CN111766174A - Method for extracting crude fat from microcapsule type fat powder and method for measuring content of crude fat - Google Patents
Method for extracting crude fat from microcapsule type fat powder and method for measuring content of crude fat Download PDFInfo
- Publication number
- CN111766174A CN111766174A CN202010627317.3A CN202010627317A CN111766174A CN 111766174 A CN111766174 A CN 111766174A CN 202010627317 A CN202010627317 A CN 202010627317A CN 111766174 A CN111766174 A CN 111766174A
- Authority
- CN
- China
- Prior art keywords
- solvent
- extraction
- hydrochloric acid
- crude fat
- fat
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N5/00—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid
- G01N5/04—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid by removing a component, e.g. by evaporation, and weighing the remainder
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention provides a method for extracting crude fat from microcapsule type fat powder, which improves the hydrolysis degree of a sample and the extraction rate of the crude fat in the microcapsule type fat powder by strictly controlling the concentration of hydrochloric acid and the sample ratio of the hydrochloric acid to the microcapsule type fat powder, further can relieve the problem of low crude fat content detection in the microcapsule type fat powder in the prior art, provides a feasible scheme for detecting the crude fat content in the microcapsule type fat powder, makes up the vacancy in detecting the crude fat content in the microcapsule type fat powder in the field of nutritional value evaluation of the existing feed, and has better industrial application value.
Description
Technical Field
The invention relates to the technical field of feed nutritive value evaluation, in particular to a method for extracting crude fat from microcapsule type fat powder and a method for measuring the content of the crude fat.
Background
Fat is one of important raw materials of animal complete feed, and has important effects in reducing animal heat stress, increasing feed energy concentration to meet energy requirement of special physiological period (such as lactating sow and weaned piglet), improving feed palatability and appearance, prolonging service life of feed granulator, reducing dust, etc. In the feed oil, the traditional raw oil (such as soybean oil, palm oil and the like) and common fat powder and the like are easy to oxidize and deteriorate in the feed production, and the fatty acid composition is unbalanced, so that the feed oil is difficult to digest and absorb by animals, the use efficiency of fat is reduced, risks are brought to animal breeding, and if researches indicate that the feed oil is difficult to digest and absorb by weaned piglets and is easy to cause piglet nutritional diarrhea.
Aiming at the problems, a new technical product, namely microcapsule type fat powder, is prepared by utilizing a microencapsulation technology from pharmaceutical and chemical engineering, taking oil as a core material and syrup, cyclodextrin, whey powder or casein as a wall material through mixing, emulsifying, enzyme digestion, homogenizing and spray drying, and can well solve the problems.
However, the method for measuring the crude fat of the microcapsule type fat powder is lack of standard, the application range of the existing standard GB/T6433-2006 for measuring the crude fat in the feed does not contain the product, and the defects of unstable detection result, poor precision, very complicated operation steps and the like exist in forced use.
Therefore, the crude fat is an important quality index of the fat powder, and the establishment and trade of the quality standard of the product are seriously affected by the deficiency of the detection method, so that the development of the extraction and detection method of the crude fat in the fat powder is urgently needed.
Disclosure of Invention
In view of the problems in the prior art, the invention provides a method for extracting crude fat from microcapsule-type fat powder, which can improve the extraction rate of the crude fat from the microcapsule-type fat powder by strictly controlling the concentration of hydrochloric acid and the sample ratio of the hydrochloric acid to the microcapsule-type fat powder; and a feasible scheme is provided for detecting the content of the crude fat in the microcapsule type fat powder, the vacancy of detecting the content of the crude fat in the microcapsule type fat powder in the field of evaluation of the nutritional value of the existing feed is made up, and the method has a good industrial application value.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides a method for extracting crude fat from microcapsule type fat powder, wherein the determination method comprises the following steps:
(1) hydrolyzing with 3-5 moL/L hydrochloric acid with mass m0Obtaining hydrolysate of the microcapsule type fat powder sample; the ratio of the hydrochloric acid to the microcapsule type fat powder sample is 15-25 mL and 1-2 g;
(2) adding a solvent into the hydrolysate obtained in the step (1) for extraction, mixing, centrifuging, separating an upper layer solvent, repeating extraction at least twice, and combining the upper layer solvents obtained by the extraction at least twice to obtain an extracting solution, wherein the solvent comprises petroleum ether;
(3) removing the solvent from the extracting solution in the step (2), and drying to obtain the extract with mass m1The crude fat sample of (2).
The method for extracting the crude fat from the microcapsule type fat powder limits at least two times of extraction by strictly controlling the concentration of hydrochloric acid and the sample ratio of the hydrochloric acid to the microcapsule type fat powder, and strictly limits extraction by adopting a solvent containing petroleum ether, wherein wall material microcapsules formed by phase change gelation of polysaccharide or protein macromolecules are broken under the condition of strong acid hydrolysis, core material grease is dissociated out, then the extraction is carried out by using an organic solvent petroleum ether, and residues are the crude fat after the solvent is removed, so that the extraction rate is high and the cost is low.
