CN116718785A - Triglyceride kit with high stability and preparation method thereof - Google Patents

Triglyceride kit with high stability and preparation method thereof Download PDF

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CN116718785A
CN116718785A CN202310780146.1A CN202310780146A CN116718785A CN 116718785 A CN116718785 A CN 116718785A CN 202310780146 A CN202310780146 A CN 202310780146A CN 116718785 A CN116718785 A CN 116718785A
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reagent
pipes
bsa
kit
naoh
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CN116718785B (en
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曹新鹏
段文杰
张迪
秦亚乔
高梦婷
张培蕾
栗静
韩萌萌
黄喜洛
姚艳辉
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Henan Jianshi Laijie Medical Technology Co ltd
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Henan Jianshi Laijie Medical Technology Co ltd
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    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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Abstract

The invention discloses a triglyceride kit with high stability and a preparation method thereof, and relates to the technical field of medical detection. According to the invention, the glycerol assay kit can meet the conventional anti-interference capability and industry technical requirement standards by changing the types of chromogens in the kit, the specific position sequences among the components and adding the specific compound surfactant and the stabilizer. The preparation method comprises the steps of preparing the first reagent and the second reagent, so that the stability of the prepared kit can reach 6 years at 2-8 ℃, the production process is easier to control, the clinical application is simpler, and the cost can be reduced.

Description

Triglyceride kit with high stability and preparation method thereof
Technical Field
The invention relates to the technical field of medical detection, in particular to a triglyceride kit with high stability and a preparation method thereof.
Background
Triglyceride (TG) is a fat molecule formed by long-chain fatty acid and glycerol, is the most abundant lipid in a human body, most tissues can supply energy by utilizing a TG decomposition product, meanwhile, tissues such as liver, fat and the like can also carry out synthesis of TG, the Triglyceride is stored in the adipose tissues, and is also a main component of blood fat, the Triglyceride in the food is digested and decomposed by pancreatic lipase after being absorbed and re-esterified into the Triglyceride, and enters the blood through a lymphatic system, under pathological conditions, the synthesized and stored Triglyceride of various tissue cells is obviously increased, and the quantitative detection of the Triglyceride content in the blood and tissue cells is a common biochemical index for clinical, clinical basis and basic medical biology research.
For stability studies of diagnostic reagents, accelerated stability experiments are often employed in the art to predict the failure period (for evaluating shelf stability, sometimes also referred to as long-term stability or expiration date) of a product. Such a method has the advantage that the investigation period is short (typically several weeks), without actually storing the reagents for 1 year, 2 years, or even longer. However, the theoretical basis for acceleration stability is the Arrhenius formula (Arrhenius equation). In order to ensure accuracy of prediction, there are many demands on the reagent in order to apply the formula. This is a prediction in an ideal state. However, biochemical reagents are complex in composition and of a wide variety, and not any reagent is perfectly suitable for accelerated stability experiments to predict its true useful life. In addition, the stability of the reagent should not be interpreted narrowly as shelf stability (long-term stability). This is because the reagent is not simply left in a warehouse with constant environmental conditions (constant temperature and humidity, direct light avoidance, no vibration) but may be exposed to a changing environment during actual operation. For example, during transportation (handling, jolting), mechanical forces are encountered and variations in temperature are encountered (although cold chain transportation may be used, constant temperature and humidity moments are unavoidable). For another example, in clinical institutions, the detection of samples is performed on large-scale equipment (biochemical analyzers). This mode of operation allows multiple samples (from tens to hundreds of samples) to be detected sequentially or simultaneously. The reagents will then be released from their stable storage environment and the packaging machine opened for use. The temperature and humidity change cannot be avoided in the process, and the sample is exposed to air after being uncapped and faces oxidation factors. Therefore, in this case, the accelerated stability test cannot evaluate the stability in other aspects.
