CN1247313A - Solid developer and its preparing process and usage - Google Patents
Solid developer and its preparing process and usage Download PDFInfo
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- CN1247313A CN1247313A CN 98117497 CN98117497A CN1247313A CN 1247313 A CN1247313 A CN 1247313A CN 98117497 CN98117497 CN 98117497 CN 98117497 A CN98117497 A CN 98117497A CN 1247313 A CN1247313 A CN 1247313A
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Abstract
A solid developer is prepared from developer (naphethydiamine hydrochloride and sulfaniline amide or sulfanilic acid) and carrier (PVC) through dissolving the developer in solvent and volatilizing the solvent while uniformly attaching the developer on carrier. The mass ratio of naphethydiamine hydrochloride to sulfaniline amide is (0.1-3) : 10. The mass ratio of PVC to developer is (1-15) : 1. Its advantages are stable and quick developing (5 min).
Description
The invention relates to a solid color developing agent and a preparation method thereof, in particular to a solid color developing agent for rapidly determining nitrite on site, a preparation method and a method for determining nitrite by using the solid color developing agent.
Nitrite is a highly toxic substance, can change hemoglobin into methemoglobin, loses oxygen delivery capacity and causes poisoning of human body, and can react with amine compounds inherent in food to form carcinogenic N-nitrosamine compounds. In the food hygiene standard, strict requirements are made on the nitrite content, and the nitrite content in the meat product is NaNO2Calculated by not more than 0.02 g/kg.
The existing nitrite is measured by a national standard method, and the method has the advantages of high sensitivity, good accuracy and the like, but has the defects of high reagent consumption, short color developing agent storage time, complex and time-consuming operation process, and is only suitable for laboratory operation.
The invention aims to provide a solid color developing agent, a manufacturing method thereof and a method for measuring nitrite by using the solid color developing agent, wherein the solid color developing agent has long and stable standing time, high color developing speed and wide color developing range and is suitable for field operation.
In order to achieve the purpose, the invention adopts the following design scheme: the solid color developing agent mainly comprises color developing agent naphthyl ethylenediamine hydrochloride, sulfanilamide or sulfanilic acid, and carrier PVC, wherein the weight ratio of naphthyl ethylenediamine hydrochloride to sulfanilamide is 0.1-3: 10, and the weight ratio of PVC to the total weight of the two color developing agents is 1-15: 1.
The preparation method of the solid developer comprises the following steps:
(1) weighing color-developing agent naphthyl ethylenediamine hydrochloride, sulfanilamide or sulfanilic acid at a ratio of 0.1-3: 10;
(2) adding a polar organic solvent which can dissolve the color developing agent but not the PVC, and completely dissolving the color developing agent;
(3) adding carrier PVC resin, and shaking uniformly, wherein the ratio of PVC to the total amount of the two color developing agents is 1-15: 1;
(4) evaporating, introducing nitrogen gas to volatilize the solvent to dryness, and obtaining the uniform solid color developing agent.
The method for determining nitrite by using the solid color developing agent comprises the following steps:
(1) weighing a sample, cutting, adding water, adding a NaOH solution, and fully oscillating and mixing;
(2) adding ZnSO47H2O, keeping boiling and oscillation during heating;
(3) addition of NH4Adding water into the AC buffer solution, and uniformly mixing;
(4) adding HCl into a centrifugal tube added with a solid color developing agent to adjust the pH value;
(5) filtering the mixed solution in the step (3), adding the filtered mixed solution into the centrifugal tube in the step (4), and closing a cover to shake;
(6) and (5) carrying out visual colorimetry on the result and the standard color gradation, and measuring the content.
The principle of the invention for measuring nitrite is as follows:
adding a uniformly mixed sample into a sodium hydroxide and zinc sulfate solution, and reacting according to the following formula:
due to the formation of Zn (OH)2The colloidal precipitate can adsorb fine particles in the solution well, can be used for removing protein, and can be heated to further remove protein and fat to obtain supernatant containing nitrite.