The mass concentration of the hydrochloric acid is 3-5 moL/L, and can be 3moL/L, 3.2moL/L, 3.5moL/L, 3.8moL/L, 4moL/L, 4.2moL/L, 4.5moL/L, 4.8moL/L or 5moL/L, and the like.
The ratio of the hydrochloric acid to the microencapsulated fat powder sample in the present invention is 15-25 mL: 1-2 g, and may be, for example, 15mL:1g, 18mL:1g, 20mL:1g, 22mL:1g, 25mL:1g, 15mL:1.2g, 18mL:1.2g, 20mL:1.2g, 21mL:1.2g, 22mL:1.2g, 15mL:1.5g, 18mL:1.5g, 20mL:1.5g, 22mL:1.5g, 25mL:1.5g, 15mL:2g, 18mL:2g, 20mL:2g, or 25mL:2 g.
Preferably, the concentration of the hydrochloric acid in the step (1) is 4 moL/L.
The invention preferably adopts hydrochloric acid with the concentration of 4moL/L, has better hydrolysis effect and is more beneficial to improving the repeatability of detection.
Preferably, the ratio of the hydrochloric acid to the microcapsule fat powder sample is 20mL: 1-2 g.
According to the invention, the ratio of the hydrochloric acid to the microcapsule type fat powder sample is preferably 20mL to 1-2 g, so that the extraction cost can be saved, and the hydrolysis of the microcapsule type fat powder sample can be better promoted.
The hydrolysis temperature in step (1) is preferably 70 to 100 ℃, and may be, for example, 70 ℃, 72 ℃, 73 ℃, 75 ℃, 78 ℃, 80 ℃, 82 ℃, 85 ℃, 88 ℃, 90 ℃, 92 ℃, 95 ℃ or 100 ℃, preferably 80 to 90 ℃.
The hydrolysis temperature is preferably 80-90 ℃, the hydrolysis effect is better, and the result is more stable.
Preferably, the hydrolysis time is 0.5-1.5 h, for example, 0.5h, 0.6h, 0.8h, 0.9h, 1.0h, 1.2h, 1.3h or 1.5h, and preferably 1 h.
Preferably, the hydrolysis is carried out under intermittent vortex conditions.
Preferably, the intermittent vortexing comprises: after mixing the hydrochloric acid with the microencapsulated fat powder sample, the mixture is first vortexed for 25 to 35 seconds, for example, 25 seconds, 26 seconds, 27 seconds, 28 seconds, 29 seconds, 30 seconds, 32 seconds, 33 seconds, 34 seconds, or 35 seconds.
Preferably, the intermittent vortex further comprises: and (3) swirling for 10-20 s at intervals of 15-25 min in the hydrolysis process, wherein the intervals can be 15min, 16min, 17min, 18min, 19min, 20min, 21min, 22min, 23min, 24min or 25min and the like, and the time for swirling for each time can be 10s, 11s, 12s, 13s, 14s, 15s, 16s, 18s or 20s and the like.
Preferably, the hydrolysis is performed in a centrifuge tube.
Preferably, the boiling point of the petroleum ether in the step (2) is 40 to 60 ℃, and may be, for example, 40 ℃, 42 ℃, 43 ℃, 45 ℃, 48 ℃, 49 ℃, 50 ℃, 52 ℃, 54 ℃, 55 ℃, 58 ℃ or 60 ℃.
Preferably, the solvent of the first extraction comprises ethanol.
The solvent in the first extraction process preferably comprises ethanol, and the presence of ethanol has defoaming effect, and can eliminate residues adhered to the wall of the petroleum ether layer and remove emulsification.
Preferably, the ethanol is ethanol with a concentration of 95 wt%.
Preferably, the volume ratio of the petroleum ether to the ethanol in the solvent for the first extraction is 3-5: 1, for example, 3:1, 3.2:1, 3.5:1, 3.8:1, 4:1, 4.2:1, 4.5:1, 4.8:1 or 5:1, and preferably 4: 1.
Preferably, the volume ratio of the solvent to the hydrochloric acid in the step (1) is 0.9-1.5: 1, for example, 0.9:1, 1.0:1, 1.1:1, 1.2:1, 1.3:1, 1.4:1 or 1.5:1, and preferably 1-1.25: 1.
Preferably, the number of times of the extraction in step (2) is two.
Preferably, the extraction is performed in a centrifuge tube.
Preferably, the mixing comprises oscillatory mixing.
Preferably, the time for the oscillating mixing is 35-45 s, for example, 35s, 36s, 38s, 40s, 42s, 44s or 45 s.
Preferably, the rotation speed of the centrifugation is 4500-5500 r/min, such as 4500r/min, 4800r/min, 4900r/min, 5000r/min, 5200r/min or 5500 r/min.
Preferably, the centrifugation time is 5-7 min, for example, 5min, 5.2min, 5.4min, 5.5min, 5.8min, 6min, 6.2min, 6.5min, 6.8min or 7 min.
