CN108663467A - A kind of preparation method of fish meal Analysis on amino acid components standard substance - Google Patents
A kind of preparation method of fish meal Analysis on amino acid components standard substance Download PDFInfo
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- CN108663467A CN108663467A CN201810645094.6A CN201810645094A CN108663467A CN 108663467 A CN108663467 A CN 108663467A CN 201810645094 A CN201810645094 A CN 201810645094A CN 108663467 A CN108663467 A CN 108663467A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
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Abstract
The invention discloses a kind of preparation methods of fish meal Analysis on amino acid components standard substance.Fish meal is cleaned first, is crushed after removal of impurities；Soak degreasing in solvent is added in fish meal after crushing；Fish meal after degreasing is spontaneously dried and is dried successively, and fish meal sample is obtained；Fish meal sample is subjected to mixing, is sampled after mixing and carries out uniformity detection；Fish meal sample after mixing is dried again, is dispensed after dry, and sample carries out irradiation inactivated after packing, is finally vacuum-packed, is preserved under the conditions of 20 DEG C after packaging.It is prepared for fish meal Analysis on amino acid components standard substance using the present invention, solves critical issue existing in the prior art, the domestic blank that there is no fish meal sample amino acid standard matter is filled up, the Development Level to improve China's health of masses, food, environment etc. standard substance is laid a good foundation.
Description
One, technical field：
The present invention relates to a kind of preparation methods of standard substance, more particularly to a kind of fish meal Analysis on amino acid components mark
The preparation method of quasi- substance.
Two, technical background：
Fish meal is rich in protein, and wherein combination of amino acids is reasonable, rich content, nutritive value are high, therefore is widely used as
Feed.But bad businessman is adulterated with the high protein raw material incorporation of the low price such as feather meal, leather powder, blood meal, meat meal tankage, shadow
The quality for ringing fish meal, to influence the growth and development of poultry and the quality of meat.As people are anti-for raw-food material quality safety
The raising and growing to the pursuit of quality of life of model consciousness, also increasingly causes related portion to the detection of raw-food material
The attention of door.Product control to protein raw material, amino acid analysis are most reliable and most effective means.Currently, amino acid analysis
Method mainly has：Using the national standard method GB/T 18246 and column front derivation reversed phase high performance liquid of automatic amino acid analyzer
Phase chromatography.It is accordingly detected according to national standard, needs corresponding forage standard substance.Currently, there is amino acid in China
Mixed solution standard substance (number：GBW (E) 100062), but the Analysis on amino acid components reference substance using fish meal as matrix
Matter is still blank.
Substrate standard substance is measuring process control, the important carrier that method confirms, arbitration and examination verification are evaluated, and is
Ensure that measurement result is reliable, promotes the important support of laboratory testing capability.When detecting feed matrix sample, matrix effect is past
Toward being one of systematic error source, influence result it is accurate whether.The actually detected sample of user should be selected to match as far as possible
Substrate standard substance reduces the operating cost in laboratory, improves the validity and reliability of testing result.Therefore, ammonia in fish meal
The development of base sour component analytical standard substance, acts not only as a kind of etalon, to determine amino acid in sample fish meal sample
Component content；Meanwhile it can also be used as a kind of standard sample, it is whether reasonable come the method that detects sample pre-treatments.This for
The pre-treating method of fish meal also plays the effect of a supervision and inspection so that the safety verification of fish meal is more reasonable and rigorous.
Currently, the pre-treatment and analysis valued methods research of fish meal are always the key of research, so there is an urgent need to develop a kind of fish
Powder matrix amino acid standard matter.
Three, invention content：
The technical problem to be solved by the present invention is to：In order to solve to there is no lacking for fish meal matrix amino acid standard matter at present
It falls into, the present invention provides a kind of preparation method of fish meal Analysis on amino acid components standard substance.
