CN105410034B - A kind of compound accelerant of cordyceps sinensis cultivation and preparation method thereof - Google Patents

A kind of compound accelerant of cordyceps sinensis cultivation and preparation method thereof Download PDF

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CN105410034B
CN105410034B CN201510838962.9A CN201510838962A CN105410034B CN 105410034 B CN105410034 B CN 105410034B CN 201510838962 A CN201510838962 A CN 201510838962A CN 105410034 B CN105410034 B CN 105410034B
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mucor
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cordyceps sinensis
mycelium
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CN105410034A (en
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汪超
刘璐
李冬生
卢忠诚
刘婷
石勇
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Henan Huanghuai Biotechnology Co.,Ltd.
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Hubei University of Technology
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract

The invention discloses compound accelerant of a kind of cordyceps sinensis cultivation and preparation method thereof, belong to agricultural plantation technology field.The present invention is broken from solution using Mucor mycelium, is subject to inositol, n-octacosanol, FOS and copper gluconate and is compounded, so as to the embryo material compound accelerant being prepared into needed for a kind of suitable cordyceps sinensis growth.This embryo material compound accelerant can substantially shorten cordyceps sinensis growth cycle, improve out the weight of cordyceps sinensis in careless, increase unit volume, and dramatically increase its cordycepin content, and processing method is simple, and cost is cheap, can widely use, and strengthens Business Economic Benefit.

Description

A kind of compound accelerant of cordyceps sinensis cultivation and preparation method thereof
Technical field
The invention belongs to agricultural plantation technology field, and in particular to a kind of cultivation embryo material of cordyceps sinensis rapidly and efficiently cultivation technique Compound accelerant and preparation method thereof.
Background technology
Cordyceps militaris also known as northern Chinese caterpillar Fungus, abbreviation cordyceps sinensis, belong to Ascomycetes Hypocreales Clavicipitaceae Cordyceps.Cordyceps sinensis It is divided into stroma, mycelium, fructification, its sub-entities is rich in a variety of essential amino acids, healthy trace elements with household and vitamin Etc. nutritional ingredient, and adenosine, cordycepin, cordycepic acid, Cordyceps sinensis polysaccharide, superoxide dismutase (SOD) isoreactivity composition, have A variety of pharmacological functions such as antitumor, disease-resistant former, anti-oxidant and enhancing body immunity, therefore, develop Cordyceps sinensis fungus Resource tool is of great significance, and the artificial cultivation of its sub-entities is the important channel of worm grass resources utilization.
In recent years, with the growth in the living standard of people and the increase to healthy food demand, cordyceps sinensis increasingly by To liking for people, Cordceps militaris has identical medical nutritious health effect with itself and natural cordyceps, and can artificial scale The advantages such as production, turn into the substitute of natural cordyceps, are used widely in industries such as health care, food, food and drink, north The artificial culture of cordyceps sinensis has succeeded in China and has entered the industrialization stage.
Mucor（Mucor）It is the one big category in Zygomycotina Zygomycetes Mucoales Mucoraceae fungi, it is a kind of aerobic Property, growth is fast, easily culture, distribution be wide, the bacterial strain of production complex enzyme, be the strain that traditional food sauce is brewageed in China, outgoing at home The industrial applications such as ferment production fermented bean curd, fermented soya bean processed, wine brewing have with a long history.Mucor can also produce protease, lipase, pectin Enzyme, renin, starch saccharification, oxalic acid, lactic acid, butanedioic acid and glycerine etc. are produced, is widely used in modern food industry.