Preferably, step (2) comprises: adding petroleum ether and 95 wt% ethanol into the hydrolysate obtained in the step (1) in a volume ratio of 3-5: 1 as a first solvent, sealing a centrifuge tube, oscillating and mixing for 35-45 s, centrifuging at 4500-5500 r/min for 5-7 min, and separating an upper layer solvent to complete first extraction; adding petroleum ether into the lower layer liquid as a second solvent, repeatedly performing second extraction, and combining the upper layer solvents obtained by the two extractions to obtain an extracting solution; the volume ratio of the first solvent to the hydrochloric acid is 1-1.25: 1; the volume ratio of the second solvent to the hydrochloric acid is 1-1.25: 1.
The invention preferably adopts a two-step extraction method for extraction, which can save the detection time and the extraction solvent and can ensure the extraction rate and the subsequent detection result.
Preferably, the removing the solvent in step (3) comprises distilling off the solvent and removing the solvent by a lipometer.
Preferably, the drying temperature is 101-105 ℃, for example, 101 ℃, 102 ℃, 103 ℃, 104 ℃ or 105 ℃.
Preferably, the drying time is 10-20 min, for example, 10min, 12min, 14min, 15min, 16min, 17min, 18min, 19min or 20 min.
Preferably, the drying is followed by a cooling step.
Preferably, the cooling is performed in a dry environment.
Preferably, the cooling is performed in a dryer.
In a second aspect, the invention provides a method for determining the content of crude fat in microencapsulated fat powder, wherein the method for determining the content of crude fat comprises the steps of extracting crude fat by using the method for extracting crude fat in microencapsulated fat powder of the first aspect, and using m as the raw fat1And m0The ratio of (A) to (B) is recorded as the crude fat content in the microcapsule-type fat powder.
The determination method provided by the invention adopts the extraction method of the first aspect of the invention to extract the crude fat, adopts a gravimetric method to determine the crude fat content, has high precision of the detection result and good repeatability, and makes up for the vacancy of detecting the crude fat content in the microcapsule type fat powder in the existing feed.
Preferably, said m1The measurement method (2) comprises: the weighing container has a mass m2Weighing containerTotal mass m with crude fat sample3By m3Minus m2Obtaining the mass m of the crude fat sample1。
Preferably, the container is dried and cooled, and the weight of the container is measured to be m2。
Preferably, the drying temperature of the container is 101-105 ℃, for example, 101 ℃, 102 ℃, 103 ℃, 104 ℃ or 105 ℃ and the like.
Preferably, the drying time of the container is 25-35 min, such as 25min, 26min, 27min, 28min, 29min, 30min, 31min, 32min, 33min, 34min or 35 min.
Preferably, the cooling of the container is performed in a dry environment.
Preferably, the cooling of the vessel is performed in a dryer.
Preferably, the container is an aluminium cylinder.
As a preferred technical scheme of the invention, the method comprises the following steps:
(1) weighing m0Adding 3-5 moL/L hydrochloric acid into a micro-capsule type fat powder sample in a centrifugal tube, sealing the centrifugal tube, performing vortex for 25-35 s, and performing hydrolysis at 80-90 ℃, wherein the hydrolysis time is 0.9-1.5 h, and the vortex is performed for 10-20 s every 15-25 min in the hydrolysis process to obtain a hydrolysis solution; wherein the ratio of the hydrochloric acid to the microcapsule fat powder sample is 20mL to 1-2 g;
drying the aluminum cylinder at 101-105 ℃ for 25-35 min, placing the dried aluminum cylinder in a dryer to cool to room temperature, and weighing the aluminum cylinder with the mass m2；
(2) Adding petroleum ether and ethanol with the concentration of 95 wt% into the hydrolysate in the step (1) in a volume ratio of 3-5: 1 as a first solvent, sealing a centrifuge tube, oscillating and mixing for 35-45 s, centrifuging at 4500-5500 r/min for 5-7 min, and absorbing an upper layer solvent into the aluminum cylinder in the step (1) to finish first extraction;
adding petroleum ether into the lower layer liquid as a second solvent, repeatedly performing second extraction, combining the upper layer solvents obtained by the two extractions into the aluminum cylinder in the step (1) to obtain an extracting solution;
the volume ratio of the first solvent to the hydrochloric acid is 1-1.25: 1; the volume ratio of the second solvent to the hydrochloric acid is 1-1.25: 1;
(3) removing the solvent from the extracting solution in the step (2), drying at 101-105 ℃ for 10-20 min, placing in a dryer, cooling to room temperature, wherein the total mass of the weighing container and the crude fat sample is m3By m3Minus m2Obtaining the mass m of the crude fat sample1In m is1And m0The ratio of (A) to (B) is recorded as the crude fat content in the microcapsule-type fat powder.
Compared with the prior art, the invention has at least the following beneficial effects:
(1) the method for extracting the crude fat from the microcapsule fat powder strictly controls the concentration of hydrochloric acid, the proportion of the hydrochloric acid to a sample, the extraction times and the solvent, greatly improves the extraction rate of the crude fat, and has the extraction rate of more than 99.5 wt%;
(2) the method for measuring the content of the crude fat in the microcapsule fat powder makes up the defect that the prior art does not have the measurement for the content of the crude fat in the microcapsule fat powder, and has high repeatability of detection;
(3) the method for measuring the content of the crude fat in the microcapsule fat powder is simple and convenient to operate and high in precision, and under the repeated condition, the difference of 2 independent measurement results does not exceed 2.02% of the arithmetic mean value.
