CN109507411B - Method and portable device for rapidly detecting nitrite in saliva - Google Patents
Method and portable device for rapidly detecting nitrite in saliva Download PDFInfo
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- 210000003296 saliva Anatomy 0.000 title claims abstract description 78
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
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- Life Sciences & Earth Sciences (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
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Abstract
本发明公开了一种快速检测唾液中亚硝酸盐的方法和便携装置，属于医学检验技术领域。该检测装置包括唾液采集部分、反应槽部分和读数部分，通过唾液采集部分收集的唾液与试剂槽内的试剂在反应槽内反应，得到的反应液进入读数部分的斜面上，通过读数窗口观测到的液面与刻度来进行读数，判断健康状况。本发明的检测方法具有安全无创，操作简便易行，检测装置采用一体化封闭设计，既避免反应试剂的外泄，也确保反应检测的准确性和结果的可靠性，并在5min内即可快速完成样本采集，检测和健康判断。
The invention discloses a method and a portable device for rapidly detecting nitrite in saliva, belonging to the technical field of medical examination. The detection device includes a saliva collection part, a reaction tank part and a reading part. The saliva collected by the saliva collection part reacts with the reagent in the reagent tank in the reaction tank, and the obtained reaction liquid enters the slope of the reading part, and the reading window is observed. The liquid level and scale can be read to judge the health status. The detection method of the invention is safe, non-invasive, simple and easy to operate, and the detection device adopts an integrated closed design, which not only avoids the leakage of the reaction reagent, but also ensures the accuracy of the reaction detection and the reliability of the result, and can be quickly completed within 5 minutes. Complete sample collection, testing and health judgment.
Description
Technical Field
The invention belongs to the field of medical examination, and particularly relates to a method and a portable device for rapidly detecting nitrite in saliva.
Background
Nitrite, a generic term for a class of inorganic compounds that are widely found in the human environment, is one of the most prevalent nitrogen-containing compounds in nature. If a person takes excessive nitrite, the nitrite oxidizes ferrous hemoglobin in blood to generate methemoglobin in vivo, so that the ferrohemoglobin loses the functions of carrying oxygen and transporting oxygen, thereby causing hypoxia of tissues. The sources of nitrite in the human body are of two types, food intake and self-production. Nitrite is used as an excellent bacteriostatic agent and preservative, and is widely used in the processing of meat products. Nitric oxide is a simple free radical gas molecule and is also a very important signal transduction molecule of human body, and the physiological, pathological and medicinal values of nitric oxide are widely accepted. Under the condition of normal oxygen, L-arginine is catalyzed and oxidized by nitric oxide synthetase to generate L-citrulline and nitric oxide at the same time. Nitrite, nitrate are the major and stable metabolites of nitric oxide. In addition, the oral symbiotic nitrate reductase also converts ingested nitrate to nitrite. Nitrite can react with amine substances in an external or in-vivo acidic environment to finally form a strong carcinogenic substance, namely nitrosamine, so that canceration of a digestive system is induced in vivo. After a large amount of nitrite enters a human body, hypoxia, enterogenous cyanosis, teratogenesis and the like are caused to harm human health. The nitrite content in humans is a marker of inflammation and possibly of the occurrence of tumors or cancer, and a high nitrate content in saliva means the occurrence of disease or inflammation and unhealthy diet. The determination of the nitrite content in saliva can be an important indicator of the body's function.
At present, the experimental methods for detecting nitrite reported at home and abroad include ion chromatography, spectrophotometry, fluorescence photometry, high performance liquid chromatography and the like. The determination of nitrite content in the national food safety standard GB 5009.33-2016 also adopts ion chromatography or spectrophotometry, and nitrite in blood or urine is usually detected by Griess kit or high performance liquid chromatography. For example, the Chinese patent application (application publication No. CN103760121A, application publication date 2014.04.30) discloses a method for detecting nitrite in blood, which uses a porous plate and an enzyme-labeling instrument to measure the absorbance of a colored complex formed by nitrite and Griess, and uses blood without nitrite as a base solution for preparing a working curve, thereby eliminating the interference of a blood base and improving the sensitivity and accuracy of detection. The method has the characteristics of high accuracy, large detection sample quantity, small detection sample quantity, high detection efficiency and the like.
