CN103901034B - A kind of detectable bar detecting microalbumin in urine and preparation method thereof - Google Patents
A kind of detectable bar detecting microalbumin in urine and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of detectable bar detecting microalbumin in urine, including substrate and filter paper, described filter paper adheres on the substrate, described filter paper immerses successively and has A liquid and B liquid, wherein: described A liquid is to comprise: the aqueous buffer solution of sodium citrate, citric acid and concentrated hydrochloric acid；Described B liquid is to comprise: the methanol of oxolane, polypropylene glycol and sulfonephthalein dyestuff or alcoholic solution.Detectable bar prepared by this case, in acid condition, sulfonephthalein dyestuff can react rapidly with the microalbumin in urine, when microalbumin concentration is within the scope of 0～200mg/L, reagent paper presents from colourless to navy blue obvious color range, it is easy to visual resolution is with machine-readable, and measurement result is accurate, and not by the interference of other ions, stable performance in urine；Meanwhile, the preparation method of this reagent strip is simple and efficient, and cost of manufacture is cheap, is suitable for large-scale promotion application.
Description
Technical field
The present invention relates to urine detection field, particularly to a kind of detectable bar detecting microalbumin in urine and preparation method thereof.
Background technology
Microalbumin refers to the albumin occurring trace in Human Urine.Albumin is the normal protein matter in a kind of blood, but minute quantity albumin only occurs in urine in physiological conditions.Under normal circumstances, the albumin in urine is few, answers < 20mg/L specific to every liter of urinaryalbumin for body metabolism.If the microalbumin found after health check-up in urine is within the scope of 20mg/L～200mg/L, just belong to microalbuminuria, microalbuminuria reflection human kidney exceptional leakage protein.Commonly used trace albumin index monitors the generation of nephropathy.The detection of trace albumin is early discovery nephropathy diagnosis index most sensitive, the most reliable.By the numerical value of Urinary microalbumin, just can accurately diagnose the state of an illness in conjunction with the statement of incidence, symptom and medical history.
Detecting microalbumin in prior art and generally adopt immunization, such as immune-gold labeled, immunofluorescence etc., but according to the reagent strip complex production process that immunization is designed, production cost is high, it is difficult to large-scale promotion application.
Summary of the invention
For the deficiencies in the prior art part, the present invention provides a kind of detectable bar detecting microalbumin in urine and preparation method thereof, compared to prior art, this preparation method is simple and efficient, and cost of manufacture is cheap, and reagent strip is easy to use, being easy to preserve, testing result is quick and precisely.
Technical scheme is summarized as follows:
A kind of detectable bar detecting microalbumin in urine, including substrate and filter paper, described filter paper adheres on the substrate, and immersing successively in described filter paper has A liquid and B liquid, wherein:
Described A liquid is to comprise: the aqueous buffer solution of 4.0wt%～4.5wt% sodium citrate, 4.2wt%～4.8wt% citric acid and 3.0wt%～3.5wt% concentrated hydrochloric acid；
Described B liquid is to comprise: the methanol of 4.0wt%～10wt% oxolane, 0.5wt%～1.5wt% polypropylene glycol and 0.03wt%～0.15wt% sulfonephthalein dyestuff or alcoholic solution.
Preferably, the detectable bar of microalbumin in described detection urine, pH=2.0～3.5 of described A liquid.
Preferably, the detectable bar of microalbumin in described detection urine, described substrate material is Corvic, or polypropylene, or polymethyl methacrylate.
Preferably, the detectable bar of microalbumin in described detection urine, described filter paper is common quantitative filter paper or common qualitative filter paper.
Preferably, the detectable bar of microalbumin in described detection urine, described sulfonephthalein dyestuff is 5 ', 5 "-dinitro-3 ', 3 "-two iodo-3,4,5,6-tetrabromophenol sulfonphthaleins.
The preparation method of a kind of detectable bar detecting microalbumin in urine, it comprises the steps:
(1) configuration A liquid: 40～45g sodium citrate, 42～48g citric acid and 30～35mL concentrated hydrochloric acid are put into container, adds water to 1000mL constant volume；
(2) configuration B liquid: 40～100mL oxolane, 5～15mL polypropylene glycol and 0.3～1.5g sulfonephthalein dyestuff put into container, adds methanol or ethanol to 1000mL constant volume；
(3) being immersed by filter paper in A liquid, take out after fully soaking, be placed in the vacuum drying oven that temperature is 70 DEG C dry, drying time is 10～20 minutes；
(4) being again dipped in B liquid by the filter paper after drying, take out after fully soaking, be placed in the vacuum drying oven that temperature is 60 DEG C dry, drying time is 5～15 minutes；
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detectable bar detecting microalbumin in urine.
