CN101021524B - Method for producing non-potassium cyanide haeomolytic agent - Google Patents
Method for producing non-potassium cyanide haeomolytic agent Download PDFInfo
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- CN101021524B CN101021524B CN2007100515760A CN200710051576A CN101021524B CN 101021524 B CN101021524 B CN 101021524B CN 2007100515760 A CN2007100515760 A CN 2007100515760A CN 200710051576 A CN200710051576 A CN 200710051576A CN 101021524 B CN101021524 B CN 101021524B
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Abstract
A produce method of hemolysis agent used by home TEK style three grouping blood corpuscle analyzer with out potassium cyanide. Using dodecyl season amine salt as the main function reagent of the hemolysis agent, using innoxious material instead of potassium cyanide to measure hematoglobin, its technology steps are: (1) prepare purify water, (2)confect buffer, (3) dissolve EDTA, (4) dissolve dodecyl season amine salt, (5) constant volume, (6) filtration and filling. The technology recipe of the production is reasonable, the raw material is easy to get, and its process is easy, the cost is low, and it did not add antiseptic.
Description
Technical field
The present invention relates to a kind of working method of non-potassium cyanide haeomolytic agent, the working method of the nontoxic hemolytic agent that particularly a kind of homemade TEK series tripartite group blood cell analyser that does not contain potassium cyanide uses.
Background technology
Tripartite group blood cell analyser is that present China popularizes the routine blood test detecting instrument that uses; This quasi-instrument and reagent are import in China for a long time, because of reagent expensive and supply of material reason such as untimely bring heavy financial burden and inconvenience for hospital and patient.Popularize in three parts of crowd's cellanalyzers that use in China; Various brands are no less than 20 kinds; Because the dilution ratio of various instruments and the difference of assay method; The agent prescription of various instruments has nothing in common with each other, and the cellanalyzer of different model and manufacturer must use special-purpose matched reagent, and this is an important difficult point of cellanalyzer reagent exploitation.
Hemolytic agent is the main reagent in the tripartite group blood cell analyser reagent; Its first effect is under the mating reaction of dilution; Make erythrocyte membrane dissolving fully in the extremely short time; Make various leucocytes at the appointed time through changing leukocytic membrane structure, all kinds of leucocyte contents of suitable release are controlled all kinds of leucocyte volume sizes and are maintained certain limit.Second effect is the mensuration of content of hemoglobin; Be after hemolytic agent makes erythroclasis discharge haemoglobin (Hb); Haemoglobin combines to form cyanohemoglobin with potassium cyanide in the hemolytic agent; (generally 530～550nm) times colorimetrics, Hb content is proportional in the variation of absorbance and the liquid, and instrument just can show Hb concentration at specific wavelength.
Therefore estimate the quality of hemolytic agent quality, not only will consider the accuracy of differential count in certain action time, also will consider the accuracy of hemoglobinometry.
The major defect of existing TEK cellanalyzer hemolytic agent is: 1, cost of products is high; 2, use hypertoxic potassium cyanide to measure haemoglobin; 3, adding antiseptic influences the classification and determination effect, increases production cost.
Summary of the invention
The working method that the purpose of this invention is to provide a kind of non-potassium cyanide haeomolytic agent, its technical recipe are rationally, raw material is easy to get, low production cost, and the hemolytic agent of production is nontoxic, do not contain antiseptic.
