CN101793777A - Reagent (kit) for measuring formaldehyde and method for measuring concentration of formaldehyde - Google Patents
Reagent (kit) for measuring formaldehyde and method for measuring concentration of formaldehyde Download PDFInfo
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- CN101793777A CN101793777A CN200910028545A CN200910028545A CN101793777A CN 101793777 A CN101793777 A CN 101793777A CN 200910028545 A CN200910028545 A CN 200910028545A CN 200910028545 A CN200910028545 A CN 200910028545A CN 101793777 A CN101793777 A CN 101793777A
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- coenzyme
- stabilizing agent
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Abstract
The invention relates to a reagent (kit) for measuring formaldehyde by using an enzyme colorimetric method as well as a method for measuring the concentration of the formaldehyde, and composition and ingredients of the reagent, belonging to the technical field of food/environmental test. The reagent (kit) comprises the main ingredients of buffer solution, coenzyme, formaldehyde dehydrogenase and stabilizing agent. The concentration of the formaldehyde is measured by mixing a sample and the reagent according a certain volume ratio to carry out a series of enzymatic reaction, placing the reactant under an ultraviolet/visible light analyzer and detecting the ascending degree of absorbance at a dominant wavelength of 340nm.
Description
Technical field
The present invention relates to a kind of formaldehyde determination reagent (box), the invention still further relates to the method for measuring concentration of formaldehyde simultaneously, belong to food/environmental test determination techniques field.
Background technology
The assay method of formaldehyde determination mainly contains spectrophotometric method, chromatography, electrochemical process, chemical titration etc.:
The main method of spectrophotometry has acetylacetone method, chromotropic acid method, MBTH method, pair pinkish red method, AHMT method etc. several.The advantage of spectrophotometric method is easy and simple to handle; Shortcoming is that sensitivity is lower, and pollution medium is arranged, and mensuration is had interference.
Chromatography mainly contains gas chromatography, high performance liquid chromatography, the chromatography of ions etc., directly utilizes chromatography less, generally is and the logotype mutually of other analytical instrument, as GC-MS, HPLC-UV etc.The chromatography method is simple, fast, directly, but shortcomings such as complex operation, reagent consumption are big, method poor selectivity.
Electrochemical process comprises oscilloscopic polarography, adsorptive voltammetry and Ge La derivative reagent method, in this method air the sensitivity of formaldehyde and accuracy all very high, be suitable for measuring trace formaldehyde in the room air.
Fluorometry is quick because of having, easy, highly sensitive, high repeatability and other advantages is subjected to people and payes attention to widely, and in particular for the analysis monitoring of trace formaldehyde in the indoor air environment, its advantage is more outstanding.Advantages such as chemiluminometry has highly sensitive, and the range of linearity is wide, and instrument and equipment is simple.
Chemical titration mainly comprises potentiometric titration, iodimetric titration and acid base titration.
Comprehensively above-mentioned, the neither one method can use self-reacting device to detect formaldehyde, and enzyme process detects the reagent of formaldehyde, the advantage of enzyme process be the specificity height interference-free, easy to operate, with low cost, can mass detection, the method sensitivity.
Summary of the invention
The technical problem to be solved in the present invention is: propose a kind of enzymic colorimetric (EnzymaticColorimetric Method) technology of utilizing, metering reduced form nicotinamide coenzyme (reduced coenzyme) is in the variation of 340nm wavelength place absorbance, measured the method for concentration of formaldehyde, simultaneously, the present invention also will provide in order to realize the formaldehyde determination reagent (box) of this method, adopt this reagent not only can be ultraviolet analyser or half, carrying out concentration of formaldehyde on the automatic clinical chemistry analyzer measures, and finding speed is fast, the accuracy height, thereby can obtain practical applying.
Concentration of formaldehyde assay method of the present invention is as follows:
Formaldehyde+coenzyme+water
Formaldehyde dehydrogenaseFormic acid+reduced coenzyme
This method is used formaldehyde dehydrogenase (formaldehyde dehydrogenase; EC 1.2.1.46) enzymatic reaction colorimetric end-point method.Formaldehyde dehydrogenase enzymolysis formaldehyde, coenzyme (not having absorption peak at the 340nm place) reduces the most at last becomes reduced coenzyme (absorption peak being arranged at the 340nm place), thereby measured the degree that reduced coenzyme rises in 340nm place absorbance, by measuring the degree that 340nm place absorbance rises, can calculate the concentration of formaldehyde.
