CN108152096A - A kind of method that tellurium selenium cobalt ions in human faecal mass is removed using lithium base chlorite - Google Patents
A kind of method that tellurium selenium cobalt ions in human faecal mass is removed using lithium base chlorite Download PDFInfo
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- CN108152096A CN108152096A CN201711214879.XA CN201711214879A CN108152096A CN 108152096 A CN108152096 A CN 108152096A CN 201711214879 A CN201711214879 A CN 201711214879A CN 108152096 A CN108152096 A CN 108152096A
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- chlorite
- caprophyl
- tellurium
- lithium base
- cobalt ions
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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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- 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
Abstract
The invention discloses a kind of methods that tellurium selenium cobalt ions in human faecal mass is removed using lithium base chlorite, tellurium, selenium, cobalt ions in caprophyl liquid is exchanged to lithium base chlorite interlayer by the present invention using lithium base chlorite, it has adsorbed tellurium, selenium, the lithium base chlorite of cobalt ions and lithium carbonate to be removed by filtration, so as to achieve the purpose that remove tellurium, selenium, cobalt ions in caprophyl liquid.The present invention can remove tellurium, selenium, cobalt ions in human faecal mass by specific adsorption, and the excrement that the excrement donor for making tellurium, selenium, cobalt ions exceeded is contributed after processing can be up to standard.
Description
Technical field
The invention belongs to the transplanting of human excrement bacterium, fermentation, detection technique fields, are related to a kind of utilization lithium base chlorite removal human excrement
Just the method for middle tellurium selenium cobalt ions.
Background technology
" caprophyl transplanting " (Fecal microbiota transplantation, FMT) refers to will be in healthy human faecal mass
Functional flora is transplanted in patient's gastrointestinal tract, rebuilds new intestinal flora, realizes the treatment of enteron aisle and parenterally disease.Caprophyl
It transplants as the effective means for rebuilding intestinal flora, has been used for Clestridium difficile infection, inflammatory bowel disease, intractable just
A variety of floras such as secret, metabolic disease, intestine immunity defect, intestinal intolerance diseases related treatment and pilot study, and recognized
To be breakthrough Advances in Medicine in recent years.
Caprophyl feeds approach and is divided into three kinds of upper digestive tract, middle alimentary canal, lower digestive tract approach.Upper digestive tract approach refers mainly to
Oral caprophyl capsule；Middle alimentary canal approach is included through nasointestinal tube, gastroscope biopsy channel hole, percutaneous endoscopic gastrostomy jejunum pipe；Under disappear
Change road approach and include Sigmoidoscope, bowel lavage, Colostomy and through scope enteron aisle tube planting operation (transendoscopic
Enteral tubing, TET) etc..Doctor should select its most suitable for the characteristics of different situations and various approach of patient
Import pathway.
The state of caprophyl transplanting can be divided into it is fresh, freeze and three kinds of forms of fermenting.Since the safety of caprophyl transplanting becomes
The overriding concern factor of clinical decision, therefore, in order to detach a good caprophyl group without germ, to contributing the requirement of excrement person
It is very high, that diarrhea does not occur, did not use antibiotic at least near 3-6 months and it is necessary to without infectious disease and gastrointestinal tract
Disease, in addition, there is the restrictive condition for including age, physiology, pathology, psychology, sincerity, time, environment etc..
Since the caprophyl that qualified excrement donor provides cannot meet clinical demand completely, so, for certain ions
Exceeded caprophyl by technical finesse, complies with the requirement of caprophyl transplanting, has very important realistic meaning.
With the variation of human habitat, tellurium in human faecal mass, selenium, cobalt ions it is exceeded increasingly common, work as excrement
When tellurium, selenium, cobalt ions are exceeded in the excrement of supplier, the requirement of caprophyl transplanting will not met.
Invention content
The purpose of the present invention is to provide a kind of methods that tellurium selenium cobalt ions in human faecal mass is removed using lithium base chlorite.
The technical solution used in the present invention is：
It is a kind of prepare without tellurium, selenium, cobalt ions caprophyl liquid method,
1) sterile NaCl solution will be added in fecal specimens, adds lithium base chlorite, excrement slurry is obtained after stirring evenly
Body；
2) by excrement slurry initial filter, filtrate is taken, under the protection of carbon dioxide gas, nitrogen, helium, neon or argon gas,
3~6min is stirred with 1800~2200rpm every 0.8~1.2h, caprophyl slurry is obtained after 4~7h；
3) caprophyl slurry by filtration is taken into filtrate, obtains thick caprophyl liquid；Thick caprophyl liquid is centrifuged, precipitation is taken, colourless nothing is resuspended to obtain
The caprophyl liquid for being free of tellurium, selenium, cobalt ions of taste.
