CN104198216A - Sampling method of fish body muscle for stable isotope analysis - Google Patents
Sampling method of fish body muscle for stable isotope analysis Download PDFInfo
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- CN104198216A CN104198216A CN201410497767.XA CN201410497767A CN104198216A CN 104198216 A CN104198216 A CN 104198216A CN 201410497767 A CN201410497767 A CN 201410497767A CN 104198216 A CN104198216 A CN 104198216A
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- 210000003205 muscle Anatomy 0.000 title claims abstract description 69
- 241000251468 Actinopterygii Species 0.000 title claims abstract description 68
- 238000005070 sampling Methods 0.000 title claims abstract description 36
- 238000000034 method Methods 0.000 title claims abstract description 22
- 238000004458 analytical method Methods 0.000 title claims abstract description 19
- 229910001220 stainless steel Inorganic materials 0.000 claims abstract description 20
- 239000010935 stainless steel Substances 0.000 claims abstract description 20
- 238000001035 drying Methods 0.000 claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000008367 deionised water Substances 0.000 claims abstract description 7
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 7
- 239000000919 ceramic Substances 0.000 claims abstract description 4
- 239000004570 mortar (masonry) Substances 0.000 claims abstract description 4
- 239000000843 powder Substances 0.000 claims abstract description 4
- 238000004140 cleaning Methods 0.000 claims abstract description 3
- 239000008280 blood Substances 0.000 claims abstract 2
- 210000004369 blood Anatomy 0.000 claims abstract 2
- 230000000155 isotopic effect Effects 0.000 claims description 15
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052799 carbon Inorganic materials 0.000 claims description 6
- 210000000988 bone and bone Anatomy 0.000 claims description 5
- 238000012545 processing Methods 0.000 claims description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 3
- 230000037396 body weight Effects 0.000 claims description 3
- 238000013461 design Methods 0.000 claims description 3
- 239000004576 sand Substances 0.000 claims description 3
- 238000012360 testing method Methods 0.000 claims description 3
- 210000001519 tissue Anatomy 0.000 claims description 3
- 241000191442 Tenualosa reevesii Species 0.000 claims description 2
- 238000007710 freezing Methods 0.000 claims description 2
- 230000008014 freezing Effects 0.000 claims description 2
- 229910000041 hydrogen chloride Inorganic materials 0.000 claims description 2
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 230000008569 process Effects 0.000 abstract description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract 1
- 238000011109 contamination Methods 0.000 abstract 1
- 230000007774 longterm Effects 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 230000004075 alteration Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 238000001514 detection method Methods 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 241001459819 Carassius gibelio Species 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000003851 biochemical process Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000000700 radioactive tracer Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
Landscapes
- Sampling And Sample Adjustment (AREA)
Abstract
The invention discloses a sampling method of a fish body muscle for stable isotope analysis. The sampling method comprises the steps: 1) designing a muscle sampling tube, 2) preparing a fish sample: ensuring a fish body is fresh and not deteriorated, 3) treating sampling tools: prefiring a stainless steel scalpel, a pair of forceps and the muscle sampling tube for sampling on an alcohol lamp for later use. 4) cutting into the fish body: cutting under a dorsal fin and above vertebrae of the fish body with the prefired scalpel, 5) taking the muscle: inserting one end of the prefired muscle sampling tube into an incision, rotating in a direction parallel to another incision of the fish body, 6) cleaning: washing away blood from the muscle with deionized water, 7) drying, 8) grinding: transferring a dried muscle sample into a prefired ceramic mortar, and grinding into and storing powder. The method is easy, and easy and simple to operate, and can effectively avoid and reduce contamination arising from a sample collecting device and a sample container in a sampling process, and the dried fish body muscle sample can be stored stably for a long term.
Description
Technical field
The present invention relates to analytical test field, more specifically relate to a kind of sampling method of the fish body back white muscles for Stable Isotopic Analysis.
