CN111413170A - Preparation method of heavy metal standard substance of crayfish matrix - Google Patents
Preparation method of heavy metal standard substance of crayfish matrix Download PDFInfo
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- CN111413170A CN111413170A CN202010297891.7A CN202010297891A CN111413170A CN 111413170 A CN111413170 A CN 111413170A CN 202010297891 A CN202010297891 A CN 202010297891A CN 111413170 A CN111413170 A CN 111413170A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2866—Grinding or homogeneising
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Abstract
The invention discloses a preparation method of a heavy metal standard substance of a crayfish matrix, which comprises the following steps: removing impurities from live crayfish, soaking, cleaning, and homogenizing to obtain lobster meat; adding a heavy metal solution into the homogenized lobster meat, and uniformly mixing the lobster meat and the heavy metal solution; freeze-drying the homogenate, and performing staged freeze-drying; grinding the freeze-dried sample into powder, sieving, packaging and sterilizing. The decontamination of live crayfish comprises removing the shell, pancreas, and shrimp line of live crayfish, leaving only the shrimp meat. Soaking and cleaning, which comprises soaking and cleaning the meat of living crayfish with normal saline. The preparation method disclosed by the invention is mechanically operated in the whole process, and large-scale mechanized production is used in the homogenizing, vacuum freeze drying and grinding stages, so that the product yield and efficiency can be greatly improved, and the preparation method is more efficient, simple, convenient and reliable. The preparation process of the invention can produce the standard substance in large batch, and the freeze-dried standard substance has stable performance and is convenient to transport and carry.
Description
Technical Field
The invention relates to a preparation method of a heavy metal standard substance of a crawfish matrix, belonging to the technical field of preparation of standard substances for detecting the content of heavy metals in aquatic products.
Background
The crayfish is named as Procambarus clarkii, is propagated in the Yangtze river delta area near Nanjing after being introduced into China from Japan, gradually spreads to the middle and lower reaches of the Yangtze river due to strong adaptability, omnivorous property, high growth speed and the like, and is deeply loved by the masses of people due to delicious taste and high nutritional value. However, since the crayfish has low requirements for the growth environment, has the characteristics of rancidity and benthic property, is easy to enrich heavy metals, and is not beneficial to the health of human bodies after being eaten, the safety detection of the crayfish is particularly important.
One typically will adjust the relative density to be greater than 4.5 g/cm3The elements of (a) are called heavy metal elements, such as: mercury, lead, nickel, cadmium, copper, chromium, and the like. When it exceeds a certain amount, it may cause biological hazards such as carcinogenicity and teratogenicity, and may also cause damage to the central nervous system, digestive system, respiratory system and various organs. In the detection process, a reference substance (which we often refer to as a standard substance), especially a standard substance related to the matrix of the detection sample, is often used to achieve more accurate measurement results, thereby reducing the matrix effect. By matrix effect is meant: the effect of other components in the sample than the target analyte on the value measured for the analyte.
The existing heavy metal standard substances in China can be divided into two types, one type is a pure heavy metal solution and a plurality of heavy metals are mixed. Still another group is based on various fish, meat, etc. Among the relevant standard substances of aquatic products in China, the standard substances of heavy metals with aquatic products as matrixes are as follows: the standard substance for analyzing the components of GBW10024 scallops, the standard substance for analyzing the components of total mercury and methyl mercury in GBW10029 fish, the standard substance for analyzing the forms of inorganic elements and organic tin in GBW10068 oysters and the like. At present, no heavy metal standard substance (namely no crayfish freeze-dried powder) taking crayfish as a matrix and no preparation process of the crayfish standard substance exist. In the prior art, the preparation process of the dried crayfish only carries out freeze-drying and dehydration treatment on the crayfish, and the preparation efficiency is low. The crayfish is increasingly favored by people, and the annual eating amount can reach 4.5 ten thousand tons, so that the heavy metal standard substance based on the crayfish matrix needs to be developed urgently.
