CN106199018A - A kind of monofilm biochemistry dry tablet for detecting blood fat and preparation method - Google Patents
A kind of monofilm biochemistry dry tablet for detecting blood fat and preparation method Download PDFInfo
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- CN106199018A CN106199018A CN201610530127.3A CN201610530127A CN106199018A CN 106199018 A CN106199018 A CN 106199018A CN 201610530127 A CN201610530127 A CN 201610530127A CN 106199018 A CN106199018 A CN 106199018A
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- dry
- filter paper
- dry tablet
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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
- G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
Abstract
A kind of monofilm biochemistry dry tablet for detecting blood fat and preparation method, belong to technical field of medical instruments, it is characterized in that: fix cholesterin detection reagent A in monolayer chromatography filter paper simultaneously and this chromatography filter paper of detectable B attaches on the cover slip.Preparation method is: it is circular that chromatography filter paper is cut into a diameter of 6 mm by (1), standby；(2) spraying of the chromatography filter paper on microscope slide 9 11 μ L cholesterin detection reagent B, cryotherapy, and lyophilizing in drying machine, repeat 35 times；Then spraying cholesterin detection reagent A to filter paper, cryotherapy, and lyophilizing in drying machine, this step repeats 35 times；(3) will transfer to coverslip, be cholesterol detection monofilm biochemistry dry tablet；(4) upper machine testing；(5) examination criteria curve and the standard color range of dry tablet are prepared.Providing the benefit that: the range of linearity is wider, repeatability, having good stability, detection limit is relatively low, and detection speed is fast, and simple to operate, patient can detect voluntarily.
Description
Technical field
The invention belongs to technical field of medical instruments, relate to the improvement of a kind of clinical vitro detection reagent.
Background technology
In blood plasma, contained lipid is referred to as blood fat, mainly include cholesterol (or T-CHOL) (TC), triglyceride (TG),
High density lipoprotein (HDL) and low density lipoprotein, LDL (LDL).Though plasma lipid content only accounts for the minimum of whole body lipid total amount
Point, but exogenous and endogenous lipid material all needs to run between each tissue through entering blood.Therefore, lipids contents can reflect
The situation of internal lipid metabolism.
The rising of epidemiology and clinical research confirmation, plasma cholesterol and triglyceride levels is with atherosclerotic
Occurring relevant, atherosclerosis, incidence rate and the serum High Density Lipoprotein Cholesterol level of coronary heart disease are negative correlation, and with
Low-density lipoprotein cholesterol level is proportionate.Dysbolism of blood fat is to cause the multiple disease such as atherosclerosis and fatty liver
Sick important risk factor.Recently as the raising of people's living standard, metabolism disorder of blood lipid becomes increasingly to increase, becomes serious
Affect the key factor of health of people level.
Analysis of blood lipid is for clinical hyperlipemia and the diagnosis of dyslipoproteinemia and atherosclerosis, cardiovascular diseases
The assessment of risks of (such as coronary heart disease) and preventing and treating significant, and infiltration applications in other many clinically relevant specialty disease
Sick research.Therefore Analysis of blood lipid and clinical practice thereof are the most increasingly paid attention to by clinical each subject.
Dry chemistry is also called solid state chemistry, is that in measuring one, required reagent in whole or in part pre-fixes in dry tablet load
In body, theoretical according to Kubelka-Munk, its reflectance and solid-phase layer thickness, the absorption coefficient of light of unit thickness and solid state reaction
Layer scattering coefficient relevant, when the scattering coefficient of solid-phase layer thickness and solid state reaction layer is fixed, reflectance only with unit thickness
The absorption coefficient of light relevant, and the absorption coefficient of light size of unit thickness is directly proportional to testing concentration, by measuring reflectance,
Testing concentration can be calculated.During mensuration, fluid sample such as serum, blood plasma, whole blood, urine, cerebrospinal fluid etc. are directly added drop-wise to solid
Surely have on the biochemical dry tablet of reagent, to detect moisture in sample as solvent, cause the examination pre-fixed in determinand and dry tablet
Agent generation chemical reaction also produces reaction signal, finally by visual or instrument detection, it is achieved testing concentration carries out qualitative, partly
Quantitatively or detection by quantitative.
Compared with wet chemistry method, Dry chemical method has instrument miniature portable, reagent simple to operate, dry is prone to preservation,
Detection speed is fast, sample consumption is little, be not required to configure any liquid reagent, and inspection work can be carried out whenever and wherever possible, detects the time
Short, easy and simple to handle, accuracy and precision are higher.This technology has only to instrument and supporting dry tablet thereof, it is not necessary to institute in wet-chemical
The liquid reagent used or any solution.Dry tablet is single use, produces without any waste liquid, it is possible to avoid polluting.And energy
Enough apply the field cannot touched at wet chemistry method, such as Clinical Laboratorys such as space station, battlefield, field or naval vessels, also can answer
For the field such as food safety, environmental monitoring.
