the problem of shortage of fossil energy can be solved to a certain extent, and the difficult problem of environmental pollution can be greatly relieved.
The fat is first hydrolyzed to glycerin and Long chain fatty acids (Long-chain fatty acids, LCFAs), and these two hydrolysates are then degraded in different ways, respectively. The glycerol is converted into various volatile fatty acids, formic acid and alcohols through an acidification reaction stage, and then is converted into methane through an acetylation reaction. The methanogenic potential of fats and oils is 90% from LCFAs, and therefore LCFAs are important components of the constituent fats and oils. LCFAs in natural oils include unsaturated long-chain fatty acids such as oleic acid (C18:1), linoleic acid (C18:2), and saturated long-chain fatty acids such as stearic acid (C18:0), palmitic acid (C16:0), myristic acid (C14:0), and lauric acid (C12:0).
LCFAs are important intermediate products in the anaerobic degradation process of grease, so that the concentration of LCFAs in anaerobic fermentation liquid is accurately measured, and the method has important significance for further analyzing the anaerobic degradation path, inhibition and accumulation change rule of the LCFAs and knowing the degradation process and methane yield of grease waste.
At present, the related limit value and detection standard of long-chain fatty acid in anaerobic fermentation liquid are not available. Most of the existing related researches are methods for measuring the content of long-chain fatty acid in the matrix such as grease, serum and the like, and the researches on the method for measuring the content of long-chain fatty acid in the matrix of the anaerobic fermentation liquid are less, so that the method for accurately measuring the content of long-chain fatty acid in the anaerobic fermentation liquid is provided, has great significance for researching the degradation process of grease waste, and can provide important technical support for researching the efficient anaerobic digestion treatment of the grease waste.
Disclosure of Invention
The present invention has been made to solve the above-mentioned problems.
Firstly, the invention provides a method for detecting long-chain fatty acid in anaerobic fermentation liquid, which comprises the following steps:
(1) The anaerobic fermentation liquid is subjected to freeze vacuum drying and dehydration treatment to obtain a sample; mixing the sample with an extractant, extracting, and separating a liquid phase;
the volume ratio of the extractant is 20-30: 1 methanol and concentrated hydrochloric acid;
(2) Detecting the liquid phase by gas chromatography-mass spectrometry;
wherein the conditions of the gas chromatography include: chromatographic column DB-5MS (30 mm. Times.0.25 μm), sample inlet temperature 300 ℃; the temperature-raising program is as follows: initial temperature: maintaining at 50deg.C for 5min, and then heating to 170deg.C at 20deg.C/min; raising the temperature to 200 ℃ at 10 ℃/min; then raising the temperature to 220 ℃ at 5 ℃/min, and keeping the temperature for 5min; raising the temperature to 250 ℃ at 2 ℃/min and keeping the temperature for 2min.
According to the invention, long-chain fatty acid in anaerobic fermentation liquid can be accurately and rapidly detected qualitatively or quantitatively by extracting the anaerobic fermentation liquid by adopting an extractant consisting of methanol and concentrated hydrochloric acid with proper proportion and then performing gas chromatography-mass spectrometry detection under the gas chromatography condition.
The long-chain fatty acid in the present invention means a fatty acid having 6 to 24 carbon atoms.
The anaerobic fermentation liquid in the invention is grease waste anaerobic fermentation liquid.
Preferably, the conditions of the gas chromatography further include: and (3) split sampling, wherein the split ratio is 10:1.
Preferably, the conditions of the gas chromatography further include: constant flow, column flow was 1.0mL/min.
Preferably, in step (1), the sample is mixed with an extractant and then extracted, a liquid phase is separated, a solid phase is extracted for the second time, and the liquid phases obtained by the two extractions are mixed for gas chromatography-mass spectrometry detection.
The long-chain fatty acid can be enriched through secondary extraction, so that the sensitivity and accuracy of detection are further improved.
Preferably, the liquid phases obtained from the two extractions are mixed and filtered with a 0.45 μm filter for gas chromatography-mass spectrometry detection.
Preferably, the extraction is ultrasonic extraction.
More preferably, the time of ultrasonic extraction is 20 to 40min, most preferably 25 to 35min.