Further, the preparation method of the lithium base chlorite is：Water, lithium metaphosphate will be added in chlorite mineral, added
Heat adds in Na to 70~80 DEG C2CO3；2.5~3.5h is stirred, deionized water centrifuge washing is used after standing 20~30h, recycles solid
Object obtains lithium base chlorite after dry.
Further, the chlorite mineral, lithium metaphosphate, Na2CO3, water amount ratio be 80~120g：8~12g：18
~22g：1L.
Further, the cation exchange capacity of gained lithium base chlorite is 40~60mmol/100g.
Further, in step 1), the amount ratio of fecal specimens, NaCl solution and lithium base chlorite is 1g: 3~7mL：
0.5~1.5g.
Further, a concentration of 0.85%~0.95%w/v of the NaCl solution.
Further, in step 1), the speed of the stirring is 1800~2200rpm, and the time is 2~4min.
Further, in step 2), the concrete operations of the initial filter are：It is 4.5~5.5mm's by excrement slurry via hole diameter
Strainer filtering removes large particulate matter.
Further, in step 3), the concrete operations of the caprophyl slurry by filtration are：Successively via hole diameter for 1.5~
2.5mm, 0.8~1.2mm, 0.4~0.6mm and 0.2~0.3mm strainer filtering.
Further, in step 3), the rotating speed of centrifugation is 5500~6500rpm, and the time is 12~18min.
The beneficial effects of the invention are as follows：
The method report of tellurium in removal human faecal mass, selenium, cobalt ions is temporarily had not seen at present, and the present invention can pass through specific adsorption
Tellurium, selenium, the cobalt ions in human faecal mass are removed, the excrement that the excrement donor for making tellurium, selenium, cobalt ions exceeded is contributed is after processing
It can be up to standard.
Specific embodiment
It is a kind of prepare without tellurium, selenium, cobalt ions caprophyl liquid method,
1) sterile NaCl solution will be added in fecal specimens, adds lithium base chlorite, excrement slurry is obtained after stirring evenly
Body；
2) by excrement slurry initial filter, filtrate is taken, under the protection of carbon dioxide gas, nitrogen, helium, neon or argon gas,
3~6min is stirred with 1800~2200rpm every 0.8~1.2h, caprophyl slurry is obtained after 4~7h；
3) caprophyl slurry by filtration is taken into filtrate, obtains thick caprophyl liquid；Thick caprophyl liquid is centrifuged, precipitation is taken, colourless nothing is resuspended to obtain
The caprophyl liquid for being free of tellurium, selenium, cobalt ions of taste.
Preferably, the preparation method of the lithium base chlorite is：Water, lithium metaphosphate, heating will be added in chlorite mineral
To 70~80 DEG C, Na is added in2CO3；2.5~3.5h is stirred, deionized water centrifuge washing is used after standing 20~30h, recycles solid
Object obtains lithium base chlorite after dry.
Preferably, the chlorite mineral, lithium metaphosphate, Na2CO3, water amount ratio be 80~120g：8~12g：18~
22g：1L.
Preferably, the temperature of the drying is 100~110 DEG C, and the time is 45~55h.
Preferably, the cation exchange capacity of gained lithium base chlorite is 40~60mmol/100g.
Preferably, in step 1), the amount ratio of fecal specimens, NaCl solution and lithium base chlorite is 1g: 3~7mL：0.5
~1.5g.
Preferably, a concentration of 0.85%~0.95%w/v of the NaCl solution.
Preferably, in step 1), the speed of the stirring is 1800~2200rpm, and the time is 2~4min.
Preferably, in step 2), the concrete operations of the initial filter are：By the filter that excrement slurry via hole diameter is 4.5~5.5mm
Net filtration removes large particulate matter.
Preferably, in step 3), the concrete operations of the caprophyl slurry by filtration are：Successively via hole diameter for 1.5~2.5mm,
The strainer filtering of 0.8~1.2mm, 0.4~0.6mm and 0.2~0.3mm.
Preferably, in step 3), the rotating speed of centrifugation is 5500~6500rpm, and the time is 12~18min.
Preferably, in step 3), it is sterile 0.85%~0.95%w/v NaCl solutions that liquid used, which is resuspended,.
