CN111551412B - Preparation method of soil sample containing cyanide and total cyanide - Google Patents
Preparation method of soil sample containing cyanide and total cyanide Download PDFInfo
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- CN111551412B CN111551412B CN202010455911.9A CN202010455911A CN111551412B CN 111551412 B CN111551412 B CN 111551412B CN 202010455911 A CN202010455911 A CN 202010455911A CN 111551412 B CN111551412 B CN 111551412B
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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
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65D—CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
- B65D81/00—Containers, packaging elements, or packages, for contents presenting particular transport or storage problems, or adapted to be used for non-packaging purposes after removal of contents
- B65D81/18—Containers, packaging elements, or packages, for contents presenting particular transport or storage problems, or adapted to be used for non-packaging purposes after removal of contents providing specific environment for contents, e.g. temperature above or below ambient
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65D—CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
- B65D85/00—Containers, packaging elements or packages, specially adapted for particular articles or materials
- B65D85/50—Containers, packaging elements or packages, specially adapted for particular articles or materials for living organisms, articles or materials sensitive to changes of environment or atmospheric conditions, e.g. land animals, birds, fish, water plants, non-aquatic plants, flower bulbs, cut flowers or foliage
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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
- G01N1/34—Purifying; Cleaning
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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
- 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
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
Abstract
The invention discloses a method for preparing a soil sample containing cyanide and total cyanide, which comprises the following steps: collecting a soil sample, and transferring in a dark place; freezing the soil sample, and acting on microorganisms to form ice crystals in microbial cells to puncture the cells so as to kill part of the microorganisms, thereby reducing the activity of soil microorganisms; vacuum low-temperature freeze drying; and (4) crushing the sample, removing impurities, and storing in a dark and sealed manner. The influence of the light effect on soil cyanide and total cyanide is reduced by shading and blocking, and the cyanide and the total cyanide in the soil can be effectively inhibited from being decomposed by microorganisms through freezing and freeze drying, so that the storage time of the soil cyanide and the total cyanide is prolonged.
Description
Technical Field
The invention belongs to the technical field of environmental detection, relates to a soil sample preservation method, and particularly relates to a soil sample preservation method containing cyanide and total cyanide.
Background
Cyanide refers to a compound containing a cyano group (-CN) in the molecule of the compound, and belongs to a highly toxic compound.
Total cyanide refers to the sum of organic cyanide, free cyanide ion, complex and simple metal cyanide. Cyanide complexes of cobalt and thiocyanide are excluded. Free cyanide refers to the sum of free cyanide ions and simple metal cyanides, excluding organic cyanides.
Mainly HCN, CN in the environment-And in the form of a complex cyanide ion. Human activities such as gold production, electroplating chemical industry, organic chemical industry and other industries bring a large amount of cyanide-containing wastewater; causing great harm to ecological environments such as water, soil and the like. Therefore, in environmental monitoring, it is very important to measure the cyanide concentration.
Currently there is no good method of preserving soil samples containing cyanide as well as total cyanide for a while. Under the existing conventional conditions, the sample storage time is only 48h, and the actual measurement operation difficulty is high. The pretreatment for measuring the cyanide concentration has a significant influence on the accuracy of the measurement result.
In the process of implementing the invention, the inventor finds that at least the following problems exist in the prior art:
1. the preservation time of the soil sample containing cyanide and total cyanide is only 48h, which is not beneficial to the actual determination work and the sample retention and reinspection.
2. The soil moisture content is greatly influenced by environmental factors and weather factors, and the existing soil sample collection means is not beneficial to realizing standardized detection.
Disclosure of Invention
In view of the above, the present invention aims to provide a soil sample preservation method capable of realizing long-term preservation.
The inventor provides a technical scheme for solving the technical problems by continuous innovation and innovation through long-term exploration and attempt, and multiple experiments and endeavors, and provides a method for preparing a soil sample containing cyanide and total cyanide, which comprises the following steps:
s1: collecting a soil sample, and transferring in a dark place;
s2: freezing the soil sample, and acting on microorganisms to form ice crystals in microbial cells to puncture the cells so as to kill part of the microorganisms, thereby reducing the activity of soil microorganisms;
s3: vacuum low-temperature freeze drying;
s4: and (4) crushing the sample, removing impurities, and storing in a dark and sealed manner.
According to a preferred embodiment of the method for preparing soil samples containing cyanide and total cyanide according to the present invention, in the step S2, the freezing temperature is lower than-10 ℃; the freezing time is 20-50 min.
According to a preferred embodiment of the method for preparing soil samples containing cyanide and total cyanide, in step S2, the freezing temperature is-20 ℃ and the freezing time is 30 min.
According to a preferred embodiment of the method for preparing soil samples containing cyanide and total cyanide, in step S3, the samples are dried in a low-temperature freeze dryer under reduced pressure for 4 hours, the vacuum degree is controlled to be 4-6Pa, and the freezing temperature is-50 ℃.
According to a preferred embodiment of the method for preparing soil samples containing cyanide and total cyanide according to the present invention, in the step S4, the crushed particle size of the samples is 1mm or less.