Background technology
Stable isotope technology is to utilize the stable isotope of the light elements such as hydrogen, nitrogen, carbon, oxygen, sulphur as tracer atom, studies various biochemical processes and mechanism, and a technology of relation between the micromechanism of molecule and character.This technology is applied to food safety detection and ecosystem energy flow widely, fish are important component parts of our food, also be consumer important in aquatic ecosystem, therefore, in aquatic product quality detection and aquatic ecosystem research, often to analyze fish muscle sample by stable isotope technology.Yet the instability in the processing of fish body muscle samples and preservation process, easily produces interference to isotope analysis.Therefore, must be convenient and swift for the fish body muscle samples sampling of Stable Isotopic Analysis, can at utmost low eliminating disturb, sample also can be preserved for a long time and keep stable.
Due to the required sample size of Stable Isotopic Analysis seldom, sample receives that the risk of pollution is relatively high.Sample collecting apparatus and sampling receptacle, and the cross pollution of sample room, all can make the reliability of Stable Isotopic Analysis reduce, and in addition, the result of Stable Isotopic Analysis also can be subject to different samplings and the interference of disposal route.Therefore, minimum sample collecting apparatus and sampling receptacle are disturbed or disturbed to this method by choosing without stable isotope, sample collecting apparatus and sampling receptacle are done to get rid of interference processing, and simplify sampling process, can effectively reduce the probability that sample is disturbed and pollutes, sample can be preserved for a long time and keep stable after low temperature drying is processed, and is conducive to improve the reliability in fish Stable Isotopic Analysis.
Summary of the invention
The object of the invention is to be to provide a kind of sampling method of the fish body muscle for Stable Isotopic Analysis, easy to implement the method, easy and simple to handle, can effectively avoid and reduce in sampling process the pollution bringing due to sample collecting apparatus and sampling receptacle, and sample cross pollution and the sampling interference bringing to Stable Isotopic Analysis lack of standardization, the fish body muscle samples after the method sampling drying can be preserved and preserve stable for a long time.
In order to achieve the above object, the present invention takes following technical measures:
A kind of sampling method of the fish body muscle for Stable Isotopic Analysis: the steps include:
1. design muscle stopple coupon: choosing internal diameter is 8-10mm, external diameter is the low carbon austenitic acid-resistant stainless steel pipe of 9-10mm, intercepting 11-12cm length, one end section and stainless-steel tube major diameter are in 90 °, the other end becomes 30 ° with stainless-steel tube major diameter, and the otch of 30 ° of one end of stainless-steel tube is sharp with sand papering;
2. prepare fish sample: choose fish sample, require fish body fresh not corrupt.Spring and autumn air temperature surpass 2h in the time of the dead sample preparation of 10-18 ℃ of macrura reevesii body, and summer, sample should not processed immediately if freezing, and winter temperature sample in the time of-1-4 ℃ preferably also will be processed within 48h after the death of fish body.Individual of sample size will guarantee to get enough back white muscles, with clear water, cleans up fish body, dries fish surface moisture, measures total length, body weight;
3. prepare sampling instrument: stainless steel scalpel, tweezers, the muscle stopple coupon of sampling use is stand-by after pre-burning on spirit lamp, the Eppendorf pipe that holds the 5ml capacity of muscle samples soaks 28-32 minute in 10% hydrogen chloride (HCl), dry after repeatedly cleaning 3-4 time with deionized water, finish label stand-by;
4. at fish body upper cut: with the scalpel of pre-burning, below fish body dorsal fin, vertebra top, makes two otch near the position in fish body the place ahead perpendicular to vertebra, and incision length is not less than 15mm, degree of depth 12-15mm, two otch spacing 39-41mm;
5. get muscle: one end of 30 ° of the muscle stopple coupons of pre-burning is inserted from an otch, be parallel to fish body to another cut-out direction slow circumvolve, muscle stopple coupon is not encountered fish body bone, until the front end of muscle stopple coupon is crossed another otch, extract muscle stopple coupon out, with the tweezers of pre-burning, take out muscle samples, do not get back white muscle Fish tissue in addition, as bone, scale etc.;
6. clean: with deionized water, muscle is rinsed well, after drying, proceeded to the Eppendorf pipe of numbering;
7. dry: at 58-62 ℃, dry to constant weight;
8. grind: the muscle samples after drying is proceeded to grind into powder in the ceramic mortar of pre-burning, cross 80 order aperture screen clothes, proceed to Eppendorf pipe and preserve or be sent to test.
