CN113109479A - Method for detecting tributyrin by high performance liquid chromatography - Google Patents
Method for detecting tributyrin by high performance liquid chromatography Download PDFInfo
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- UYXTWWCETRIEDR-UHFFFAOYSA-N Tributyrin Chemical compound CCCC(=O)OCC(OC(=O)CCC)COC(=O)CCC UYXTWWCETRIEDR-UHFFFAOYSA-N 0.000 title claims abstract description 255
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 17
- 238000001514 detection method Methods 0.000 claims abstract description 26
- 239000012488 sample solution Substances 0.000 claims abstract description 22
- 239000000523 sample Substances 0.000 claims abstract description 20
- 239000012224 working solution Substances 0.000 claims abstract description 16
- 239000011550 stock solution Substances 0.000 claims abstract description 15
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 105
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 28
- 239000012071 phase Substances 0.000 claims description 25
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- 239000012528 membrane Substances 0.000 claims description 7
- 238000005303 weighing Methods 0.000 claims description 7
- 239000012074 organic phase Substances 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 5
- 238000007865 diluting Methods 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 239000011259 mixed solution Substances 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 2
- 238000012360 testing method Methods 0.000 description 14
- 230000014759 maintenance of location Effects 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 239000012086 standard solution Substances 0.000 description 5
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000010998 test method Methods 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000011097 chromatography purification Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- -1 glycerin fatty acid esters Chemical class 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
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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
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
- G01N30/74—Optical detectors
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
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- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a method for detecting tributyrin by high performance liquid chromatography, which comprises the following steps: 1. preparing a mobile phase; 2. preparing a tributyrin standard stock solution; 3. preparing a tributyrin standard working solution; 4. detecting by high performance liquid chromatography; 5. drawing a standard curve; 6. preparing a tributyrin sample solution; 7. and (3) detecting a tributyrin sample. The method has the advantages of simple and convenient operation, short detection time, high precision and accuracy and good stability.
Description
Technical Field
The invention relates to the technical field of feed detection, in particular to a method for detecting tributyrin by high performance liquid chromatography.
Background
The tributyrin is a precursor of butyric acid, can naturally pass through the stomach, provides quick and efficient butyric acid for the intestinal tract, can promote the growth and repair of intestinal mucosal cells of cultured animals, relieves intestinal inflammation, strengthens the barrier function of the intestinal tract, and improves the production performance of the cultured animals. With the banning of antibiotics in the feed, tributyrin as a green, safe and effective feed additive is widely applied to the feed and breeding industries. However, the detection of tributyrin has no corresponding national standard, row standard and landmark, and related literature reports can hardly be found, so that the development of the high performance liquid chromatography detection method of tributyrin has important value for the standard application of tributyrin in the market.
At present, most feed enterprises for producing tributyrin adopt a saponification titration method to detect the content of tributyrin products. The method has the advantages of complex pretreatment operation, long time consumption for detecting a sample, low detection precision and accuracy, easy counterfeiting, easy misunderstanding of other glycerin fatty acid esters as tributyrin and many defects.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a method for detecting tributyrin by high performance liquid chromatography, which has the advantages of simple and convenient operation, short detection time, high precision and accuracy and good stability.