The Chinese patent application No. CN201410697066.0 discloses a reagent for detecting serum triglyceride with strong stability, which comprises 4-hydroxyethyl piperazine ethane sulfonic acid-sodium glutamate buffer solution, lipoprotein lipase, glycerol kinase, peroxidase, glycerol-3-phosphate oxidase, bovine serum albumin, 4-aminoantipyrine, lauryl sulfate triethanolamine, coenzyme FAD, polyvinyl alcohol, sodium perbenzoate and the like. Although the lauryl sulfuric acid triethanolamine is added into the reagent, the stability of lipoprotein lipase can be effectively protected; the coenzyme FAD and the polyvinyl alcohol are added into the reagent, so that active groups of enzymes in the glycerol-3-phosphate oxidase can be effectively protected to ensure that the enzymes are not easy to inactivate, thereby improving the stability of the enzymes; sodium benzoate is added into the reagent, so that the anti-interference capability and stability of the reagent on reducing substances can be improved. But the problems of reagent discoloration and blank rise are not solved.
Disclosed in chinese patent application No. CN201610865116.0 is a kit and method for assaying triacetin, the kit comprising a reagent set 1 and a reagent set 2, the reagent set 1 comprising a magnesium salt, adenosine triphosphate, glycerol kinase, glycerophosphate oxidase, peroxidase and a chromogen, the reagent set 2 comprising lipoprotein lipase and 4-aminoantipyrine, wherein the chromogen comprises N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3, 5-Dimethoxyaniline (DAOS). Although interference can be counteracted by using DAOS chromogens and controlling the amount of tool enzyme used, stability tends to be generally short shelf life.
Therefore, it is necessary to provide a triglyceride kit with high stability and a preparation method thereof to solve the above problems.
Disclosure of Invention
The invention aims at: solves the problems that the existing triglyceride measuring kit has more types of stabilizers and can improve the anti-interference capability, but the stability is always general.
The invention adopts the following technical scheme for realizing the purposes:
a triglyceride kit with high stability comprises a first reagent and a second reagent;
the first reagent comprises PIPES 20mmol-200mmol/L, naOH 0.2-10g/L, mgCl 2 0.5-5g/L, 0.5-5g/L of compound surfactant, 0.1-5g/L of 4-chlorophenol, 0.5-5KU/L, BSA (0.3-3) g/L of ascorbate oxidase and 0.3-2g/L of preservative;
the PH of the first reagent is more than or equal to 6.80;
the second reagent comprises PIPES 30-300mmol/L, naOH, PIPES 0.2-10g/L, ATP, PIPES 0.5-10g/L, BSA, sucrose 5-50g/L, LPL, sucrose 5-30 KU/L, GK 2-20KU/L, POD, sucrose 2-20KU/L, GPO, PIPES 5-50KU/L, 4-AAP 0.2-3g/L, EDTA, PIPES 0.2-5g/L, preservative 0.3-3g/L;
the PH of the second reagent is more than or equal to 6.80.
Further, the kit is used for measuring triglyceride content in a sample, wherein the sample is serum or plasma.
Further, PIPES 20mmol/L, naOH 0.2.2 g/L, mgCl in the first reagent 2 3g/L, 0.2g/L of compound surfactant, 0.2g/L of 4-chlorophenol, 0.5KU/L, BSA 0.5.5 g/L of ascorbyl oxidase and 1g/L of preservative;
ph=6.90 of the first reagent;
PIPES 300mmol/L, naOH 5g/L, ATP g/L, BSA g/L, sucrose 25g/L, LPL KU/L, GK KU/L, POD 10KU/L, GPO KU/L,4-AAP 1.5g/L, EDTA 3g/L and preservative 0.4g/L;
the PH of the second reagent=7.00.
Further, PIPES 200mmol/L, naOH 6g/L and MgCl in the first reagent 2 0.5g/L, 2g/L of compound surfactant, 2g/L of 4-chlorophenol, 5KU/L of ascorbyl oxidase, 1g/L of BSA and 2g/L of preservative;
ph=7.50 of the first reagent;
the second reagent comprises PIPES 30mmol/L, naOH 0.9g/L, ATP 10g/L, BSA 5g/L, sucrose 50g/L, LPL 15KU/L, GK 10KU/L, POD 20KU/L, GPO 40KU/L,4-AAP 0.6g/L, EDTA 1g/L and preservative 0.5g/L;
the PH of the second reagent=7.00.