Experiment shows, takes all factors into consideration from the accuracy of measurement result and economy two aspects of preparation cost, no matter is single agent, two agent or three doses, and the formaldehyde determination reagent of the present invention (box) of following composition relation is comparatively desirable:
Damping fluid 100mmol/L
Stabilizing agent 500mmol/L
Coenzyme 3mmol/L
Formaldehyde dehydrogenase 10000U/L
Formaldehyde determination reagent of the present invention (box) can be single agent, comprising:
Damping fluid, stabilizing agent, coenzyme, formaldehyde dehydrogenase.
Reagent (box) can be dry powder, and use the back that is dissolved in water before use; Also can be mixed with liquid reagent, directly use.
Also above-mentioned single agent reagent can be made into following pair of agent reagent:
Reagent 1
Damping fluid, stabilizing agent, coenzyme.
Reagent 2
Damping fluid, stabilizing agent, formaldehyde dehydrogenase.
Coenzyme, the position of formaldehyde dehydrogenase in reagent 1 or reagent 2 can not limit.Reagent (box) can be dry powder, and use the back that is dissolved in water before use; Also can be mixed with liquid reagent, directly use.
No matter be single agent, two agent or three doses, the present invention measures the method for concentration of formaldehyde, and its coenzyme can be NADP
+, NAD
+Or thio-NAD
+In a kind of.
Embodiment
The present invention is further illustrated below in conjunction with examples of implementation.
Embodiment one
The formaldehyde determination reagent of present embodiment is single reagent, comprising:
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 500mmol/L
Coenzyme 3mmol/L
Formaldehyde dehydrogenase 10000U/L
Reagent divides the bottle of packing into after all dissolving and preparing, and carries out freeze drying, makes powdered reagent; Before the use, add pure water, use after redissolving.
On automatic clinical chemistry analyzer, set: 37 ℃ of temperature, 10 minutes reaction time, initial absorbance≤0.1, test predominant wavelength 340nm, test commplementary wave length 405nm, the volume ratio of tested formaldehyde sample and reagent is 1/25, the Direction of Reaction is positive reaction (reaction of rising), about about 0 minute of time delay is about 5 minutes detection times.
After adding sample and reagent, make them mixed and have reaction, reactant places under the Biochemical Analyzer the most at last, detects the degree that predominant wavelength 340nm absorbance rises, thereby calculates the concentration of formaldehyde.
Embodiment two
The formaldehyde determination reagent of present embodiment is double reagent, comprising:
Reagent 1
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 50mmol/L
Coenzyme 3mmol/L
Reagent 2
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 50mmol/L
Formaldehyde dehydrogenase 10000U/L
Reagent divides the bottle of packing into after all dissolving and preparing, and makes liquid double reagent, can directly use.
On automatic clinical chemistry analyzer, set: 37 ℃ of temperature, 10 minutes reaction time, initial absorbance≤0.1, test predominant wavelength 340nm, test commplementary wave length 405nm, the volume ratio of tested formaldehyde sample and reagent 1, reagent 2 is 2/20/5, the Direction of Reaction is positive reaction (reaction of rising), about about 0 minute of time delay is about 5 minutes detection times.
After adding sample and reagent, make them mixed and have reaction, reactant places under the Biochemical Analyzer the most at last, detects the degree that predominant wavelength 340nm absorbance rises, thereby calculates the concentration of formaldehyde.
The applicant adopts other assay methods of putting down in writing in the above summary of the invention all can reach purpose of the present invention through experimental verification, in view of situation such as determination step and above embodiment roughly the same, do not separately enumerate.
In a word, experimental results show that: adopt assay method of the present invention can draw required measurement result by general biochemical analyzer fully---the blank reagent absorbance changes (Δ A/min)≤0.0005; Absorbance time response curve should be the rising curve until terminal point; Reagent can be surveyed effectively, and (R 〉=0.99) linear range can reach 30mmol/L; The inaccuracy of reagent test, its relative deviation be no more than ± and 3%; The coefficient of variation (CV)≤2% of the precision of reagent test (repeatability); The sensitivity of reagent can reach 0.12 ± 0.06 Δ A/mmol/L; Reagent is preserved down at 2-8 ℃, and activity can be stablized 1 year;---the present invention is highly sensitive, degree of accuracy good, the linear range broadness, and stationary phase is long, is enough to easy to utilize.
