CN101713728B - Method for measuring content of hemachrome - Google Patents
Method for measuring content of hemachrome Download PDFInfo
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- CN101713728B CN101713728B CN2009101852807A CN200910185280A CN101713728B CN 101713728 B CN101713728 B CN 101713728B CN 2009101852807 A CN2009101852807 A CN 2009101852807A CN 200910185280 A CN200910185280 A CN 200910185280A CN 101713728 B CN101713728 B CN 101713728B
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- hemachrome
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Abstract
The invention relates to a method for measuring the content of hemachrome. The invention provides a method for solving oxidation interference when the hemachrome is measured by a spectrophotometric method. The main content of the method comprises the following steps: (1) preparing a standard test solution of the hemachrome; (2) manufacturing a standard curve of the hemachrome; (3) preparing a sample solution containing the hemachrome; and (4) measuring the content of the hemachrome of sample solution. The invention reduces interference factors in the process of measuring the hemachrome by adding sodium D-isoascorbate in the solution, increases the accuracy of measuring the content of the hemachrome by the spectrophotometric method and provides a more accurate and reliable method for measuring the content of the hemachrome.
Description
(1) technical field
The present invention relates to a kind of assay method of content of hemachrome.
(2) background technology
Protoheme extracts from animal blood, the Fe2+ compound of protoporphyrin IX---metalloporphyrin coordination compound, it is the prothetic group of various hemoproteins, it and different protein combination can form haemoglobin, myoglobins, cromoci, hydrogen peroxidase and peroxidase etc.Protoheme constitutes the iron porphyrin compound by porphyrin and a part are ferrous, molecular formula C34H30FeN4O4, and relative molecular mass 614.48, structural formula is:
Protoheme be modern medicine generally acknowledge prevent and treat hypoferric anemia absorptivity height, effective biological source of iron, no iron taste, not stimulating gastrointestinal; Protoheme still is the important source material of antineoplastic, now has been used to treat diseases such as liver cancer, carcinoma of urinary bladder, melanoma, retina tumor; In food service industry, protoheme can replace cooked meat product to jeopardize the colour former nitrite and the synthetic food color of health, makes meat products attractive in appearance more tempting, reduces the nitrite carcinogenesis.Therefore, in recent years, protoheme was all obtaining application aspect food, medicine and the fine chemistry industry, and more and more came into one's own.Products such as heme sweets with blood nourishing function, beverage, food, medicine are developed and are developed in China's research in this field of throwing oneself in the recent period in succession.
The assay method of relevant protoheme has multiple, pyridine hemochromogen absorption measurement method, ultraviolet spectroscopy, polarography and electrode method as fluorescence method, protoheme, these assay methods each have characteristics separately, but certain shortcoming is arranged also, the use cost of these methods is all very high, and many laboratories do not have these equipment yet.Present because visible spectrophotometry has easily row easy and simple to handle, quick, and equipment is simply accepted by numerous researchers, be more a kind of method of use in protoheme is measured.But disturbed by other composition, measured value generally has and departs from.Main interference factors in the mensuration process is oxidation, protoheme color instability, easily oxidized and brown stain.The interference of oxidation when the present invention is intended to solve visible spectrophotometry mensuration protoheme, thus assurance provided for accurately measuring content of hemachrome.
(3) summary of the invention
The objective of the invention is to determine accurately the content of protoheme.
Purpose of the present invention is achieved by following technical solution:
(1) preparation of protoheme standard test solution: precision takes by weighing 105 ℃ of Powdered protoheme standard items 20.0mg that are dried to constant weight, take by weighing sodium isoascorbate 10mg again,, in the brown volumetric flask of 100mL, shake up with 0.1mol/L NaOH solution constant volume, get protoheme standard test solution.
(2) making of protoheme typical curve: precision is measured protoheme standard test solution 1mL, 2mL, 3mL, 4mL, 5mL, promptly contain 200,400,600,800,1000 μ g protohemes, with 0.1mol/LNaOH solution constant volume in the brown volumetric flask of 100ml, shake up, with 0.1mol/L NaOH solution is blank, measure its light absorption value at the 400nm place, concentration with protoheme is horizontal ordinate, light absorption value is an ordinate, make regression treatment, get regression equation: A=aC+b, wherein A is that light absorption value, C are protoheme concentration.
