CN102998385A - Method for simultaneously measuring contents of two organic acids in corn stalk fermentation broth - Google Patents
Method for simultaneously measuring contents of two organic acids in corn stalk fermentation broth Download PDFInfo
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- CN102998385A CN102998385A CN2012104786156A CN201210478615A CN102998385A CN 102998385 A CN102998385 A CN 102998385A CN 2012104786156 A CN2012104786156 A CN 2012104786156A CN 201210478615 A CN201210478615 A CN 201210478615A CN 102998385 A CN102998385 A CN 102998385A
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Abstract
The invention relates to a method for simultaneously measuring contents of two organic acids in corn stalk fermentation broth, and belongs to the technical field of chemical analysis detection. The method comprises the following steps of: preparing a mobile phase with certain concentration and pH value from monopotassium phosphate and acetonitrile as isocratic eluent; detecting by a high performance liquid chromatography, and determining the content of lactic acid and acetic acid in the corn stalk fermentation broth. The method is simple, rapid, and high in accuracy. The analysis result of the content of the lactic acid and the acetic acid can be simultaneously obtained just by sampling once under the same chromatographic condition; mixing a solution of the monopotassium phosphate and the acetonitrile at a volume ratio of 94:6 is adopted as a liquid chromatography mobile phase; and the sample can be well separated, and the detection time is also shortened.
Description
Technical field
The present invention relates to a kind of method of measuring simultaneously two kinds of organic acid contents in the Fermented Solution of Maize Straws, belong to chemical analysis detection technique field.
Background technology
Agricultural crop straw quantity is large, distribution is wide, is the secondary product of agricultural production, also is important living resources.Wherein, annual maize straw output is 22 * 10
9Ton, the plantation amount is large, in field burning or accumulation, has both wasted resource, again contaminated environment.Can be with maize straw as feed or as papermaking, but undressed maize straw, and the absorption of unsuitable herbivorous animal also should not be stored simultaneously.In order to address the above problem, maize straw is under anaerobic fermented, strengthened its absorptivity, improved storage time.Organic acid is the principal ingredient that maize straw under anaerobic ferments and, and comprises lactic acid, acetic acid, propionic acid etc.By analyzing organic acid content in the Fermented Solution of Maize Straws, have great importance for research fodder maize stalk quality.
At present, the organic acid analytical approach mainly contains acid base titration, thin-layered chromatography, vapor-phase chromatography and high performance liquid chromatography etc.Acid base titration is only applicable to macro-analysis, and can only measure total acid content; Thin-layered chromatography can be measured micro-organic acid, but quantitative accuracy is poor and sensitivity is low; Vapor-phase chromatography needs that organic acid is carried out saponification derives, and complex operation and error ratio are larger; High performance liquid chromatography is multiple organic acid in the working sample simultaneously, and have simple to operate, accuracy is high and the advantage of favorable reproducibility, is the more widely method of using at present.But adopt present method can't realize also in the Fermented Solution of Maize Straws that lactic acid separates fast and accurately with acetic acid and measures.
Summary of the invention
The objective of the invention is: the problem that is difficult to separate detection for lactic acid and acetic acid, a kind of method of analyze measuring simultaneously two kinds of organic acid contents in the Fermented Solution of Maize Straws is provided, and the method can realize separating and mensuration of lactic acid and acetic acid in the Fermented Solution of Maize Straws quickly and accurately.
Technical scheme of the present invention is: a kind of method of measuring simultaneously two kinds of organic acid contents in the Fermented Solution of Maize Straws may further comprise the steps:
(1) then the standard solution of difference making lactic acid, acetic acid variable concentrations gradient mixes constant volume and is mixed with mixed standard solution;
(2) preparation of testing sample: the maize straw of fermentation is extruded fermentation liquor with squeezer, after precipitation, centrifuging, with 0.45 μ m membrane filtration, stand-by;
(3) with each standard solution of step (1) and step (2) testing sample through the high performance liquid chromatograph analysis, chromatographiccondition is as follows:
Chromatographic column: anti-phase C-18 chromatographic column;
Sampling volume: 1 μ L;
Flow velocity: 1mL/min;
Detect wavelength: 215nm;
Column temperature: room temperature;
Mobile phase: the Lin acid dihydride Jia – acetonitrile mixed solution of 0.02mol/L volume ratio 94:6 is 2.3 with the phosphoric acid adjust pH;
Type of elution: isocratic elution;
(4) with lactic acid and two kinds of organic acid content of acetic acid in the external standard method quantitative analysis Fermented Solution of Maize Straws.