The technical scheme adopted for realizing the above purpose of the invention is as follows:
a method for detecting tributyrin by high performance liquid chromatography comprises the following steps:
1. preparing a mobile phase;
2. preparing a tributyrin standard stock solution:
accurately weighing a tributyrin standard substance, dissolving the tributyrin standard substance with acetonitrile, and preparing into a tributyrin standard stock solution;
3. preparing a glycerol tributyrate standard working solution:
taking a tributyrin standard stock solution, and diluting the tributyrin standard stock solution into a series of tributyrin standard working solutions with gradient concentrations by using acetonitrile;
4. and (3) high performance liquid chromatography detection:
chromatographic conditions are as follows:
the chromatographic column adopts a Wondasil C18-WR chromatographic column or the chromatographic column with equivalent performance, the mobile phase is a mixed solution of an organic phase and a water phase, the organic phase is acetonitrile or methanol, the water phase is water, the volume percentage of the organic phase is 70-90%, the volume percentage of the water phase is 10-30%, the detection wavelength of an ultraviolet detector is 210-240nm, and the flow rate is 0.8-1.2 ml/min;
detecting a series of tributyrin standard working solutions with gradient concentrations to obtain peak areas of tributyrin in each standard working solution;
5. drawing a standard curve:
taking the concentration of tributyrin as an abscissa and the peak area as an ordinate, drawing according to the detected data, fitting to obtain a standard curve, and obtaining a functional relation y ═ k of the standard curve according to the standard curve1x+b1；
6. Preparing a tributyrin sample solution:
dissolving a tributyrin sample by using acetonitrile to prepare a tributyrin sample solution;
7. detection of tributyrin sample:
and (4) detecting the tributyrin sample solution by adopting the conditions in the step (4) to obtain the peak area of the tributyrin, substituting the peak area of the tributyrin into a function relation of a standard curve to obtain the concentration of the tributyrin sample solution, and multiplying the concentration of the tributyrin sample solution by the volume of the tributyrin sample solution to obtain the mass of the tributyrin sample.
Furthermore, the column length of the chromatographic column is 150-250mm, the inner diameter is 4.6mm, and the particle size is 5 μm.
Further, the column temperature of the chromatographic column is 25-40 ℃.
Further, the concentration of the tributyrin standard working solution is 0.1-1.0 mg/ml.
Further, the tributyrin standard working solution and the tributyrin sample solution are filtered by a 0.22-micron organic filter membrane before detection.
Compared with the prior art, the invention has the advantages and beneficial effects that:
1. the method adopts the high performance liquid chromatography to detect the content of the tributyrin, and the sample pretreatment operation is convenient, rapid, safe and efficient.
2. The method adopts the high performance liquid chromatography to detect the content of the tributyrin, has good separation effect, high precision and accuracy, good repeatability and stability and short detection time, greatly shortens the detection time of the tributyrin and improves the production efficiency.
3. The instrument required by the method for detecting the content of the tributyrin is a high performance liquid chromatograph with an ultraviolet detector, can be rapidly popularized and applied in the feed industry, and has very wide prospect.
Drawings
FIG. 1 is a standard curve obtained in example 1.
FIG. 2 is a chromatogram of the sixth test of precision.
Detailed Description
The present invention will be described in detail with reference to specific examples.
The reagents required for the following examples and experiments were:
tributyrin standard: the molecular formula is as follows: c15H26O6(ii) a The content is as follows: not less than 99.0 percent; the manufacturer and the brand are as follows: sigma T8626; CAS number: 60-01-5.
Acetonitrile: carrying out chromatographic purification; methanol: carrying out chromatographic purification; water: meets the three-stage water specified in GB/T6682.
The following examples and experiments used instruments and equipment:
high performance liquid chromatography (with ultraviolet detector); an electronic analytical balance; the sensory quantity is 0.0001 g; an ultrasonic cleaner; a solvent filter; microporous filter membrane: pore size 0.22 μm, organic system; an injector: 1 ml; volumetric flask: 50ml, 25ml and 10 ml; a liquid transferring gun: 100ul, 1 ml.
First, selection of test conditions
1. Selection of length of chromatographic column
Test tests show that the content of tributyrin can be detected when the length of the chromatographic column is 150mm, 200mm and 250mm, and the detection result of tributyrin is not influenced.
2. Selection of sample solvent
The tributyrin is easy to dissolve in alcohol, clear solution can be obtained by using methanol and ethanol, and when a selection test of maximum wavelength is carried out, the peak area of the tributyrin is the largest at 210nm, methanol and ethanol have terminal absorption influence at the position, the tributyrin is also easy to dissolve in acetonitrile, and the acetonitrile has no terminal absorption influence at the position of 210nm, so the acetonitrile is preferably used as a solvent.