Drawings
FIG. 1 is a graph showing the results of crude fat detection according to the present invention for FA-4 at different temperatures and hydrochloric acid concentrations.
FIG. 2 is a graph showing the results of crude fat detection according to the present invention for FA-5 at different temperatures and hydrochloric acid concentrations.
Detailed Description
The technical scheme of the invention is further explained by the specific implementation mode in combination with the attached drawings.
The present invention is described in further detail below. The following examples are merely illustrative of the present invention and do not represent or limit the scope of the claims, which are defined by the claims.
The accuracy of the mass weighing in the following examples was 0.1 mg.
First, an embodiment
Example 1
The embodiment provides a method for extracting crude fat from microcapsule type fat powder, which comprises the following steps:
(1) weighing a 1-2 g microcapsule type fat powder sample in mass, adding 20mL of hydrochloric acid of 4moL/L into a centrifugal tube, screwing down a cover of the centrifugal tube, performing vortex for 30s, and performing hydrolysis under the condition of water bath of a water bath kettle at 85 ℃, wherein the hydrolysis time is 1h, and the vortex is performed for 15s every 20min in the hydrolysis process to obtain a hydrolysate;
(2) adding 20mL of petroleum ether and 5mL of ethanol with the concentration of 95 wt% into the hydrolysate in the step (1) as a first solvent, screwing down a centrifugal tube cover, oscillating and mixing for 40s, centrifuging for 6min at 5000r/min, and absorbing an upper layer of solvent into the aluminum tube in the step (1) to finish first extraction;
adding 20mL of petroleum ether serving as a second solvent into the lower layer liquid, repeatedly performing secondary extraction, and combining the upper layer solvents obtained by the two extractions into the aluminum cylinder in the step (1) to obtain an extracting solution;
(3) distilling the extracting solution obtained in the step (2) to remove the solvent, drying at 103 +/-2 ℃ for 15min, and cooling to room temperature in a drier to obtain the extract with mass m1Crude fat sample mass of (2).
Example 2
This example provides a process for extracting crude fat from microencapsulated fat powder, which is the same as in example 1 except that step (2) is performed for a third extraction with 20mL of petroleum ether.
Example 3
This embodiment provides a method for determining the content of crude fat in microencapsulated fat powder, which comprises the following steps:
(1) weighing a 1-2 g microcapsule type fat powder sample in mass, adding 20mL of hydrochloric acid of 4moL/L into a centrifugal tube, screwing down a cover of the centrifugal tube, performing vortex for 30s, and performing hydrolysis under the condition of water bath of a water bath kettle at 85 ℃, wherein the hydrolysis time is 1h, and the vortex is performed for 15s every 20min in the hydrolysis process to obtain a hydrolysate;
drying the aluminum cylinder at 103 +/-2 ℃ for 30min, cooling the dried aluminum cylinder to room temperature in a drier, and weighing the aluminum cylinder with the mass m2；
(2) Adding 20mL of petroleum ether and 5mL of ethanol with the concentration of 95 wt% into the hydrolysate in the step (1) as a first solvent, screwing down a centrifugal tube cover, oscillating and mixing for 40s, centrifuging for 6min at 5000r/min, and absorbing an upper layer of solvent into the aluminum tube in the step (1) to finish first extraction;
adding 20mL of petroleum ether serving as a second solvent into the lower layer liquid, repeatedly performing secondary extraction, and combining the upper layer solvents obtained by the two extractions into the aluminum cylinder in the step (1) to obtain an extracting solution;
(3) distilling the extracting solution obtained in the step (2) to remove the solvent, drying at 103 +/-2 ℃ for 15min, placing in a dryer for cooling to room temperature, and weighing the total mass of the aluminum cylinder and the crude fat sample to be m3By m3Minus m2Obtaining the mass m of the crude fat sample1In m is1And m0The ratio of (A) to (B) is recorded as the crude fat content in the microcapsule-type fat powder.
Example 4
This example provides a method for measuring the crude fat content in microencapsulated fat powder, which is the same as in example 3 except that 3moL/L hydrochloric acid was used in step (1).
Example 5
This example provides a method for measuring the crude fat content in microencapsulated fat powder, which is the same as in example 3 except that 5moL/L hydrochloric acid was used in step (1).
Example 6
This example provides a method for measuring the crude fat content in microencapsulated fat powder, which is the same as in example 3 except that 6moL/L hydrochloric acid was used in step (1).
Example 7
This example provides a method for measuring the crude fat content in microencapsulated fat powder, which is the same as in example 3 except that the "85 ℃ water bath condition" in step (1) is replaced with the "70 ℃ water bath condition".
Example 8
This example provides a method for measuring the crude fat content in microencapsulated fat powder, which is the same as in example 3 except that "85 ℃ water bath conditions" is replaced with "100 ℃ heating conditions" in step (1).
Example 9
This example provides a method for measuring the crude fat content in microencapsulated fat powder, which is the same as in example 3 except that the "85 ℃ water bath condition" in step (1) is replaced with the "80 ℃ water bath condition".