At present, dry chemistry product great majority are dependent on import, as accounted for the maximum Johnson & Johnson of the U.S. of the market share, Switzerland Roche
With the major company such as kyoto, Japan, and domestic dry chemistry research is started late, application state external mature technology mostly, only at surface structure
On improved, in addition to better simply dry chemistry Urine test paper and analyser, also do not have more dry chemistry instrument and reagent to carry
Body product occurs.This situation causes the cost of dry chemical method inspection and raises, and is unfavorable for that dry analysis technology is cured in China
Learn the promotion and application in inspection., there is preparation complexity, high in cost of production shortcoming in the development of multi-layer film structure dry tablet.
Summary of the invention
It is an object of the invention to: a kind of monofilm biochemistry dry tablet for detecting blood fat and preparation method are provided, solve
The problem that the dry tablet operation of preparation is complicated, has the range of linearity wider simultaneously, repeatability, has good stability, and detection limit is relatively low, inspection
Degree of testing the speed is fast, characteristic simple to operate.
The technical scheme is that a diameter of 6 mm circle thickness be 0.3 mm, flow velocity be 130 mm/30 min,
Water carrying capacity is to fix cholesterin detection reagent A(cholesteryl esterase, Ascorbic Acid Oxidation in the monolayer chromatography filter paper of 10 μ L simultaneously
Enzyme, ESPAS(ADPS)) and detectable B(cholesterol oxidase, 4-amino peace is for neighbour woods), this chromatography filter paper is attached to lid
On slide.
Wherein, described cholesteryl esterase >=300 U/L in cholesterin detection reagent A, ascorbic acid oxidase >=
1000 U/L, ESPAS(ADPS) 100 mg/L；In detectable B cholesterol oxidase >=500 U/L, 4-amino peace for than
Adjacent woods 150 mg/L.
The method of the present invention:
(1) chromatography filter paper is cut into a diameter of 6 mm circular, standby；
(2) the rectangle chromatography filter paper in (1) is placed on microscope slide, first the spraying of the chromatography filter paper on microscope slide 9-11
μ L cholesterin detection reagent B, freezing 10 min of ultralow temperature (-80 DEG C), and-30 DEG C of lyophilizing in vacuum freeze drier, freeze
The dry time is about 30-40 min, and this step repeats 3-5 time；Then 9-11 μ L cholesterol detection is sprayed to same chromatography filter paper
Reagent A, freezing 10 min of ultralow temperature (-80 DEG C), and-30 DEG C of lyophilizing in vacuum freeze drier, freeze-drying time is about
30-40 min, this step repeats 3-5 time；The different blood lipids index of detection, detectable sample-adding amount, freeze-drying time and sample-adding number of times
Different；
(3) chromatography filter paper being fixed with detectable in (2) is transferred to 24 × 24 mm coverslip central authorities, is cholesterol inspection
Survey monofilm biochemistry dry tablet；
(4) upper machine testing: use full spectrum Dry-type biochemical analyser that dry tablet biochemical prepared by (3) is detected, detect wavelength
Being 550 nm, educate temperature time 1 s, detect time 240 s, sample-adding measures 10 μ l, and standard substance are 2 times of serial dilutions；
(5) examination criteria curve and the standard color range of dry tablet are prepared
Using the blood fat dry tablet of above-mentioned preparation and full spectrum Dry-type biochemical analyser, Criterion product concentration is relatively strong with reflection light
Functional relationship between degree, as the foundation of cholesterol detection concentration.Light relative intensity is reflected with standard solution biochemistry dry tablet
Meansigma methods is vertical coordinate Y, is abscissa X with testing concentration in standard substance (unit is as mmol/L), the cholesterol (TC) of foundation,
Triglyceride (TG) and the Dry-type biochemical assay standard curve of high density lipoprotein (HDL-C), as shown in Figures 2 to 4,
Meet logarithmic function relation.The detection limit of three kinds of monofilm dry tablets of the Dry-type biochemical assay set up is respectively
0.0517 mmol/L, 0.113 mmol/L, 0.249 mmol/L, the range of linearity be respectively 3.44 ~ 25.83 mmol/L, 2.46
~12.3 mmol/L、1~3.24 mmol/L.The biochemical dry tablet prepared with the standard substance addition of gradient dilution, with serum as blank
Comparison, cholesterol standards is that 2 times of gradient dilutions add in serum, drips on prepared dry tablet, treat anti-after sample blending
Should terminate i.e. to prepare the standard color range of different colours, can be used for the semi-quantitative analysis of sample detection.