Further, the kit also comprises one or two of a quality control product and a calibrator.
Further, the compound surfactant is one or a mixture of at least two of tritonx-100, PEG6000, CHAPS, THESIT, tween 20, PEG8000, emulgen B-66 and Emulgen A90.
Further, the preservative is one or a mixture of at least two of PC300, PC950, sodium azide, thimerosal and sodium benzoate.
Further, the triton x-100 concentration is 2g/L.
Further, the PC950 concentration was 0.8ml/L.
A preparation method of a triglyceride kit with high stability comprises the following steps:
s1, preparing a first reagent: weighing triton x-100 with a 100ml beaker, adding water, heating and dissolving on an electromagnetic oven, and naturally cooling at 18-40 ℃ for standby;
adding 800ml of water into a 1L beaker, weighing PIPES and NaOH, putting into water, stirring for dissolution, adjusting the pH value to 7.00+/-0.05, and sequentially weighing 4-chlorophenol and MgCl 2 Ascorbic acid oxygenDissolving the lipase, proclin950 and BSA in the above solution, stirring to dissolve, adding cooled triton-100 into the solution, and mixing to 1L to obtain the first reagent.
S2, preparing a second reagent: adding 800ml of water into a 1L beaker, putting PIPES and NaOH into the water, stirring and dissolving, adjusting the pH value to 7.00+/-0.05, sequentially measuring ATP, BSA, sucrose, LPL, GK, GPO, POD, 4-AAP, EDTA, proclin950 and BSA, dissolving in the above solutions, stirring and dissolving, fixing the volume to 1L, and uniformly mixing to obtain a second reagent.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the invention, the glycerol assay kit can meet the conventional anti-interference capability and industry technical requirement standard by changing the types of chromogens in the reagent, the specific position sequence among the components and adding the specific composite surfactant, BSA and EDTA, and the stability of the kit can reach 6 years at 2-8 ℃.
2. The preparation method provided by the invention is more convenient to prepare, the clinical application is simpler, and the cost can be reduced.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
A stable triglyceride kit comprising a first reagent and a second reagent:
the first reagent comprises PIPES 20mmol-200mmol/L, naOH 0.2-10g/L, mgCl 2 0.5-5g/L, 0.5-5g/L of compound surfactant, 0.1-5g/L of 4-chlorophenol, 0.5-5KU/L, BSA, 0.3-3g/L of ascorbate oxidase and 0.3-2g/L of preservative;
the PH of the first reagent is more than or equal to 6.80;
the second reagent comprises PIPES 30-300mmol/L, naOH, PIPES 0.2-10g/L, ATP, PIPES 0.5-10g/L, BSA, sucrose 5-50g/L, LPL, sucrose 5-30 KU/L, GK 2-20KU/L, POD, sucrose 2-20KU/L, GPO, PIPES 5-50KU/L, 4-AAP 0.2-3g/L, EDTA, PIPES 0.2-5g/L, preservative 0.3-3g/L;
the PH of the second reagent is more than or equal to 6.80.
Specifically, the kit further comprises one or two of a quality control product and a calibrator.
Specifically, the kit is used for measuring triglyceride content in a sample, wherein the sample is serum or plasma;
specifically, the compound surfactant is one or a mixture of at least two of tritonx-100, PEG6000, CHAPS, THESIT, tween 20, PEG8000, emulgen B-66 and Emulgen A90.
Specifically, the preservative is one or a mixture of at least two of PC300, PC950, sodium azide, merthiolate and sodium benzoate.
Specifically, the triton x-100 concentration is 2g/L, the 4-chlorophenol concentration is 2g/L, and the PC950 concentration is 0.8ml/L.
A preparation method of a triglyceride kit with high stability comprises the following steps:
s1, preparing a first reagent: weighing triton x-100 with a 100ml beaker, adding water, heating and dissolving on an electromagnetic oven, and naturally cooling at 18-40 ℃ for standby;
adding 800ml of water into a 1L beaker, weighing PIPES and NaOH, putting into water, stirring for dissolution, adjusting the pH value to 7.00+/-0.05, and sequentially weighing 4-chlorophenol and MgCl 2 Dissolving ascorbate oxidase, proclin950 and BSA in the above solution, stirring to dissolve, adding cooled triton-100 into the solution, and mixing to 1L to obtain the first reagent.