Preferably, the conditions of the mass spectrum include: EI ionization mode, ionization energy 70eV, ion source temperature 230 ℃, transmission line temperature 270 ℃, quadrupole mass analyzer temperature: the solvent delay time is 2min at 150 ℃, the full scanning and the selective ion scanning are carried out, and the full scanning range is m/z30.00-400.00.
Preferably, the qualitative analysis is performed using a combination of retention time and mass spectrometry retrieval.
Preferably, the quantitative detection is achieved by establishing a standard curve.
Preferably, the weight to volume ratio of the sample to the extractant is 1g: 45-55 mL.
The extraction effect is better under the proportion.
Compared with the prior art, the invention has the beneficial effects that:
the invention establishes a gas chromatography-mass spectrometry detection method for long-chain fatty acid in anaerobic fermentation liquid. The detection method is simple and quick, quantitative and qualitative is accurate, and the detection requirement of long-chain fatty acid in anaerobic fermentation liquid can be met.
Drawings
FIG. 1 is a chromatogram of the total ion flow of 33 long chain fatty acid methyl esters.
Fig. 2 is a standard graph.
FIG. 3 is a chromatogram of the total ion flow of long chain fatty acid methyl esters in a test sample.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The examples are not intended to identify the particular technology or conditions, and are either conventional or are carried out according to the technology or conditions described in the literature in this field or are carried out according to the product specifications. The reagents and instruments used, etc. are not identified to the manufacturer and are conventional products available for purchase by regular vendors. The anaerobic fermentation liquid in the following examples is an anaerobic fermentation liquid of grease waste.
Example 1
The embodiment provides a method for detecting long-chain fatty acid in anaerobic fermentation liquid, which comprises the following steps:
(1) Sample pretreatment: freezing and preserving anaerobic fermentation liquid in a condition below-18 ℃ and performing vacuum freeze drying and dehydration treatment by adopting a freeze dryer; adding 0.5g of sample into a centrifuge tube, adding 24mL of methanol and 1mL of concentrated hydrochloric acid, and performing ultrasonic extraction for 30min; placing the sample in a centrifuge, centrifuging at 3000rpm for 10min, separating liquid phase and solid phase, and immediately sealing the supernatant; performing secondary extraction on the first centrifugal solid phase, combining the two centrifugal liquid phases, and rapidly recording the volume of the combined centrifugal liquid phases; sample preparation is completed after membrane filtration by a 0.45 mu m filter membrane, and gas chromatography-mass spectrometry detection is carried out;
(2) Establishing a standard curve: taking 50 mu L of long-chain fatty acid methyl ester standard substance, dissolving with methanol, fixing the volume, preparing 20.0 mu g/mL standard stock solution, and sealing and storing at 4 ℃; measuring a certain volume of standard stock solution, diluting with methanol to prepare a series of standard working solutions, and detecting by gas chromatography-mass spectrometry; drawing a standard curve by taking the peak area of the quantitative ion of the target as an ordinate and the corresponding concentration as an abscissa;
(3) Detection by gas chromatography-mass spectrometry:
the conditions of the gas chromatography were: the chromatographic column DB-5MS (30 mm multiplied by 0.25 mu m) is adopted, the constant flow is carried out, and the column flow is 1.0mL/min; sample injection volume 1.0. Mu.L; split sample injection, wherein the split ratio is 10:1; the temperature of the sample inlet is 300 ℃; heating to an initial temperature of 50 ℃, maintaining for 5min, and then heating to 170 ℃ at 20 ℃/min; raising the temperature to 200 ℃ at 10 ℃/min; then raising the temperature to 220 ℃ at 5 ℃/min, and keeping the temperature for 5min; raising the temperature to 250 ℃ at 2 ℃/min, and keeping for 2min;
the conditions of mass spectrometry were: EI ionization mode, ionization energy 70eV, ion source temperature 230 ℃, transmission line temperature 270 ℃, quadrupole mass analyzer temperature: the solvent delay time is 2min at 150 ℃, the full scanning and the selective ion scanning are carried out, and the full scanning range is m/z 30.00-400.00;
(4) Calculating by an external standard method:
and drawing a standard curve according to the concentration of the added standard solution and the area of the quantitative ion peak to obtain a standard curve equation. And (3) carrying out qualitative analysis by adopting a combination mode of retention time and mass spectrum retrieval, and selecting ion peak area for quantification. During quantification, characteristic ions of an internal standard substance are extracted first, and peak areas are obtained through integration; and extracting characteristic ions of the substance to be analyzed, integrating to obtain peak areas, substituting the obtained peak areas into a standard curve for calculation, and obtaining the concentration of fatty acid methyl ester in the sample according to the calculation result. And then converting by the following formula to obtain the concentration of the fatty acid in the sample.