For example, the Chinese patent application (application publication No. CN 103235093A, application publication No. 2013.08.07) discloses a method for detecting nitrate and nitrite in blood, urine and tissue, wherein nitrate ions and an ion pair reagent form an ion pair under a weak acid condition, and the nitrite and a Griess reagent undergo a diazotization coupling reaction to form an azo compound, so that the nitrate and nitrite which cannot be retained on a chromatographic column and have no ultraviolet absorption can be subjected to liquid chromatography analysis at different wavelengths simultaneously, and mutual interference is eliminated. The method has great applicability to the detection of nitrate and nitrite in blood, urine and tissues with great matrix interference and low content, and the detection limits of the nitrate and the nitrite are 0.2 mu g/mL and 0.05 mu g/mL respectively. The method has the characteristics of simple operation, quick, effective and accurate measurement and the like.
For another example, the Chinese patent application (application publication No. CN 104280383A, application publication No. 2015.01.14) discloses a method for rapidly detecting nitrite in blood and urine, which comprises the steps of carrying out diazotization coupling reaction on nitrite and Griess reagent to form azo compound, carrying out cloud point extraction, depositing colored azo compound at the bottom of a centrifugal tube, comparing the red color of the tube bottom with a standard color scale, and judging the content according to the color depth. However, because the enrichment times of cloud point extraction are large, and the blood and urine are decolorized and deproteinized, the interference is greatly eliminated, and the detection limit of nitrite in the system can reach 0.01 mu g/mL. The invention has the characteristics of simplicity, rapidness, high sensitivity, field detection and the like.
For another example, the chinese invention patent application (application publication No. CN 104458712A, application publication date: 2015.03.25) discloses a high-sensitivity rapid detection method for nitrite in blood and urine, which utilizes the diazo coupling reaction between nitrite and Griess reagent to form azo compound, uses nonionic surfactant including fatty alcohol polyoxyethylene ether AEO series to perform cloud point extraction, and has the characteristics of mild operation conditions, low cloud point temperature, short phase separation time, high extraction efficiency, and the like. The detection limit of nitrite in the system can reach 0.005 mu g/mL. The invention has the characteristics of simplicity, rapidness, high sensitivity, field detection and the like.
Although the detection methods reported in the patent documents have uniqueness, the detection processes all need professional personnel and professional instruments, and the operation process is complicated, long in time consumption, high in cost and limited to a certain extent.
Generally, the method can be applied to the field and can give out a detection result within 30min, namely, the method can be regarded as a field rapid detection method. The existing method for rapidly detecting nitrite mainly determines the content of nitrite according to the color reaction of the nitrite and a color developing agent, generally adopts the forms of test paper, a kit and the like, but lacks a portable rapid and accurate detection device. Although the test paper can achieve the purpose of rapidness and portability, the test paper can only detect the content of the nitrite qualitatively and semi-quantitatively, and a portable method and a portable device which can conveniently detect and accurately quantify are clinically and particularly needed to be applied to clinical and individual self-detection. In recent years, the relationship between nitrate and nitrite and inflammation and disease in the human body has been receiving increased attention and has been reported. Saliva, a body fluid that is abundant and easy to collect, is most suitable as a first choice for detecting diseases and inflammations. With the increasing severity of health and safety problems, how to realize the rapid, sensitive, simple, convenient and accurate detection of the content of nitrite in saliva by using a portable device has a very important significance for the healthy life of people.
Disclosure of Invention
The invention aims to provide a method and a portable device for rapidly detecting nitrite in saliva, which are simple and convenient to operate, reliable in detection, and capable of realizing integration of saliva collection, sealing detection and early warning display.
In order to achieve the above objects of the invention, the inventor finally obtains the following method and portable device for rapidly detecting nitrite in saliva through a great deal of experimental research and design, wherein the method for rapidly detecting nitrite in saliva comprises the following steps:
saliva collection part: the saliva collection, the bubble removal and the liquid drainage are realized through a saliva collector; the reaction tank part: saliva collected by the saliva collecting part, a Griess A reagent and a Griess B reagent in the reagent tank enter a reaction tank C to react to form reaction liquid; reading part: reaction liquid enters the reading inclined plane for reading, the reading device 4 comprises a cylindrical cavity 16, one end of the cylindrical cavity 16 is provided with a reading window 17, the other end of the cylindrical cavity is provided with a transparent inclined plane cover 13, and the inner side of the transparent inclined plane cover 13 is provided with a reading display inclined plane 12.
