CN102925536A - Preparation method for quality control materials of glucose-6-phosphate dehydrogenase activity assay and preparation - Google Patents
Preparation method for quality control materials of glucose-6-phosphate dehydrogenase activity assay and preparation Download PDFInfo
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- CN102925536A CN102925536A CN2012104346264A CN201210434626A CN102925536A CN 102925536 A CN102925536 A CN 102925536A CN 2012104346264 A CN2012104346264 A CN 2012104346264A CN 201210434626 A CN201210434626 A CN 201210434626A CN 102925536 A CN102925536 A CN 102925536A
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Abstract
The invention provides a preparation method for quality control materials of glucose-6-phosphate dehydrogenase activity assay and a preparation. The method includes the following steps: (1) preparing an imidazole buffer solution; (2) preparing the quality control materials of high concentration, middle concentration and low concentration of glucose-6-phosphate dehydrogenase; and (3) packaging separately and storing. The preparation has the advantages of being capable of being used as an indoor quality control material of the glucose-6-phosphate dehydrogenase activity assay, capable of monitoring if corresponding solvent goes bad in circulation, storing and use processes, capable of monitoring operation conditions of a monitoring device, and meeting requirements of laws and actual work. The quality control materials of the high concentration, the middle concentration and the low concentration of the glucose-6-phosphate dehydrogenase are good in uniformity, difference among tubes is smaller than 2%, the quality control materials of the high concentration, the middle concentration and the low concentration of the glucose-6-phosphate dehydrogenase all contain preservative and enzyme protective agents, and under the condition of low-temperature storing, the period of validity is larger than one year.
Description
[technical field]
The present invention relates to a kind of Internal Quality Control thing, relate in particular to a kind of Internal Quality Control thing of measuring the glucose-6-phosphate dehydrogenase (G6PD) activity.
[background technology]
Glucose-6-phosphate dehydrogenase (G6PD) (glucose-6-phosphatase dehydrogenase, G-6-PD) deficiency disease is a kind of heredopathia due to the remarkable shortage of this enzyme in the red corpuscle, is commonly called as favism.Its main clinical manifestation is hemolytic anemia.
The mechanism of this disease generation haemolysis is not yet fully clear, think at present: glucose-6-phosphate dehydrogenase (G6PD) is necessary desaturase in the metabolism of Red Blood Cells Glucose pentose phosphate shunt, it makes G6P disengage H+, thereby make oxidized form of nicotinamide-adenine dinucleotide I (NADP+) be reduced into DPNH I (NADPH), NADPH is oxidation resistant important substance in the red corpuscle, it can make the intracellular Sleep-promoting factor B of red corpuscle (GSSG) be reduced into reduced glutathion (GSH) and keep the activity of catalase (Catalase, Cat).Cat is the reductase enzyme that hydrogen peroxide (H2O2) is reduced into water.The Main Function of GSH is: (integrity of oxyphorase SH), zymoprotein and membranin avoids H2O2 to containing-oxidation of SH material 1. to protect red corpuscle to include sulfydryl.2. jointly make H2O2 be reduced into water with gsh hydrogen peroxide (GSHPX).During glucose-6-pbosphate dehydrogenase deficiency, NADPH generates not enough, and Cat and GSH reduce.Therefore, when body is subject to oxide compound when infringement, the H2O2 that oxygenizement produces can not in time be reduced into water, too much H2O2 act on contain-oxidative damage all occurs in oxyphorase, membranin and zymoprotein hyperamization Lactoferrin and the membranin of SH.Oxyphorase is oxidized to methemoglobin and distortion globin corpusculum (Heinzbody), and the excessively oxygen injury of erythrocyte membrane then causes the oxidation of membrane lipid and membranin sulfydryl, and above-mentioned effect finally causes oxidative damage and the haemolysis of cell.
Measuring the Glucose-6-phosphate dehydrogenate activity is this sick Main Means of diagnosis.