To solve the above-mentioned problems, the technical solution adopted by the present invention is that：
The present invention provides a kind of preparation method of fish meal Analysis on amino acid components standard substance, and the preparation method includes
Following steps：
A, impurity elimination：Fish meal is subjected to screening and removing impurities first；
B, it crushes：Fish meal after step a removal of impurities is crushed, grinding and sieving obtains the fish that fineness is 60~80 mesh
Powder；
C, degreasing：It will be impregnated in fish meal complete wetting to solvent that step b is crushed, in soaking process fully
Stirring, soaking time are 12~15 hours, are filtered after immersion, filter solvent；By the fish meal obtained after filtering according to same
Immersion process carry out second and impregnate, be filtered after immersion, filter solvent, obtain the fish meal of degreasing；
D, drying：The fish meal obtained after step c degreasings is naturally shady and cool 4~6 hours dry, it is put into after natural drying true
It is dried in empty drying box, fish meal sample is obtained after drying；
E, mixing：The fish meal sample that step d is obtained is placed in mixing machine and is sufficiently mixed, incorporation time is 5~20
Minute, the sample of different piece in mixing machine is sampled after mixing, uniformity detection is carried out after sampling；
F, it dries again：Fish meal sample after step e mixings is placed in drying box and is dried, it is dry to fish meal sample
Moisture ＜ 1% in product；
G, it dispenses：Fish meal sample after step f dryings is sub-packed in vial, and covers tightly threaded cap, sealing sample rule
Lattice are 5g/ bottles；
H, irradiation inactivated：The sample of packing is used60Co gamma-ray irradiations inactivate, and dose intensity is 7kGy~10kGy,
The irradiation inactivated time is 2 hours；
I, the sample after step h irradiation inactivateds is vacuum-packed using aluminum foil compound bag, after packaging under the conditions of -20 DEG C
It preserves.
According to the preparation method of above-mentioned fish meal Analysis on amino acid components standard substance, the braiding of 20 mesh is used in step a
Sieve carries out screening and removing impurities to fish meal.
According to the preparation method of above-mentioned fish meal Analysis on amino acid components standard substance, fineness described in step b is 80
Mesh.
According to the preparation method of above-mentioned fish meal Analysis on amino acid components standard substance, solvent described in step c is anhydrous
Any one of ether, petroleum ether, acetone, n-hexane, methanol and ethyl alcohol.
According to the preparation method of above-mentioned fish meal Analysis on amino acid components standard substance, the solvent be anhydrous ether or
Petroleum ether.
It is naturally shady and cool dry described in step d according to the preparation method of above-mentioned fish meal Analysis on amino acid components standard substance
Dry is that the fish meal tiling after degreasing is placed in enamel tray to be dried under the conditions of natural environment non-direct sunlight；
Drying temperature is 60~80 DEG C when the drying, and drying time is 1~2 hour.
According to the preparation method of above-mentioned fish meal Analysis on amino acid components standard substance, mixing machine described in step e uses
V-Mixer or flat mixer；
The uniformity detection：Mixed three samples are extracted, with reference to GB/T 5918-2008《Feed product mixes
The measurement of the uniformity》Uniformity Preliminary detection is carried out, then homogeneity statistical inspection is carried out to testing result with method of analysis of variance.
According to the preparation method of above-mentioned fish meal Analysis on amino acid components standard substance, drying box described in step f is true
Empty drying box or air dry oven, drying temperature are 80~105 DEG C, and drying time is 8~12 hours.
According to the preparation method of above-mentioned fish meal Analysis on amino acid components standard substance, vial described in step g is band
The brown bottle of screw thread mouth.
According to the preparation method of above-mentioned fish meal Analysis on amino acid components standard substance, step h middle dosage intensity is
7kGy。
The positive beneficial effect of the present invention：
1, the fish meal substrate standard substance prepared using technical solution of the present invention can ensure fish closer to actual sample
The better magnitude tracing of amino acid detection project in powder, it is as a result relatively reliable and comparable, solve detection caused by matrix difference
Result difference problem, improves magnitude tracing level, and the judgement for fish meal quality provides foundation.
2, the fish meal substrate standard substance prepared using technical solution of the present invention is easy to get, is at low cost.It is prepared by the present invention
Standard substance stability it is good, can under the conditions of -20 DEG C long-term preservation (at least a year), have preferable uniformity and stabilization
Property.
3, the present invention is prepared for fish meal Analysis on amino acid components standard substance, solves key existing in the prior art
Problem has filled up the domestic blank that there is no fish meal amino acid standard matter, to improve the side such as China's health of masses, food, environment
The Development Level of face standard substance is laid a good foundation.
4, the present invention has chosen fish meal as matrix, and the substrate standard substance protein concentration content is moderate, user
Just, quality is good.
5, the development of fish meal Analysis on amino acid components standard substance, in the control of enterprise product quality, the system of product standard
Important meaning is suffered from terms of the fixed, examination and test of products and the quality control of certification authority and evaluation, Laboratory Accreditation, the standard
Sound development of the development of substance to China's feed industry improves Animal product quality and plays positive impetus.