The growth and development of cordyceps sinensis need efficient nutriment, use rice to make culture medium for primary raw material at present, The yield of cordyceps sinensis is restricted, and the cordycepin content in cordyceps sinensis is relatively low, and cultivation period is longer.Chinese Patent Application No. " 201410079301.8 green Cordceps militaris industrialization production method " discloses the method and concrete norm of cordyceps sinensis industrialization production； Chinese Patent Application No. 201310192693.4 " silkworm chrysalis northern Chinese caterpillar Fungus industrial cultivation technique " is to cordyceps sinensis industrialized production mistake Terms and conditions and equipment requirement in journey are optimized；A kind of " the Guangdong Cordyceps militaris of Chinese Patent Application No. 201310266374.3 The method that fructification large-scale planting method " discloses the selection and submerged fermentation strain of cordyceps species；Chinese patent Shen Please number 201310068907.7 " a kind of cordyceps sinensis quick cultivation technologies of artificial culture " to cordyceps species carried out hybridization and Domestication；Chinese Patent Application No. 200310119105.0 " northern Chinese caterpillar fungus culture liquid " and Chinese Patent Application No. 201110263224.8 " a kind of Cordceps militaris quick cultivation technologies ", by add carbon source, nitrogen source, inorganic matter and some vitamin come Promote the growth of cordyceps sinensis, the invention that the above is cultivated to cordyceps sinensis simply from cultivation condition, strain improvement and adds nutrient solution to optimize The growth of cordyceps sinensis, the growth cycle of cordyceps sinensis is not set substantially to shorten, efficient industrialization, and improve the content of wherein cordycepin.
To sum up, to shorten cultivation period, effectively saved there is an urgent need to a kind of embryo material compound accelerant of high-efficient simple at present Time, and increase economic efficiency, increase active component content, make the efficient industrialization of production of cordyceps sinensis.
The content of the invention
It is an object of the invention to be crushed using Mucor mycelium from solution, it is subject to inositol, n-octacosanol, ketose Compounded with copper gluconate, so as to the embryo material compound accelerant being prepared into needed for a kind of suitable cordyceps sinensis growth.This embryo material Compound accelerant can substantially shorten cordyceps sinensis growth cycle, improve out the weight of cordyceps sinensis in careless, increase unit volume, and make its worm Careless cellulose content dramatically increases, and processing method is simple, and cost is cheap, can widely use, and strengthens Business Economic Benefit.
In order to realize the purpose of the present invention, inventor is studied by lot of experiments, is finally obtained following technical scheme：
A kind of growing mixed accelerator of cordyceps sinensis, described embryo material compound accelerant is by microorganism Mucor bacterial cell disruption from solution Concentrate and inositol, n-octacosanol, FOS and copper gluconate compounding form, described FOS is 1~3 Fructosyl is combined the ketose of generation by β (2-1) glycosidic bond with the fructosyl in sucrose.
A kind of preparation method of the growing mixed accelerator of cordyceps sinensis, this method comprise the following steps：
（1）Mucor is cultivated on YPG liquid culture mediums, cultivation temperature is 30 ~ 35 DEG C, originates pH6.0 ~ 7.0, rotating speed is 150-200r/min, 40 ~ 60h of time, isolated Mucor mycelium；
（2）Mucor mycelium is crushed to self-dissolving under the following conditions to obtain crushing from solution：Mycelia weight and water volume ratio For 1:1~1:5（m/v）, each 30s ~ 60s of ultrasonication condition, 30s ~ 60s is spaced, circulated 15 ~ 30 times, 30 after mycelia is broken ~ 35 DEG C, stand 6 ~ 12h；
（3）To step（2）The Mucor of gained is broken to add n-octacosanol, inositol, FOS, glucose from solution Sour copper is simultaneously well mixed, and obtains embryo material compound accelerant.Wherein n-octacosanol is according to the matter crushed with Mucor mycelium from solution Measure volume ratio 0.5% ~ 2% to add, inositol adds according to the broken mass volume ratio 1% ~ 5% from solution of Mucor mycelium, oligomeric Fructose according to the addition of the broken mass volume ratio 0.05 ‰ from solution of Mucor mycelium ~ 0.2 ‰, copper gluconate according to hair Add broken mass volume ratio 0.05 ‰ ~ 0.1 ‰ from solution of mould filament.