Preferably, the preparation method of the detectable bar of microalbumin in described detection urine, pH=2.0～3.5 of described A liquid.
Preferably, the preparation method of the detectable bar of microalbumin in described detection urine, described rectangle paper slip is of a size of 0.55 × 0.80cm.
The invention has the beneficial effects as follows: detectable bar prepared by this case, in acid condition, sulfonephthalein dyestuff can react rapidly with the microalbumin in urine, when microalbumin concentration is within the scope of 0～200mg/L, reagent paper presents from colourless to navy blue obvious color range, it is easy to visual resolution is with machine-readable, and measurement result is accurate, and not by the interference of other ions, stable performance in urine；Meanwhile, the preparation method of this reagent strip is simple and efficient, and cost of manufacture is cheap, is suitable for large-scale promotion application.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further detail, to make those skilled in the art can implement according to this with reference to description word.
The preparation method of a kind of detectable bar detecting microalbumin in urine, it comprises the steps:
(1) configuration A liquid: 40～45g sodium citrate, 42～48g citric acid and 30～35mL concentrated hydrochloric acid are put into container, adds water to 1000mL constant volume, obtain the aqueous buffer solution of pH=2.0～3.5.
(2) configuration B liquid: 40～100mL oxolane, 5～15mL polypropylene glycol (be preferably PPG2000) and 0.3～1.5g sulfonephthalein dyestuff are put into container, addition methanol or ethanol to 1000mL constant volume；The purity of methanol or ethanol does not limit, it will be preferred that absolute methanol or dehydrated alcohol, it is also possible to be the purity of 80% (v/v%) or lower (but as far as possible with anhydrous system).Sulfonephthalein dyestuff is preferably 5 ', and 5 "-dinitro-3 ', 3 " and-two iodo-3,4,5,6-tetrabromophenol sulfonphthaleins.
(3) being immersed by filter paper in A liquid, take out after fully soaking, be placed in the vacuum drying oven that temperature is 70 DEG C dry, drying time is 10～20 minutes.
(4) being again dipped in B liquid by the filter paper after drying, take out after fully soaking, be placed in the vacuum drying oven that temperature is 60 DEG C dry, drying time is 5～15 minutes.
(5) dried filter paper and substrate are fixed, be cut into the rectangle paper slip being of a size of 0.55 × 0.80cm, make a kind of detectable bar detecting microalbumin in urine.This substrate material can be Corvic, or polypropylene, or polymethyl methacrylate；And filter paper can be common quantitative filter paper or common qualitative filter paper.
Embodiment 1
The preparation method that the present embodiment provides a kind of detectable bar detecting microalbumin in urine, it comprises the steps:
(1) configuration A liquid: 40g sodium citrate, 42g citric acid and 30mL concentrated hydrochloric acid are put into container, adds water to 1000mL constant volume；
(2) configuration B liquid: 40mL oxolane, 5mL polypropylene glycol and 0.3g sulfonephthalein dyestuff put into container, adds methanol to 1000mL constant volume；
(3) being immersed by filter paper in A liquid, take out after fully soaking, be placed in the vacuum drying oven that temperature is 70 DEG C dry, drying time is 10 minutes；
(4) being again dipped in B liquid by the filter paper after drying, take out after fully soaking, be placed in the vacuum drying oven that temperature is 60 DEG C dry, drying time is 5 minutes；
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detectable bar detecting microalbumin in urine.
Embodiment 2
The preparation method that the present embodiment provides a kind of detectable bar detecting microalbumin in urine, it comprises the steps:
(1) configuration A liquid: 45g sodium citrate, 48g citric acid and 35mL concentrated hydrochloric acid are put into container, adds water to 1000mL constant volume；
(2) configuration B liquid: 100mL oxolane, 15mL polypropylene glycol and 1.5g sulfonephthalein dyestuff put into container, adds ethanol to 1000mL constant volume；
(3) being immersed by filter paper in A liquid, take out after fully soaking, be placed in the vacuum drying oven that temperature is 70 DEG C dry, drying time is 20 minutes；
(4) being again dipped in B liquid by the filter paper after drying, take out after fully soaking, be placed in the vacuum drying oven that temperature is 60 DEG C dry, drying time is 15 minutes；
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detectable bar detecting microalbumin in urine.