Four, it illustrates：
17 kinds of amino acid mixed solution standard substance chromatograms after Fig. 1 derivatization reactions；
Fig. 2 derivative reagent blank chromatograms；
The chromatogram of fish meal Analysis on amino acid components standard substance sample of the present invention after Fig. 3 derivatization reactions.
Five, specific implementation mode：
The present invention is further explained with reference to embodiments, but is not intended to limit the range of technical solution of the present invention protection.
Embodiment 1：
The preparation method of fish meal Analysis on amino acid components standard substance of the present invention, the detailed step of the preparation method are as follows：
A, impurity elimination：Commercially available fish meal is subjected to screening and removing impurities using the wire gauze sieve of 20 mesh first；
B, it crushes：Fish meal after step a removal of impurities is crushed, the fish meal that fineness is 80 mesh is obtained after crushing；
C, degreasing：It impregnates, impregnates in the fish meal complete wetting to solvent anhydrous ether obtained after step b is crushed
Time be 12 hours, in soaking process every 4 hours be sufficiently stirred, be filtered after immersion, filter the anhydrous second of solvent
Ether；
The fish meal obtained after first time is filtered carries out second according to same immersion process and impregnates, and is carried out after immersion
Filtering, filters solvent anhydrous ether, obtains the fish meal of degreasing；
D, drying：The fish meal obtained after step c degreasings is spontaneously dried 6 hours, and (natural drying is：Fish meal tiling is set
It is dried under the conditions of natural environment non-direct sunlight in enamel tray), it is put into vacuum drying chamber and carries out after natural drying
Drying, drying temperature are 80 DEG C, and drying time is (fish meal tiling to be placed in enamel tray when drying, is placed into vacuum for 1 hour
Drying box is dried), fish meal sample is obtained after drying；
E, mixing：The fish meal sample that step d is obtained is placed in V-Mixer and is sufficiently mixed, incorporation time 10
Minute, the sample of different piece in mixing machine is sampled after mixing, uniformity detection is carried out after sampling；
F, it dries again：Fish meal sample after step e mixings is placed in drying box and is dried, drying temperature 80
DEG C, drying time is 12 hours, moisture ＜ 1% in gained fish meal sample after drying；
G, it dispenses：Fish meal sample after step f dryings is sub-packed in the vial of threaded mouth, and covers tightly threaded cap,
It is 5g/ bottles to seal sample specification, is divided into and fills 500 bottles；
H, irradiation inactivated：The sample of packing is used60Co gamma-ray irradiations inactivate, dose intensity 7kGy, irradiation inactivated
Time is 2 hours；
I, the sample after step h irradiation inactivateds is vacuum-packed using aluminum foil compound bag, after packaging under the conditions of -20 DEG C
It preserves.
It is the dependence test to 1 gained fish meal Analysis on amino acid components standard substance sample of embodiment below：
(1) related test results of each processing stage fish meal sample
The commercially available fish meal of raw material that the present invention uses and by dry (moisture in gained fish meal sample after drying again again
Content ＜ 1%) afterwards the Related Component testing result of fish meal sample refer to table 1.
The commercially available fish meal of 1 raw material of table and by again dry after fish meal sample Related Component testing result
| Protein (%) | Fatty (%) | Moisture (%) | |
| Commercially available fish meal | 68.1 | 9.1 | 9.6 |
| Fish meal sample after drying again | 77.1 | 6.0 | 1.0 |
During the preparation method of fish meal Analysis on amino acid components standard substance of the present invention, the gained sample after packing
Testing result before and after irradiation inactivated refers to table 2 and table 3.
The testing result of gained sample before irradiation inactivated after packing of table 2
| Title | Total number of bacteria (a/g) | Total number of molds (a/g) |
| Fish meal | 10000 | <10 |
The testing result of gained sample after irradiation inactivated after packing of table 3
It indicates：From the above data, it uses60The dose intensity of Co γ be 7kGy x ray irradiation xs inactivate 2 hours be best spoke
According to dosage.