Preferably, Griess reagent a: sulfanilic acid + 10% acetic acid
Griess B reagent: naphthylamine + 10% acetic acid
The portable device for rapidly detecting nitrite in saliva comprises a device body, a saliva collecting part, a reaction groove part and a reading part; the saliva collecting part comprises a saliva collector 1 and a collector 5, and a hydrophilic material 6 is arranged at the lower part of the saliva collector 1; the reaction groove part comprises a buckle cover 3 and a reaction groove plate 2, a reagent groove A8, a reagent groove B9 and a reaction groove C7 are arranged on the reaction groove plate 2, the reagent groove A8 and the reagent groove B9 can be respectively communicated with a reaction groove C7 through a channel 18 and a channel 19, a one-way valve 21 is arranged on the channel 18, a one-way valve 22 is arranged on the channel 19, and a bulge 11 corresponding to the position of the reaction groove C7 is arranged on the buckle cover 3; the reading display part comprises a liquid inlet channel 20 which can be communicated with the reaction tank C7 and a reading device 4, a one-way valve 23 is arranged on the channel 20, the reading device 4 comprises a cylindrical cavity 16, a reading window 17 is arranged at one end of the cylindrical cavity 16, a transparent inclined surface cover 13 is arranged at the other end of the cylindrical cavity, a reading display inclined surface 12 is arranged on the inner side of the transparent inclined surface cover 13, and the reaction liquid enters the reading device 4 for reading.
The collector 5 is a funnel-shaped collector.
The hydrophilic material 6 is konjac glucomannan or dimethyl sulfoxide or polytetrafluoroethylene or nitric acid-acetic acid mixed cellulose ester, the hydrophilic material 6 is distributed at the bottom of the funnel-shaped saliva collector, and the bottom of the collector 5 is communicated with the reaction tank C7.
A sealing valve 10 is arranged at the connecting part of the collector 5 and the reaction tank C7.
The cylindrical cavity 16, the reading display inclined plane 12 and the reading window 17 form a closed space, and the transparent inclined plane cover 13, the reading display inclined plane 12 and the reading window 17 are made of transparent plastics.
The reading shows that a thin layer space 15 of 0.2mm is arranged between the inclined plane 12 and the transparent inclined plane cover 13, and the top end of the transparent inclined plane cover 13 is provided with an air hole 14 with the aperture of 10 mu m.
The reading shows that the inclined plane 12 and the cylindrical cavity 16 axially form an included angle of 45 degrees.
The reading display inclined plane 12 is provided with three scales inside the contact part of the cylindrical cavity 16.
In conclusion, the invention provides a method for rapidly detecting nitrite in saliva and a portable device, which comprises a saliva collecting device, a reaction groove plate block and a data reading device, wherein the method adopts the color development reaction of Griess reagent and nitrite, uses refraction to replace the reading of a spectrophotometer, completes the accurate determination of the nitrite in the saliva in the integrated portable device, and provides early warning instructions. Compared with the prior art, the invention has the following advantages:
safe and non-invasive: the saliva content is rich, the collection is convenient, the method is completely different from the traumatic property of blood collection, the pain and fear of the detected object to acupuncture are avoided, and the exposure probability and risk of the detection personnel can be reduced;
the method is simple and easy to implement: the detection can be finished only by simply pulling, shaking, buckling and reading, and can be finished by the public without the participation of professional detection personnel;
the design is sealed to the integral type: the harm of the leakage of the reaction reagent to human bodies or the environment is avoided in the whole detection process, and the certainty of the volumes of the reaction reagent, the sample and the detection liquid is also ensured;
the method is rapid and accurate: the detection can be finished within 5 minutes, the reading scale is accurate and corresponds to the content of the nitrite, and the organism condition is reflected in time.
Drawings
FIG. 1 is a schematic structural diagram of a detecting device according to the present invention;
FIG. 2 is a schematic view of the internal structure of FIG. 1;
FIG. 3 is a schematic view of a portion of the structure of FIG. 2;
FIG. 4 is a schematic structural view of the saliva collecting device in FIG. 1;
FIG. 5 is a schematic view of the reading device of FIG. 1;
FIG. 6 is a schematic view showing the structure of the slope scale distribution in reading.
Wherein, the reference numbers of each part in fig. 1 to 6 are as follows:
the device comprises a saliva collector 1, a reaction tank plate block 2, a buckle cover 3, a reading device 4, a collector 5, a hydrophilic material 6, a reaction tank C7, a reagent tank A8, a reagent tank B9, a sealing valve 10 at the joint of the saliva collector and the reaction tank, a bulge 11, a reading display inclined plane 12, a transparent inclined plane cover 13, an air hole 14, a thin-layer space 15 between the inclined plane and the inclined plane cover, a cylindrical cavity 16, a reading window 17, a channel 18 for connecting the reagent tank A and the reaction tank C, a channel 19 for connecting the reagent tank B and the reaction tank C, a channel 20 for connecting the reaction tank C and the reading device, a one-way valve 21 in the channel 18, a one-way valve 22 in the channel 19 and a one-way valve 23 in the channel 20.