（4）The cordyceps sinensis prepared growth embryo material compound accelerant is added in original embryo material, sterilizes, is inoculated with, culture It can harvest within 49 days.
Preferably, the preparation method of the growing mixed accelerator of cordyceps sinensis as described above, wherein described Mucor is Prey grace hair Mould CICC 3115.
The inventors discovered that Mucor mycelia, under conditions of nutrition is lacked, the various enzyme systems inside mycelium start to decompose Cell membrane and large biological molecule.Ultrasound accelerates the broken of cell membrane, and intracellular nutriment flows out, while in the presence of enzyme, born of the same parents Internal protein, nucleic acid, polysaccharide further hydrolyze.Amino acid, protein, the core that mycelium discharges in broken autolytic process Acid and carbohydrate have very high nutritive value.Form in the dry of thalline main component and there are about 50% protein, 6% nucleic acid, 16% Sugar, 11% amino acid, 6% lipid.Mucor mycelia is broken from nutrients such as the substantial amounts of enzyme of dissolution, polypeptide, amino acid, carbohydrates The flavor substance such as matter and nucleic acid, Mucor is in broken the autolytic process not only a large amount of amino acid of dissolution, protein, nucleic acid, carbohydrate in addition, Also dissolution oxalic acid, lactic acid, butanedioic acid and glycerine etc..
N-octacosanol is a kind of naturally occurring unitary higher aliphatic, is universally acknowledged anti-fatigue effect thing Matter.As a kind of natural alcohol, n-octacosanol has good biological degradability, easily absorbs and utilizes for animal body.Inositol is wide It is general to be distributed in animal and plant body, it is the growth factor of animal, microorganism, a kind of material being widely present in food, knot Structure is similar to glucose.FOS is a kind of natural active matter.Sugariness is 0.3-0.6 times of sucrose；Both sucrose is maintained Pure sweetening characteristics, it is and salubriouser than sucrose sweet taste, there is regulation gut flora, breed Bifidobacterium, promote the absorption of calcium, adjust Save blood fat, immunological regulation, the novel sweetener of the healthcare function such as anti-caries tooth, be described as most potential new after the antibiotic epoch Generation additive-growth-promoting material；Copper is the composition of a variety of important enzyme systems of intracellular, and can promote the absorption and utilization of iron, grape Saccharic acid copper is easily utilized by the microorganism as organic matter.
Compared with prior art, the present invention is sufficiently used caused mycelia in Mucor incubation, and mycelia is carried out Broken wall and self-dissolving processing, obtain the abundant protein of intracellular, polypeptide, nucleic acid, sugar, lipid, organic acid, polyalcohol, after concentrated With inositol, n-octacosanol, FOS and copper gluconate, compounded according to a certain percentage.Grown in common cordyceps sinensis This built agent is added in required embryo material, cordyceps sinensis growth can be effectively facilitated, shorten growth cycle, improve out careless and cordyceps sinensis Cellulose content, increase the economic benefit of enterprise, and process simply, lower cost, can widely use.
Embodiment
With reference to embodiments further the present invention will be described, but present disclosure be not limited solely to it is following Embodiment.Mucor employed in each embodiment is Prey grace Mucor CICC 3115, from Chinese industrial Microbiological Culture Collection Administrative center.
Embodiment 1：
（1）The spore suspensions of Mucor CICC 3115 are seeded in 115 DEG C of sterilizing 15min YPG liquid culture mediums, culture Temperature is 30 DEG C, originates pH6.0, rotating speed 150r/min, time 60h.
（2）After Mucor culture terminates, filter out mycelium and clean, then carry out freezing ultrasonication to filtering out mycelia, Mycelia weight and water volume ratio（m/v）For 1:5, ultrasonication condition is each 30s, is spaced 60s, is circulated 30 times, mycelia crushes Afterwards at 35 DEG C, 6h is stood, is obtained broken from solution.