(2) present invention prepares inspection and the definite value of gained fish meal Analysis on amino acid components standard substance sample
The present invention carries out analysis measurement using pre-column derivatization-ultra-performance liquid chromatography, and specific determination condition is such as
Under：
Instrument：Ultra performance liquid chromatography matches ultraviolet-visible detector (Waters, the U.S.)；
Liquid phase chromatogram condition：
Chromatographic column：AccQ-TAGTM ULTRA C18(2.1 × 100mm, 1.7 μm)；
Mobile phase：A：AccQTag eluents A (10 times of dilution), B：AccQTag eluents B；
Flow velocity：0.7mL/min；
Column temperature：55℃；
Ultraviolet detection wavelength：260nm；
Sample size：1μL；
According to the gradient that mobile phase is arranged in table 4, separate run times 9.5min.
The liquid phase gradient table of 4 derivatization method of table
Fish meal Analysis on amino acid components standard substance prepared by the present invention uses pre-column derivatization-ultra performance liquid chromatography
The processing procedure that method is analyzed is as follows：
1) pre-treatment of Analysis on amino acid components standard substance：
Sour water solution：Fish meal Analysis on amino acid components standard substance prepared by the 0.3g embodiment of the present invention 1 is taken to be added to
10mL, a concentration of 6mol/L hydrochloric acid solution in, then hydrolyzed 22 hours under the conditions of 110 DEG C；50mL appearances are filled into after hydrolysis
In measuring bottle, with water constant volume, the hydrolyzation sample of fish meal Analysis on amino acid components standard substance is made.
2) preparation of AccQ-Fluor derivative reagents：
AccQ-Fluor derivating agent powder is diluted first, the acetonitrile that 1mL is drawn using pipettor is added to derivative
It is sealed in reagent bottle, the quality of agent powder derived from derivative reagent bottle is 1mg, vortex mixed 15 seconds after addition；Then
It is heated in the baking oven for being preheating to 55 DEG C, to dissolve all derivative reagent powder, heating time is 8 minutes, after heating for dissolving
The AccQ-Fluor derivative reagents prepared.
3) fish meal Analysis on amino acid components standard substance prepared by 17 kinds of amino acid mixed solution standard substances and the present invention
The derivatization reaction of sample：
The derivatization reaction of 17 kinds of amino acid mixed solution standard substances (by amino acid converting for stable AQC derivatives)：
Baking oven is opened and constant temperature is to 55 DEG C, 10 μ L, 17 kinds of amino acid mixed solution standard substances is taken to inject clean derivative tube bottom
Portion adds in 70 μ L AccQ-Fluor Buffer1 borate buffers to derivative pipe, vortex mixed, finally draws that 20 μ L are above-mentioned to be matched
The derivative reagent of system is added under vorticity in derivative pipe, and keeps vortex mixed 15 seconds, and gained after vortex mixed is spread out
Production object is transferred in derivative dedicated micro-sampling bottle, obtains 17 kinds of amino acid mixed solution reference substances of derivatization reaction
Matter.
The derivatization reaction of fish meal Analysis on amino acid components standard substance sample prepared by the present invention：With above-mentioned 17 kinds of amino
The derivatization reaction of sour mixed solution standard substance.
The derivative reagent used is purchased from the AccQ-Fluor reagent packets of Waters companies production.
By 17 kinds of amino acid mixed solution standard substances, the fish meal aminoacid ingredient of the present invention point after above-mentioned derivatization reaction
Analysis standard substance sample, which is placed in 55 DEG C of thermostatic drying chambers, heats 10min, then cools to room temperature, to be measured.According to above-mentioned limit
The determination condition of fixed pre-column derivatization-ultra-performance liquid chromatography is to 17 kinds of amino acid mixed solution marks after derivatization reaction
Fish meal Analysis on amino acid components standard substance sample of the present invention after quasi- substance, derivative reagent blank, derivatization reaction carries out respectively
Detection, obtained chromatogram refer to attached drawing 1-3.
The chromatogram of 17 kinds of amino acid mixed solution standard substances after derived from attached drawing 1 has inside including reagent peak
18 chromatographic peaks, are followed successively by according to the sequencing of retention time：His, Ser, Arg, Gly, Asp, Glu, Thr, Ala, Pro,
Deriv (derivative reagent peak), Cys, Lyr, Tyr, Met, Val, Ile, Leu, Phe.
Pass through the fish of the present invention after control 17 kinds of amino acid mixed solution standard substances, derivative reagent blank, derivatization reactions
The chromatogram of powder Analysis on amino acid components standard substance sample, it may be determined that amino acid in the standard substance prepared by the present invention
Type and its content.