According to a preferred embodiment of the method for preparing soil samples containing cyanide and total cyanide, the impurity removal is sieving, magnetic adsorption and electrostatic adsorption.
According to a further embodiment of the method for preparing soil samples containing cyanide and total cyanide, the method further comprises adjusting the pH value of the soil samples to be alkaline before S2.
According to a preferred embodiment of the method for preparing soil samples containing cyanide and total cyanide according to the invention, the pH is not lower than 12.
According to a preferred embodiment of the method for preparing soil samples containing cyanide and total cyanide according to the invention, sodium hydroxide solution is added to the soil samples.
According to a preferred embodiment of the method for preparing soil samples containing cyanide and total cyanide, the soil samples are stored in a brown glass bottle in a sealed and dark manner and are stored at room temperature or below.
Compared with the prior art, one of the technical solutions has the following advantages:
a) in one embodiment of the preparation method of the soil sample containing cyanide and total cyanide, the influence of the illumination effect on the soil cyanide and the total cyanide is reduced by shading and blocking, and the cyanide and the total cyanide in the soil can be effectively inhibited from being decomposed by microorganisms through freezing and freeze drying, so that the storage time of the soil cyanide and the total cyanide is prolonged.
b) In one embodiment of the method for preparing the soil sample containing cyanide and total cyanide, the soil sample with more standardized characteristics can be obtained by sieving, electrostatic adsorption, magnetic adsorption and other treatments, and the influence of metal impurities and organic impurities on the measurement result is avoided.
c) In one embodiment of the method for preparing the soil sample containing cyanide and total cyanide, the soil is adjusted to be alkaline, so that cyanide does not overflow in the freeze drying process, and the precision and the accuracy of the detection result are fully ensured.
d) In one embodiment of the method for preparing the soil sample containing cyanide and total cyanide, the soil sample is frozen at the temperature of-20 ℃ to form ice crystals in microbial cells to puncture the cells so as to kill part of microbes, so that the microbial activity of the soil is reduced, and then water is rapidly sublimated in a vacuum low-temperature freeze drying mode to further kill and inactivate the microbes so as to stabilize the cyanide and the total cyanide in the soil.
e) By measuring the contents of cyanide and total cyanide in the soil sample prepared by the method, compared with the contents of cyanide and total cyanide in a fresh sample, the method does not cause loss of cyanide and total cyanide in the soil during treatment, and can obtain a long-term stable preservation effect.
Drawings
FIG. 1 is a graph showing trends in cyanide and total cyanide recovery in soil samples under different temperature storage conditions.
FIG. 2 is a graph showing the trend of cyanide and total cyanide recovery after low temperature vacuum drying of a soil sample after freezing.
FIG. 3 is a graph showing the trend of cyanide and total cyanide recovery after vacuum drying of frozen soil samples at room temperature.
FIG. 4 is a graph showing the tendency of cyanide and total cyanide recovery rate of soil samples prepared by the method of the present invention under different storage temperature conditions.
Detailed Description
The following description will be given with reference to specific examples.
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the embodiments of the present invention will be described in detail and completely with reference to the accompanying drawings. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. Thus, the following detailed description of embodiments of the invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention.
Example 1
Currently there is no good method of preserving soil samples containing cyanide as well as total cyanide for a while. Under the existing conventional conditions, the sample retention time is at most 48h, the measurement work of cyanide and total cyanide is greatly influenced, and the requirement on detection time is very urgent.
This example makes it possible to prolong the storage time of soil cyanides and total cyanides, and it has been demonstrated by tests that the contents of cyanides and total cyanides in soil samples do not differ significantly from the contents of cyanides and total cyanides in fresh soil samples after 17 days of storage.
The method for preparing the soil sample containing cyanide and total cyanide comprises the following steps:
s1: and collecting a soil sample and transferring in a dark place.
Collecting soil samples according to the standard requirements of HJ745-2015, processing in dark place, and quickly transporting to a laboratory for processing.
Soil cyanide and total cyanide are greatly influenced by illumination, and the decomposition of the soil cyanide and the total cyanide can be reduced by light-shielding treatment, such as loading and transporting collected soil samples by using containers such as a cassette, a brown glass bottle and the like.
The inventor finds that the decomposition effect of bacteria in soil cyanide and total cyanide soil is large, and the reduction/inactivation of bacteria is the key to prolonging the storage time of soil cyanide and total cyanide. Specifically, the method comprises a freezing treatment and a freeze-drying treatment, and the inactivation treatment is carried out on the microorganisms.
S2: freezing soil sample, acting on microorganism, and making the microorganism form ice crystal in the microorganism cell to pierce the cell and make part of microorganism die, so as to reduce soil microorganism activity. Three major factors for the survival of bacteria are water, temperature and nutrients, wherein the nutrients are the target substances which are required to be preserved.
Transferring the sample into a container, and freezing for 20-50min at the temperature lower than-10 ℃; preferably, the freezing temperature in the embodiment is-20 ℃, and the freezing market is selected for 30 min.