With reference to specific embodiment, the present invention is further illustrated.
Illustrate embodiments of the present invention below by way of specific specific example, those skilled in the art can be by this specification
Disclosed content understands other advantages and effect of the present invention easily.The present invention can also pass through in addition different specific realities
The mode of applying is embodied or practiced, the various details in this specification can also be based on different viewpoints with application, without departing from
Various modifications or alterations are carried out under the spirit of the present invention.
It should be clear that in the following example not specifically dated process equipment or device using conventional equipment in the art or
Device；All pressure values and range are all referring to absolute pressure.
In addition, it should also be understood that, one or more method and step mentioned in the present invention does not repel before and after the combination step
Other methods step can also be inserted into there may also be other methods step or between the step of these are specifically mentioned, unless separately
It is described；It should also be understood that the combination connection relation between one or more equipment/device mentioned in the present invention is not repelled
The front and rear two equipment/devices specifically mentioned there may also be other equipment/device or at these of the unit equipment/device it
Between can also be inserted into other equipment/device, unless otherwise indicated.Moreover, unless otherwise indicated, the number of various method steps is only
Differentiate the convenient tool of various method steps rather than the ordering for limitation various method steps or the enforceable model of the restriction present invention
It encloses, relativeness is altered or modified, and in the case where changing technology contents without essence, when being also considered as, the present invention is enforceable
Scope.
Wherein, tellurium, selenium, concentration of the cobalt ions in caprophyl liquid by inductively coupled plasma spectrometer (ICP,
Optima2000DV, the U.S.) test (test method bibliography Xiang Cai, Guang-Jian Dai, Shao-Zao Tan,
Yu Ouyang,You-Sheng Ouyang,Qing-Shan Shi.Synergistic antibacterial zinc ions
and cerium ions loadedα-zirconium phosphate.Materials Letters.2012,67(1):199-
201.)。
Embodiment 1 is a kind of to remove the method that tellurium, selenium, cobalt ions prepare caprophyl liquid in human faecal mass using lithium base chlorite
(1) preparation of lithium base chlorite：100g chlorite minerals (commercial product, grain size are 0.5~2.0mm) are added to
In 1L water, 10g lithium metaphosphates are added in, are heated to 75 DEG C, add in 20g Na2CO3；Electromagnetism 60rpm stirs 3h, and standing is spent afterwards for 24 hours
Ionized water centrifuge washing 4 times removes bottom insoluble matter, recycles solids；At 105 DEG C lithium base chlorite is obtained after dry 48h；Institute
The cation exchange capacity (CEC) of the lithium base chlorite obtained is 50mmol/100g.
(2) 40 ± 0.1g lithium bases chlorite (grain size is 0.5~2.0mm) is taken, adds in the sterile 0.9%NaCl solution of 100mL,
After the stainless steel filtering net of 500r/min stirrings 5min, 0.25mm filter out moisture, lithium base chlorite slurry is obtained, it is spare.
(3) 50 ± 1g donor fresh excretas are placed in blender, add in the sterile 0.9%NaCl solution of 250mL, add
The lithium base chlorite slurry (grain size is 0.5~2.0mm) that upper step is got ready, 2000r/min homogeneous 3min obtain excrement slurry；
(4) stainless steel filtering net filtering of the excrement slurry through 5.0mm is removed into large particulate matter therein, after obtaining initial filter
Excrement slurry；
(5) the excrement slurry after initial filter is transported to laboratory and is handled in 2h after acquisition on ice；
(6) the excrement slurry after initial filter is transferred in nitrogen bioengineering kitchen, is passed through carbon dioxide gas, and in homogenizer
In every 1h 5min is stirred with 2000r/min, caprophyl slurry is obtained after 6h；
(7) caprophyl slurry filters step by step by the stainless steel filtering net of diameter 2.0mm, 1.0mm, 0.5mm and 0.25mm, to go
Except the lithium base chlorite, lithium carbonate, unabsorbed swill and the finely ground particle substance that have adsorbed tellurium, selenium, cobalt ions, thick excrement is obtained
Bacterium solution；
(8) thick caprophyl liquid is centrifuged into 15min with the rate of 6000r/min, and caprophyl precipitation is resuspended in 250mL's
In 0.9%NaCl solution, the caprophyl liquid 250mL of nearly colorless and odorless is obtained.
Measure tellurium in gained caprophyl liquid, selenium, cobalt ions total content are that zero (instrument can't check, tellurium, selenium, cobalt ions total content
＜ 4ppb), sodium ions content 155mmol/L.