Disclosure of Invention
The invention aims to overcome the defects of no crayfish matrix heavy metal standard substance and no preparation method in the prior art, and provides a preparation method of the crayfish matrix heavy metal standard substance, which has the following technical scheme:
the preparation method of the heavy metal standard substance of the crayfish matrix comprises the following steps:
removing impurities from live crayfish, soaking, cleaning, and homogenizing to obtain lobster meat;
adding a heavy metal solution into the homogenized lobster meat, and uniformly mixing the lobster meat and the heavy metal solution;
freeze-drying the homogenate, and performing staged freeze-drying;
grinding the freeze-dried sample into powder, sieving, packaging and sterilizing.
Further, decontamination of live crayfish comprises removing the shell, pancreas, and shrimp line from live crayfish leaving only the meat portion of the crayfish.
Further, the soaking and cleaning step comprises soaking and cleaning the meat of the live crayfish with physiological saline.
Preferably, the normal saline is 0.85% sodium chloride solution, the soaking time is 10 min, and the washing is repeated 3 times.
Further, homogenizing includes mincing the meat of the cleaned live crayfish.
Preferably, the lobster meat of the cleaned live crayfish is crushed by a knife grinder, the speed of the knife grinder is 1500 r/min, positive and negative rotation is set, and the mixture is homogenized for 10 min.
Further, the method for adding the heavy metal solution to the homogenized lobster meat comprises the following steps: the standard solution was weighed into the homogenized lobster meat using an electronic balance.
Further, the freeze drying adopts a vacuum freeze dryer, the staged freeze drying adopts a gradient freeze drying program, and the gradient freeze drying program comprises the following steps:
freezing at-20 deg.C for 1h20min;
drying at-10 deg.C under 200 μ bar vacuum degree for 24h30min;
drying for 24h30min at 0 deg.C;
drying again at 20 deg.C for 10h30 min.
Further, the sample after freeze-drying is ground by an ultracentrifugal grinder.
Compared with the prior art, the invention has the following beneficial effects:
the heavy metal standard substance of the crayfish matrix prepared by the preparation method can provide a fixed value reference for detection of heavy metals in crayfish. The preparation method disclosed by the invention is mechanically operated in the whole process, and large-scale mechanized production is used in the homogenizing, vacuum freeze drying and grinding stages, so that the product yield and efficiency can be greatly improved, and the preparation method is more efficient, simple, convenient and reliable. The invention can make up for domestic blank, and the prepared standard substance can greatly reduce the gap between the standard substance and the detection sample. The crayfish matrix prepared by the invention is a detection process for reducing a sample to the maximum extent by adding the heavy metal solution into the lobster meat, thereby eliminating the matrix effect in detection. The preparation process of the invention can produce the standard substance in large batch, and the freeze-dried standard substance has stable performance and is convenient to transport and carry.
Drawings
FIG. 1 is a flow chart of the preparation method of the heavy metal standard substance of crawfish matrix of the present invention.
Detailed Description
The invention is further described below with reference to the accompanying drawings. The following examples are only for illustrating the technical solutions of the present invention more clearly, and the protection scope of the present invention is not limited thereby.
Example 1
As shown in figure 1 of the drawings, in which,
the preparation method of the heavy metal standard substance of the crayfish matrix comprises the following steps:
removing impurities from live crayfish, soaking, cleaning, and homogenizing to obtain lobster meat;
adding a heavy metal solution into the homogenized lobster meat, and uniformly mixing the lobster meat and the heavy metal solution;
freeze-drying the homogenate, and performing staged freeze-drying;
grinding the freeze-dried sample into powder, sieving, packaging and sterilizing.
Specifically, in this embodiment, the decontamination of the live crayfish comprises removing dead crayfish from the crayfish, removing the shell, pancreas, shrimp line, etc. from the live crayfish, and leaving only the shrimp meat.
In this embodiment, the immersion cleaning specifically includes immersing and cleaning the peeled meat of the living crayfish with a physiological saline solution.
In this example, the normal saline is a 0.85% sodium chloride solution, the soaking time is 10 min, and the washing is repeated 3 times.
In particular, in this example, the homogenizing includes mincing the meat of the cleaned live crayfish.
In the preferred embodiment, the cleaned lobster meat of live crayfish is ground by a knife grinder at 1500 r/min, and the machine is rotated 4 times in the forward and reverse directions to homogenize the lobster meat for 10 min. Adopt sword formula grinding appearance to homogenize lobster meat for homogenate is more abundant. The knife grinder in this example is a HM300 type knife grinder manufactured by beijing griffoldman instruments and devices ltd.