3. Selection of detection wavelength
And scanning the tributyrin standard solution and the tributyrin sample solution at the full wavelength within the range of 200-400nm by using an ultraviolet-visible spectrophotometer to determine the wavelength corresponding to the maximum absorption peak of the tributyrin, wherein the result shows that the tributyrin has the maximum absorption peak at 210nm and the interference of impurities and baseline noise is less, so that 210nm is preferably used as the detection wavelength.
4. Selection of mobile phase:
weighing a certain amount of tributyrin standard, dissolving with acetonitrile, and detecting wavelength by using a Wondasil C18-WR chromatographic column (5um, 4.6mm multiplied by 250 mm): 210nm, column temperature 30 ℃, flow rate: 1ml/min, measured in different mobile phases, the results obtained are shown in Table 1:
TABLE 1 Effect of different flows on the retention time and Peak area
The results show that: when methanol and water (the volume percentage content is 80 percent of methanol and 20 percent of water) are used as mobile phases, the baseline noise is obvious, and the up-and-down fluctuation is large; when acetonitrile and water (the volume percentage content is acetonitrile 90 percent and water is 10 percent) are taken as a mobile phase, the retention time of tributyrin is too short, unknown small peaks are connected with tributyrin peaks, and the separation degree is poor; when acetonitrile and water (the volume percentage content is acetonitrile 80 percent and water 20 percent) and acetonitrile and water (the volume percentage content is acetonitrile 70 percent and water 30 percent) are used as mobile phases, the separation degree is good, the base line is stable, and considering that when the acetonitrile and water (the volume percentage content is acetonitrile 80 percent and water 20 percent) are used as the mobile phases, the retention time is relatively earlier, and the detection time can be saved during large-scale detection, so the acetonitrile and water (the volume percentage content is acetonitrile 80 percent and water 20 percent) are preferably used as the mobile phases.
5. Selection of flow rate:
weighing a certain amount of tributyrin standard, dissolving with acetonitrile, and detecting wavelength by using a Wondasil C18-WR chromatographic column (5um, 4.6mm multiplied by 250 mm): 210nm, column temperature 30 ℃, mobile phase: the results of acetonitrile and water (volume percentage: acetonitrile 80% and water 20%) measured at different flow rates are shown in Table 2:
TABLE 2 Effect of choice of flow Rate on Retention time and Peak area
As a result, the larger the flow rate, the shorter the retention time and the smaller the peak area, and when the flow rate is 1.0ml/min, the retention time, the peak area and the resolution are all preferable, so that the flow rate is preferably 1.0 ml/min.
Example 1
1. Preparing a mobile phase:
measuring a certain amount of acetonitrile (the volume percentage of the acetonitrile is 80%), measuring a certain amount of water (the volume of the water accounts for 20%), pouring the measured acetonitrile and ultrapure water into a wire-mouth bottle, and putting the wire-mouth bottle into an ultrasonic cleaner for ultrasonic treatment for 20 min;
2. preparing a tributyrin standard stock solution:
accurately weighing 25mg (accurate to 0.0001g) of tributyrin standard, placing the 25mg into a 25ml volumetric flask, adding about 20ml of acetonitrile, placing the volumetric flask into an ultrasonic cleaner for dissolving for 20min, cooling to room temperature, diluting to a scale with the acetonitrile, and fully shaking up to obtain 1.0mg/ml tributyrin standard stock solution;
3. preparing a glycerol tributyrate standard working solution:
taking a tributyrin standard stock solution, and diluting the tributyrin standard stock solution into tributyrin standard working solutions with the concentrations of 0.1mg/ml, 0.2mg/ml, 0.4mg/ml, 0.6mg/ml, 0.8mg/ml and 1.0mg/ml by using acetonitrile;
4. and (3) high performance liquid chromatography detection:
chromatographic conditions are as follows:
the chromatographic column adopts a Wondasil C18-WR chromatographic column, and the parameters are as follows: the column length is 250mm, the inner diameter is 4.6mm, and the granularity is 5 mu m;
the mobile phase is a mixed solution of acetonitrile and water, the acetonitrile accounts for 80 percent by volume, and the water accounts for 20 percent by volume;
the detection wavelength of the ultraviolet detector is 210nm, the flow rate is 1ml/min, the sample injection amount is 20 mu l, and the column temperature is 30 ℃;