（3）To step（2）The Mucor of gained is broken to add n-octacosanol, inositol, FOS from solution（For 1~3 Individual fructosyl is combined the ketose of generation by β (2-1) glycosidic bond with the fructosyl in sucrose）, copper gluconate and mix Uniformly, embryo material compound accelerant is obtained；Wherein n-octacosanol is according to the mass volume ratio 1% crushed with Mucor mycelium from solution Addition, inositol according to Mucor mycelium is broken is added from the mass volume ratio 1.5% of solution, FOS according to mucor The broken mass volume ratio 0.05 ‰ from solution of filament adds, and copper gluconate is according to the matter crushed with Mucor mycelium from solution Volume ratio 0.06 ‰ is measured to add.
（4）The cordyceps sinensis prepared growth embryo material compound accelerant is added in original embryo material and puddled uniformly, sterilizing, is connect Kind, culture can harvest for 49 days.
Embodiment 2：
（1）The spore suspensions of Mucor CICC 3115 are seeded in 115 DEG C of sterilizing 15min YPG liquid culture mediums, culture Temperature is 30 DEG C, originates pH6.0, rotating speed 150r/min, time 60h.
（2）After Mucor culture terminates, filter out mycelium and clean.Then freezing ultrasonication is carried out to filtering out mycelia, Mycelia weight and water volume ratio（m/v）For 1:4, ultrasonication condition is each 30s, is spaced 60s, is circulated 30 times, mycelia crushes Afterwards at 35 DEG C, 6h is stood, is obtained broken from solution.
（3）To step（2）The Mucor of gained is broken to add n-octacosanol, inositol, FOS from solution（Sugarcane fruit three Sugar）, copper gluconate and be well mixed, obtain embryo material compound accelerant；Wherein n-octacosanol crushes according to Mucor mycelium Added from the mass volume ratio 1.5% of solution, inositol according to Mucor mycelium is broken adds from the mass volume ratio 3.0% of solution Add, FOS adds according to the broken mass volume ratio 0.09 ‰ from solution of Mucor mycelium, copper gluconate according to The broken mass volume ratio 0.08 ‰ from solution of Mucor mycelium adds.
（4）The cordyceps sinensis prepared growth embryo material compound accelerant is added in original embryo material and puddled uniformly, sterilizing, is connect Kind, culture can harvest for 49 days.
Embodiment 3：
（1）The spore suspensions of Mucor CICC 3115 are seeded in 115 DEG C of sterilizing 15min YPG liquid culture mediums, culture Temperature is 30 DEG C, originates pH6.0, rotating speed 150r/min, time 60h.
（2）After Mucor culture terminates, filter out mycelium and clean.Then freezing ultrasonication is carried out to filtering out mycelia, Mycelia weight and water volume ratio（m/v）For 1:3, ultrasonication condition is each 30s, is spaced 60s, is circulated 30 times, mycelia crushes Afterwards at 35 DEG C, 6h is stood, is obtained broken from solution.
（3）To step（2）The Mucor of gained is broken to add n-octacosanol, inositol, FOS from solution（Sugarcane fruit three Sugar）, copper gluconate and be well mixed, obtain embryo material compound accelerant；Wherein n-octacosanol crushes according to Mucor mycelium Added from the mass volume ratio 2.0% of solution, inositol according to Mucor mycelium is broken adds from the mass volume ratio 4.5% of solution Add, FOS adds according to the broken mass volume ratio 0.13 ‰ from solution of Mucor mycelium, copper gluconate according to The broken mass volume ratio 0.1 ‰ from solution of Mucor mycelium adds.
Comparative example 1：（Thalline is not added with from solution）
（1）It is subject to n-octacosanol, inositol, FOS in the original embryo material grown to cordyceps sinensis（Ketose）, Portugal Grape saccharic acid copper is simultaneously well mixed, and wherein n-octacosanol is according to 1.0% addition of the original embryo material quality grown with cordyceps sinensis, flesh Alcohol adds according to the mass volume ratio 1.5% of the original embryo material grown with cordyceps sinensis, and FOS is original according to being grown with cordyceps sinensis Embryo material mass volume ratio 0.05 ‰ add, copper gluconate according to the original embryo material grown with cordyceps sinensis mass volume ratio 0.06 ‰ additions.