(4) uniformity to the fish meal Analysis on amino acid components standard substance prepared by the present invention, study on the stability, with
And the analysis of definite value and uncertainty：
1) uniformity testing：
According to the requirement of national standard reference material technology specification (JJG 1006-1994), carried out using variance analysis equal
Even property experiment.25 samples are extracted from fish meal Analysis on amino acid components standard substance sample prepared by the present invention for uniform
Property examine, each sample measures 3 progress variance analyses, judges that each group is surveyed by the comparison of between-group variance and intra-class variance
Whether there is or not system deviations between magnitude.
By the above uniformity testing, gained inventive samples uniformity testing is the results detailed in Table 5.
The Evaluation for Uniformity result of fish meal Analysis on amino acid components standard substance prepared by 5 present invention of table
According to detection evaluation result：Analysis on amino acid components standard substance sample prepared by the judgement present invention is uniform.
2) study on the stability：
It is by multiple samples (a collection of sample) that are of the invention while preparing, under the same conditions that long-time stability, which are investigated,
It is measured with the continuity of time.In 24 months, to the fish meal Analysis on amino acid components standard substance for preparing of the present invention into
7 study on the stability are gone, respectively on the day of sample preparation, after 1 month, after 2 months, after 3 months, after 6 months, 12 months
3 are randomly selected to the standard substance of preparation with after 24 months afterwards, every is surveyed 3 times, is positioned at -20 DEG C and 4 DEG C and is preserved, into
Row long-time stability are investigated.
Line fitting approach is selected to evaluate its stability.The time is represented with X, the characteristic of standard substance is represented with Y
Value, fitting is in alignment, then has slope β1
Intercept is calculated by following formula：
The standard deviation of straight line can be calculated by following formula：
When | β1| ＜ t0.95,n-2·s(β1) when slope be inapparent, be judged to not observe unstability.
Short-term stability, which is investigated, uses synchronism stability Journal of Sex Research, is introduced into so that the measurement in all stability studies exists
Repeatability occurs, i.e., only carries out a wheel calibration.Analysis on amino acid components standard substance is in transportational process, reality in fish meal of the present invention
Test the stability under the conditions of room temperature in use etc., respectively by standard substance -20 DEG C, 4 DEG C, room temperature, 40 DEG C place 1 day, 2 days,
4 days, 7 days, 15 days and 30 days, the content of amino acid in monitoring standard substance carried out estimation of stability to result.
It is investigated by the above long-time stability, gained long-time stability are investigated the results detailed in Table 6 and table 7.
The long-time stability evaluation result (4 DEG C) of 6 fish meal Analysis on amino acid components standard substance of the present invention of table
The long-time stability evaluation result (- 20 DEG C) of 7 fish meal Analysis on amino acid components standard substance of the present invention of table
Long-time stability investigate result：Analysis result shows that standard substance of the present invention is good in 2 year internal stabilities.
It is investigated by the above short-term stability, gained short-term stability is investigated the results detailed in Table 8 and table 9.
Study on the stability result (the room temperature of 8 fish meal Analysis on amino acid components standard substance of the present invention of table：20±5℃)
The study on the stability result (40 DEG C) of 9 fish meal Analysis on amino acid components standard substance of the present invention of table
Short-term stability investigates result：The above analysis result shows standard substance of the present invention under traffic condition in 30 days
It is stable, meets actual shipment requirement.
4) definite value of standard substance：
According to ISO directive/guides 35《The rule and statistical method of standard substance/standard sample definite value》It is required that the present invention
The valued methods of the fish meal Analysis on amino acid components standard substance of preparation use multiple laboratory cooperation valued methods.It invites
Henan Hai Ruizheng detection techniques Co., Ltd, agricultural product quality supervision and inspection center of the Ministry of Agriculture, Henan Province's feed product matter
Amount supervision and inspection station (save veterinary drug and supervise institute), Henan are herded through food and feed detection Co., Ltd, Henan Province's grain and oil feed product
The common definite value of Quality Supervision and Inspection Center.
The organization for participating in cooperation definite value, the analytical instrument used and detection foundation are respectively：
10 definite value laboratory summary sheet of table
The unit of every participation cooperation definite value all randomly selects fish meal Analysis on amino acid components standard prepared by the present invention
Substance 11, wherein one is used as practice sample, other 10 samples to carry out pre-treatments, each treated, and increment repeats survey
It is 2 times fixed.The pre-treatment of sample is all made of the sour processing method in national standard method GB/T 18246-2000.The measurement of sample
Method is all made of national standard method GB/T 18246-2000, and each laboratory is due to instrument model difference, specific measurement method
It is operated by instrument specification.