S2, preparing a second reagent: adding 800ml of water into a 1L beaker, putting PIPES and NaOH into the water, stirring and dissolving, adjusting the pH value to 7.00+/-0.05, sequentially measuring ATP, BSA, sucrose, LPL, GK, GPO, POD, 4-AAP, EDTA, proclin950 and BSA, dissolving in the above solutions, stirring and dissolving, fixing the volume to 1L, and uniformly mixing to obtain a second reagent.
Based on the above-described kit and preparation method, experiments were performed by six sets of examples, respectively.
Example 1
This example is a control, designated control 1
PIPES 200mmol/L, naOH 6g/L and MgCl in the first reagent 2 0.5g/L Tween 20 g/L, 4-chlorophenol 0.6g/L, ascorbyl oxidase 5KU/L, BSA 1g/L, and PC 950.8 ml/L;
first reagent ph=7.00;
the second reagent comprises PIPES 30mmol/L, naOH 0.9g/L, ATP 10g/L, BSA 5g/L, sucrose 50g/L, LPL 15KU/L, GK 10KU/L, POD 20KU/L, GPO 40KU/L,4-AAP 0.6g/L, EDTA 1g/L and P9500.8 ml/L;
the PH of the second reagent=7.20.
Example 2
This example is a control, designated control 2
The PIPES 200mmol/L, naOH 6g/L and MgCl in the first reagent 2 0.5g/L, CHAPS 2g/L, 4-chlorophenol 2g/L, ascorbyl oxidase 5KU/L, GPO 10KU/L, BSA 3g/L, PC 950.8 ml/L;
ph=7.00 of the first reagent;
the second reagent comprises PIPES 30mmol/L, naOH 0.9g/L, ATP 10g/L, BSA 5g/L, sucrose 50g/L, LPL 15KU/L, GK 10KU/L, POD 20KU/L,4-AAP 0.6g/L, EDTA 1g/L and PC 950.8 ml/L;
the PH of the first reagent=7.00.
Example 3
This example is a control, designated control 3
The PIPES 200mmol/L, naOH 6g/L and MgCl in the first reagent 2 0.5g/L, 4g/L of 4-chlorophenol, 5KU/L of ascorbyl oxidase;
ph=7.00 of the first reagent;
the second reagent comprises PIPES 30mmol/L, naOH 0.9g/L, ATP 10g/L, sucrose 50g/L, LPL 15KU/L, GK 10KU/L, POD 20KU/L, GPO 40KU/L and 4-AAP 0.6g/L;
the PH of the second reagent=7.00.
Example 4
This example is a test example of the kit configuration of the present invention, and is designated as test example 1
The PIPES 200mmol/L, naOH 6g/L and MgCl in the first reagent 2 0.5g/L, triton x-100.2 g/L, 4-chlorophenol 2g/L, ascorbyl oxidase 5KU/L, BSA 1g/L, PC9500.8ml/L;
ph=7.50 of the first reagent;
the second reagent comprises PIPES 30mmol/L, naOH 0.9g/L, ATP 10g/L, BSA 5g/L, sucrose 50g/L, LPL 15KU/L, GK 10KU/L, POD 20KU/L, GPO 40KU/L,4-AAP 0.6g/L, EDTA 1g/L and PC9500.8ml/L;
the PH of the second reagent=7.00.
Example 5
This example is a test example of the kit configuration of the present invention, and is designated as test example 2
The first reagent comprises PIPES 20mmol/L, naOH 0.2g/L and MgCl 2 3g/L, CHAPS 0.2g/L, 4-chlorophenol 0.2g/L, ascorbyl oxidase 0.5KU/L, BSA 0.5g/L, thimerosal 0.8ml/L;
ph=6.90 of the first reagent;
the second reagent comprises PIPES 300mmol/L, naOH 5g/L, ATP 5g/L, BSA 3g/L, sucrose 25g/L, LPL 30KU/L, GK 20KU/L, POD 10KU/L, GPO 25KU/L,4-AAP 1.5g/L, EDTA 3g/L and PC 300.4 g/L;
the PH of the second reagent=7.00.