The Ministry of Health in 2006 promulgate " clinical laboratory test capacity management method " the 25 regulation of j: " clinical laboratory test capacity should carry out indoor quality control to the Clinical laboratory test of carrying out, the rendering quality control chart ".At present, the existing commercialization Quality Control thing of most detection by quantitative (such as gpt activity mensuration etc.), but the glucose-6-phosphate dehydrogenase (G6PD) determination of activity does not have commercialization Quality Control thing, as not voluntarily preparation just be difficult to carry out Internal Quality Control.
[summary of the invention]
The object of the present invention is to provide a kind of compound method and preparation of glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing
The present invention is achieved through the following technical solutions:
The compound method of described glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing comprises the steps:
(1) preparation imidazole buffer: it is an amount of to weigh imidazoles, two water disodium ethylene diamine tetraacetate (EDTANa22H20), enzymatic protective reagent, sodium azide, is dissolved in the 900ml distilled water, and with 1mol/L vinegar acid for adjusting pH to 7.4, adding distil water is diluted to 1L.
(2) the high, medium and low concentration Quality Control of preparation glucose-6-phosphate dehydrogenase (G6PD) thing: get three parts of above-mentioned imidazole buffers, add respectively an amount of glucose-6-phosphate dehydrogenase (G6PD) to final concentration and be about 6000U/L, 3000U/L, 1000U/L.
(3) packing and preservation: glucose-6-phosphate dehydrogenase (G6PD) Quality Control thing divided be filled to the disposable plastic centrifuge tube, put-20 ℃ of Refrigerator stores.
The composition of high concentration glucose-6-phosphate dehydrogenase enzyme assay Quality Control thing preparation is
Imidazoles (g) 3.4 to 34;
EDTANa22H2O (g) 0.93 to 9.3;
Enzymatic protective reagent (g) 5 to 20;
Sodium azide (g) 1 to 10;
G-6-PD(U)5000-7000；
1mol/L acetic acid is transferred pH to 7.4;
H2O(ml)1000；
The composition of middle concentration glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing preparation is
Imidazoles (g) 3.4 to 34;
EDTANa22H2O (g) 0.93 to 9.3;
Enzymatic protective reagent (g) 5 to 20;
Sodium azide (g) 1 to 10;
G-6-PD(U)2000-4000；
1mol/L acetic acid is transferred pH to 7.4;
H2O(ml)1000；
The composition of low concentration glucose-6-phosphate dehydrogenase enzyme assay Quality Control thing preparation is
Imidazoles (g) 3.4 to 34;
EDTANa22H2O (g) 0.93 to 9.3;
Enzymatic protective reagent (g) 5 to 20;
Sodium azide (g) 1 to 10;
G-6-PD(U)500-1500；
1mol/L acetic acid is transferred pH to 7.4;
H2O(ml)1000。
Beneficial effect of the present invention is: the present invention can be used as the Internal Quality Control thing of glucose-6-phosphate dehydrogenase (G6PD) determination of activity; both can monitor corresponding reagent in circulation; preserve; whether occur in the use procedure going bad; but the operation conditions of Monitoring equipment equipment also; this is the requirement of rules; especially real work in the urgent need to; the glucose-6-phosphate dehydrogenase (G6PD) of the present invention's preparation is high; in; lower concentration Quality Control thing homogeneity is good; difference is less than 2% between pipe; the glucose-6-phosphate dehydrogenase (G6PD) of the present invention's preparation is high; in; all contain sanitas and enzymatic protective reagent in the lower concentration Quality Control thing; under the cryopreservation condition, validity period was greater than 1 year.
[embodiment]
The below is described further the present invention according to embodiment:
Embodiment 1:
The compound method of described glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing comprises the steps:
(1) preparation imidazole buffer: it is an amount of to weigh imidazoles, two water disodium ethylene diamine tetraacetate (EDTANa22H2O), enzymatic protective reagent, sodium azide, is dissolved in the 900ml distilled water, and with 1mol/L vinegar acid for adjusting pH to 7.4, adding distil water is diluted to 1L.
