CN101329229A - Cyanogen-free leucocyte tri-grouping environment protection type haemolysin for blood cell analysis - Google Patents
Cyanogen-free leucocyte tri-grouping environment protection type haemolysin for blood cell analysis Download PDFInfo
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- CN101329229A CN101329229A CNA2008101387270A CN200810138727A CN101329229A CN 101329229 A CN101329229 A CN 101329229A CN A2008101387270 A CNA2008101387270 A CN A2008101387270A CN 200810138727 A CN200810138727 A CN 200810138727A CN 101329229 A CN101329229 A CN 101329229A
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Abstract
The invention relates to hemolysin used for blood cell analysis, in particular to the environmental-protection typed hemolysin of no-cyanogen leucocyte three-classification used for blood cell analysis. The hemolysin comprises the components as follows (by density): 0.5-5.0g/L of sodium lauryl sulphate (SLS), 3.0-10.0g/L of quaternary ammonium salt, 0.5-5.0g/L of nonionic surfactant, 0.5-3.0g/L of alkali metal salt, and residual quantity of deionized water. The hemolysin of the invention can quickly dissolve erythrocyte and release haemoglobin; the hemolysin and the haemoglobin form stable haemoglobin derivatives so as to lead the concentration to be exact and easy to be measured; furthermore, the leucocyte three-classification measurement is carried out simultaneously. The hemolysin of the invention can be highly relative to the HiCN mensuration recommended by International Council for Standardization in Hematology (ICSH) and can carry out the leucocyte three-classification exactly and quickly.
Description
Technical field
The present invention relates to the hemolysin that a kind of blood cell analysis is used, be specifically related to a kind of no cyamelide cell tri-grouping environment protection type hemolysin of blood cell analysis that is used for.
Background technology
The mensuration of Arneth's count and hemoglobin concentration is to analyze the important indicator of blood of human body in the medical diagnosis.Leucocyte in the blood has five types: lymphocyte, monocyte, neutrophil cell, eosinophil and basophilic granulocyte, back three is referred to as granulocyte, therefore, leucocyte being divided into three subgroups is: lymphocyte, monocyte, granulocyte.
Hemolysin is the key reagents of hemoglobin testing content (Hb) and white blood cell count(WBC) (WBC) or leukocyte differential count, belongs to one of the most frequently used reagent of clinical examination, is used in cellanalyzer with dilution etc.
Measure hemoglobin concentration and Arneth's count and all need the dilute blood sample, and need special hemolysin, on the one hand, hemolysin makes red blood cell (RBC) dissolving rapidly, discharge haemoglobin and combine the stable haemoglobin dervative of formation with hemolysin, this derivant has maximum absorption band under specific wavelength, by measuring its absorbance, thereby multiply by the content that its volumetric molar concentration calculates the haemoglobin in the blood sample; Lysed erythrocyte carries out white blood cell count(WBC) simultaneously, in order to avoid disturb leukocytic detection (because the erythrocyte number in the blood is far away more than quantity of leucocyte).On the other hand, in order to guarantee the detection of three subgroups of leucocyte, leukocytic volume also needs change to a certain degree, the leucocyte film of this moment needs dissolving to a certain degree, so that discharge endochylema, keep nuclear good working condition simultaneously, keep certain volume so that hive off mensuration.
1996, (the International Council for Standardizationin Hematology of ICSH, ICSH) recommend cyanmethemoglobin (HiCN) determination method as international hemoglobinometry standard method, its principal character is: at crest λ=540nm, during trough λ=504nm, have tangible crest, trough, though this method can more accurately be measured Hb, but its maximum shortcoming is exactly, the prussiate that uses has severe toxicity, to user's health and environment structure threat.
The patent No. is that 00106117.8 document discloses a kind of cyanogen-less hemolysin, and this hemolysin is made up of cetyl trimethyl ammonium bromide, Triton X-100, medical grade ethanol.
The patent No. is that 03140211.9 document discloses a kind of cyanide-free hemolysin and using method thereof, and this hemolysin comprises a kind of cationic surfactant, a kind of anionic surfactant, a kind of non-ionic surfactant.
The patent No. is that 200610078645.2 document discloses a kind of cyanogen-less hemolysin that is used for cellanalyzer and preparation method thereof, and it consists of: quaternary ammonium salt, imidazoles, alkali metal chlorizated salt, water.
The patent No. is that 200610012725.8 document discloses a kind of environment-friendly non-cyanogen hemolytic agent, and this hemolysin is mixed by damping fluid and stock solution, and wherein each set of dispense ratio of damping fluid is: dipotassium hydrogen phosphate 15.63g/L, citric acid 3.78g/L, water are surplus; Each set of dispense of stock solution is than being DTAC 100.00g/L, and sodium hydrogen phosphate 15.63g/L, citric acid 3.78g/L, water are surplus.
Above-mentioned patent is applicable to mensuration Hb and can not accurately carries out leucocyte three simultaneously and hive off.The present invention can with (the International Council for Standardization in Hematology of ICSH, ICSH) cyanmethemoglobin of Tui Jianing (HiCN) determination method height correlation can be carried out leucocyte three accurately and rapidly again and be hived off.