Detailed Description
The core of the invention is to provide a portable device for rapidly detecting nitrite in saliva so as to improve the detection speed, the detection convenience and the on-site detection accuracy.
The present invention will be described in further detail with reference to the drawings and the detailed description, but the present invention is not limited thereto.
As shown in fig. 1-6, a method for rapidly detecting nitrite in saliva comprises the following steps:
saliva collection part: the saliva collector 1 is used for realizing the saliva enrichment, the bubble removal and the liquid drainage; the reaction tank part: saliva collected by the saliva collecting part, a Griess A reagent and a Griess B reagent in the reagent tank enter a reaction tank C to react to form reaction liquid; a reading display section: reaction liquid enters a reading device 4 for reading, the reading device 4 comprises a cylindrical cavity 16, one end of the cylindrical cavity 16 is provided with a reading window 17, the other end of the cylindrical cavity is provided with a transparent inclined surface cover 13, and the inner side of the transparent inclined surface cover 13 is provided with a reading display inclined surface 12.
A portable device for rapidly detecting nitrite in saliva comprises a device body, a saliva collecting part, a reaction groove part and a reading display part; the saliva collecting part comprises a saliva collector 1 and a funnel-shaped collector 5, wherein the lower part of the saliva collector 1 is provided with a konjak glucomannan or dimethyl sulfoxide or polytetrafluoroethylene or nitric acid-acetic acid mixed cellulose ester hydrophilic material 6, and the hydrophilic material 6 is distributed at the bottom of the funnel-shaped saliva collector 5 and is communicated with the reaction tank C7. A sealing valve 10 is arranged at the connecting part of the collector 5 and the reaction tank C7; the reaction groove part comprises a buckle cover 3 and a reaction groove plate 2, a reagent groove A8, a reagent groove B9 and a reaction groove C7 are arranged on the reaction groove plate 2, the reagent groove A8 and the reagent groove B9 can be respectively communicated with a reaction groove C7 through a channel 18 and a channel 19, a one-way valve 21 is arranged on the channel 18, a one-way valve 22 is arranged on the channel 19, and a bulge 11 corresponding to the position of the reaction groove C7 is arranged on the buckle cover 3; the reading display part comprises a liquid inlet channel 20 which can be communicated with the reaction tank C7 and a reading device 4, a one-way valve 23 is arranged on the channel 20, the reading device 4 comprises a cylindrical cavity 16, a reading window 17 is arranged at one end of the cylindrical cavity 16, a transparent inclined surface cover 13 is arranged at the other end of the cylindrical cavity, a reading display inclined surface 12 is arranged on the inner side of the transparent inclined surface cover 13, the cylindrical cavity 16, the reading display inclined surface 12 and the reading window 17 form a closed space, and the transparent inclined surface cover 13, the reading display inclined surface 12 and the reading window 17 are made of transparent plastics. The reaction solution enters a reading device 4 for reading.
A0.2 mm thin layer space 15 is formed between the reading display inclined plane 12 and the transparent inclined plane cover 13, three scales are arranged inside the reading display inclined plane 12 and the cylindrical cavity 16 in a contact mode, and the scales are marked by Roman numbers from bottom to top.
As shown in fig. 4, the saliva collecting device 1 provided by the embodiment of the present invention is composed of a funnel-shaped collector 5 and a hydrophilic material 6, wherein the hydrophilic material 6 is uniformly and completely distributed on the bottom of the collector 5.
As shown in fig. 3, the reaction chamber plate 2 according to the embodiment of the present invention is schematically constructed in an initial operation state, and includes a reagent chamber A8, a reagent chamber B9, and a reaction chamber C7. Wherein the upper surface of the reaction tank C7 is provided with a one-way sealing valve 10 which is a joint with the saliva collector 1. The surface of the buckle cover 3 is provided with a bulge 11 made of silica gel, and the bulge is positioned right above the sealing valve 10. The reaction tank C7 is connected to and not communicated with the reagent tank A8, the reagent tank B9 and the reading device 4 via the channel 18, the channel 19 and the channel 20. The passages 18, 19 and 20 are provided with check valves 21, 22 and 23, respectively.
As shown in fig. 5, the reading device 4 according to the embodiment of the present invention has a schematic structural diagram, and the reading display slope 12 and the transparent slope cover 13 are provided, and the top end of the transparent slope cover 13 is provided with an air hole 14 with an aperture of 10 μm. The readings show that the inclined plane 12 and the cylindrical cavity 16 form an included angle of 45 degrees in the axial direction. The readings show a thin layer space 15 of 0.2mm between the ramp 12 and the transparent ramp cover 13.