Example 10
This example provides a method for measuring the crude fat content in microencapsulated fat powder, which is the same as in example 3 except that the "85 ℃ water bath condition" in step (1) is replaced with the "90 ℃ water bath condition".
Example 11
This embodiment provides a method for determining the content of crude fat in microencapsulated fat powder, which comprises the following steps:
(1) weighing a 1-2 g microcapsule type fat powder sample in mass, adding 15mL of hydrochloric acid of 4moL/L into a centrifugal tube, screwing down a cover of the centrifugal tube, performing vortex for 25s, and performing hydrolysis under the condition of water bath of a water bath kettle at 85 ℃, wherein the hydrolysis time is 0.9h, and the vortex is performed for 20s every 15min in the hydrolysis process to obtain a hydrolysate;
drying the aluminum cylinder at 105 ℃ for 25min, cooling the aluminum cylinder to room temperature in a dryer after drying, and weighing the aluminum cylinder with the mass m2；
(2) Adding 15mL of petroleum ether and 5mL of ethanol with the concentration of 95 wt% into the hydrolysate in the step (1) as a first solvent, screwing down a centrifugal tube cover, oscillating and mixing for 35s, centrifuging at 4500r/min for 5min, and absorbing an upper layer solvent into the aluminum tube in the step (1) to finish first extraction;
adding 15mL of petroleum ether serving as a second solvent into the lower layer liquid, repeatedly performing second extraction, and combining the upper layer solvents obtained by the two extractions into the aluminum cylinder in the step (1) to obtain an extracting solution;
(3) distilling the extracting solution obtained in the step (2) to remove the solvent, drying at 103 ℃ for 10min, placing in a dryer for cooling to room temperature, and weighing the aluminum cylinder and the crude fat sample to obtain m3By m3Minus m2Obtaining the mass m of the crude fat sample1In m is1And m0The ratio of (A) to (B) is recorded as the crude fat content in the microcapsule-type fat powder.
Example 12
This embodiment provides a method for determining the content of crude fat in microencapsulated fat powder, which comprises the following steps:
(1) weighing a 1-2 g microcapsule fat powder sample in mass, adding 25mL of 4moL/L hydrochloric acid into a centrifugal tube, screwing down a centrifugal tube cover, performing vortex for 45s, and performing hydrolysis under the condition of water bath of a water bath kettle at 80 ℃, wherein the hydrolysis time is 1.5h, and the vortex is performed for 10s every 15min in the hydrolysis process to obtain a hydrolysate;
drying the aluminum cylinder at 101 ℃ for 35min, cooling the aluminum cylinder to room temperature in a dryer after drying, and weighing the aluminum cylinder with the mass m2；
(2) Adding 25mL of petroleum ether and 4mL of 95 wt% ethanol serving as a first solvent into the hydrolysate obtained in the step (1), screwing down a centrifugal tube cover, oscillating and mixing for 45s, centrifuging at 5500r/min for 7min, and absorbing an upper layer solvent into the aluminum tube obtained in the step (1) to finish first extraction;
adding 25mL of petroleum ether serving as a second solvent into the lower layer liquid, repeatedly performing second extraction, and combining the upper layer solvents obtained by the two extractions into the aluminum cylinder in the step (1) to obtain an extracting solution;
(3) distilling the extracting solution obtained in the step (2) to remove the solvent, drying at 105 ℃ for 20min, placing in a dryer for cooling to room temperature, and weighing the total mass of the aluminum cylinder and the crude fat sample to be m3By m3Minus m2Obtaining the mass m of the crude fat sample1In m is1And m0The ratio of (A) to (B) is recorded as the crude fat content in the microcapsule-type fat powder.
Second, comparative example
Comparative example 1
This comparative example provides a method for extracting crude fat from microencapsulated fat powder, which is the same as in example 1 except that the first extraction is carried out using only 20mL of petroleum ether and 5mL of ethanol in step (2) and the second extraction is not carried out.
Comparative example 2
This comparative example provides a method for determining the crude fat content in microencapsulated fat powder, which employs the method in GB/T6433/ISO6492:1999 for determining the crude fat content in microencapsulated fat powder.
Third, test and results
In order to detect the detection effect of the method provided by the invention on different microcapsule type fat powders, 5 samples are purchased from different manufacturers (the standard crude fat content is required to be more than or equal to 50 wt%), and are respectively named as FA-1, FA-2, FA-3, FA-4 and FA-5.
1. For the extraction methods provided in example 1, example 2 and comparative example 1, the extraction rate of each extraction for the sample FA-4 was measured, and the extraction rate of each extraction at a hydrochloric acid concentration of 3moL/L was additionally measured, and the results are shown in table 1, where the unit in table 1 is: wt%.
TABLE 1
As can be seen from table 1, not only more than 99.5 wt% of the crude fat can be extracted by using 2 extractions, but also the solvent amount and the operation steps can be saved, which is a preferable step in consideration of the higher extraction rate of 2 extractions in example 1 with respect to one extraction in comparative example 2, and the simpler extraction steps of two extractions in example 1 with respect to three extractions in example 2, and the difference in extraction rate from example 2 is small.