The invention has the beneficial effects as follows: the range of linearity is wider, repeatability, have good stability, detection limit relatively low, detect speed
Hurry up, simple to operate, patient can detect voluntarily, and this biochemistry dry tablet is single-layer membrane structure, it is possible to effectively reduces cost, therefore can be used for
Clinical quickly detection, has good economic benefit and application prospect.
Detect with accuracy with betweenrun precision in batch
Should the most once prepare each 20 of three kinds of blood fat dry tablets, and detect serum sample, every sample is interior on the same day
Replication 20 times, calculates testing result meansigma methods, relative standard deviation and the coefficient of variation, and batch interior repeatability of surveyed data is such as
Shown in Fig. 6.
Should the most once prepare each 20 of three kinds of blood fat dry tablets, and detect serum sample, every sample replication every day 2
Secondary, measure 10 days altogether, calculate testing result meansigma methods, relative standard deviation and the coefficient of variation, repeated between the criticizing of surveyed data
As shown in Figure 7.
Shown by the above results, three monofilm biochemistry dry tablets of the blood fat prepared respectively batch in, batch between imprecision and
Inaccuracy is respectively less than 5%, in allowed band.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of preparation method of the present invention；
Fig. 2 is the standard curve that Dry-type biochemical of the present invention analyzes method mensuration cholesterol (TG)；
Fig. 3 is the standard curve that Dry-type biochemical of the present invention analyzes method mensuration triglyceride (TC)；
Fig. 4 is the standard curve that Dry-type biochemical of the present invention analyzes method mensuration high density lipoprotein (HDL-C)；
Fig. 5 is the structure chart of biochemistry dry tablet of the present invention；
Fig. 6 is batch interior meansigma methods of repeated experiment, relative standard deviation, coefficient of variation table；
Fig. 7 be batch between the meansigma methods of repeated experiment, relative standard deviation, coefficient of variation table.
Detailed description of the invention
Below in conjunction with the accompanying drawings the present invention is described further:
Embodiment 1
The biochemical dry tablet of preparation uses a diameter of 6 mm circle thickness to be 0.3 mm, flow velocity is 130 mm/30 min, water carrying capacity is
The monolayer chromatography filter paper of 10 μ L.By being fixed in chromatography filter paper by lipids detection liquid biochemical reagents, establish lipids detection
The preparation method of monofilm biochemistry dry tablet, prepares the standard color range of lipids detection monofilm biochemistry dry tablet, and uses full spectrum
Dry-type biochemical analyser, sets up dry tablet reflection light relative intensity (Y) bent with the quantitative measurement standard of blood fat standard concentration (X)
Line, and determine the relevant parameter of mensuration, such as incubative time, sample-adding amount etc..So far lipids detection monofilm dry type is established raw
Change assay method.
Illustrate as a example by cholesterol below；
(1) chromatography filter paper is cut into a diameter of 6 mm circular, standby；
(2) the rectangle chromatography filter paper in (1) is placed on microscope slide, first the spraying of the chromatography filter paper on microscope slide 9-11
μ L cholesterin detection reagent B, freezing 10 min of ultralow temperature (-80 DEG C), and-30 DEG C of lyophilizing in vacuum freeze drier, freeze
The dry time is about 30-40 min, and this step repeats 3-5 time；Then 9-11 μ L cholesterol detection is sprayed to same chromatography filter paper
Reagent A, freezing 10 min of ultralow temperature (-80 DEG C), and-30 DEG C of lyophilizing in vacuum freeze drier, freeze-drying time is about
30-40 min, this step repeats 3-5 time；The different blood lipids index of detection, detectable sample-adding amount, freeze-drying time and sample-adding number of times
Different；
(3) chromatography filter paper being fixed with biochemical reagents of preparation in (2) is transferred to 24 × 24mm coverslip central authorities, is preparation
The monofilm biochemistry dry tablet of cholesterol detection.
(4) upper machine testing: use full spectrum Dry-type biochemical analyser that dry tablet biochemical prepared by (3) is detected, detection
Wavelength is 550 nm, educates temperature time 1 s, detects time 240 s, and sample-adding measures 10 μ l, and standard substance are 2 times of serial dilutions；
(5) examination criteria curve and the standard color range of dry tablet are prepared
Using the blood fat dry tablet of above-mentioned preparation and full spectrum Dry-type biochemical analyser, Criterion product concentration is relatively strong with reflection light
Functional relationship between degree, as the foundation of cholesterol detection concentration.Light relative intensity is reflected with standard solution biochemistry dry tablet
Meansigma methods is vertical coordinate Y, is abscissa X with testing concentration in standard substance (unit is as mmol/L), the cholesterol (TC) of foundation,
Triglyceride (TG) and the Dry-type biochemical assay standard curve of high density lipoprotein (HDL-C), as shown in Figures 2 to 4,
Meet logarithmic function relation.The detection limit of three kinds of monofilm dry tablets of the Dry-type biochemical assay set up is respectively
0.0517 mmol/L, 0.113 mmol/L, 0.249 mmol/L, the range of linearity be respectively 3.44 ~ 25.83 mmol/L, 2.46
~12.3 mmol/L、1~3.24 mmol/L.The biochemical dry tablet prepared with the standard substance addition of gradient dilution, obtains corresponding dry tablet
Standard color range, can be used for measuring samples concentration and carries out sxemiquantitative.