filtering the tributyrin standard working solution with the series of gradient concentrations by using a 0.22-micrometer organic filter membrane, detecting on the machine, repeatedly detecting each concentration twice, and recording the peak area corresponding to each concentration;
5. drawing a standard curve:
taking the concentration of tributyrin as an abscissa and the peak area as an ordinate, drawing according to detected data, and fitting to obtain a standard curve, wherein as shown in fig. 1, a function relation y of the standard curve is obtained according to the standard curve, wherein the function relation y is 780885x +6263.9, and the result shows that the concentration of tributyrin and the corresponding peak area present a good linear relation, which indicates that the linear relation of the concentration of tributyrin standard solution and the peak area is good when the concentration of tributyrin standard solution is 0.1-1.0 mg/ml;
6. preparing a tributyrin sample solution:
weighing a proper amount of tributyrin sample (about equivalent to 0.02g of tributyrin and accurate to 0.0001g), placing the sample in a 25ml volumetric flask, adding about 20ml of mobile phase, ultrasonically vibrating for 20min to dissolve the tributyrin sample, cooling to room temperature, fixing the volume to the scale with acetonitrile, shaking up, standing to obtain tributyrin sample solution, and filtering with a 0.22 mu m organic filter membrane before loading;
7. detection of tributyrin sample:
and (4) detecting the tributyrin sample solution by adopting the conditions in the step (4) to obtain the peak area of the tributyrin, substituting the peak area of the tributyrin into a function relation of a standard curve to obtain the concentration of the tributyrin sample solution, and multiplying the concentration of the tributyrin sample solution by the volume of the tributyrin sample solution to obtain the mass of the tributyrin sample.
First, precision test
The test method comprises the following steps: preparing 1.025mg/ml of tributyrin standard solution by using the method of the step 2 in the example 1, and continuously performing machine detection on the tributyrin standard solution for 8 times by using the conditions of the step 4 in the example 1;
and (3) test results:
the results of the test are shown in table 3:
TABLE 3 results of the repeatability tests
As can be seen from Table 3, the Relative Standard Deviation (RSD) of retention time was 0.01%, and the Relative Standard Deviation (RSD) of peak area was 0.36%, thus indicating that tributyrin detected by the high performance liquid chromatography of the present invention was excellent in reproducibility and precision.
Test two, recovery test
The test method comprises the following steps:
a1.025 mg/ml tributyrin standard stock solution was prepared as in step 2 of example 1, and 6 parts of a 20mg tributyrin standard sample (to the nearest 0.0001g) were weighed out and treated as follows per part of tributyrin standard: placing in a 50ml volumetric flask, respectively adding 10ml of 1mg/ml tributyrin standard stock solution and about 30ml of mobile phase, ultrasonically vibrating for 20min to dissolve the sample, cooling to room temperature, fixing the volume to scale with the mobile phase solution, shaking, standing, filtering with 0.22 μm organic filter membrane, and detecting on a machine;
and (3) test results:
the results of the assay are shown in table 4:
TABLE 4 recovery test results
As can be seen from Table 4, the recovery rate of tributyrin is 99.34% -100.96%, and the relative standard deviation RSD of the recovery rate is 0.63%, thereby showing that the tributyrin detected by the high performance liquid chromatography of the invention has good accuracy.
Third, stability test
The test method comprises the following steps:
weighing 20mg of tributyrin standard sample (accurate to 0.0001g), placing in a 50ml volumetric flask, adding about 20ml of mobile phase, ultrasonically vibrating for 20min to dissolve the sample, cooling to room temperature, fixing the volume to the scale with the mobile phase solution, shaking up, standing, refrigerating at 4 ℃, respectively detecting on a machine for 0, 2, 4, 8, 12, 24, 48 and 72 hours, and filtering with a 0.22 μm organic filter membrane before the first detection on the machine;
and (3) test results:
the results of the assay are shown in table 5:
TABLE 5 stability test results
As can be seen from Table 5, the relative standard deviation RSD of the peak area obtained by detection is 0.59% within 0-72h, thereby showing that the tributyrin detected by the high performance liquid chromatography of the invention has good stability.