Example 6
This example is a test example of the kit configuration of the present invention, and is designated as test example 3
The PIPES of the first reagent is 100mmol/L, naOH of 3g/L and MgCl 2 1.6g/L, THESIT 1.6g/L, 4-chlorophenol 2g/L, ascorbyl oxidase 2.2KU/L, BSA 1.3g/L, sodium azide 1.1ml/L;
ph=7.20 of the first reagent;
in the second reagent, PIPES 160mmol/L, naOH 2.1g/L, ATP 0.5g/L, BSA 0.3g/L, sucrose 6g/L, LPL 3KU/L, GK 2KU/L, POD 4KU/L, GPO 7KU/L,4-AAP 1.5g/L, EDTA 4.8g/L and sodium benzoate 1.9g/L;
the PH of the second reagent=7.30.
Tests were performed for examples 1-6 above, and divided into three steps:
the first step: setting parameters of a Hitachi 7600 full-automatic biochemical analyzer:
the analysis method comprises the following steps: a two-point end point method;
the reaction direction is as follows: positive;
dominant wavelength: 505nm, negative wavelength: 700nm;
temperature: 37 degrees;
sample size: 3ul;
reagent 1:240ul;
reagent 2:60ul;
measuring points 10-32;
a 37-degree acceleration kit for measuring the measured values of Langdao complex quality control level two and level three (the 37-degree acceleration is 7 days, the attenuation degree of the enzyme is seen, and the stability of the reaction kit is measured);
HN1530-1496U N target: 1.06mmol/L (0.89-1.23), corresponding to the data in the left column below each control and test case in tables 1 and 2;
HE1532-1223UE target value: 2.92mmol/L (2.46-3.38), corresponding to the data in the right column below each control and test case in tables 1 and 2;
TABLE 1
TABLE 2
As can be seen from the comparison of tables 1 and 2: under the condition of adding a proper protective agent and a stabilizing agent in the verification of the accelerated destruction reagent for 47 days, the quality control value of the reagent is stable and reduced slightly compared with the quality control value of the test group combined according to a specific position, and about 3-4% of the reagent can be accepted and can be solved as a linear guarantee;
the reagent of the comparative example 3 without the protective agent and the stabilizer has been deviated by more than 10% in the fifth day of acceleration, is not in accordance with the technical requirements of manufacturers, and has been deviated by more than 18-20% in the seventh day;
the reagent, which was not arranged in the specific order, was in comparative example 2, had crashed on the third day of acceleration and had lost the enzyme activity completely on the fifth day;
the reagents, which were not arranged in the particular order, in comparative example 1, were not in compliance with the manufacturer's specifications by a deviation of more than 10% on the fifth day of acceleration, and were in compliance with the manufacturer's specifications by a deviation of 20% on the seventh day.
And a second step of: on-board reagent stability: determining Langdao quality control (28 days 2-8 degree instrument reagent cabin, quality control determination, stability of reagent kit after opening bottles, stability in instrument reagent cabin after opening bottles, accuracy of measured value)
TABLE 3 Table 3
TABLE 4 Table 4
As can be seen from the comparison of tables 3 and 4: the reagent in test examples 1-3 was bottled for 28 days to determine Langdao quality control, and the reagent kit was used for 28 days with a 2-8 degree instrument, so that the reagent kit had no problem and the deviation of the measured value was about 1.5%;
the Langdao quality control product is measured after the reagent of comparative example 1 is opened for 28 days, the deviation of the measured value of a reagent bin of a 2-8 degree instrument is about 30 percent, and the technical requirements of manufacturers are not met;
the Langdao quality control product is measured after the reagent of comparative example 2 is opened for 7 days, the deviation of the measured value of a reagent bin of a 2-8 degree instrument is about 30 percent, and the technical requirements of manufacturers are not met;
control 3 the Langdao quality control was measured 28 days after the reagent was opened, the deviation of the measured value was about 8-9% in the reagent bin of the 2-8 degree instrument, and the stability of the open bottle was not satisfied, but the stability of the pot life was not satisfied.