Adding ammonium acetate buffer solution into the supernatant, and under an acidic condition, carrying out diazo coupling reaction on the sulfanilamide nitrite and the N-1-naphthyl ethylenediamine hydrochloride toobtain reddish purple azo pigment, and comparing and quantifying with a standard color scale.
the invention is further illustrated by the following examples and figures.
FIG. 1 is a graph showing the effect of color development time on absorbance
FIG. 2 is a graph showing the effect of heating time on absorbance
FIG. 3 is a graph showing the effect of acidity on absorbance
FIG. 4 is a graph showing the effect of buffer dosage on absorbance
Example 1
Weighing a color-developing agent, namely naphthyl ethylenediamine hydrochloride, sulfanilamide or sulfanilic acid according to the mass ratio of 1: 10, placing the weighed color-developing agent in a round-bottom flask, adding a proper amount of organic solvent, namely acetonitrile, or adding a carrier PVC (polyethylene resin with the granularity of 80-100 meshes) which is 10 times of the color-developing agent after the color-developing agent is completely dissolved by using an organic solvent capable of dissolving the color-developing agent but not dissolving the PVC, shaking the mixture uniformly, treating the mixture by using a rotary evaporator, introducing nitrogen at the same time, volatilizing the solvent to be dry to form a uniform solid color-developing agent, storing the solid color-developing agent in a sealed manner, and storing the solid color-developing agent for a long time.
The following tests were carried out on the formulation of the developer: 5mL of nitrite standard solution of 2. mu.g/mL is taken out of the colorimetric tube, and the color developing agent is added according to the proportioning data of the following table (Table 1) and the absorbance is measured.
TABLE 1 formulation of N-1-naphthyl ethylenediamine hydrochloride and sulfanilamide
N-1 naphthyl ethylenediamine hydrochloride (g) | 0.1 | 0.02 | 0.05 | 0.10 | 0.15 | 0.20 | 0.30 |
Sulfanilamide or p-aminobenzene Sulfonic acid (g) | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 |
Absorbance (A) | 0.339 | 0.341 | 0.398 | 0.403 | 0.403 | 0.402 | 0.398 |
As can be seen from the above table, the absorbance obtained is the greatest when the ratio of N-1-naphthyl ethylenediamine hydrochloride to sulfanilamide is 1: 10.
In the ratio of the color-developing agent to the PVC, the following experiment was carried out by placing 5mL of a 2. mu.g/mL nitrite standard solution in a colorimetric tube, adding 1.0g of the color-developing agent and a predetermined amount of PVC, adjusting the acidity, and measuring the absorbance (see Table 2)
TABLE 2 ratio of developer to PVC
Developer (g) | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 |
PVC(g) | 1.0 | 2.0 | 5.0 | 6.0 | 7.0 | 8.0 | 10.0 | 15.0 |
Absorbance of the solution | 0.400 | 0.402 | 0.399 | 0.339 | 0.403 | 0.400 | 0.405 | 0.403 |
As can be seen from the above table, the absorbance is the greatest when the developer is mixed with PVC at a ratio of 1: 10.
Example 2
Taking about 1g of nitrite to be detected on site, putting the sample in a 15mL test tube with scales, cutting the sample by using a cutting rod, adding water to 5mL scales, shaking up, adding 0.5mol/L NaOH solution 1mL, and fully oscillatingMixing, adding 120g/L ZnSo47H2After 1mL of O solution is oscillated and matched, the O solution is heated on an alcohol lamp for 1.5min, the boiling phenomenon is kept when the O solution is heated, the O solution is oscillated and mixed constantly, 2mL of ammonium acetate buffer solution is added, water is added to the O solution to reach the scale of 15mL, the O solution and the ammonium acetate buffer solution are mixed evenly, hydrochloric acid (1+1) is added into 1mL of a plastic centrifuge tube into which 0.02g of solid color developing agent is added to reach the scale of 0.5mL, the mixture is filtered at 1mL of the plastic centrifuge tube by a filter, and the content of the O solution and a pre-prepared standard color scale are subjected to visual colorimetry after the O solution is closed.
When the sample to be detected is pretreated, the concentration of NaOH is controlled to be 0.3-1.0mol/L, the dosage of the NaOH is controlled to be 1.5-0.5mL, and the ZnSO4·7H2The concentration of O is 60-120g/L, the dosage is 2-1mL, and the heating time is 1.5-2 min.