2. The test was conducted by the methods provided in example 3 and comparative example 1 using FA-4 and FA-5 as test samples, each sample was measured for crude fat content using two parallel samples, and precision was represented by the absolute difference between the measured values of the two parallel samples, and the average value of the crude fat content and the precision results are shown in Table 2.
TABLE 2
As can be seen from Table 2, the method provided in example 3 has a high crude fat content, which is closer to the crude fat content in the microencapsulated fat powder and has a precision of 0.5 wt% or less, while the method provided in comparative example 1 has a precision of 1 wt% or more for the microencapsulated fat powder, thereby indicating that the method provided by the present invention can better measure the crude fat content and has higher repeatability for the microencapsulated fat powder.
3. For the method provided in example 3, 5 samples were taken and aligned between three laboratory development rooms, according to GB/T6379.2-2004 measurement method and accuracy of results (accuracy and precision) part 2: basic method for determining the reproducibility and reproducibility of standard measurement methods, reproducibility variance, repeatability variance + standard inter-chamber variance, i.e.: SR2＝Sr2+SL2The standard deviation of reproducibility was calculated, and the allowable difference in reproducibility and reproducibility of the method was calculated and determined based on this, and the results are shown in Table 3 (unit: wt% in Table 3).
TABLE 3
As can be seen from Table 3, the method provided in example 3 has better detection effect in different laboratories and different samples, and the original data are tested by Cochran (Cochran) and have good precision and no outlier. The average value is checked by Grubbs, the results are consistent, and the precision and the content of crude fat are high or low, or the entrapment rate is high or low, and no correlation exists. It can be further seen from Table 2 that the repeatability limit is between 0.66 wt% and 0.99 wt% and the reproducibility limit is between 0.55 wt% and 2.02 wt%. Therefore, the method has the advantages that the repeated allowable difference is that the difference between the results of two independent measurements is not more than 2.02%, and the detection effect is good.
4. For example 3 and examples 4-10, the crude fat detection results for FA-4 and FA-5 at different temperatures and hydrochloric acid concentrations were examined, wherein the detection results for FA-4 are shown in FIG. 1, and the detection results for FA-5 are shown in FIG. 2.
As can be seen from fig. 1 and 2, the results of hydrolysis at two temperatures of 80 ℃ and 90 ℃ show that the samples are more stable than 70 ℃ and 100 ℃ and closer to the theoretical value regardless of the encapsulation efficiency, the results of combination of the results with the acid concentrations of 3moL/L and 4moL/L are better in precision and correctness, and the hydrolysis effect of the 4moL/L acid concentration combination on the samples with high encapsulation efficiency is the best, so that the results of detection in examples 3 and 9-10 are better when the temperature range of 80-90 ℃ is combined with the hydrochloric acid concentration of 4moL/L for the microcapsule-type fat powder than in examples 4-8, and the results of detection are higher for the samples with different contents.
In conclusion, the method for extracting the crude fat from the microcapsule-type fat powder improves the hydrolysis degree of a sample and the extraction rate of the crude fat by strictly controlling the concentration of hydrochloric acid and the sample ratio of the hydrochloric acid to the microcapsule-type fat powder, and the extraction rate of two times of extraction is more than 99.5 wt%; the method effectively solves the problem of low crude fat content in a microcapsule type fat powder sample measured in the prior art, provides a feasible scheme for detecting the crude fat content in the microcapsule type fat powder, makes up the vacancy of detecting the crude fat content in the microcapsule type fat powder in the existing feed nutritive value evaluation field, and has the repeatability limit of 0.66-0.99 wt%, the reproducibility limit of 0.55-2.02 wt%, good detection effect and high repeatability.
The applicant declares that the present invention illustrates the detailed structural features of the present invention through the above embodiments, but the present invention is not limited to the above detailed structural features, that is, it does not mean that the present invention must be implemented depending on the above detailed structural features. It should be understood by those skilled in the art that any modifications of the present invention, equivalent substitutions of selected components of the present invention, additions of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
Claims (10)
1. A method for extracting crude fat from microcapsule type fat powder is characterized in that the determination method comprises the following steps:
(1) hydrolyzing with 3-5 moL/L hydrochloric acid with mass m0Obtaining hydrolysate from the microcapsule type fat powder sample, wherein the proportion of the hydrochloric acid to the microcapsule type fat powder sample is 15-25 mL and 1-2 g;
(2) adding a solvent into the hydrolysate obtained in the step (1) for extraction, mixing, centrifuging, separating an upper layer solvent, repeating extraction at least twice, and combining the upper layer solvents obtained by the extraction at least twice to obtain an extracting solution, wherein the solvent comprises petroleum ether;
(3) removing the solvent from the extracting solution in the step (2), and drying to obtain the extract with mass m1The crude fat sample of (2).
2. The extraction method according to claim 1, wherein the concentration of the hydrochloric acid in the step (1) is 4 moL/L;
preferably, the ratio of the hydrochloric acid to the microcapsule fat powder sample is 20mL: 1-2 g.