Summary of the invention
The objective of the invention is to overcome the prior art deficiency, provide a kind of can with (the International Council for Standardization in Hematology of ICSH, ICSH) recommend cyanmethemoglobin (HiCN) determination method height correlation, can carry out the no cyamelide cell tri-grouping environment protection type hemolysin of blood cell analysis that leucocyte three hives off again accurately and rapidly.
For reaching above purpose, technical scheme of the present invention is:
The no cyamelide cell tri-grouping environment protection type hemolysin of blood cell analysis comprises following component:
1, NaLS (SLS), its content range is 0.5～5.0g/l.With one of composition of no cyamelide cell tri-grouping environment protection type hemolysin, its principle is as blood cell analysis, in the blood various Hb all can with low concentration SLS effect, generation SLS-Hb brownish red compound, its absorption curve crest is at 538nm.The advantage of using NaLS (SLS) to measure hemoglobin concentration is simple to operate, is colour-stable, and accuracy and accuracy meet the requirements, and the own environmental protection of SLS, nuisanceless.Use NaLS (SLS) as one of composition of hemolysin, must grasp its concentration, the NaLS of high concentration (SLS) is excessive to leukocytic destruction.
Further, described NaLS (SLS), preferred concentration range for is: 1.0～2.0g/L.
2, quaternary ammonium salt, the surface-active contents scope is 3.0～10.0g/L.Further, the preferred C of quaternary ammonium salt
12～C
18Long chain alkyl ammonium salt.The effect of quaternary ammonium salt is a lysed erythrocyte, is convenient to discharge haemoglobin (Hb) and combines with SLS, dissolves red blood cell simultaneously and is beneficial to white blood cell count(WBC).
3, non-ionic surfactant, content range are 0.5～5.0g/L.It is solubilizing lipids that non-ionic surfactant mainly acts on, and accelerates erythrocytic dissolving, and effectively prevents the muddiness of blood measuring liquid.
4, alkali metal salt, content range are 0.5～3.0g/L.The effect of alkali metal salt is the ionic strength of regulating hemolysin, and increases the activity of surfactant.
5, surplus is a deionized water.
This is invented described blood cell analysis and cooperates dilution to use with no cyamelide cell tri-grouping environment protection type hemolysin, and lysed erythrocyte is beneficial to white blood cell count(WBC); And make the leucocyte perforation, and endochylema flows out, and cell membrane shrinks (detection kernel), in the leucocyte after the shrinkage, the lymphocyte minimum, monocyte is big slightly, and neutrophil cell exists with the maxicell form on the contrary; Combine simultaneously, form stable SLS-Hb brownish red compound fast, be used to measure hemoglobin concentration with the haemoglobin that discharges in the red blood cell.
Hemolysin of the present invention is lysed erythrocyte fast, and discharges haemoglobin, and hemolysin and haemoglobin form stable haemoglobin dervative, makes accurately and easily mensuration of concentration, and carries out leucocyte three mensuration of hiving off simultaneously.Hemolysin of the present invention can with (the International Council forStandardization in Hematology of ICSH, ICSH) recommend cyanmethemoglobin (HiCN) determination method height correlation, can carry out leucocyte three accurately and rapidly again and hive off.
Description of drawings:
Fig. 1: the hemolysin haemoglobin dervative curve map of the embodiment of the invention 1
Fig. 2: the hemolysin of the embodiment of the invention 1 and HiCN manual method are measured hemoglobin concentration comparison diagram as a result
Fig. 3: the hemolysin of the embodiment of the invention 1 and microscopy total white blood cells be comparison diagram as a result
Fig. 4: the hemolysin of the embodiment of the invention 1 and microscopy lymphocyte be comparison diagram as a result
Fig. 5: the hemolysin of the embodiment of the invention 1 and microscopy monocyte be comparison diagram as a result
Fig. 6: the hemolysin of the embodiment of the invention 1 and microscopy granulocyte be comparison diagram as a result
Embodiment:
Embodiment 1:
Prepare this hemolysin in following ratio:
(1) NaLS 1.7g
(2) cetyl trimethyl ammonium bromide 5.8g
(3) polyoxyethylene nonylphenol ether 0.8g
(4) potassium chloride 1.4g
(5) deionized water 1000ml
NaLS, cetyl trimethyl ammonium bromide, polyoxyethylene nonylphenol ether, potassium chloride are mixed according to the above ratio, add deionized water, be stirred under the room temperature evenly, use filtrator to filter to 1000ml.
Utilize spectrophotometer to carry out the haemoglobin dervative determination experiment with no cyamelide cell tri-grouping environment protection type hemolysin blood cell analysis of the present invention, the result as shown in Figure 1.
Blood cell analysis of the present invention is applied to blood analyser with no cyamelide cell tri-grouping environment protection type hemolysin:
(1) measure hemoglobin concentration, result and cyanmethemoglobin (HiCN) determination method relatively;
(2) carry out leucocyte three and hive off, result and manual method sediments microscope inspection are relatively.
Test result is seen Fig. 2 to Fig. 6.