According to above-mentioned experiment, gained definite value is the results detailed in Table 11.
Each laboratory of 11 fish meal Analysis on amino acid components standard substance of the present invention of table value data that is averaged summarizes
| Amino acid | Laboratory 1 | Laboratory 2 | Laboratory 3 | Laboratory 4 | Laboratory 5 | Laboratory 6 | Asserting value |
| Asparatate | 6.305 | 6.348 | 6.173 | 6.382 | 6.383 | 6.166 | 6.29 |
| Threonine | 2.965 | 2.953 | 2.831 | 2.794 | 2.927 | 3.140 | 2.94 |
| Serine | 2.629 | 2.596 | 2.637 | 2.583 | 2.558 | 2.652 | 2.61 |
| Glutamic acid | 9.798 | 9.766 | 9.591 | 9.653 | 9.803 | 9.776 | 9.73 |
| Glycine | 4.408 | 4.409 | 4.314 | 4.542 | 4.451 | 4.367 | 4.42 |
| Alanine | 4.415 | 4.416 | 4.417 | 4.444 | 4.479 | 4.519 | 4.45 |
| Cystine | 0.579 | 0.563 | 0.519 | 0.615 | 0.568 | 0.515 | 0.56 |
| Valine | 3.396 | 3.447 | 3.177 | 3.473 | 3.371 | 3.123 | 3.33 |
| Methionine | 2.103 | 2.063 | 1.993 | 2.031 | 2.069 | 2.163 | 2.07 |
| Isoleucine | 2.874 | 2.869 | 2.890 | 2.986 | 2.754 | 3.094 | 2.91 |
| Leucine | 5.245 | 5.198 | 5.161 | 5.268 | 5.243 | 5.250 | 5.23 |
| Tyrosine | 2.112 | 2.051 | 2.037 | 2.127 | 2.101 | 2.175 | 2.10 |
| Phenylalanine | 2.920 | 2.944 | 3.018 | 3.027 | 2.974 | 2.912 | 2.97 |
| Lysine | 5.885 | 5.939 | 5.621 | 5.812 | 5.877 | 5.635 | 5.79 |
| Histidine | 1.840 | 1.815 | 1.822 | 2.052 | 1.822 | 1.687 | 1.84 |
| Arginine | 4.376 | 4.454 | 4.565 | 4.422 | 4.389 | 4.245 | 4.41 |
| Proline | 2.834 | 2.782 | 2.848 | 2.853 | 2.822 | 2.604 | 2.79 |
Cooperation definite value conclusion：The result that unit by participating in cooperation definite value is provided meets the requirements, this project
Definite value is finally the average value that cooperation definite value laboratory provides result, as shown in table 11.
5) analysis on Uncertainty：
The uncertainty source of fish meal Analysis on amino acid components standard substance prepared by the present invention is made of six parts：Ammonia
The uncertainty of base acid mixed solution standard substance introducing, balance weigh the uncertainty introduced, peak area ratio measures introducing
The uncertainty and stability that uncertainty, the uniformity difference that uncertainty, value data statistics introduce introduce introduce not
Degree of certainty.Analysis on Uncertainty is the results detailed in Table 12 and table 13.
The uncertainty result of 12 fish meal Analysis on amino acid components standard substance of the present invention of table
The combined standard uncertainty of fish meal Analysis on amino acid components standard substance of the present invention is multiplied by Coverage factor k=2 and obtains
To expanded uncertainty.Gained expanded uncertainty is the results detailed in Table 13.
The extension of 13 fish meal Analysis on amino acid components standard substance of the present invention of table does not know result
Definite value conclusion：The standard value and uncertainty that the substrate standard substance of the present invention finally determines are as shown in table 13.
Fish meal Analysis on amino acid components standard substance prepared by the present invention can be feed factory, testing agency etc. provide with
The consistent standard substance of sample to be tested matrix is in determined amino acid laboratory to improve the accuracy of determined amino acid
Portion's quality control provides reference material；Definite value for feed factory quality-control product provides reference material.