3. The extraction method according to claim 1 or 2, wherein the hydrolysis temperature in step (1) is 70 to 100 ℃, preferably 80 to 90 ℃;
preferably, the hydrolysis time is 0.5-1.5 h, preferably 1 h;
preferably, the hydrolysis is carried out under intermittent vortexing conditions;
preferably, the intermittent vortexing comprises: mixing hydrochloric acid with a microcapsule type fat powder sample, and then carrying out primary vortex for 25-35 s;
preferably, the intermittent vortex further comprises: swirling for 10-20 s every 15-25 min in the hydrolysis process;
preferably, the hydrolysis is performed in a centrifuge tube.
4. The extraction method according to any one of claims 1 to 3, wherein the petroleum ether in the step (2) has a boiling point of 40 to 60 ℃;
preferably, the solvent of the first extraction comprises ethanol;
preferably, the ethanol is ethanol with a concentration of 95 wt%;
preferably, the volume ratio of the petroleum ether to the ethanol in the solvent for the first extraction is 3-5: 1, preferably 4: 1;
preferably, the volume ratio of the solvent to the hydrochloric acid in the step (1) is 0.9-1.5: 1, and preferably 1-1.25: 1.
5. The extraction method according to any one of claims 1 to 4, wherein the number of times of extraction in step (2) is two;
preferably, the extraction is performed in a centrifuge tube;
preferably, the mixing comprises oscillatory mixing;
preferably, the time of the oscillating mixing is 35-45 s;
preferably, the rotating speed of the centrifugation is 4500-5500 r/min;
preferably, the centrifugation time is 5-7 min.
6. The extraction method according to any one of claims 1 to 5, wherein the step (2) comprises: adding petroleum ether and 95 wt% ethanol into the hydrolysate obtained in the step (1) in a volume ratio of 3-5: 1 as a first solvent, sealing a centrifuge tube, oscillating and mixing for 35-45 s, centrifuging at 4500-5500 r/min for 5-7 min, and separating an upper layer solvent to complete first extraction;
adding petroleum ether into the lower layer liquid as a second solvent, repeatedly performing second extraction, and combining the upper layer solvents obtained by the two extractions to obtain an extracting solution;
the volume ratio of the first solvent to the hydrochloric acid is 1-1.25: 1; the volume ratio of the second solvent to the hydrochloric acid is 1-1.25: 1.
7. The extraction process according to any one of claims 1 to 6, wherein the solvent removal in step (3) comprises distillation of the solvent and aliphatometer removal of the solvent;
preferably, the drying temperature is 101-105 ℃;
preferably, the drying time is 10-20 min;
preferably, said drying comprises a cooling step;
preferably, the cooling is performed in a dry environment;
preferably, the cooling is performed in a dryer.
8. A method for measuring the content of crude fat in microcapsule-type fat powder, which is characterized in that the method for measuring adopts the method for extracting crude fat in microcapsule-type fat powder of any one of claims 1 to 7 to extract the crude fat, and m is used1And m0The ratio of (A) to (B) is recorded as the crude fat content in the microcapsule-type fat powder.
9. The method according to claim 8, wherein m is1The measurement method (2) comprises: the weighing container has a mass m2The total mass of the weighing container and the crude fat sample is m3By m3Minus m2Obtaining the mass m of the crude fat sample1。
Preferably, the container is dried and cooled, and the weight of the container is measured to be m2；
Preferably, the drying temperature of the container is 101-105 ℃;
preferably, the drying time of the container is 25-35 min;
preferably, the cooling of the container is performed in a dry environment;
preferably, the cooling of the container is performed in a dryer;
preferably, the container is an aluminium cylinder.
10. An assay method according to claim 8, wherein the method comprises the steps of:
(1) weighing m0Adding 3-5 moL/L hydrochloric acid into a micro-capsule type fat powder sample in a centrifugal tube, sealing the centrifugal tube, performing vortex for 25-35 s, and performing hydrolysis at 80-90 ℃, wherein the hydrolysis time is 0.9-1.5 h, and the vortex is performed for 10-20 s every 15-25 min in the hydrolysis process to obtain a hydrolysis solution; wherein the ratio of the hydrochloric acid to the microcapsule fat powder sample is 20mL to 1-2 g;
drying the aluminum cylinder at 101-105 ℃ for 25-35 min, placing the dried aluminum cylinder in a dryer to cool to room temperature, and weighing the aluminum cylinder with the mass m2；
(2) Adding petroleum ether and ethanol with the concentration of 95 wt% into the hydrolysate in the step (1) in a volume ratio of 3-5: 1 as a first solvent, sealing a centrifuge tube, oscillating and mixing for 35-45 s, centrifuging at 4500-5500 r/min for 5-7 min, and absorbing an upper layer solvent into the aluminum cylinder in the step (1) to finish first extraction;
adding petroleum ether into the lower layer liquid as a second solvent, repeatedly performing second extraction, combining the upper layer solvents obtained by the two extractions into the aluminum cylinder in the step (1) to obtain an extracting solution;
the volume ratio of the first solvent to the hydrochloric acid is 1-1.25: 1; the volume ratio of the second solvent to the hydrochloric acid is 1-1.25: 1;