And a third step of: anti-jamming capability test for test cases
Test examples 1-3 were all more stable on-board, but test example 1 was the most stable on-board than test examples 2 and 3, so test example 1 was selected for tamper resistance testing
Interference evaluation: the interfering substances were diluted in a gradient with ultrapure water and the concentrations of the interfering substances were adjusted to 1:9 to a control serum (i.e., normal human mixed serum without interfering substances), the final concentration of each interfering substance is shown in the first line of data in tables 3 and 4;
respectively measuring detection results of blank serum and serum added with interference substances with different concentrations by using a test case kit, measuring each serum sample for 3 times to obtain a mean value, and calculating deviation ((mean-blank control)/blank control 100%) of the mean value and a blank control serum measurement value, namely the interference degree;
if the absolute value of the interference degree is smaller than 10%, the interference degree is marked as anti-interference, otherwise, the interference degree is marked as non-anti-interference;
typical interfering substances in serum are ascorbic acid, bilirubin, hemoglobin, etc.;
TABLE 5 test case reagent hemoglobin interference test
Table 6 test example reagent ascorbic acid interference test
Concentration of interfering substance Blank control 0.03g/L 0.06g/L 0.12g/L 0.25g/L 0.5g/L
Test one 1.92 1.90 1.90 1.88 1.85 1.77
Test II 1.93 1.91 1.89 1.89 1.83 1.78
Test three 1.94 1.90 1.89 1.87 1.82 1.76
Mean value of 1.93 1.90 1.89 1.88 1.83 1.77
Interference level -1.55％ -2.07％ -2.58％ -5.18％ -8.30％
Whether or not to resist interference Anti-interference Anti-interference Anti-interference Anti-interference Anti-interference
In conclusion, the detection kit provided by the invention has good anti-interference performance and stability, has good sensitivity, effectively overcomes various defects in the prior art, and has high industrial utilization value.
The present invention is not limited to the preferred embodiments, but the patent protection scope of the invention is defined by the claims, and all equivalent structural changes made by the specification and the drawings are included in the scope of the invention.

Claims (10)

1. A strong stability triglyceride kit comprising a first reagent and a second reagent, characterized in that:
the first reagent comprises PIPES 20mmol-200mmol/L, naOH 0.2-10g/L, mgCl 2 0.5-5g/L, 0.5-5g/L of compound surfactant, 0.1-5g/L of 4-chlorophenol, 0.5-5KU/L, BSA (0.3-3) g/L of ascorbate oxidase and 0.3-2g/L of preservative;
the PH of the first reagent is more than or equal to 6.80;
the second reagent comprises PIPES 30-300mmol/L, naOH, PIPES 0.2-10g/L, ATP, PIPES 0.5-10g/L, BSA, sucrose 5-50g/L, LPL, sucrose 5-30 KU/L, GK 2-20KU/L, POD, sucrose 2-20KU/L, GPO, PIPES 5-50KU/L, 4-AAP 0.2-3g/L, EDTA, PIPES 0.2-5g/L, preservative 0.3-3g/L;
the PH of the second reagent is more than or equal to 6.80.
2. A stable triglyceride kit according to claim 1, wherein: the kit is used for measuring triglyceride content in a sample, wherein the sample is serum or plasma.
3. A stable triglyceride kit according to claim 1, wherein: PIPES 20mmol/L, naOH 0.2.2 g/L, mgCl in the first reagent 2 3g/L, 0.2g/L of compound surfactant, 0.2g/L of 4-chlorophenol, 0.5KU/L, BSA 0.5.5 g/L of ascorbyl oxidase and 1g/L of preservative;
ph=6.90 of the first reagent;
PIPES 300mmol/L, naOH 5g/L, ATP g/L, BSA g/L, sucrose 25g/L, LPL KU/L, GK KU/L, POD 10KU/L, GPO KU/L,4-AAP 1.5g/L, EDTA 3g/L and preservative 0.4g/L;
the PH of the second reagent=7.00.