Blank control group：Operating procedure is only added without excrement and lithium with the present embodiment above-mentioned steps (1)~step (8)
Base chlorite, obtains the liquid 250mL of nearly colorless and odorless, measure tellurium, selenium, cobalt ions total content in liquid be zero (instrument can't check,
Tellurium, selenium, cobalt ions total content ＜ 4ppb), sodium ions content is zero (being shown in Table 1).
Contrast groups 1 (have excrement without chlorite)：Operating procedure is with the present embodiment above-mentioned steps (1)~step (8), only not
Lithium base chlorite is added in, obtains the liquid 250mL of nearly colorless and odorless, measuring tellurium, selenium, cobalt ions content in liquid is respectively
0.7mmol/L, 2.9mmol/L, 0.5mmol/L, tellurium, selenium, cobalt ions total content are 4.1mmol/L, and sodium ions content is
151.5mmol/L。
Contrast groups 2 (have excrement to have chlorite)：Operating procedure only adds with the present embodiment above-mentioned steps (1)~step (8)
Enter chlorite and be not added with lithium base chlorite, obtain the liquid 250mL of nearly colorless and odorless, measure tellurium in liquid, selenium, cobalt ions content difference
For 0.6mmol/L, 2.7mmol/L, 0.3mmol/L, tellurium, selenium, cobalt ions total content are 3.6mmol/L, and sodium ions content is
151.5mmol/L (is shown in Table 1).
The testing result of 1 different disposal group ions content of table
Note：aMmol/L, mM/l；
bN/A:Instrument can't check, content ＜ 4ppb (ppb:μ g/L (micrograms per litre).
From table 1 it follows that by the exceeded donor fresh excreta of lithium base chlorite treated tellurium, selenium, cobalt ions
No longer containing tellurium, selenium, cobalt ions, the excrement that the excrement donor for making tellurium, selenium, cobalt ions exceeded is contributed can reach after processing
Mark, and the content of sodium ion is held essentially constant.
Embodiment 2 is a kind of to remove the method that tellurium, selenium, cobalt ions prepare caprophyl liquid in human faecal mass using lithium base chlorite
(1) preparation of lithium base chlorite：120g chlorite minerals (commercial product, grain size are 0.5~2.0mm) are added to
In 1L water, 8g lithium metaphosphates are added in, are heated to 80 DEG C, add in 22g Na2CO3；Electromagnetism 60rpm stirs 3.5h, is used after standing 20h
Deionized water centrifuge washing 4 times removes bottom insoluble matter, recycles solids；At 100 DEG C lithium base chlorite is obtained after dry 45h；
The cation exchange capacity (CEC) of the lithium base chlorite of gained is 60mmol/100g.
(2) 50 ± 1g donor fresh excretas are placed in blender, add in the sterile 0.95%NaCl solution of 150mL, then add
50g lithium bases chlorite (grain size is 0.5~2.0mm), 2200r/min homogeneous 4min obtain excrement slurry；
(3) stainless steel filtering net filtering of the excrement slurry through 5.0mm is removed into large particulate matter therein, after obtaining initial filter
Excrement slurry；
(4) the excrement slurry after initial filter is transported to laboratory and is handled in 2h after acquisition on ice；
(5) the excrement slurry after initial filter is transferred in nitrogen bioengineering kitchen, is passed through nitrogen gas, and every in homogenizer
5min is stirred with 2000r/min every 1.2h, caprophyl slurry is obtained after 4h；
(6) caprophyl slurry filters step by step by the stainless steel filtering net of diameter 2.0mm, 1.0mm, 0.5mm and 0.25mm, to go
Except the lithium base chlorite, lithium carbonate, unabsorbed swill and the finely ground particle substance that have adsorbed tellurium, selenium, cobalt ions, thick excrement is obtained
Bacterium solution；
(7) thick caprophyl liquid is centrifuged into 18min with the rate of 6500r/min, and caprophyl precipitation is resuspended in 250mL's
In 0.9%NaCl solution, the caprophyl liquid 250mL that nearly colorless and odorless is free of tellurium, selenium, cobalt ions is obtained.