(3) contain the preparation of the sample solution of protoheme: precision takes by weighing 105 ℃ of samples that contain protoheme that are dried to the 20mg of constant weight and puts into beaker, take by weighing sodium isoascorbate 10mg again, add the dissolving of 50mL 0.1mol/L NaOH solution, transfer to then in the brown volumetric flask of 100mL, be settled to scale.Use the funnel filtering solution, discard filtrate just.
(4) mensuration of sample liquid content of hemachrome: filtrate is measured light absorption value at the 400nm place.Light absorption value substitution typical curve formula is obtained the concentration (C of protoheme in the sample liquid
0), then by the content of protoheme in the following formula calculation sample.
In the formula: C
0-for supplying protoheme concentration in the test agent liquid, μ g/mL;
W-is the weight for test agent, g.
(4) description of drawings
Accompanying drawing: protoheme typical curve
(5) specific embodiments
Embodiment one
(1) preparation of protoheme standard test solution: precision takes by weighing 105 ℃ of Powdered protoheme standard items 20.0mg that are dried to constant weight, take by weighing sodium isoascorbate 10mg again,, in the brown volumetric flask of 100mL, shake up with 0.1mol/L NaOH solution constant volume, get protoheme standard test solution;
(2) making of protoheme typical curve: precision is measured protoheme standard test solution 1mL, 2mL, 3mL, 4mL, 5mL, promptly contain 200,400,600,800,1000 μ g protohemes, with 0.1mol/L NaOH solution constant volume in the brown volumetric flask of 100ml, shake up, with 0.1mol/LNaOH solution is blank, measure its light absorption value at the 400nm place, concentration with protoheme is horizontal ordinate, light absorption value is an ordinate, make regression treatment, get regression equation: A=0.0775C-0.0026, R2=0.9998 (A is a light absorption value, and C is a protoheme concentration);
(3) contain the preparation of the sample solution of protoheme: precision takes by weighing 105 ℃ of samples that contain protoheme that are dried to the 20mg of constant weight and puts into beaker, take by weighing sodium isoascorbate 10mg again, add the dissolving of 50mL 0.1mol/L NaOH solution, transfer to then in the brown volumetric flask of 100mL, be settled to scale.Use the funnel filtering solution, discard filtrate just;
(4) mensuration of sample liquid content of hemachrome: filtrate is measured light absorption value at the 400nm place.Light absorption value substitution typical curve formula is obtained the concentration (C of protoheme in the sample liquid
1), then by the content of protoheme in the following formula calculation sample.(the results are shown in Table 1)
In the formula: C
1-for supplying protoheme concentration in the test agent liquid, μ g/mL;
W-is the weight for test agent, g.
The mensuration of content of hemachrome in the table 1 haemoglobin powder
Embodiment two
(1) preparation of protoheme standard test solution: precision takes by weighing 105 ℃ of Powdered protoheme standard items 20.0mg that are dried to constant weight, take by weighing sodium isoascorbate 10mg again,, in the brown volumetric flask of 100mL, shake up with 0.1mol/L NaOH solution constant volume, get protoheme standard test solution;
(2) making of protoheme typical curve: precision is measured protoheme standard test solution 1mL, 2mL, 3mL, 4mL, 5mL, promptly contain 200,400,600,800,1000 μ g protohemes, with 0.1mol/LNaOH solution constant volume in the brown volumetric flask of 100ml, shake up, with 0.1mol/LNaOH solution is blank, measure its light absorption value at the 400nm place, concentration with protoheme is horizontal ordinate, light absorption value is an ordinate, make regression treatment, get regression equation: A=0.0775C-0.0026, R2=0.9998 (A is a light absorption value, and C is a protoheme concentration);
(3) contain the preparation of the sample solution of protoheme: precision takes by weighing 105 ℃ of rough protoheme samples that are dried to the 20mg of constant weight and puts into beaker, take by weighing sodium isoascorbate 10mg again, add the dissolving of 50mL 0.1mol/LNaOH solution, transfer to then in the brown volumetric flask of 100mL, be settled to scale.Use the funnel filtering solution, discard filtrate just;
(4) mensuration of sample liquid content of hemachrome: filtrate is measured light absorption value at the 400nm place.Light absorption value substitution typical curve formula is obtained the concentration (C of protoheme in the sample liquid
2), then by the content of protoheme in the following formula calculation sample.(the results are shown in Table 2)
In the formula: C
2-for supplying protoheme concentration in the test agent liquid, μ g/mL;
D-is an extension rate, D=3 among the embodiment;
W-is the weight for test agent, g.