Technical solution of the present invention has following advantage:
1, under same chromatographic condition, only need advance sample one time, can obtain simultaneously lactic acid and acetic acid content analysis result, simple, fast, accuracy is high.
2, determine that liquid chromatogram mobile phase is the mixed solution of potassium dihydrogen phosphate and acetonitrile, volume ratio is potassium dihydrogen phosphate: acetonitrile=94:6, can make sample obtain good separation, and shorten detection time.
Description of drawings
Fig. 1 is the lactic acid standard diagram;
Fig. 2 is the acetic acid standard diagram;
Fig. 3 is the mixed sample standard diagram;
Fig. 4 is the Fermented Solution of Maize Straws collection of illustrative plates;
Fig. 5 is the standard regression curve of lactic acid;
Fig. 6 is the standard regression curve of acetic acid.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is described in further detail.
Embodiment 1:
1, the preparation of organic acid standard solution: accurately take by weighing lactic acid (ω %=99.8%), acetic acid (ω %=99.5%) each 11.824g, 7.3945g, with ultrapure water respectively constant volume to obtain concentration in the volumetric flask of 25ml be 0.4720g/mL and 0.2943g/mL; Then dilute making lactic acid concentration and be 0.1463,0.1768,0.2233,0.3031g/mL, acetic acid concentration is 0.0763,0.0936,0.1211, the solution of 0.1724g/mL.Get again 1.40mL lactic acid standard solution and 0.70mL acetic acid standard solution in the 25mL volumetric flask, be diluted to scale with ultrapure water, shake up, be mixed with mixed standard solution (mixed sample).
2, the preparation of solution to be measured: the maize straw of fermentation is extruded fermentation liquor with squeezer; Then take out the fermentation liquor of 5～10mL, precipitation, centrifuging; At last, filter through the organic filter membrane of 0.45 μ m (new Asia, Shanghai City purifies device factory), stand-by.
3, liquid-phase chromatographic analysis condition:
(1) chromatographic column: 150mm * 4.6mm, the anti-phase C-18 chromatographic column of the Shimadzu Shim-Pack VP-ODS of 5 μ m;
(2) sampling volume: 1 μ L;
(3) flow velocity: 1mL/min;
(4) column temperature: room temperature;
(5) detect wavelength: 215nm;
(6) mobile phase: the potassium dihydrogen phosphate of 0.02mol/L volume ratio 94:6-acetonitrile mixed solution is 2.3 with the phosphoric acid adjust pH;
(7) type of elution: isocratic elution.
4, get respectively 1 μ L standard solution and solution to be measured, be injected into liquid chromatograph through perfusor and analyze mensuration, obtained by chromatographic work station: the standard solution chromatogram of lactic acid, acetic acid and mixed sample is seen Fig. 1-3, and the retention time that draws lactic acid, acetic acid is respectively 2.422 and 2.592min; The chromatogram of Fermented Solution of Maize Straws is seen Fig. 4; The standard regression curve of lactic acid and acetic acid is seen Fig. 5,6.
5, the result calculates:
According to the chromatogram peak height, utilize external standard method by two kinds of organic acid content of lactic acid and acetic acid in the regression equation calculation sample of each standard solution:
Embodiment 2:
Present embodiment is that from the different of embodiment 1 mobile phase is in the step 3: the potassium dihydrogen phosphate of 0.02mol/L volume ratio 98:2-acetonitrile mixed solution, all the other steps are identical.
Present embodiment is obtained by chromatographic work station: the retention time that draws lactic acid, acetic acid is respectively 2.872 and 2.940min; Can find out that by the chromatogram of Fermented Solution of Maize Straws lactic acid and acetic acid overlaps, fail well separately.
Claims (5)
1. a method of measuring simultaneously two kinds of organic acid contents in the Fermented Solution of Maize Straws is characterized in that, may further comprise the steps:
(1) then the standard solution of difference making lactic acid, acetic acid variable concentrations gradient mixes constant volume and is mixed with mixed standard solution;
(2) preparation of testing sample: the maize straw of fermentation is extruded fermentation liquor with squeezer, through precipitation, centrifuging, finally by the organic membrane filtration of 0.45 μ m, stand-by;
(3) with each standard solution of step (1) and step (2) testing sample through the high performance liquid chromatograph analysis, analysis condition is as follows:
Chromatographic column: anti-phase C-18 chromatographic column;
Sampling volume: 1 μ L;
Flow velocity: 1mL/min;
Detect wavelength: 215nm;
Column temperature: room temperature;
Mobile phase: the 0.02mol/L volume ratio is potassium dihydrogen phosphate-acetonitrile of 94:6, is 2.3 with the phosphoric acid adjust pH;
Type of elution: isocratic elution;
(4) with lactic acid and two kinds of organic acid content of acetic acid in the external standard method quantitative analysis Fermented Solution of Maize Straws.