X LCFA ＝X FAME ×F FAME-FA
Wherein:
X LCFA -fatty acid content in milligrams per kilogram (mg/kg);
X FAME -fatty acid methyl ester content in milligrams per kilogram (mg/kg);
F FAME-FA -conversion coefficient of fatty acid methyl ester into fatty acid.
The test results are shown in table 1, the total ion flow chromatograms of 33 long chain fatty acid methyl esters are shown in fig. 1, the standard curves (using four long chain fatty acid methyl esters as examples) are shown in fig. 2, and the total ion flow chromatograms of the long chain fatty acid methyl esters in the test samples are shown in fig. 3.
TABLE 1
The peak time, ion information and detection limit of the sample are shown in table 2.
TABLE 2
Example 2
The embodiment provides a method for detecting long-chain fatty acid in anaerobic fermentation liquid, which is different from embodiment 1 only in steps:
and adding a long-chain fatty acid methyl ester standard substance into the sample to prepare a standard concentration of 40.0 mug/mL, wherein the ultrasonic time is 10 minutes and 50 minutes respectively. Taking C10:00 as an example, the peak areas of C10:00 at different ultrasound times are compared.
The results are shown in Table 3.
TABLE 3 Table 3
Comparative example 1
The comparative example provides a method for detecting long-chain fatty acid in anaerobic fermentation liquid, and the steps are only different from those of the embodiment 1:
the concentrated hydrochloric acid in the extractant is replaced with equal volumes of sulfuric acid and potassium hydroxide. A standard substance of long-chain fatty acid methyl ester is added into a sample to prepare a standard concentration of 40.0 mug/mL, and the peak area of C10:00 under the condition of different extractant with equal volume is compared by taking C10:00 as an example.
The results are shown in Table 4.
TABLE 4 Table 4
Comparative example 2
The comparative example provides a method for detecting long-chain fatty acid in anaerobic fermentation liquid, and the steps are only different from those of the embodiment 1:
the volume ratio of the extractant methanol to the concentrated hydrochloric acid is 10:1. A standard substance of long-chain fatty acid methyl ester is added into a sample to prepare a standard concentration of 40.0 mug/mL, and the peak area of C10:00 under the condition of different extractant with equal volume is compared by taking C10:00 as an example.
The results are shown in Table 5.
TABLE 5
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (10)
1. The method for detecting the long-chain fatty acid in the anaerobic fermentation liquid is characterized by comprising the following steps of:
(1) The anaerobic fermentation liquid is subjected to freeze vacuum drying and dehydration treatment to obtain a sample; mixing the sample with an extractant, extracting, and separating a liquid phase;
the volume ratio of the extractant is 20-30: 1 methanol and concentrated hydrochloric acid;
(2) Detecting the liquid phase by gas chromatography-mass spectrometry;
wherein the conditions of the gas chromatography include: chromatographic column DB-5MS (30 mm. Times.0.25 μm), sample inlet temperature 300 ℃; the temperature-raising program is as follows: initial temperature: maintaining at 50deg.C for 5min, and then heating to 170deg.C at 20deg.C/min; raising the temperature to 200 ℃ at 10 ℃/min; then raising the temperature to 220 ℃ at 5 ℃/min, and keeping the temperature for 5min; raising the temperature to 250 ℃ at 2 ℃/min and keeping the temperature for 2min.
2. The method of detecting according to claim 1, wherein the conditions of the gas chromatography further comprise: and (3) split sampling, wherein the split ratio is 10:1.