(2) preparation glucose-6-phosphate dehydrogenase (G6PD) high density Quality Control thing: get three parts of above-mentioned imidazole buffers, add respectively an amount of glucose-6-phosphate dehydrogenase (G6PD) to final concentration and be about 6000U/L.
(3) packing and preservation: glucose-6-phosphate dehydrogenase (G6PD) Quality Control thing divided be filled to the disposable plastic centrifuge tube, put-20 ℃ of Refrigerator stores.
The composition of high concentration glucose-6-phosphate dehydrogenase enzyme assay Quality Control thing preparation is
Imidazoles (g) 3.4;
EDTANa2·2H2O(g)0.93；
Enzymatic protective reagent (g) 5;
Sodium azide (g) 1;
G-6-PD(U)5000；
1mol/L acetic acid is transferred pH to 7.4;
H2O(ml)1000；
Use high value Quality Control thing of the present invention to carry out glucose-6-phosphate dehydrogenase (G6PD) determination of activity Internal Quality Control 1 year, cumulative mean value is: 6017U/L, the accumulation standard deviation is: 175.7U/L, accumulation CV value is: 2.92%.
Embodiment 2:
The compound method of described glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing comprises the steps:
(1) preparation imidazole buffer: it is an amount of to weigh imidazoles, two water disodium ethylene diamine tetraacetate (EDTANa22H2O), enzymatic protective reagent, sodium azide, is dissolved in the 900ml distilled water, and with 1mol/L vinegar acid for adjusting pH to 7.4, adding distil water is diluted to 1L.
(2) concentration Quality Control thing in the preparation glucose-6-phosphate dehydrogenase (G6PD): get three parts of above-mentioned imidazole buffers, add respectively an amount of glucose-6-phosphate dehydrogenase (G6PD) to final concentration and be about 3000U/L.
(3) packing and preservation: glucose-6-phosphate dehydrogenase (G6PD) Quality Control thing divided be filled to the disposable plastic centrifuge tube, put-20 ℃ of Refrigerator stores.
The composition of middle concentration glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing preparation is
Imidazoles (g) 18;
EDTANa2·2H2O(g)4.5；
Enzymatic protective reagent (g) 12;
Sodium azide (g) 5;
G-6-PD(U)3000；
1mol/L acetic acid is transferred pH to 7.4;
H2O(ml)1000；
Use intermediate value Quality Control thing of the present invention to carry out glucose-6-phosphate dehydrogenase (G6PD) determination of activity Internal Quality Control 1 year, cumulative mean value is: 2976U/L, the accumulation standard deviation is: 103.3U/L, accumulation CV value is: 3.47%.
Embodiment 3:
The compound method of described glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing comprises the steps:
(1) preparation imidazole buffer: it is an amount of to weigh imidazoles, two water disodium ethylene diamine tetraacetate (EDTANa22H2O), enzymatic protective reagent, sodium azide, is dissolved in the 900ml distilled water, and with 1mol/L vinegar acid for adjusting pH to 7.4, adding distil water is diluted to 1L.
(2) preparation glucose-6-phosphate dehydrogenase (G6PD) lower concentration Quality Control thing: get three parts of above-mentioned imidazole buffers, add respectively an amount of glucose-6-phosphate dehydrogenase (G6PD) to final concentration and be about 1000U/L.
(3) packing and preservation: glucose-6-phosphate dehydrogenase (G6PD) Quality Control thing divided be filled to the disposable plastic centrifuge tube, put-20 ℃ of Refrigerator stores.
The composition of low concentration glucose-6-phosphate dehydrogenase enzyme assay Quality Control thing preparation is
Imidazoles (g) 34;
EDTANa2·2H2O(g)9.3；
Enzymatic protective reagent (g) 20;
Sodium azide (g) 10;
G-6-PD(U)1500；
1mol/L acetic acid is transferred pH to 7.4;
H2O(ml)1000。
Use low value Quality Control thing of the present invention to carry out glucose-6-phosphate dehydrogenase (G6PD) determination of activity Internal Quality Control 1 year, cumulative mean value is: 1038U/L, the accumulation standard deviation is: 50.3U/L, accumulation CV value is: 4.85%.