The fish of cultivating under same experimental conditions, the fish body muscle samples of using this sampling method to obtain and preserve, the aberration rate between the different fish bodies of Stable Isotopic Analysis result is far smaller than the variation between existing national standard (GB/T18932.1-2002) disposal route.
the present invention compared with prior art, has the following advantages and effect:
?minimum sample collecting apparatus and sampling receptacle are disturbed or disturbed to this method by choosing without stable isotope, sample collecting apparatus and sampling receptacle are done to get rid of interference processing, and simplified sampling process, can effectively reduce the probability that sample is disturbed and pollutes, sample can be preserved for a long time and keep stable after low temperature drying is processed, and is conducive to improve the reliability in fish Stable Isotopic Analysis.
The hybridized prussian carp of cultivating under same experimental conditions, use this sampling method to obtain and preserve the wherein muscle samples of 5 fishes, the average variation of obtaining sample Stable Isotopic Analysis result by the method is far smaller than with the variation between existing national standard (GB/T18932.1-2002) disposal route.The average aberration rate 0.13 ‰ of the carbon stable isotope between 5 fish bodies wherein, lower than the mean difference 0.4 ‰ of Different Nutrition inter-stage carbon isotope in food web; Article 5, between fish body, the average aberration rate of nitrogen stable isotope is 0.97 ‰, lower than the mean difference 3.4 ‰ of Different Nutrition inter-stage nitrogen stable isotope.
Accompanying drawing explanation
Fig. 1 is a kind of fish body back white muscles stainless steel stopple coupon schematic diagram.
Wherein, 1 is muscle stopple coupon internal diameter, and 2 is muscle stopple coupon external diameter, 3 is stainless-steel tube cross section and 30 ° of angles of body major diameter, 4 is stainless-steel tube cross section and 90 ° of angles of body major diameter, and 5 is fish body muscle stopple coupon (integral body), and 6 is the length of fish body muscle stopple coupon.
Fig. 2 is a kind of fish body back otch schematic diagram.
Wherein 7 is incision length, and 8 is the distance between two otch, and 9 is incision site and direction position.
Embodiment
embodiment1:
Below in conjunction with concrete diagram, to of the present invention, be elaborated:
A kind of sampling method of the fish body muscle for Stable Isotopic Analysis: the steps include:
1. design muscle stopple coupon: choose muscle stopple coupon internal diameter 1 and be 8 or 9 or 10mm, muscle stopple coupon external diameter 2 is 9 or 9.5 or the low carbon austenitic acid-resistant stainless steel pipe of 10mm, the length 6 of intercepting fish body muscle stopple coupon is 9 or 10 or a section of 11cm, 90 ° 4 of one end, stainless-steel tube cross section and stainless-steel tube cross section and body major diameter, 30 ° 3 of the other end and stainless-steel tube cross section and body major diameter, the otch of stainless-steel tube cross section and 30 ° of 3 one end of body major diameter is sharp with sand papering, forms fish body muscle stopple coupon 5;
2. prepare fish sample: choose fish sample, require fish body fresh not corrupt, Individual Size guarantees to get enough back white muscles, with clear water, cleans up fish body, dries fish surface moisture, measures total length, body weight;
3. prepare sampling instrument: stainless steel scalpel, tweezers, the muscle stopple coupon of sampling use is stand-by after pre-burning on spirit lamp, the Eppendorf pipe that holds the 5ml capacity of muscle samples soaks 28 or 29 or 30 or 31 or 32 minutes in 10% HCl, with deionized water, repeatedly clean 3 or 4 times and be dried afterwards, finish label stand-by;
4. at fish body upper cut: with the scalpel of pre-burning below fish body dorsal fin, vertebra top, makes two otch near incision site and the direction position 9 in fish body the place ahead perpendicular to vertebra, and incision length 7 is not less than 15mm, the degree of depth 12 or 13 or 14 or 15mm, two otch spacings 8 are 40mm;
5. get muscle: one end that the fish body muscle stopple coupon of pre-burning 5 cross sections are tilted is inserted from an otch, be parallel to fish body to another cut-out direction slow circumvolve, fish body muscle stopple coupon 5 is not encountered fish body bone, until the front end of fish body muscle stopple coupon 5 is crossed another otch, extract muscle stopple coupon out, with the tweezers of pre-burning, take out muscle samples, do not get back white muscle other Fish tissue in addition, as bone, scale etc.;
6. clean: with deionized water, muscle is rinsed well, after drying, proceeded to the Eppendorf pipe of numbering;
7. dry: at 58 or 59 or 60 or 61 or 62 ℃, dry to constant weight;
8. grind: the muscle samples after drying is proceeded to grind into powder in the ceramic mortar of pre-burning, cross 80 order aperture screen clothes, proceed to Eppendorf pipe and preserve or be sent to test.