Claims (5)
1. A method for detecting tributyrin by high performance liquid chromatography is characterized by comprising the following steps:
1.1, preparing a mobile phase;
1.2, preparing a tributyrin standard stock solution:
accurately weighing a tributyrin standard substance, dissolving the tributyrin standard substance with acetonitrile, and preparing into a tributyrin standard stock solution;
1.3, preparing a tributyrin standard working solution:
taking a tributyrin standard stock solution, and diluting the tributyrin standard stock solution into a series of tributyrin standard working solutions with gradient concentrations by using acetonitrile;
1.4, detecting by high performance liquid chromatography:
chromatographic conditions are as follows:
the chromatographic column adopts a Wondasil C18-WR chromatographic column, the mobile phase is a mixed solution of an organic phase and a water phase, the organic phase is acetonitrile or methanol, the water phase is water, the volume percentage of the organic phase is 70-90%, the volume percentage of the water phase is 10-30%, the detection wavelength of an ultraviolet detector is 210-240nm, and the flow rate is 0.8-1.2 ml/min;
detecting a series of tributyrin standard working solutions with gradient concentrations to obtain peak areas of tributyrin in each standard working solution;
1.5, drawing a standard curve:
taking the concentration of tributyrin as an abscissa and the peak area as an ordinate, drawing according to the detected data, fitting to obtain a standard curve, and obtaining a functional relation y ═ k of the standard curve according to the standard curve1x+b1；
1.6, preparing a tributyrin sample solution:
dissolving a tributyrin sample by using acetonitrile to prepare a tributyrin sample solution;
1.7, detection of a tributyrin sample:
and (3) detecting the tributyrin sample solution by adopting the conditions of the step 1.4 to obtain the peak area of the tributyrin, substituting the peak area of the tributyrin into a function relation of a standard curve to obtain the concentration of the tributyrin sample solution, and multiplying the concentration of the tributyrin sample solution by the volume of the tributyrin sample solution to obtain the mass of the tributyrin sample.
2. The method for detecting tributyrin by high performance liquid chromatography according to claim 1, characterized in that: the chromatographic column has column length of 150-250mm, inner diameter of 4.6mm and particle size of 5 micron.
3. The method for detecting tributyrin by high performance liquid chromatography according to claim 1, characterized in that: the temperature of the chromatographic column is 25-40 ℃.
4. The method for detecting tributyrin by high performance liquid chromatography according to claim 1, characterized in that: the concentration of the tributyrin standard working solution is 0.1-1.0 mg/ml.
5. The method for detecting tributyrin by high performance liquid chromatography according to claim 1, characterized in that: and filtering the tributyrin standard working solution and the tributyrin sample solution by using a 0.22 mu m organic filter membrane before detection.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6783949B1 (en) * | 1999-04-30 | 2004-08-31 | Aventis Pharma Deutschland Gmbh | Processes for determining whether a test substance contains lipases or lipase inhibitors |
| CN108642105A (en) * | 2018-04-18 | 2018-10-12 | 江南大学 | A kind of method of enzyme process inclusion tributyrin |
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- 2021-04-19 CN CN202110417883.6A patent/CN113109479A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6783949B1 (en) * | 1999-04-30 | 2004-08-31 | Aventis Pharma Deutschland Gmbh | Processes for determining whether a test substance contains lipases or lipase inhibitors |
| CN108642105A (en) * | 2018-04-18 | 2018-10-12 | 江南大学 | A kind of method of enzyme process inclusion tributyrin |
Non-Patent Citations (2)
| Title |
|---|
| 中国药典委员会: "《中华人民共和国药典 2020年版 四部》", 31 May 2020, 中国医药科技出版社 * |
| 浙江高昇生物科技有限公司: "混合型饲料添加剂 三丁酸甘油酯", 《Q/ZGS 浙江高昇生物科技有限公司企业标准》 * |
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