As shown in fig. 6, the distribution structure diagram of the internal scale lines of the reading display inclined plane 12 provided by the embodiment of the present invention is that the reading display inclined plane 12 is connected to the cylindrical cavity 16, three scales are provided inside, and the scale lines from bottom to top are marked with roman numerals "i, ii, iii", which respectively mean "normal", "early warning" and "inflammation";
the refraction principle of the present invention is that light rays enter the reaction liquid in the reading display inclined plane 12 to generate refraction, and the sine ratio of the incident angle is constant, and the ratio is the refractive index. The content of soluble chromogenic substance in the reaction solution is in direct proportion to the refractive index in the common environment, and the refractive index of the reaction solution is measured, so that the concentration of nitrite is calculated.
Meanwhile, a green background material can be arranged in the reading display inclined plane 12, the color of the reaction liquid can be highlighted when the reaction liquid enters the reading display inclined plane 12, and the refractive index formed by the content of the soluble chromogenic substance in the reaction liquid and the green background material form color superposition, so that the scale interface is clearer and is easy to distinguish.
As shown in FIG. 1, the device according to the present invention is schematically constructed before use, wherein the saliva collector 1 is connected to the reaction plate 2, the reaction plate 2 and the reading device 4 are partially connected to each other through the transparent inclined cover 13 and the cylindrical cavity 16, and wherein the reaction well C7 is connected to and not connected to the reagent well A8, the reagent well B9 and the reading device 4.
As shown in FIG. 1, the embodiment of the present invention provides an overall schematic view of the apparatus after use, the reaction chamber plate 2 is covered with the cover 3, the reaction chamber plate 2 is connected with the reading device 4, wherein the reaction chamber C7 is communicated with the reagent chamber A8, the reagent chamber B9 and the reading device 4.
After saliva collection is finished, the saliva with determined volume immediately enters a reaction tank C7, after the saliva collector 1 is pulled out, the connecting part of the funnel-shaped collector 5 and the reaction tank C7 is immediately sealed, and the connecting part is provided with a sealing valve 10; before the reaction occurs, the inside of the reaction tank C7 is relatively vacuum; the reaction tank plate part comprises: reagent tank a: 1mL of Griess A reagent, and reagent tank B: griess B reagent 1 mL.
Pulling out the saliva collector 1, immediately initiating the communication of the grooves A8, B9 and C7, enabling the Griess A reagent and the Griess B reagent in the reagent groove A, B to enter the reaction groove C, and reacting with saliva in the reaction groove C;
after the saliva collector 1 was pulled out, the device was shaken for 15s and allowed to stand for 3 min. Covering a buckle cover 3, wherein the buckle cover 3 is a cover plate with buckles at the edge, in the covering process, the pressing and buckling of a bayonet are caused, so that the reaction tank C7 is communicated with the reading device 4, the buckle cover 3 is provided with a bulge 11 above the reaction tank C7 correspondingly, and the reaction liquid with determined volume can enter the reading inclined plane thin layer space 15 by covering the buckle cover 3; after the reaction liquid enters a reading inclined plane thin layer space 15, a reading display inclined plane 12 needs to be aligned to a light source, and then the position of a scale mark is observed through a reading window 17, so that the nitrite content in a detected person is known to be at the level of normal, early warning and inflammation;
the invention is simple and easy to operate, can finish detection only by simple pulling, shaking, cover buckling and reading, does not need the participation of professional detection personnel, and can be finished by the public; through the integrated closed design, the harm of the leakage of the reaction reagent to human bodies or environment is avoided in the whole detection process, and the certainty of the volumes of the reaction reagent, the sample and the detection liquid is also ensured; the invention has the advantages of rapid and accurate detection, completion of detection within 5 minutes, accurate reading scale corresponding to the content of nitrite, and timely response of organism conditions.
In conclusion, the invention provides a portable device for rapidly detecting nitrite in saliva, which comprises a saliva collecting device, a reaction groove plate block and a data reading device, and the portable device is capable of integrally, portably, rapidly and accurately detecting the content of nitrite in saliva, indirectly reflecting the inflammation condition of a human body, and realizing primary inflammation screening and convenient disease detection of the public.
Claims (7)
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| CN114504509A (en) * | 2020-11-17 | 2022-05-17 | 戴梓钊 | Portable psychological stress detector |
| CN115944323B (en) * | 2022-12-07 | 2023-09-29 | 杭州莱和生物技术有限公司 | Sampling and collecting device for saliva drug detection and collecting method thereof |
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