Comparative example 2：（It is not added with n-octacosanol）
（1）The spore suspensions of Mucor CICC 3115 are seeded in 115 DEG C of sterilizing 15min YPG liquid culture mediums, culture Temperature is 30 DEG C, originates pH6.0, rotating speed 150r/min, time 60h.
（2）After Mucor culture terminates, filter out mycelium and clean.Then freezing ultrasonication is carried out to filtering out mycelia, Mycelia weight and water volume ratio（m/v）For 1:4, ultrasonication condition is each 30s, is spaced 60s, is circulated 30 times, mycelia crushes Afterwards at 35 DEG C, 6h is stood, is obtained broken from solution.
（3）To step（2）The Mucor of gained is broken to add inositol, FOS from solution（Ketose）, glucose Sour copper is simultaneously well mixed, and obtains embryo material compound accelerant；Its mysoinositol is according to the quality volume crushed with Mucor mycelium from solution Than 3.0% addition, FOS adds according to the broken mass volume ratio 0.09 ‰ from solution of Mucor mycelium, gluconic acid Copper adds according to the broken mass volume ratio 0.08 ‰ from solution of Mucor mycelium.
（4）The cordyceps sinensis prepared growth embryo material compound accelerant is added in original embryo material and puddled uniformly, sterilizing, is connect Kind, cultivate 49 days and harvest.
Comparative example 3：（It is not added with inositol）
（1）The spore suspensions of Mucor CICC 3115 are seeded in 115 DEG C of sterilizing 15min YPG liquid culture mediums, culture Temperature is 30 DEG C, originates pH6.0, rotating speed 150r/min, time 60h.
（2）After Mucor culture terminates, filter out mycelium and clean.Then freezing ultrasonication is carried out to filtering out mycelia, Mycelia weight and water volume ratio（m/v）For 1:3, ultrasonication condition is each 30s, is spaced 60s, is circulated 30 times, mycelia crushes Afterwards at 35 DEG C, 6h is stood, is obtained broken from solution.
（3）To step（2）The Mucor of gained is broken to add n-octacosanol, FOS from solution（Ketose）、 Copper gluconate is simultaneously well mixed, and obtains embryo material compound accelerant；Wherein n-octacosanol crushes self-dissolving according to Mucor mycelium The mass volume ratio 2.0% of liquid adds, FOS according to Mucor mycelium is broken adds from the mass volume ratio 0.13 ‰ of solution Add, copper gluconate adds according to the broken mass volume ratio 0.1 ‰ from solution of Mucor mycelium.
（4）The cordyceps sinensis prepared growth embryo material compound accelerant is added in original embryo material and puddled uniformly, sterilizing, is connect Kind, cultivate 49 days and harvest.
Comparative example 4：（It is not added with FOS）
（1）The spore suspensions of Mucor CICC 3115 are seeded in 115 DEG C of sterilizing 15min YPG liquid culture mediums, culture Temperature is 30 DEG C, originates pH6.0, rotating speed 150r/min, time 60h.
（2）After Mucor culture terminates, filter out mycelium and clean.Then freezing ultrasonication is carried out to filtering out mycelia, Mycelia weight and water volume ratio（m/v）For 1:3, ultrasonication condition is each 30s, is spaced 60s, is circulated 30 times, mycelia crushes Afterwards at 35 DEG C, 6h is stood, is obtained broken from solution.
（3）To step（2）The Mucor of gained is broken to be added n-octacosanol, inositol, copper gluconate from solution and mixes Close uniformly, obtain embryo material compound accelerant；Wherein n-octacosanol is according to the mass volume ratio crushed with Mucor mycelium from solution 2.0% addition, inositol according to Mucor mycelium is broken is added from the mass volume ratio 4.5% of solution, copper gluconate according to The broken mass volume ratio 0.1 ‰ from solution of Mucor mycelium adds.