The according to the above description announcement of book and instruction, those skilled in the art in the invention can also carry out suitable change and modification to above-mentioned embodiment.Therefore, the embodiment that discloses and describe above the present invention is not limited to also should fall in the protection domain of claim of the present invention modifications and changes more of the present invention.In addition, although used some specific terms in this specification sheets, these terms do not consist of any restriction to the present invention just for convenience of description.
Claims (2)
1. the compound method of a glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing is characterized in that: comprise the steps
(1) preparation imidazole buffer: it is an amount of to weigh imidazoles, two water disodium ethylene diamine tetraacetate (EDTANa22H2O), enzymatic protective reagent, sodium azide, is dissolved in the 900ml distilled water, and with 1mol/L vinegar acid for adjusting pH to 7.4, adding distil water is diluted to 1L;
(2) the high, medium and low concentration Quality Control of preparation glucose-6-phosphate dehydrogenase (G6PD) thing: get three parts of above-mentioned imidazole buffers, add respectively an amount of glucose-6-phosphate dehydrogenase (G6PD) to final concentration and be about 6000U/L, 3000U/L, 1000U/L;
(3) packing and preservation: glucose-6-phosphate dehydrogenase (G6PD) Quality Control thing divided be filled to the disposable plastic centrifuge tube, put-20 ℃ of Refrigerator stores.
2. glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing preparation, it is characterized in that: the composition of high concentration glucose-6-phosphate dehydrogenase enzyme assay Quality Control thing preparation is
Imidazoles (g) 3.4 to 34;
EDTANa22H2O (g) 0.93 to 9.3;
Enzymatic protective reagent (g) 5 to 20;
Sodium azide (g) 1 to 10;
G-6-PD(U)5000-7000；
1mol/L acetic acid is transferred pH to 7.4;
H2O(ml)1000；
The composition of middle concentration glucose-6-phosphate dehydrogenase (G6PD) determination of activity Quality Control thing preparation is
Imidazoles (g) 3.4 to 34;
EDTANa22H2O (g) 0.93 to 9.3;
Enzymatic protective reagent (g) 5 to 20;
Sodium azide (g) 1 to 10;
G-6-PD(U)2000-4000；
1mol/L acetic acid is transferred pH to 7.4;
H2O(ml)1000；
The composition of low concentration glucose-6-phosphate dehydrogenase enzyme assay Quality Control thing preparation is
Imidazoles (g) 3.4 to 34;
EDTANa22H2O (g) 0.93 to 9.3;
Enzymatic protective reagent (g) 5 to 20;
Sodium azide (g) 1 to 10;
G-6-PD(U)500-1500；
1mol/L acetic acid is transferred pH to 7.4;
H2O(ml)1000。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104880350A (en) * | 2015-05-22 | 2015-09-02 | 宁波美康生物科技股份有限公司 | Preparation method for leucine aminopeptidase quality control product |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5716797A (en) * | 1995-01-13 | 1998-02-10 | Wako Pure Chemical Industries, Ltd. | Stable two-part reagent for the measurement of creatine kinase activity |
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2012
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5716797A (en) * | 1995-01-13 | 1998-02-10 | Wako Pure Chemical Industries, Ltd. | Stable two-part reagent for the measurement of creatine kinase activity |
Non-Patent Citations (2)
| Title |
|---|
| 刘和录等: "５种定性试验质控物的研制与应用", 《国际检验医学杂志》 * |
| 贾璋林等: "葡萄糖-6-磷酸脱氢酶活性浓度检测法的初步研究", 《检验医学与临床》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104880350A (en) * | 2015-05-22 | 2015-09-02 | 宁波美康生物科技股份有限公司 | Preparation method for leucine aminopeptidase quality control product |
| CN104880350B (en) * | 2015-05-22 | 2017-11-28 | 美康生物科技股份有限公司 | A kind of preparation method of leucine aminopeptidase quality-control product |
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Application publication date: 20130213 |