2. a kind of method of measuring simultaneously two kinds of organic acid contents in the Fermented Solution of Maize Straws according to claim 1, it is characterized in that, the compound method of lactic acid and acetic acid standard solution is in the step (1): accurately take by weighing lactic acid (ω %=99.8%), acetic acid (ω %=99.5%) each 11.824g, 7.3945g, with ultrapure water respectively constant volume to obtain concentration in the volumetric flask of 25ml be 0.4720g/mL and 0.2943g/mL; Then dilute making lactic acid concentration and be 0.1463,0.1768,0.2233,0.3031g/mL, acetic acid concentration is 0.0763,0.0936,0.1211, the solution of 0.1724g/mL.
3. a kind of method of measuring simultaneously two kinds of organic acid contents in the Fermented Solution of Maize Straws according to claim 1, it is characterized in that, the compound method of mixed standard solution is in the step (1): get 1.40mL lactic acid standard solution and 0.70mL acetic acid standard solution in the 25mL volumetric flask, be diluted to scale with ultrapure water, shake up, be mixed with mixed standard solution.
4. a kind of method of measuring simultaneously two kinds of organic acid contents in the Fermented Solution of Maize Straws according to claim 1 is characterized in that, anti-phase C-18 chromatographic column is 150mm * 4.6mm in the step (3), the VP-ODS post of 5 μ m.
5. a kind of method of measuring simultaneously two kinds of organic acid contents in the Fermented Solution of Maize Straws according to claim 1 is characterized in that, according to the chromatogram peak height, utilizes external standard method by the content of lactic acid and acetic acid in the regression equation calculation sample of each standard solution.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107505407A (en) * | 2017-07-19 | 2017-12-22 | 贵州茅台酒股份有限公司 | It is a kind of while determine lactic acid in white wine fermented grain, the method for acetic acid content |
| CN109374791A (en) * | 2018-12-24 | 2019-02-22 | 宜昌人福药业有限责任公司 | A kind of remaining method of acid in high effective liquid chromatography for measuring remifentanil hydrochloride raw material |
| CN114674941A (en) * | 2021-12-31 | 2022-06-28 | 南通奥凯生物技术开发有限公司 | Method for rapidly detecting acetic acid in feed |
-
2012
- 2012-11-22 CN CN2012104786156A patent/CN102998385A/en active Pending
Non-Patent Citations (5)
| Title |
|---|
| 努尔古丽等: "高效液相色谱法定量测定采油微生物发酵菌液中的有机酸", 《分析测试技术与仪器》 * |
| 吕旭聪等: "反相高效液相色谱法同时快速测定黄酒和葡萄酒中有机酸的含量", 《食品与发酵工业》 * |
| 牟英等: "高效液相色谱法测定1,3-丙二醇发酵液中的有机酸", 《分析化学》 * |
| 范宁伟等: "玉米发酵液中乳酸的分离提纯", 《吉林化工学院学报》 * |
| 郑志等: "反相高效液相色谱法测定发酵液中乳酸的含量", 《食品科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107505407A (en) * | 2017-07-19 | 2017-12-22 | 贵州茅台酒股份有限公司 | It is a kind of while determine lactic acid in white wine fermented grain, the method for acetic acid content |
| CN109374791A (en) * | 2018-12-24 | 2019-02-22 | 宜昌人福药业有限责任公司 | A kind of remaining method of acid in high effective liquid chromatography for measuring remifentanil hydrochloride raw material |
| CN109374791B (en) * | 2018-12-24 | 2022-06-03 | 宜昌人福药业有限责任公司 | Method for determining acid residue in remifentanil hydrochloride raw material by using high performance liquid chromatography |
| CN114674941A (en) * | 2021-12-31 | 2022-06-28 | 南通奥凯生物技术开发有限公司 | Method for rapidly detecting acetic acid in feed |
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Application publication date: 20130327 |