Embodiment 2：
The preparation method of fish meal Analysis on amino acid components standard substance of the present invention, the detailed step of the preparation method are as follows：
A, impurity elimination：Commercially available fish meal is subjected to screening and removing impurities using the wire gauze sieve of 20 mesh first；
B, it crushes：Fish meal after step a removal of impurities is crushed, the fish meal that fineness is 80 mesh is obtained after crushing；
C, degreasing：It will be impregnated in fish meal complete wetting to solvent anhydrous ether that step c is crushed, when immersion
Between be 15 hours, in soaking process every 3 hours be sufficiently stirred, be filtered after immersion, filter solvent anhydrous ether；
The fish meal obtained after first time is filtered carries out second according to same immersion process and impregnates, and is carried out after immersion
Filtering, filters solvent anhydrous ether, obtains the fish meal of degreasing；
D, drying：By the fish meal obtained after step c degreasings, naturally 4 hours shady and cool dry (natural drying is：After degreasing
Fish meal tiling be placed in enamel tray be dried under the conditions of natural environment non-direct sunlight), be put into vacuum after natural drying
It is dried in drying box, drying temperature is 80 DEG C, and drying time is (fish meal tiling to be placed in enamel tray when drying in 1 hour
In, it is placed into vacuum drying chamber and is dried), fish meal sample is obtained after drying；
E, mixing：The fish meal sample that step d is obtained is placed in V-Mixer and is sufficiently mixed, incorporation time 15
Minute, the sample of different piece in mixing machine is sampled after mixing, uniformity detection is carried out after sampling；
F, it dries again：Fish meal sample after step e mixings is placed in drying box and is dried, drying temperature 80
DEG C, drying time is 12 hours, moisture ＜ 1% in gained fish meal sample after drying；
G, it dispenses：Fish meal sample after step f dryings is sub-packed in the vial of threaded mouth, and covers tightly threaded cap,
It is 5g/ bottles to seal sample specification, is divided into and fills 500 bottles；
H, irradiation inactivated：The sample of packing is used60Co gamma-ray irradiations inactivate, dose intensity 8kGy, irradiation inactivated
Time is 2 hours；
I, the sample after step h irradiation inactivateds is vacuum-packed using aluminum foil compound bag, after packaging under the conditions of -20 DEG C
It preserves.
The dependence test of the present embodiment products obtained therefrom fish meal Analysis on amino acid components standard substance is the same as embodiment 1.Through surveying
Examination, the present embodiment products obtained therefrom are fully able to be used as aminoacid ingredient standard substance.
Embodiment 3：
The preparation method of fish meal Analysis on amino acid components standard substance of the present invention, the detailed step of the preparation method are as follows：
A, impurity elimination：Commercially available fish meal is subjected to screening and removing impurities using the wire gauze sieve of 20 mesh first；
B, it crushes：Fish meal after step a removal of impurities is crushed, the fish meal that fineness is 60 mesh is obtained after crushing；
C, degreasing：It impregnates, impregnates in the fish meal complete wetting to solvent anhydrous ether obtained after step c is crushed
Time be 14 hours, in soaking process every 3 hours be sufficiently stirred, be filtered after immersion, filter the anhydrous second of solvent
Ether；
The fish meal obtained after first time is filtered carries out second according to same immersion process and impregnates, and is carried out after immersion
Filtering, filters solvent anhydrous ether, obtains the fish meal of degreasing；
D, drying：The fish meal obtained after step c degreasings is spontaneously dried 5 hours, and (natural drying is：By the fish after degreasing
Powder tiling is placed in enamel tray to be dried under the conditions of natural environment non-direct sunlight), vacuum drying is put into after natural drying
It is dried in case, drying temperature is 80 DEG C, and drying time is (fish meal tiling to be placed in enamel tray when drying, is put for 1 hour
Vacuum drying chamber is set to be dried), fish meal sample is obtained after drying；
E, mixing：The fish meal sample that step d is obtained is placed in V-Mixer and is sufficiently mixed, incorporation time 12
Minute, the sample of different piece in mixing machine is sampled after mixing, uniformity detection is carried out after sampling；
F, it dries again：Fish meal sample after step e mixings is placed in drying box and is dried, drying temperature 105
DEG C, drying time is 8 hours, moisture ＜ 1% in gained fish meal sample after drying；
G, it dispenses：Fish meal sample after step f dryings is sub-packed in the vial of threaded mouth, and covers tightly threaded cap,
It is 5g/ bottles to seal sample specification, is divided into and fills 500 bottles；
H, irradiation inactivated：The sample of packing is used60Co gamma-ray irradiations inactivate, and dose intensity 7.5kGy, irradiation is gone out
Live time is 2 hours；
I, the sample after step h irradiation inactivateds is vacuum-packed using aluminum foil compound bag, after packaging under the conditions of -20 DEG C
It preserves.