Claims (2)
1., for detecting the monofilm biochemistry dry tablet of blood fat, it is characterized in that: a diameter of 6 mm circle thickness be 0.3 mm,
Flow velocity is 130 mm/30 min, water carrying capacity is to fix cholesterin detection reagent A i.e. gallbladder in the monolayer chromatography filter paper of 10 μ L simultaneously
Sterin esterase, ascorbic acid oxidase, ESPASADPS and detectable B cholesterol oxidase, 4-amino peace, should for neighbour woods
Chromatography filter paper attaches on the cover slip；
Wherein, described cholesteryl esterase >=300 U/L in cholesterin detection reagent A, ascorbic acid oxidase >=1000 U/
L, ESPASADPS100 mg/L；Cholesterol oxidase >=500 U/L, 4-amino peace in detectable B is for neighbour woods 150
mg/L。
2., for detecting a preparation method for the monofilm biochemistry dry tablet of blood fat, its method is:
(1) chromatography filter paper is cut into a diameter of 6 mm circular, standby；
(2) the rectangle chromatography filter paper in (1) is placed on microscope slide, first the spraying of the chromatography filter paper on microscope slide 9-11
μ L cholesterin detection reagent B, at-80 DEG C of cryotherapy 10 min, and-30 DEG C of lyophilizing in vacuum freeze drier, freeze
The dry time is about 30-40 min, and this step repeats 3-5 time；Then 9-11 μ L cholesterol detection is sprayed to same chromatography filter paper
Reagent A, at-80 DEG C of cryotherapy 10 min, and-30 DEG C of lyophilizing in vacuum freeze drier, freeze-drying time is about
30-40 min, this step repeats 3-5 time；The different blood lipids index of detection, detectable sample-adding amount, freeze-drying time and sample-adding number of times
Different；
(3) chromatography filter paper being fixed with detectable in (2) is transferred to 24 × 24 mm coverslip central authorities, is cholesterol inspection
Survey monofilm biochemistry dry tablet；
(4) upper machine testing: use full spectrum Dry-type biochemical analyser that dry tablet biochemical prepared by (3) is detected, detect wavelength
Being 550 nm, educate temperature time 1 s, detect time 240 s, sample-adding measures 10 μ l, and standard substance are 2 times of serial dilutions；
(5) examination criteria curve and the standard color range of dry tablet are prepared
Using the blood fat dry tablet of above-mentioned preparation and full spectrum Dry-type biochemical analyser, Criterion product concentration is relatively strong with reflection light
Functional relationship between degree, as the foundation of cholesterol detection concentration；Light relative intensity is reflected with standard solution biochemistry dry tablet
Meansigma methods is vertical coordinate Y, and in standard substance, testing concentration is abscissa X, and the cholesterol TC of foundation, triglyceride TG are with highly dense
The Dry-type biochemical assay standard curve of degree lipoprotein HDL-C, meets logarithmic function relation；The Dry-type biochemical set up is divided
The detection limit of three kinds of monofilm dry tablets that analysis method measures is respectively 0.0517 mmol/L, 0.113 mmol/L, 0.249 mmol/
L, the range of linearity is respectively 3.44 ~ 25.83 mmol/L, 2.46 ~ 12.3 mmol/L, 1 ~ 3.24 mmol/L；Use gradient dilution
Standard substance add the biochemical dry tablet of preparation, with serum as blank, cholesterol standards is that 2 times of gradient dilutions add serum
In, drip after sample blending on prepared dry tablet, question response terminates i.e. to prepare the standard color range of different colours, can be used for sample
The semi-quantitative analysis that product examine is surveyed.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106645763A (en) * | 2017-01-03 | 2017-05-10 | 长沙中生众捷生物技术有限公司 | Total cholesterol detection reagent and total cholesterol detection paper |
| CN107402209A (en) * | 2017-09-15 | 2017-11-28 | 光景生物科技(苏州)有限公司 | It is a kind of to be used to detect test strips of LDL-C and preparation method thereof in serum |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106645763A (en) * | 2017-01-03 | 2017-05-10 | 长沙中生众捷生物技术有限公司 | Total cholesterol detection reagent and total cholesterol detection paper |
| CN107402209A (en) * | 2017-09-15 | 2017-11-28 | 光景生物科技(苏州)有限公司 | It is a kind of to be used to detect test strips of LDL-C and preparation method thereof in serum |
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