3. The method of detecting according to claim 1, wherein the conditions of the gas chromatography further comprise: constant flow, column flow was 1.0mL/min.
4. The method according to claim 1, wherein in the step (1), the sample is mixed with an extractant and then extracted, a liquid phase is separated, a solid phase is subjected to secondary extraction, and the liquid phases obtained by the two extractions are mixed for gas chromatography-mass spectrometry detection.
5. The method according to claim 4, wherein the liquid phase obtained by the two extractions is mixed and filtered with a 0.45 μm filter for gas chromatography-mass spectrometry detection.
6. The method of claim 1, wherein the extraction is ultrasonic extraction.
7. The method of claim 1, wherein the conditions of the mass spectrum comprise: EI ionization mode, ionization energy 70eV, ion source temperature 230 ℃, transmission line temperature 270 ℃, quadrupole mass analyzer temperature: the solvent delay time is 2min at 150 ℃, the full scanning and the selective ion scanning are carried out, and the full scanning range is m/z30.00-400.00.
8. The method of claim 7, wherein the qualitative analysis is performed by a combination of retention time and mass spectrometry.
9. The method of claim 7, wherein the quantitative determination is achieved by establishing a standard curve.
10. The method according to claim 1, wherein the weight to volume ratio of the sample to the extractant is 1g: 45-55 mL.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6057161A (en) * | 1997-10-27 | 2000-05-02 | Varian, Inc. | Method for determination of drugs of abuse in biological samples |
| US20030175851A1 (en) * | 2000-06-29 | 2003-09-18 | Pind Peter Frode | Method and equipment for monitoring syntrophic relations in a biological process fluid |
| CN102539609A (en) * | 2011-12-31 | 2012-07-04 | 青岛啤酒股份有限公司 | Detection method for measuring free fatty acid in beer through methyl esterification in bottle-headspace SPME (solid-phase micro-extraction)-gas chromatography and mass spectrum |
| CN113176358A (en) * | 2021-05-11 | 2021-07-27 | 苏州帕诺米克生物医药科技有限公司 | Pretreatment method and detection method of micro sample for metabonomics and lipidomics research |
| CN115112807A (en) * | 2022-08-18 | 2022-09-27 | 青岛排水有限公司 | Method for rapidly determining volatile fatty acid in sludge |
-
2023
- 2023-11-15 CN CN202311518426.1A patent/CN117517517A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6057161A (en) * | 1997-10-27 | 2000-05-02 | Varian, Inc. | Method for determination of drugs of abuse in biological samples |
| US20030175851A1 (en) * | 2000-06-29 | 2003-09-18 | Pind Peter Frode | Method and equipment for monitoring syntrophic relations in a biological process fluid |
| CN102539609A (en) * | 2011-12-31 | 2012-07-04 | 青岛啤酒股份有限公司 | Detection method for measuring free fatty acid in beer through methyl esterification in bottle-headspace SPME (solid-phase micro-extraction)-gas chromatography and mass spectrum |
| CN113176358A (en) * | 2021-05-11 | 2021-07-27 | 苏州帕诺米克生物医药科技有限公司 | Pretreatment method and detection method of micro sample for metabonomics and lipidomics research |
| CN115112807A (en) * | 2022-08-18 | 2022-09-27 | 青岛排水有限公司 | Method for rapidly determining volatile fatty acid in sludge |
Non-Patent Citations (3)
| Title |
|---|
| 刘诺: "餐厨垃圾厌氧发酵产酸产氢及恶臭控制技术研究", 中国博士学位论文全文数据库工程科技Ⅰ辑, no. 6, 15 June 2020 (2020-06-15), pages 027 - 38 * |
| 延莎 等: "不同菌种发酵对藜麦蛋白质特性及脂质构成的影响", 中国农业科学, vol. 53, no. 10, 31 December 2020 (2020-12-31), pages 2045 - 2054 * |
| 曾蕴仪: "强化泔脚废油脂厌氧发酵产奇数碳 VFAs 效能及机理研究", 中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑, no. 2, 15 February 2021 (2021-02-15), pages 027 - 1394 * |
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