Claims (5)
1. the method for measurement of concentration of the formaldehyde of an enzymic colorimetric, its method is as follows:
Formaldehyde+coenzyme+water
Formaldehyde dehydrogenaseFormic acid+reduced coenzyme
The end reaction thing is placed under ultraviolet analyser or half, the automatic clinical chemistry analyzer, detect the degree that predominant wavelength 340nm absorbance rises, calculate the concentration measurement result of formaldehyde.
2. a formaldehyde determination reagent (box), principal ingredient comprises:
Damping fluid 20---500mmol/L
Stabilizing agent 1---4000mmol/L
DPN diphosphopyridine nucleotide---6mmol/L
Formaldehyde dehydrogenase 1000---80000U/L
The concentration of reagent composition not necessarily is only limited to above-mentioned scope; Effect is better in this scope, and outside this scope, reagent still can reagentia.It is characterized in that: reagent (box) can be dry powder, and use the back that is dissolved in water before use; Also can be mixed with liquid reagent, directly use.
3. according to the described formaldehyde determination reagent of claim 2 (box), it is characterized in that:
Form single agent reagent by damping fluid, stabilizing agent, coenzyme, formaldehyde dehydrogenase.
4. according to the described formaldehyde determination reagent of claim 2 (box), it is characterized in that:
Form two agent reagent by damping fluid, stabilizing agent, coenzyme, formaldehyde dehydrogenase; Reagent 1 is made up of damping fluid, stabilizing agent, coenzyme; Reagent 2 is made up of damping fluid, stabilizing agent, formaldehyde dehydrogenase.Coenzyme, the position of formaldehyde dehydrogenase in reagent 1 or reagent 2 can not limit.
5. according to the described formaldehyde determination reagent of claim 2 (box), it is characterized in that: also comprise stabilizing agent 1-4000mmol/L or 0.1%-100% volume ratio.Described stabilizing agent is: ammonium sulfate (AmmoniaSulfate), glycerine (Glycerol), propylene glycol (Propylene Glycol), ethylene glycol (Ethylene glycol) and at least one of the preservatives.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910028545A CN101793777A (en) | 2009-02-04 | 2009-02-04 | Reagent (kit) for measuring formaldehyde and method for measuring concentration of formaldehyde |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910028545A CN101793777A (en) | 2009-02-04 | 2009-02-04 | Reagent (kit) for measuring formaldehyde and method for measuring concentration of formaldehyde |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101793777A true CN101793777A (en) | 2010-08-04 |
Family
ID=42586588
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910028545A Pending CN101793777A (en) | 2009-02-04 | 2009-02-04 | Reagent (kit) for measuring formaldehyde and method for measuring concentration of formaldehyde |
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|---|---|
| CN (1) | CN101793777A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107389670A (en) * | 2017-06-28 | 2017-11-24 | 防城港市质量技术监督局 | A kind of kit for determining formaldehyde in air |
| WO2019242430A1 (en) * | 2018-06-22 | 2019-12-26 | 科之杰新材料集团有限公司 | Method of determining formaldehyde content in concrete additive |
-
2009
- 2009-02-04 CN CN200910028545A patent/CN101793777A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107389670A (en) * | 2017-06-28 | 2017-11-24 | 防城港市质量技术监督局 | A kind of kit for determining formaldehyde in air |
| WO2019242430A1 (en) * | 2018-06-22 | 2019-12-26 | 科之杰新材料集团有限公司 | Method of determining formaldehyde content in concrete additive |
| CN110632242A (en) * | 2018-06-22 | 2019-12-31 | 科之杰新材料集团有限公司 | Method for testing formaldehyde content in concrete admixture |
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| PB01 | Publication | ||
| DD01 | Delivery of document by public notice |
Addressee: Aijie Biological Science & Technology Co., Ltd., Suzhou City Wang Erzhong Document name: Notification that Application Deemed to be Withdrawn |
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| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20100804 |