In sum and according to the measurement result analysis, blood cell analysis of the present invention with no cyamelide cell tri-grouping environment protection type hemolysin haemoglobin dervative absorption curve maximum absorbance at the 538nm place, compare according to international standard cyanmethemoglobin (HiCN) determination method maximum absorbance 540nm and hemoglobin concentration result, hemolysin of the present invention meets the hemoglobin concentration requirement fully.
Use leucocyte three grouping result and the manual method sediments microscope inspection leucocyte three grouping result height correlations of hemolysin of the present invention, hemolysin of the present invention meets the leucocyte three mensuration requirement of hiving off fully.
Embodiment 2:
Prepare this hemolysin in following ratio:
(1) NaLS 2.0g
(2) Tetradecyl Trimethyl Ammonium Bromide 6.0g
(3)Triton?X-100 1.0g
(4) potassium chloride 2.0g
(5) deionized water 1000ml
The preparation method is identical with embodiment 1.
Blood cell analysis of the present invention with no cyamelide cell tri-grouping environment protection type hemolysin haemoglobin dervative absorption curve maximum absorbance at the 538nm place, compare according to international standard cyanmethemoglobin (HiCN) determination method maximum absorbance 540nm and hemoglobin concentration result, hemolysin of the present invention meets the hemoglobin concentration requirement fully.
Use leucocyte three grouping result and the manual method sediments microscope inspection leucocyte three grouping result height correlations of hemolysin of the present invention, hemolysin of the present invention meets the leucocyte three mensuration requirement of hiving off fully.
Embodiment 3:
Prepare this hemolysin in following ratio:
(1) NaLS 2.0g
(2) DTAB 8.0g
(3) Tween-80 4.0g
(4) potassium chloride 0.9g
(5) deionized water 1000ml
The preparation method is identical with embodiment 1.
Blood cell analysis of the present invention with no cyamelide cell tri-grouping environment protection type hemolysin haemoglobin dervative absorption curve maximum absorbance at the 538nm place, compare according to international standard cyanmethemoglobin (HiCN) determination method maximum absorbance 540nm and hemoglobin concentration result, hemolysin of the present invention meets the hemoglobin concentration requirement fully.
Use leucocyte three grouping result and the manual method sediments microscope inspection leucocyte three grouping result height correlations of hemolysin of the present invention, hemolysin of the present invention meets the leucocyte three mensuration requirement of hiving off fully.
Claims (3)
1. a blood cell analysis is characterized in that comprising following component with no cyamelide cell tri-grouping environment protection type hemolysin:
NaLS, its content range is 0.5～5.0g/L;
Quaternary ammonium salt, content range are 3.0～10.0g/L;
Non-ionic surfactant, content range are 0.5～5.0g/L;
Alkali metal salt, content range are 0.5～3.0g/L;
Surplus is a deionized water.
2. blood cell analysis according to claim 1 is characterized in that the concentration range of described NaLS is: 1.0～2.0g/L with no cyamelide cell tri-grouping environment protection type hemolysin.
3. blood cell analysis according to claim 1 is characterized in that with no cyamelide cell tri-grouping environment protection type hemolysin described quaternary ammonium salt is C
12～C
18Long chain alkyl ammonium salt.
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| CNA2008101387270A CN101329229A (en) | 2008-07-30 | 2008-07-30 | Cyanogen-free leucocyte tri-grouping environment protection type haemolysin for blood cell analysis |
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| CNA2008101387270A CN101329229A (en) | 2008-07-30 | 2008-07-30 | Cyanogen-free leucocyte tri-grouping environment protection type haemolysin for blood cell analysis |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101907628A (en) * | 2009-06-08 | 2010-12-08 | 武汉中太生物技术有限公司 | Reagent kit for measuring content of C-reactive protein in whole blood and measuring method thereof |
| CN104634953A (en) * | 2015-02-28 | 2015-05-20 | 韩冰 | Environment-friendly and non-toxic hemolytic agent |
| CN108132343A (en) * | 2017-12-12 | 2018-06-08 | 山东兰桥医学科技有限公司 | A kind of leucocyte four divides cluster analysis hemolytic agent |
| CN109596834A (en) * | 2018-10-18 | 2019-04-09 | 东软威特曼生物科技(南京)有限公司 | A kind of surface activator composition applied to external diagnosis reagent |
| 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 |
-
2008
- 2008-07-30 CN CNA2008101387270A patent/CN101329229A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101907628A (en) * | 2009-06-08 | 2010-12-08 | 武汉中太生物技术有限公司 | Reagent kit for measuring content of C-reactive protein in whole blood and measuring method thereof |
| CN104634953A (en) * | 2015-02-28 | 2015-05-20 | 韩冰 | Environment-friendly and non-toxic hemolytic agent |
| CN108132343A (en) * | 2017-12-12 | 2018-06-08 | 山东兰桥医学科技有限公司 | A kind of leucocyte four divides cluster analysis hemolytic agent |
| CN109596834A (en) * | 2018-10-18 | 2019-04-09 | 东软威特曼生物科技(南京)有限公司 | A kind of surface activator composition applied to external diagnosis reagent |
| 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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Open date: 20081224 |