（4）The cordyceps sinensis prepared growth embryo material compound accelerant is added in original embryo material and puddled uniformly, sterilizing, is connect Kind, cultivate 49 days and harvest.
Comparative example 5：（It is not added with copper gluconate）
（1）The spore suspensions of Mucor CICC 3115 are seeded in 115 DEG C of sterilizing 15min YPG liquid culture mediums, culture Temperature is 30 DEG C, originates pH6.0, rotating speed 150r/min, time 60h.
（2）After Mucor culture terminates, filter out mycelium and clean.Then freezing ultrasonication is carried out to filtering out mycelia, Mycelia weight and water volume ratio（m/v）For 1:3, ultrasonication condition is each 30s, is spaced 60s, is circulated 30 times, mycelia crushes Afterwards at 35 DEG C, 6h is stood, is obtained broken from solution.
（3）To step（2）The Mucor of gained is broken to add n-octacosanol, inositol, FOS from solution（Sugarcane fruit three Sugar）And be well mixed, obtain embryo material compound accelerant；Wherein n-octacosanol is according to the quality crushed with Mucor mycelium from solution Volume ratio 2.0% is added, inositol according to Mucor mycelium is broken is added from the mass volume ratio 4.5% of solution, FOS is pressed Added according to the broken mass volume ratio 0.13 ‰ from solution of Mucor mycelium.
（4）The cordyceps sinensis prepared growth embryo material compound accelerant is added in original embryo material and puddled uniformly, sterilizing, is connect Kind, cultivate 49 days and harvest.
The cordyceps sinensis preparating example of the present invention, its attributional analysis such as following table：
The Indexes Comparison of cordyceps sinensis sample obtained by 1 each embodiment of table
。

Claims (4)

  1. A kind of 1. compound accelerant of cordyceps sinensis cultivation, it is characterised in that：The compound accelerant is by Mucor bacterial cell disruption from solution Concentrate forms with inositol, n-octacosanol, FOS and copper gluconate compounding.
  2. A kind of 2. compound accelerant of cordyceps sinensis cultivation as claimed in claim 1, it is characterised in that：FOS is sugarcane fruit three Sugar.
  3. A kind of 3. compound accelerant of cordyceps sinensis cultivation as claimed in claim 1, it is characterised in that：Mucor is Prey grace Mucor CICC 3115。
  4. A kind of 4. preparation method of the compound accelerant of cordyceps sinensis cultivation as claimed in claim 1, it is characterised in that：The compound rush Enter agent to prepare in accordance with the following steps：
    （1）Mucor is cultivated on YPG liquid culture mediums, cultivation temperature is 30 ~ 35 DEG C, originates pH6.0 ~ 7.0, rotating speed 150- 200r/min, 40 ~ 60h of time, isolated Mucor mycelium；
    （2）Step（1）Mucor mycelium crush self-dissolving and obtain and broken be from solution, condition：According to mycelia weight and the body of water Product ratio 1:1~1:5 are well mixed the two, each 30s ~ 60s of ultrasonication condition, are spaced 30s ~ 60s, circulate 15 ~ 30 times, bacterium At 30 ~ 35 DEG C after silk is broken, 6 ~ 12h is stood；
    （3）To step（2）The Mucor of gained is broken to add n-octacosanol, inositol, FOS, copper gluconate from solution And be well mixed, obtain embryo material compound accelerant；
    Described n-octacosanol is added according to the broken mass volume ratio 0.5% ~ 2% from solution of Mucor mycelium, and inositol is pressed Added according to the broken mass volume ratio 1% ~ 5% from solution of Mucor mycelium, FOS is according to broken certainly with Mucor mycelium The mass volume ratio 0.05 ‰ of solution ~ 0.2 ‰ are added, and copper gluconate is according to the mass body crushed with Mucor mycelium from solution Product is than 0.05 ‰ ~ 0.1 ‰ additions.
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