In this example, the method of adding the heavy metal solution to the homogenized lobster meat is specifically as follows: the standard solution was weighed into the homogenized lobster meat using an electronic balance.
In this embodiment, specifically, the adding of heavy metal includes dividing the well-homogenized lobster meat into 3 batches, wherein the first batch is not added with heavy metal solution and is used as the background of the lobster meat, the second batch is added with low-concentration heavy metal solution, and the third batch is added with high-concentration heavy metal solution. The invention provides 3 gradient concentrations which can be used by users to make standard curves. Homogenizing again after adding, and fully and uniformly mixing the heavy metal solution and the lobster meat. The standard solution is weighed by an electronic balance and then added into the shrimp meat, and the weighing method is more accurate. The heavy metals added are: chromium, cadmium, copper, mercury, lead, arsenic; the reagents used in this example were:
sodium chloride: chemical agents of the national drug group, ltd;
GBW (E)080117 mercury component analysis standard substance in water 100 mug/m L, U = 0.8% (k =2), China measurement institute.
GBW (E)080119 analysis standard substance for cadmium components in water, 100 mug/m L, U = 0.8% (k =2) in China measurement institute;
GBW (E)080119 water chromium component analysis standard substance 100 mug/m L, U = 0.8% (k =2), China measurement institute;
GBW (E)080129 standard substance for analyzing lead components in water, 100 mug/m L, U = 0.8% (k =2), China measurement institute;
GBW (E)080117 standard substance for analyzing arsenic content in water, 100 mug/m L, U = 0.8% (k =2), China measurement institute;
GBW (E)080122 water copper component analysis standard substance 100 mug/m L, Urel = 0.8% (k =2), China measurement institute.
In this embodiment, the vacuum freeze dryer is used for freeze drying, and the gradient freeze-drying procedure is used for staged freeze drying, and the gradient freeze-drying procedure is as follows:
freezing at-20 deg.C for 1h20min;
drying at-10 deg.C under 200 μ bar vacuum degree for 24h30min;
drying for 24h30min at 0 deg.C;
the freeze-drying comprises freeze-drying the obtained homogenate in a vacuum freeze-dryer, the vacuum freeze-dryer used in the present example is a L yoBeta vacuum freeze-dryer produced by spanish thailand, the present example is provided with a gradient freeze-drying procedure for freeze-drying the sample, the specific procedure is as follows:
| phases | Procedure for measuring the movement of a moving object | Temperature of | Degree of vacuum | Time of day |
| 1 | Freezing | -20℃ | / | 1h20min |
| 2 | Drying | -10℃ | 200μbar | 24h30min |
| 3 | Secondary drying | 0℃ | / | 24h30min |
| 4 | Drying again | 20℃ | / | 10h30min |
Set up gradient temperature control procedure in vacuum freeze-drying process for the sample water content further reduces, and stage nature freeze-drying makes its water content little, easily preserves the transportation. For better transportation and storage and convenience for test, the lobster meat is subjected to freeze-drying treatment and then ground into powder, so that a tester can weigh samples conveniently.
And grinding the freeze-dried sample by using an ultracentrifugal grinder.
Specifically, grinding and sieving (i.e. sample sieving), and grinding the freeze-dried sample in an ultracentrifugal grinder, wherein the aperture of the sieve is 0.5 μm; the pulverized sample was allowed to pass through a 0.5 μm sieve. In the grinding and sieving stage, the traditional pure manual grinding and sieving is abandoned, an ultracentrifugal grinding instrument (the ultracentrifugal grinding instrument: FM200, manufactured by Beijing Graydeman instruments and Equipment Co., Ltd.) is introduced, and the grinding efficiency and the grinding quantity are greatly improved.
Subpackaging and sterilizing, specifically, subpackaging the ground sample into brown penicillin bottles, wherein each bottle contains 2 g of the ground sample, covering with a chitin glue soft cover, and sealing with an aluminum cover. The subsequent sterilization route was irradiation with Co60 at a dose of 18 kGy. In the embodiment, the standard substance (namely the dried lobster meat powder) is weighed by an electronic balance, so that the subpackaging is more accurate.