The present invention realizes that like this raw material of production mainly is dodecyl quaternary amine and ethylenediamine tetraacetic acid, and its processing step is:
1, pure water preparation: pure water adopts the tap water preparation, and step comprises (1) electrodialysis desalination; (2) yin, yang ion exchange column desalination; (3) 0.22 microns ultra considered the filtering particulate, made pure electrical conductivity of water less than 5 μ s/cm;
2, damping fluid preparation: preparation 1/30M, the Na of PH=7.0
2HPO
4-KH
2PO
4100 liters of damping fluids, fully mix even, 0.22 micron ultra-filtration filters;
3, ethylenediamine tetraacetic acid dissolving: restrain ethylenediamine tetraacetic acids with 10～20 liters of Na with 25～200
2HPO
4-KH
2PO
4The damping fluid heating for dissolving is fully mixed evenly, cools off subsequent use;
4, dodecyl quaternary amine dissolving: restrain the dodecyl quaternary amines with 10～20 liters of Na with 500～4000
2HPO
4-KH
2PO
4The damping fluid heating for dissolving fully stirs and spares, and cools off subsequent use;
5, constant volume: it is mixed to dissolve subsequent use edta solution and dodecyl quaternary amine solution and remaining phosphate buffer, less than the pure water constant volume to 100 of 5 μ s/cm liter, fully mixes even with conductivity;
6, filter can: the hemolytic agent that the constant volume mixing is good, filter with 0.22 micron ultra worry, use the plastic bottle can, sealing, packing;
7, wrappage are handled: (1) 5L flexible packaging or 1L plastic bottle soaked 6～12 hours with 5～30% liquid detergents; (2) conductivity is washed 3～4 times less than the pure pond of 5 μ s/cm, and requiring does not have till the foam; (3) swing and wash completely, water is emptied, add the formaldehyde of 50～1000 μ l in each packing, seal for use;
8, to require be in the clean production environment of 300,000 grades clean room or relative closure, to carry out to process of producing product.Must be after all production equipments and pipeline (comprising pure water cation exchange column equipment and pipeline afterwards) preparation every day with 5 ~ 30%NaOH solution soaking; Emit NaOH solution before the preparation; With conductivity less than the purified rinse water of 5 μ s/cm 3～4 times, measure the wash water pH value near neutrality till.Last filtration can must be carried out as much as possible in confined conditions, and can finishes and packs at once.
This product is the asepsis environment-protecting product, and its lymph can reach very relevant (r＞0.96) with the neutrophil leucocyte testing result with the microscopic examination result, t check, P＞0.05, there was no significant difference.
Hemoglobinometry adopts nontoxic ethylenediamine tetraacetic acid to replace the potassium cyanide of severe toxicity, and this reagent is at content of hemoglobin 0～300g/l concentration range, linear R
2=0.9951, with 20 parts basic, normal, high value blood examination results and the high iron processes testing result of classical cyaniding, correlativity r=0.9995, t check, P＞0.05, there was no significant difference.
Good effect of the present invention is: 1, utilize the principal ingredient of dodecyl quaternary amine as hemolytic agent; The action intensity of dodecyl quaternary amine pair cell is suitable; Can select preparation in very wide concentration range; And the action intensity to different cells in these concentration ranges has tangible differentiation, can select suitable activity according to the requirement of cell count and classification; 2, through ethylenediamine tetraacetic acid being measured the scanning of the absorption peak of haemoglobin, can know that it has tangible absorption peak at the 537nm place.Through ethylenediamine tetraacetic acid being measured the linear determination of haemoglobin; Can know ethylenediamine tetraacetic acid mensuration haemoglobin; Detection accuracy can reach clinical requirement in the concentration range of the haemoglobin of 0～300g/l, can substitute potassium cyanide and be used for hemoglobinometry.The product asepsis environment-protecting receives consumer's favor more; 3, this product utilization GMP quality management principles in conjunction with practical experience, has been formulated scientific and reasonable production technology, and product does not add antiseptic, and the shelf-life can reach 2 years; 4, simple, the easy realization of industrialization of the production technology of this method.