FIG. 1 is a comparison of soil cyanide and total cyanide content at different temperature storage conditions. As can be seen from fig. 1, the recovery of cyanide and total cyanide after long-term storage of the sample can be significantly improved by only freezing. The bacterial activity can be reduced by freezing treatment, but the bacteria can not be completely inactivated by freezing preservation only, and the aim of preserving the soil sample for a long time cannot be achieved.
S3: and (5) carrying out vacuum low-temperature freeze drying.
The sample was dried in a cryo-lyophilizer under reduced pressure for 4 hours. Drying conditions are as follows: the vacuum degree in the whole drying process is controlled at 4-6Pa, and the freezing temperature is-50 ℃. In this example, the degree of vacuum was controlled to 5 Pa.
FIG. 2 is a graph showing the recovery effect of cyanide and total cyanide after low-temperature vacuum drying of a frozen soil sample, and FIG. 3 is a graph showing the recovery effect of cyanide and total cyanide after vacuum drying of a frozen soil sample at room temperature.
Experiments prove that water is rapidly sublimated in a vacuum low-temperature freeze drying mode, so that microorganisms are further killed and inactivated, and cyanides and total cyanides in soil are stabilized. Comparing fig. 2 and fig. 3, it can be seen that the low-temperature freeze vacuum drying can effectively preserve cyanides and total cyanides in the soil, while the room-temperature vacuum drying has a proper temperature in the whole drying process, so that the bacteria in the soil still decompose the cyanides and the total cyanides in the soil in a large amount even under the vacuum condition, and the recovery rate of the cyanides and the total cyanides is low.
S4: and (4) crushing the sample, removing impurities, and storing in a dark and sealed manner.
And (4) taking out the soil sample after drying to prepare a sample with the particle size of 1mm, and storing the sample at room temperature in a dark place. By adopting the treatment of sieving, electrostatic adsorption, magnetic adsorption and the like, the soil sample with more standardized characteristics can be obtained, and the influence of metal impurities such as iron and the like, and organic impurities such as tree roots, grass roots, paper scraps and the like on the measurement result is avoided.
This embodiment freezes, the low temperature freeze-drying back with the soil sample, and the testing result of cyanide and total cyanide in the sample is fine, and whole low temperature freeze-drying process can not cause the loss to cyanide and total cyanide in to soil, and need not low temperature and isolated oxygen after the freeze-drying, only need the light-resistant can be preserved for a long time. The influence of different storage temperatures on the pattern samples is shown in fig. 4, and the pattern samples are stored in a brown glass bottle in a sealed manner in the dark and at room temperature or below.
The method can effectively prolong the storage time of the soil sample containing cyanide and total cyanide, the preparation process can not cause any influence on the form and concentration of the cyanide and the total cyanide, and the storage of the prepared soil sample is not influenced by temperature. The instruments and equipment used in the method are convenient and easy to obtain, and the operation is simple.
As a further preferred embodiment, the pH of the soil sample is adjusted to alkaline before freezing of the soil sample, said pH being not less than 12. For example, after a soil sample is collected, a sodium hydroxide solution is added to the soil sample to adjust the pH value, so that cyanide and total cyanide are prevented from overflowing.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.
Claims (7)
1. A preparation method of a total cyanide soil sample comprises the steps of sample collection and impurity removal, and is characterized by comprising the following steps:
s1: collecting a soil sample, and transferring in a dark place; adjusting the pH value of the sample to be alkaline;
s2: freezing the soil sample, and acting on microorganisms to form ice crystals in microbial cells to puncture the cells so as to kill part of the microorganisms, thereby reducing the activity of soil microorganisms; the freezing temperature is lower than-10 ℃; freezing for 20-50 min;
s3: vacuum low-temperature freeze drying; putting the sample into a low-temperature freeze dryer, and drying for 4 hours under reduced pressure, wherein the vacuum degree is controlled to be 4-6Pa, and the freezing temperature is-50 ℃;
s4: and (4) crushing the sample, removing impurities, and storing in a dark and sealed manner.
2. The method for preparing a total cyanide soil sample according to claim 1, wherein the freezing temperature is-20 ℃ and the freezing time is 30min in step S2.
3. The method for preparing a total cyanide soil sample according to claim 1, wherein in step S4, the sample pulverized particle size is 1mm or less.
4. The method for preparing a total cyanide soil sample according to claim 3, wherein the removing impurities is sieving, using magnetic adsorption, using electrostatic adsorption.
5. The method of preparing a total cyanide soil sample according to claim 1, wherein the pH is not less than 12.
6. The method of preparing a total cyanide soil sample of claim 5, wherein a sodium hydroxide solution is added to the soil sample.
7. The method for preparing a total cyanide soil sample according to claim 1, wherein the dark sealed storage is performed by using a brown glass bottle and the storage is performed at room temperature or below.
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| CN109174949B (en) * | 2018-10-15 | 2021-01-12 | 重庆大学 | Method for restoring soil polluted by cyanide by low-temperature heat treatment technology |
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