(3) removing the solvent from the extracting solution in the step (2), drying at 101-105 ℃ for 10-20 min, placing in a dryer, cooling to room temperature, wherein the total mass of the weighing container and the crude fat sample is m3By m3Minus m2Obtaining the mass m of the crude fat sample1In m is1And m0The ratio of (A) to (B) is recorded as the crude fat content in the microcapsule-type fat powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010627317.3A CN111766174A (en) | 2020-07-01 | 2020-07-01 | Method for extracting crude fat from microcapsule type fat powder and method for measuring content of crude fat |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010627317.3A CN111766174A (en) | 2020-07-01 | 2020-07-01 | Method for extracting crude fat from microcapsule type fat powder and method for measuring content of crude fat |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111766174A true CN111766174A (en) | 2020-10-13 |
Family
ID=72723536
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010627317.3A Pending CN111766174A (en) | 2020-07-01 | 2020-07-01 | Method for extracting crude fat from microcapsule type fat powder and method for measuring content of crude fat |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111766174A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1052508A2 (en) * | 1999-05-04 | 2000-11-15 | Ronald J Komarek | A method for determining fat (crude) in feed, food, and other materials utilizing filter media encapsulation |
| CN103278420A (en) * | 2013-05-24 | 2013-09-04 | 西南大学 | Method for rapidly measuring content of crude fat in food |
| CN103760058A (en) * | 2014-01-03 | 2014-04-30 | 南通市产品质量监督检验所 | Method for determining fat by acid hydrolysis |
| CN105466805A (en) * | 2015-12-25 | 2016-04-06 | 天津市傲景农业科技发展有限公司 | Method for detecting fat content of black bean milk |
| CN108254459A (en) * | 2017-12-27 | 2018-07-06 | 嘉必优生物技术（武汉）股份有限公司 | The detection method of BaP in a kind of microcapsules pulvis |
-
2020
- 2020-07-01 CN CN202010627317.3A patent/CN111766174A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1052508A2 (en) * | 1999-05-04 | 2000-11-15 | Ronald J Komarek | A method for determining fat (crude) in feed, food, and other materials utilizing filter media encapsulation |
| CN103278420A (en) * | 2013-05-24 | 2013-09-04 | 西南大学 | Method for rapidly measuring content of crude fat in food |
| CN103760058A (en) * | 2014-01-03 | 2014-04-30 | 南通市产品质量监督检验所 | Method for determining fat by acid hydrolysis |
| CN105466805A (en) * | 2015-12-25 | 2016-04-06 | 天津市傲景农业科技发展有限公司 | Method for detecting fat content of black bean milk |
| CN108254459A (en) * | 2017-12-27 | 2018-07-06 | 嘉必优生物技术（武汉）股份有限公司 | The detection method of BaP in a kind of microcapsules pulvis |
Non-Patent Citations (2)
| Title |
|---|
| 张丽英: "《饲料分析及饲料质量检测技术》", 30 September 2016 * |
| 杨宝亮等: "《最新实用医学》", 30 June 2009 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Boyer et al. | Determination of vitamin A and carotene in milk. A rapid extraction procedure | |
| CN111766174A (en) | Method for extracting crude fat from microcapsule type fat powder and method for measuring content of crude fat | |
| CN101034065A (en) | Reagent kit for quickly testing protein content of milk powder and method for making same | |
| Botsoglou et al. | Rapid gas chromatographic method for simultaneous determination of cholesterol and α-tocopherol in eggs | |
| CN103698328B (en) | A kind of reduced sugar direct titrimetric method for improving titration precision | |
| Entenman | [56] Preparation and determination of higher fatty acids | |
| CN105572260B (en) | One kind measure dissolving pulp in α-, β-and gamma cellulose content method | |
| CN110426488A (en) | A kind of method of GB 5009.34-2016 national food safety standard detection sulfur dioxide in food | |
| CN106387296B (en) | Production method of soybean protein isolate with high extraction rate | |
| CN110028602A (en) | A method of reducing impurity content in commercial polyethylene alcohol | |
| US3062623A (en) | Reagent for the determination of fat in blood serum | |
| CN108918345A (en) | The alcoholic strength detection method of grape wine or fruit wine | |
| CN108387412A (en) | The pre-treating method of detection aquatic products Malachite Green, crystal violet and its metabolite | |
| Ua Conaill et al. | An automated method for the determination of estrogens in pregnancy | |
| CN109632786A (en) | A method of accurately detecting phytosterol ester content using microplate reader | |
| CN110749684A (en) | Rapid detection method for organic acid in enzyme | |
| Steyermark | Microdetermination of alkoxyl groups | |
| CN110455946A (en) | The rapid detection method of 1,3-Dioleic acid-2-palmitoyl triglyceride in a kind of dairy products | |
| CN109679776A (en) | A kind of the medicament cleaning solution and cleaning method of oil well drilling cuttings | |
| CN105716993A (en) | Method for measuring fat content in frozen drink | |
| Long et al. | A rapid automatic analyzer and its methodology for effective bentonite content based on image recognition technology | |
| CN108613960A (en) | The Raman spectra detection process of phthalic ester plasticizer | |
| CN108303412A (en) | A kind of assay method of concentrated bone soup peroxide value | |
| Peterson et al. | Optimization of a batch type ethyl ester process | |
| RU2805412C1 (en) | Method for determining the content of magnesium fluoride in sellaite and fluorite-sellaite products |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201013 |