Claims (1)
1. for a sampling method for the fish body muscle of Stable Isotopic Analysis, the steps include:
1) design muscle stopple coupon: choosing internal diameter is 8-10mm, external diameter is the low carbon austenitic acid-resistant stainless steel pipe of 9-10mm, intercepting 11-12cm length, one end section and stainless-steel tube major diameter are in 90 °, the other end becomes 30 ° with stainless-steel tube major diameter, the otch sand papering of 30 ° of one end of stainless-steel tube;
2) prepare fish sample: choose fish sample, fish body is fresh not corrupt, spring and autumn air temperature do not surpass 2h in the time of the dead sample preparation of 10-18 ℃ of macrura reevesii body, sample in summer freezing processing not, winter temperature sample in the time of-1-4 ℃ is processed within 48h after the death of fish body, and individual of sample size is got back white muscles, with clear water, cleans up fish body, dry fish surface moisture, measure total length, body weight;
3) prepare sampling instrument: stainless steel scalpel, tweezers, the muscle stopple coupon of sampling use is stand-by after pre-burning on spirit lamp, the Eppendorf pipe that holds the 5ml capacity of muscle samples soaks 28-32 minute in 10% hydrogen chloride, dry after repeatedly cleaning 3-4 time with deionized water, finish label stand-by;
4) at fish body upper cut: with the scalpel of pre-burning, below fish body dorsal fin, vertebra top, makes two otch near the position in fish body the place ahead perpendicular to vertebra, and incision length is not less than 15mm, degree of depth 12-15mm, two otch spacing 39-41mm;
5) get muscle: one end of 30 ° of the muscle stopple coupons of pre-burning is inserted from an otch, be parallel to fish body to another cut-out direction rotation, muscle stopple coupon is not encountered fish body bone, until the front end of muscle stopple coupon is crossed another otch, extract muscle stopple coupon out, with the tweezers of pre-burning, take out muscle samples, do not get back white muscle Fish tissue in addition;
6) clean: with deionized water, rinse out the blood above muscle, after drying, proceed to the Eppendorf pipe of numbering;
7) dry: at 58-62 ℃, dry to constant weight;
8) grind: the muscle samples after drying is proceeded to grind into powder in the ceramic mortar of pre-burning, cross 80 order aperture screen clothes, proceed to Eppendorf pipe and preserve or be sent to test.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201410497767.XA CN104198216B (en) | 2014-09-24 | 2014-09-24 | A kind of sampling method of the fish body muscle for Stable Isotopic Analysis |
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| CN201410497767.XA CN104198216B (en) | 2014-09-24 | 2014-09-24 | A kind of sampling method of the fish body muscle for Stable Isotopic Analysis |
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| CN104198216A true CN104198216A (en) | 2014-12-10 |
| CN104198216B CN104198216B (en) | 2016-08-17 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105628781A (en) * | 2015-12-24 | 2016-06-01 | 张敏 | Isotopic tracing method for fate characteristics of organic substances in aquaculture pond |
| CN116519439A (en) * | 2023-06-30 | 2023-08-01 | 交通运输部天津水运工程科学研究所 | Fish stable isotope sample pretreatment integrated equipment |
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