Embodiment
Embodiment one, be example with homemade TEK-II tripartite group blood cell analyser hemolytic agent, its processing step is:
1, pure water preparation: pure water adopts the tap water preparation, and step comprises (1) electrodialysis desalination; (2) yin, yang ion exchange column desalination; (3) 0.22 microns ultra considered the filtering particulate, made pure electrical conductivity of water less than 5 μ s/cm;
2, damping fluid preparation: preparation 1/30M, the Na of PH=7.0
2HPO
4-KH
2PO
4100 liters of damping fluids, fully mix even, 0.22 micron ultra-filtration filters;
3, ethylenediamine tetraacetic acid dissolving: restrain ethylenediamine tetraacetic acids with 10～20 liters of Na with 100
2HPO
4-KH
2PO
4The damping fluid heating for dissolving is fully mixed evenly, cools off subsequent use;
4, dodecyl quaternary amine dissolving: restrain the dodecyl quaternary amines with 10～20 liters of Na with 2000
2HPO
4-KH
2PO
4The damping fluid heating for dissolving fully stirs and spares, and cools off subsequent use;
5, constant volume preparation: will dissolve subsequent use edta solution and dodecyl quaternary amine solution and mix, and less than the pure water constant volume to 100 of 5 μ s/cm liter, fully mix even with conductivity with remaining phosphate buffer;
6, filter can: the hemolytic agent that the constant volume mixing is good, filter with 0.22 micron ultra worry, with 5L flexible packaging or the can of 1L plastic bottle, sealing, packing;
7, wrappage are handled: (1) 5L flexible packaging or 1L plastic bottle soaked 6～12 hours with 10% liquid detergent; (2) conductivity is washed 3～4 times less than the pure pond of 5 μ s/cm, and requiring does not have till the foam; (3) swing and wash completely, water is emptied, add the formaldehyde of 100～300 μ l in each packing, seal for use;
8, to require be in the clean production environment of 300,000 grades clean room or relative closure, to produce to process of producing product.Must be after all production equipments and pipeline (comprising pure water cation exchange column equipment and pipeline afterwards) preparation every day with 5 ~ 30%NaOH solution soaking; Emit NaOH solution before the preparation; With conductivity less than the purified rinse water of 5 μ s/cm 3～4 times, measure the wash water pH value near neutrality till.Last filtration can must be carried out at airtight condition as much as possible, and can finishes and packs at once.
Claims (1)
1. the working method of a non-potassium cyanide haeomolytic agent, the raw material of production mainly is dodecyl quaternary ammonium salt and ethylenediamine tetraacetic acid, it is characterized in that processing step is:
1) pure water preparation: pure water adopts the tap water preparation, and step comprises (1) electrodialysis desalination; (2) yin, yang ion exchange column desalination; (3) 0.22 microns ultra-filtration filters remove particulate, make pure electrical conductivity of water less than 5 μ s/cm;
2) damping fluid preparation: preparation 1/30M, the Na of PH=7.0
2HPO
4-KH
2PO
4100 liters of damping fluids, fully mix even, 0.22 micron ultra-filtration filters;
3) ethylenediamine tetraacetic acid dissolving: restrain ethylenediamine tetraacetic acids with 10～20 liters of steps 2) Na of preparation with 25～200
2HPO
4-KH
2PO
4The damping fluid heating for dissolving is fully mixed evenly, cools off subsequent use;
4) dodecyl quaternary ammonium salt dissolving: restrain the dodecyl quaternary ammonium salts with 10～20 liters of Na with 500～4000
2HPO
4-KH
2PO
4The damping fluid heating for dissolving fully stirs and spares, and cools off subsequent use;
5) 100 liters of Na of preparation constant volume: will dissolve subsequent use edta solution and dodecyl quaternary ammonium salt solution and step 2)
2HPO
4-KH
2PO
4Damping fluid is remaining Na after step 3) is used
2HPO
4-KH
2PO
4Damping fluid is mixed, less than the pure water constant volume to 100 of 5 μ s/cm liter, fully mixes even with conductivity;
6) filter can: the hemolytic agent that the constant volume mixing is good, with 0.22 micron ultra-filtration filters, use the plastic bottle can, sealing, packing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100515760A CN101021524B (en) | 2007-02-12 | 2007-02-12 | Method for producing non-potassium cyanide haeomolytic agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100515760A CN101021524B (en) | 2007-02-12 | 2007-02-12 | Method for producing non-potassium cyanide haeomolytic agent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101021524A CN101021524A (en) | 2007-08-22 |
| CN101021524B true CN101021524B (en) | 2012-07-04 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007100515760A Expired - Fee Related CN101021524B (en) | 2007-02-12 | 2007-02-12 | Method for producing non-potassium cyanide haeomolytic agent |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103471980B (en) * | 2013-08-23 | 2015-08-26 | 深圳中科强华科技有限公司 | A kind of chip type blood cell analyzer and method |
| CN110954489A (en) * | 2019-11-15 | 2020-04-03 | 中山市创艺生化工程有限公司 | Cyanide-free hemolytic agent and application thereof |
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2007
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