Embodiment 3
The preparation method that the present embodiment provides a kind of detectable bar detecting microalbumin in urine, it comprises the steps:
(1) configuration A liquid: 42.5g sodium citrate, 45g citric acid and 32.5mL concentrated hydrochloric acid are put into container, adds water to 1000mL constant volume；
(2) configuration B liquid: 70mL oxolane, 10mL polypropylene glycol and 0.9g sulfonephthalein dyestuff put into container, adds methanol to 1000mL constant volume；
(3) being immersed by filter paper in A liquid, take out after fully soaking, be placed in the vacuum drying oven that temperature is 70 DEG C dry, drying time is 15 minutes；
(4) being again dipped in B liquid by the filter paper after drying, take out after fully soaking, be placed in the vacuum drying oven that temperature is 60 DEG C dry, drying time is 10 minutes；
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detectable bar detecting microalbumin in urine.
Embodiment 4
The preparation method that the present embodiment provides a kind of detectable bar detecting microalbumin in urine, it comprises the steps:
(1) configuration A liquid: 41.7g sodium citrate, 45.1g citric acid and 32mL concentrated hydrochloric acid are put into container, adds water to 1000mL constant volume；
(2) configuration B liquid: by 5 ', the 5 of 50mL oxolane, 10mL polypropylene glycol and 0.58g "-dinitro-3 ', 3 " and-two iodo-3,4,5,6-tetrabromophenol sulfonphthaleins put into container, add ethanol to 1000mL constant volume；
(3) being immersed by filter paper in A liquid, take out after fully soaking, be placed in the vacuum drying oven that temperature is 70 DEG C dry, drying time is 15 minutes；
(4) being again dipped in B liquid by the filter paper after drying, take out after fully soaking, be placed in the vacuum drying oven that temperature is 60 DEG C dry, drying time is 15 minutes；
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detectable bar detecting microalbumin in urine.
Embodiment 5
The preparation method of standard color comparison card:
(1) the bovine serum albumin standard solution of 0mg/L, 20mg/L, 80mg/L, 150mg/L is configured；
(2) being dipped in above-mentioned four kind bovine serum albumin standard solution respectively by the detectable bar that above-described embodiment 1～4 prepares, take out, dry after 2 seconds, after 1 minute, colour developing is completely；
(3) the colour developing situation according to each reagent paper prepares colorimetric card, and the color range of colorimetric card is that white arrives navy blue four color ranges.
Embodiment 6
The using method of microalbumin detectable bar:
Detectable bar embodiment 1～4 prepared immerses in liquid to be measured, take out after 2 seconds, dry, after 1 minute, colour developing is completely, compare with standard color comparison card subsequently, with the concentration that the concentration corresponding to the color on the immediate standard color comparison card of color on detectable bar is liquid to be measured, if between two color ranges, then can estimate reading microalbumin concentration.
Embodiment 7
Urinary microalbumin detectable bar using method on instrument:
(1) the bovine serum albumin standard solution of 0mg/L, 20mg/L, 80mg/L, 150mg/L, 200mg/L is configured；
(2) the detectable bar that embodiment 1～4 prepares is dipped in above-mentioned five kind bovine serum albumin standard solution respectively；
(3) reflectance that Urine Analyzer measures 1 minute is used, according to 5 point-rendering standard curves；
(4) when using, the microdose urine protein detectable bar prepared according to embodiment is immersed in liquid to be measured and takes out, using Urine Analyzer to measure reflectance after 1 minute, the concentration that the reflectance recorded is corresponding with on standard curve is liquid microalbumin concentration to be measured.
Although embodiment of the present invention are disclosed as above, but listed utilization that it is not restricted in description and embodiment, it can be applied to various applicable the field of the invention completely, for those skilled in the art, it is easily achieved other amendment, therefore, under the general concept limited without departing substantially from claim and equivalency range, the present invention is not limited to specific details.
Claims (8)
1. detecting a detectable bar for microalbumin in urine, including substrate and filter paper, described filter paper adheres on the substrate, it is characterised in that described filter paper immerses A liquid successively and takes out dry and immerse B liquid and take out dry, wherein:
Described A liquid is to comprise: the aqueous buffer solution of 4.0wt%～4.5wt% sodium citrate, 4.2wt%～4.8wt% citric acid and 3.0wt%～3.5wt% concentrated hydrochloric acid；
Described B liquid is to comprise: the methanol of 4.0wt%～10wt% oxolane, 0.5wt%～1.5wt% polypropylene glycol and 0.03wt%～0.15wt% sulfonephthalein dyestuff or alcoholic solution.