Embodiment 3 is a kind of to remove the method that tellurium, selenium, cobalt ions prepare caprophyl liquid in human faecal mass using lithium base chlorite
(1) preparation of lithium base chlorite：80g chlorite minerals (commercial product, grain size are 0.5~2.0mm) are added to
In 1L water, 12g lithium metaphosphates are added in, are heated to 70 DEG C, add in 18g Na2CO3；Electromagnetism 60rpm stirs 2.5h, is used after standing 30h
Deionized water centrifuge washing 4 times removes bottom insoluble matter, recycles solids；At 110 DEG C lithium base chlorite is obtained after dry 55h；
The cation exchange capacity (CEC) of the lithium base chlorite of gained is 40mmol/100g.
(2) 50 ± 1g donor fresh excretas are placed in blender, add in the sterile 0.85%NaCl solution of 350mL, then add
65g lithium bases chlorite (grain size is 0.5~2.0mm), 2200r/min homogeneous 4min obtain excrement slurry；
(3) stainless steel filtering net filtering of the excrement slurry through 5.0mm is removed into large particulate matter therein, after obtaining initial filter
Excrement slurry；
(4) the excrement slurry after initial filter is transported to laboratory and is handled in 2h after acquisition on ice；
(5) the excrement slurry after initial filter is transferred in nitrogen bioengineering kitchen, is passed through nitrogen gas, and every in homogenizer
5min is stirred with 2000r/min every 1.2h, caprophyl slurry is obtained after 4h；
(6) caprophyl slurry filters step by step by the stainless steel filtering net of diameter 2.0mm, 1.0mm, 0.5mm and 0.25mm, to go
Except the lithium base chlorite, lithium carbonate, unabsorbed swill and the finely ground particle substance that have adsorbed tellurium, selenium, cobalt ions, thick excrement is obtained
Bacterium solution；
(7) thick caprophyl liquid is centrifuged into 12min with the rate of 6500r/min, and caprophyl precipitation is resuspended in 250mL's
In 0.9%NaCl solution, the caprophyl liquid 250mL that nearly colorless and odorless is free of tellurium, selenium, cobalt ions is obtained.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (10)
1. it is a kind of prepare without tellurium, selenium, cobalt ions caprophyl liquid method, it is characterised in that：
1) sterile NaCl solution will be added in fecal specimens, adds lithium base chlorite, excrement slurry is obtained after stirring evenly；
2) by excrement slurry initial filter, filtrate is taken, under the protection of carbon dioxide gas, nitrogen, helium, neon or argon gas, every
0.8~1.2h stirs 3~6min with 1800~2200rpm, and caprophyl slurry is obtained after 4~7h；
3) caprophyl slurry by filtration is taken into filtrate, obtains thick caprophyl liquid；Thick caprophyl liquid is centrifuged, takes precipitation, colorless and odorless is resuspended to obtain
Without tellurium, selenium, cobalt ions caprophyl liquid.
2. according to the method described in claim 1, it is characterized in that, the preparation method of the lithium base chlorite is：By chlorite
Water, lithium metaphosphate are added in mineral, is heated to 70~80 DEG C, adds in Na2CO3；2.5~3.5h is stirred, is used after standing 20~30h
Deionized water centrifuge washing recycles solids, lithium base chlorite is obtained after dry.
3. according to the method described in claim 2, it is characterized in that, the chlorite mineral, lithium metaphosphate, Na2CO3, water use
Amount is than being 80~120g：8~12g：18~22g：1L.
4. according to the method described in claim 2, it is characterized in that, the cation exchange capacity of gained lithium base chlorite for 40~
60mmol/100g。
5. according to the method described in claim 1, it is characterized in that, in step 1), fecal specimens, NaCl solution and lithium base green mud
The amount ratio of stone is 1g: 3~7mL：0.5~1.5g.
6. according to the method described in claim 1, it is characterized in that, a concentration of 0.85%~0.95%w/ of the NaCl solution
v。
7. according to the method described in claim 1, it is characterized in that, in step 1), the speed of the stirring for 1800~
2200rpm, time are 2~4min.
8. according to the method described in claim 1, it is characterized in that, in step 2), the concrete operations of the initial filter are：By excrement
The strainer filtering that slurry via hole diameter is 4.5~5.5mm removes large particulate matter.
9. according to the method described in claim 1, it is characterized in that, in step 3), the concrete operations of the caprophyl slurry by filtration
For：Via hole diameter is the strainer filtering of 1.5~2.5mm, 0.8~1.2mm, 0.4~0.6mm and 0.2~0.3mm successively.
10. according to the method described in claim 1, it is characterized in that, in step 3), the rotating speed of centrifugation is 5500~6500rpm,
Time is 12~18min.
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