4. A stable triglyceride kit according to claim 1, wherein: PIPES 200mmol/L, naOH g/L, mgCl in the first reagent 2 0.5g/L, composite surface active2g/L of agent, 2g/L of 4-chlorophenol, 5KU/L, BSA g/L of ascorbyl oxidase and 2g/L of preservative;
ph=7.50 of the first reagent;
PIPES 30mmol/L, naOH 0.9g/L, ATP g/L, BSA g/L, sucrose 50g/L, LPL KU/L, GK KU/L, POD KU/L, GPO KU/L,4-AAP 0.6g/L, EDTA g/L and preservative 0.5g/L;
the PH of the second reagent=7.00.
5. A stable triglyceride kit according to claim 1, wherein: the kit also comprises one or two of a quality control product and a calibrator.
6. A stable triglyceride kit according to claim 1, wherein: the compound surfactant is one or a mixture of at least two of triton x-100, PEG6000, CHAPS, THESIT, tween 20, PEG8000, emulgen B-66 and Emulgen A90.
7. The robust triglyceride kit of claim 6, wherein: the preservative is one or a mixture of at least two of PC300, PC950, sodium azide, merthiolate and sodium benzoate.
8. The robust triglyceride kit of claim 7, wherein: the concentration of triton-100 is 2g/L.
9. The robust triglyceride kit of claim 6, wherein: the PC950 concentration was 0.8ml/L.
10. A method for preparing a stable triglyceride kit comprising the kit of any one of claims 1-9, comprising the steps of:
s1, preparing a first reagent: weighing triton x-100 with a 100ml beaker, adding water, heating and dissolving on an electromagnetic oven, and naturally cooling at 18-40 ℃ for standby;
adding 800ml of water into a 1L beaker, weighing PIPES and NaOH, putting into water, stirring for dissolution, adjusting the pH value to 7.00+/-0.05, and sequentially weighing 4-chlorophenol and MgCl 2 Dissolving ascorbate oxidase, proclin950 and BSA in the above solution, stirring to dissolve, adding cooled triton-100 into the solution, fixing the volume to 1L, and uniformly mixing to obtain a first reagent;
s2, preparing a second reagent: adding 800ml of water into a 1L beaker, putting PIPES and NaOH into the water, stirring and dissolving, adjusting the pH value to 7.00+/-0.05, sequentially measuring ATP, BSA, sucrose, LPL, GK, GPO, POD, 4-AAP, EDTA, proclin950 and BSA, dissolving in the above solutions, stirring and dissolving, fixing the volume to 1L, and uniformly mixing to obtain a second reagent.
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Publication number Priority date Publication date Assignee Title
JP2001252096A (en) * 2000-03-13 2001-09-18 Internatl Reagents Corp Reagent for assaying triglyceride
CN104498586A (en) * 2014-11-28 2015-04-08 山东博科生物产业有限公司 Single reagent serum triglyceride detection reagent with strong stability
CN106399460A (en) * 2016-09-29 2017-02-15 四川迈克生物科技股份有限公司 Kit and method for determining triglyceride
CN108467882A (en) * 2018-03-30 2018-08-31 潍坊市康华生物技术有限公司 A kind of triglyceride detection kit
CN108949903A (en) * 2017-05-17 2018-12-07 广州市伊川生物科技有限公司 A kind of triglyceride determination kit and its measuring method
CN111235222A (en) * 2020-01-17 2020-06-05 上海高踪医疗器械科技有限公司 Triglyceride detect reagent box

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001252096A (en) * 2000-03-13 2001-09-18 Internatl Reagents Corp Reagent for assaying triglyceride
CN104498586A (en) * 2014-11-28 2015-04-08 山东博科生物产业有限公司 Single reagent serum triglyceride detection reagent with strong stability
CN106399460A (en) * 2016-09-29 2017-02-15 四川迈克生物科技股份有限公司 Kit and method for determining triglyceride
CN108949903A (en) * 2017-05-17 2018-12-07 广州市伊川生物科技有限公司 A kind of triglyceride determination kit and its measuring method
CN108467882A (en) * 2018-03-30 2018-08-31 潍坊市康华生物技术有限公司 A kind of triglyceride detection kit
CN111235222A (en) * 2020-01-17 2020-06-05 上海高踪医疗器械科技有限公司 Triglyceride detect reagent box

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