2. the detectable bar of microalbumin in detection urine according to claim 1, it is characterised in that pH=2.0～3.5 of described A liquid.
3. the detectable bar of microalbumin in detection urine according to claim 1, it is characterised in that described substrate material is Corvic, or polypropylene, or polymethyl methacrylate.
4. the detectable bar of microalbumin in detection urine according to claim 1, it is characterised in that described filter paper is common quantitative filter paper or common qualitative filter paper.
5. the detectable bar of microalbumin in detection urine according to claim 1, it is characterised in that described sulfonephthalein dyestuff is 5 ', 5 "-dinitro-3 ', 3 " and-two iodo-3,4,5,6-tetrabromophenol sulfonphthaleins.
6. the preparation method detecting the detectable bar of microalbumin in urine, its feature comprises the steps:
(1) configuration A liquid: 40～45g sodium citrate, 42～48g citric acid and 30～35mL concentrated hydrochloric acid are put into container, adds water to 1000mL constant volume；
(2) configuration B liquid: 40～100mL oxolane, 5～15mL polypropylene glycol and 0.3～1.5g sulfonephthalein dyestuff put into container, adds methanol or ethanol to 1000mL constant volume；
(3) being immersed by filter paper in A liquid, take out after fully soaking, be placed in the vacuum drying oven that temperature is 70 DEG C dry, drying time is 10～20 minutes；
(4) being again dipped in B liquid by the filter paper after drying, take out after fully soaking, be placed in the vacuum drying oven that temperature is 60 DEG C dry, drying time is 5～15 minutes；
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detectable bar detecting microalbumin in urine.
7. the preparation method of the detectable bar of microalbumin in detection urine according to claim 6, it is characterised in that pH=2.0～3.5 of described A liquid.
8. the preparation method of the detectable bar of microalbumin in detection urine according to claim 6, it is characterised in that described rectangle paper slip is of a size of 0.55 × 0.80cm.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| RU2692062C1 (en) * | 2018-06-19 | 2019-06-20 | Евгений Александрович Ширшин | Method of producing a color pattern and a method of analyzing colorimetric test strips using said pattern |
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| CN105388146B (en) * | 2015-10-20 | 2018-08-17 | 北京大学人民医院 | Kit that is a kind of while detecting sodium in urine, creatinine and microalbumin |
| CN105651765A (en) * | 2015-12-28 | 2016-06-08 | 东北农业大学 | Rapid protein test paper, and production method and detection method thereof |
| CN106596913A (en) * | 2017-01-18 | 2017-04-26 | 重庆普西医疗设备有限公司 | Preparation method of quantitative analysis test paper for urine |
| CN106645676A (en) * | 2017-01-18 | 2017-05-10 | 重庆普西医疗设备有限公司 | Multi-term urine analysis test paper and preparation method thereof |
| CN108120714B (en) * | 2017-12-20 | 2020-11-27 | 吉林省汇酉生物技术股份有限公司 | Reagent for detecting microalbumin in urine and application thereof |
| CN108226147B (en) * | 2017-12-27 | 2021-06-04 | 李宏奎 | Urine microalbumin detection reagent strip and preparation method thereof |
| CN113376150A (en) * | 2021-06-10 | 2021-09-10 | 吉林基蛋生物科技有限公司 | Urine microalbumin dry chemical detection test paper and preparation method thereof |
| CN114034693A (en) * | 2021-11-04 | 2022-02-11 | 广州达安基因股份有限公司 | Combined test paper for detecting urine creatinine urine microalbumin semiquantitative, preparation method and application thereof |
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| JP4576499B2 (en) * | 2004-01-23 | 2010-11-10 | アークレイ株式会社 | Protein measurement method |
| CN1854738A (en) * | 2005-04-20 | 2006-11-01 | 北京怡成生物电子技术有限公司 | Microdose urine protein testing chip and its tester |
| CN101750413B (en) * | 2009-11-09 | 2011-05-04 | 河南工业大学 | Rapid detection method for protein content of foods |
| CN103472060B (en) * | 2013-10-10 | 2016-01-20 | 中国科学院苏州生物医学工程技术研究所 | A kind of urine calcium ion Test paper and preparation method thereof |
| CN103630532B (en) * | 2013-11-06 | 2016-04-13 | 中国科学院苏州生物医学工程技术研究所 | A kind of urine creatinine detection reagent strip and preparation method thereof |
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| RU2692062C1 (en) * | 2018-06-19 | 2019-06-20 | Евгений Александрович Ширшин | Method of producing a color pattern and a method of analyzing colorimetric test strips using said pattern |
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