The preparation method disclosed by the invention is mechanically operated in the whole process, and large-scale mechanized production is used in the homogenizing, vacuum freeze-drying and grinding stages, so that the product yield and efficiency can be greatly improved, 100 kg of lobsters are used each time, 6.6 kg of lobster meat can be peeled off, 800 g of lobster meat dry powder is ground, 2 g of lobster meat dry powder is subpackaged in each bottle, and 400 bottles of standard substances can be produced at one time.
Electronic balance M L303, Metler-Tollido, Switzerland (China) Co.
The heavy metal standard substance of the crayfish matrix prepared by the preparation method can provide a fixed value reference for detection of heavy metals in crayfish. The preparation method disclosed by the invention is mechanically operated in the whole process, and large-scale mechanized production is used in the homogenizing, vacuum freeze drying and grinding stages, so that the product yield and efficiency can be greatly improved, and the preparation method is more efficient, simple, convenient and reliable. The invention can make up for domestic blank, and the prepared standard substance can greatly reduce the gap between the standard substance and the detection sample. The crayfish matrix prepared by the invention is a detection process for reducing a sample to the maximum extent by adding the heavy metal solution into the lobster meat, thereby eliminating the matrix effect in detection. The preparation process of the invention can produce the standard substance in large batch, and the freeze-dried standard substance has stable performance and is convenient to transport and carry.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.
Claims (9)
1. The preparation method of the heavy metal standard substance of the crayfish matrix is characterized by comprising the following steps:
removing impurities from live crayfish, soaking, cleaning, and homogenizing to obtain lobster meat;
adding a heavy metal solution into the homogenized lobster meat, and uniformly mixing the lobster meat and the heavy metal solution;
freeze-drying the homogenate, and performing staged freeze-drying;
grinding the freeze-dried sample into powder, sieving, packaging and sterilizing.
2. The method of claim 1, wherein the depurating of the live crayfish comprises removing shells, pancreas and shrimp threads of the live crayfish to leave only a shrimp meat portion.
3. The method for preparing crayfish-based heavy metal standard substance according to claim 1, wherein the immersion cleaning comprises immersing and cleaning the meat of a living crayfish with a physiological saline solution.
4. The method for preparing crayfish matrix heavy metal standard substance according to claim 3, wherein the normal saline is 0.85% sodium chloride solution, the soaking time is 10 min, and the washing is repeated 3 times.
5. The method of claim 1, wherein the homogenizing comprises mincing the meat of the cleaned live crayfish.
6. The method for preparing crayfish matrix heavy metal standard substance according to claim 5, wherein the lobster meat of the cleaned live crayfish is ground by a knife grinder, the speed of the knife grinder is 1500 r/min, the rotation is set to be positive and negative, and the mixture is homogenized for 10 min.
7. The method for preparing heavy metal standard substance of crayfish matrix according to claim 1, wherein the method for adding heavy metal solution to the homogenized lobster meat comprises: the standard solution was weighed into the homogenized lobster meat using an electronic balance.
8. The method for preparing crayfish matrix heavy metal standard substance according to claim 1, wherein the freeze drying is performed by a vacuum freeze dryer, the staged freeze drying is performed by a gradient freeze drying procedure, and the gradient freeze drying procedure is as follows:
freezing at-20 deg.C for 1h20min;
drying at-10 deg.C under 200 μ bar vacuum degree for 24h30min;
drying for 24h30min at 0 deg.C;
drying again at 20 deg.C for 10h30 min.
9. The method for preparing crayfish-based heavy metal standard substance according to claim 1, wherein the freeze-dried sample is ground by an ultracentrifugal grinder.
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Cited By (1)
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| CN116380587A (en) * | 2023-04-07 | 2023-07-04 | 云南省农业科学院质量标准与检测技术研究所 | Lentinus edodes matrix standard substance containing heavy metals and chemical forms thereof and preparation method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116380587B (en) * | 2023-04-07 | 2024-01-23 | 云南省农业科学院质量标准与检测技术研究所 | Preparation method of lentinus edodes matrix standard substance containing heavy metals and chemical forms thereof |
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