The dependence test of the present embodiment products obtained therefrom fish meal Analysis on amino acid components standard substance is the same as embodiment 1.Through surveying
Examination, the present embodiment products obtained therefrom are fully able to be used as Analysis on amino acid components standard substance.
Claims (10)
1. a kind of preparation method of fish meal Analysis on amino acid components standard substance, which is characterized in that the preparation method include with
Lower step：
A, impurity elimination：Fish meal is subjected to screening and removing impurities first；
B, it crushes：Fish meal after step a removal of impurities is crushed, grinding and sieving obtains the fish meal that fineness is 60~80 mesh；
C, degreasing：It will impregnate in fish meal complete wetting to solvent that step b is crushed, be sufficiently stirred in soaking process,
Soaking time is 12~15 hours, is filtered after immersion, filters solvent；The fish meal obtained after filtering is impregnated according to same
Method carries out second and impregnates, and is filtered after immersion, filters solvent, obtains the fish meal of degreasing；
D, drying：The fish meal obtained after step c degreasings is naturally shady and cool 4~6 hours dry, vacuum drying is put into after natural drying
It is dried in case, fish meal sample is obtained after drying；
E, mixing：The fish meal sample that step d is obtained is placed in mixing machine and is sufficiently mixed, incorporation time is 5~20 minutes,
The sample of different piece in mixing machine is sampled after mixing, uniformity detection is carried out after sampling；
F, it dries again：Fish meal sample after step e mixings is placed in drying box and is dried, it is dry to water in fish meal sample
Divide content ＜ 1%；
G, it dispenses：Fish meal sample after step f dryings is sub-packed in vial, and covers tightly threaded cap, sealing sample specification is
5g/ bottles；
H, irradiation inactivated：The sample of packing is used60Co gamma-ray irradiations inactivate, and dose intensity is 7kGy~10kGy, and irradiation is gone out
Live time is 2 hours；
I, the sample after step h irradiation inactivateds is vacuum-packed using aluminum foil compound bag, is preserved under the conditions of -20 DEG C after packaging.
2. the preparation method of fish meal Analysis on amino acid components standard substance according to claim 1, it is characterised in that：Step
Screening and removing impurities are carried out to fish meal using the wire gauze sieve of 20 mesh in a.
3. the preparation method of fish meal Analysis on amino acid components standard substance according to claim 1, it is characterised in that：Step
Fineness described in b is 80 mesh.
4. the preparation method of fish meal Analysis on amino acid components standard substance according to claim 1, it is characterised in that：Step
Solvent described in c is any one of anhydrous ether, petroleum ether, acetone, n-hexane, methanol and ethyl alcohol.
5. the preparation method of fish meal Analysis on amino acid components standard substance according to claim 4, it is characterised in that：It is described
Solvent is anhydrous ether or petroleum ether.
6. the preparation method of fish meal Analysis on amino acid components standard substance according to claim 1, it is characterised in that：Step
Naturally shady and cool drying described in d is that the fish meal tiling after degreasing is placed in enamel tray under the conditions of natural environment non-direct sunlight
It is dried；
Drying temperature is 60~80 DEG C when the drying, and drying time is 1~2 hour.
7. the preparation method of fish meal Analysis on amino acid components standard substance according to claim 1, it is characterised in that：Step
Mixing machine described in e uses V-Mixer or flat mixer；
The uniformity detection：Mixed three samples are extracted, with reference to GB/T 5918-2008《Feed product mixture homogeneity
Measurement》Uniformity Preliminary detection is carried out, then homogeneity statistical inspection is carried out to testing result with method of analysis of variance.
8. the preparation method of fish meal Analysis on amino acid components standard substance according to claim 1, it is characterised in that：Step
Drying box described in f is vacuum drying chamber or air dry oven, and drying temperature is 80~105 DEG C, and drying time is 8~12 small
When.
9. the preparation method of fish meal Analysis on amino acid components standard substance according to claim 1, it is characterised in that：Step
Vial described in g is the brown bottle of threaded mouth.
10. the preparation method of fish meal Analysis on amino acid components standard substance according to claim 1, it is characterised in that：Step
Rapid h middle dosage intensity is 7kGy.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111397995A (en) * | 2020-06-04 | 2020-07-10 | 国家烟草质量监督检验中心 | Preparation method of tobacco powder monitoring sample suitable for conventional component inspection of tobacco chemistry |
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