The mensuration of content of hemachrome in the rough protoheme sample of table 2
Claims (1)
1. the assay method of a content of hemachrome is characterized in that may further comprise the steps:
(1) preparation of protoheme standard test solution: precision takes by weighing 105 ℃ of Powdered protoheme standard items 20.0mg that are dried to constant weight, take by weighing sodium isoascorbate 10mg again,, in the brown volumetric flask of 100mL, shake up with 0.1mol/L NaOH solution constant volume, get protoheme standard test solution;
(2) making of protoheme typical curve: precision is measured protoheme standard test solution 1mL, 2mL, 3mL, 4mL, 5mL, promptly contain 200,400,600,800,1000 μ g protohemes, with 0.1mol/L NaOH solution constant volume in the brown volumetric flask of 100ml, shake up, with 0.1mol/L NaOH solution is blank, measure its light absorption value at the 400nm place, concentration with protoheme is horizontal ordinate, light absorption value is an ordinate, make regression treatment, get regression equation: A=aC+b, wherein A is that light absorption value, C are protoheme concentration;
(3) contain the preparation of the sample solution of protoheme: precision takes by weighing 105 ℃ of samples that contain protoheme that are dried to the 20mg of constant weight and puts into beaker, take by weighing sodium isoascorbate 10mg again, add the dissolving of 50mL 0.1mol/LNaOH solution, transfer to then in the brown volumetric flask of 100mL, be settled to scale, use the funnel filtering solution, discard filtrate just;
(4) mensuration of sample liquid content of hemachrome: filtrate is measured light absorption value at the 400nm place, light absorption value substitution typical curve formula is obtained the concentration (C of protoheme in the sample liquid
0), then by the content of protoheme in the following formula calculation sample,
In the formula: C
0-for supplying protoheme concentration in the test agent liquid, μ g/mL;
W-is the weight for test agent, g.
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| JP6612050B2 (en) | 2014-08-28 | 2019-11-27 | シスメックス株式会社 | Hematology analyzer, diagnosis support method, and computer program |
| CN112485246B (en) * | 2020-11-19 | 2024-04-26 | 重庆瑞卡威生物科技有限公司 | Single epithelial tissue tumor cell seepage free heme chromogenic solution |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101353350A (en) * | 2008-09-09 | 2009-01-28 | 浙江大学 | Method for purifying haemachrome |
| CN101371145A (en) * | 2005-10-28 | 2009-02-18 | 惠普开发有限公司 | Systems and methods for measuring glycated hemoglobin |
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| CN101371145A (en) * | 2005-10-28 | 2009-02-18 | 惠普开发有限公司 | Systems and methods for measuring glycated hemoglobin |
| CN101353350A (en) * | 2008-09-09 | 2009-01-28 | 浙江大学 | Method for purifying haemachrome |
Non-Patent Citations (4)
| Title |
|---|
| JP特开2007-248276A 2007.09.27 |
| 周淡宜.薄层扫描法测定血红素含量.《药物生物技术》.2002,第9卷(第5期),期刊第285-287页. * |
| 徐晓峰.光谱分析法测量血液中二价铁离子的含量.《光谱学与光谱分析》.2008,第28卷(第12期),期刊第2927-2930页. * |
| 林观样等.原子吸收分光光度法测定血红素口服液中铁的含量.《海峡药学》.2006,第18卷(第6期),期刊第52-53页. * |
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Assignee: Huaibei Enbi Feed Co.,Ltd. Assignor: Anhui Baodi Meat Products Co., Ltd. Contract record no.: 2011340000151 Denomination of invention: Method for measuring content of hemachrome Granted publication date: 20110427 License type: Exclusive License Open date: 20100526 Record date: 20110720 |
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