In this experiment, the following selections were made for the solid developer amounts:
the measurement was carried out by weighing different amounts of the solid color developing agent into 12 10mL colorimetric tubes, adding 1mL (2. mu.g/mL) of the nitrite standard solution, adjusting pH to 2.5 with HCl (1+1), and measuring the amount of the nitrite standard solution, the data are shown in Table 3
TABLE 3 data table of solid developer dosage and absorbance
Reference numerals | Amount of developer g | Absorbance value |
1 | 0.0000 | 0.000 |
2 | 0.0010 | 0.051 |
3 | 0.0020 | 0.066 |
4 | 0.0049 | 0.066 |
5 | 0.0068 | 0.065 |
6 | 0.0095 | 0.063 |
7 | 0.0123 | 0.067 |
8 | 0.0166 | 0.069 |
9 | 0.0181 | 0.067 |
10 | 0.0286 | 0.071 |
11 | 0.0409 | 0.067 |
12 | 0.0525 | 0.070 |
As is clear from the table, 0.02g is preferred under the experimental conditions. If the amount of the powder developer is controlled between 0.01 and 0.04g, the nitrite content in the sample can be effectively measured, wherein the nitrite content in the sample is less than 0.06g/kg (the allowable value is 0.02g/kg according to the national standard and NaNO is used as the NaNO2Meter).
Selection of homogeneous and heterogeneous conditions:
in order to determine whether the sample is homogeneous or not, the following experiment was performed. After the samples are respectively weighed, the samples are not homogenized and are homogenized by an FS-2 type adjustable high-speed disperser, the operation is carried out according to the experimental steps, the absorbance of the samples is measured (six times of parallel measurement), the following table is obtained, and the two groups of data are subjected to t test.
TABLE 4 measurement data of homogeneity and inhomogeneity
Homogenizing | Is not homogeneous | |||
Reference numerals | NO2Concentration X1 | X12 | NO2Concentration X2 | X22 |
1 | 0.048 | 0.0023 | 0.049 | 0.0024 |
2 | 0.048 | 0.0023 | 0.048 | 0.0023 |
3 | 0.049 | 0.0024 | 0.047 | 0.0022 |
4 | 0.048 | 0.0023 | 0.049 | 0.0024 |
5 | 0.048 | 0.0023 | 0.48 | 0.0023 |
6 | 0.042 | 0.0018 | 0.047 | 0.0022 |
The data are statistically processed, and the two groups of data have no significant difference, and the measured result is not influenced by the uniformity and the nonuniformity, so that the sample is directly sampled and then cut in a test tube by a cutting rod during field measurement.
Determination of color development time:
since the experiment is based on the diazo coupling reaction, the degree of color development of the diazo coupling is greatly influenced by time. The time is too short, and the color reaction is not completely carried out; if the time is too long, the stability of the developer is lowered and the absorbance is lowered, so that it is necessary to select an appropriate color developing time. 1mL of 2.0. mu.g/mL nitrite standard use solution was taken in a 10mL colorimetric tube, and diazo coupling solid reagent was added thereto, the pH was adjusted to 2.5 with hydrochloric acid (1+1), and the mixture was left for 1 to 11min to conduct colorimetry, whereby Table 5 was obtained.
TABLE 5 Table of relationship between color development time and absorbance
Number of measurements | 1 | 2 | 3 | 5 | 7 | 9 | 11 |
1 | 0.253 | 0.254 | 0.254 | 0.254 | 0.255 | 0.254 | 0.254 |
2 | 0.240 | 0.241 | 0.241 | 0.241 | 0.241 | 0.241 | 0.241 |
3 | 0.254 | 0.255 | 0.255 | 0.255 | 0.256 | 0.256 | 0.255 |
As can be seen from Table 5 and FIG. 1, the absorbance values after 2min of the color development time were large and stable, and the measurement effect was good. When the nitrite content in the sample is less than 0.06g/kg, the absorbance values all reach the maximum value and are stable as long as the color development time exceeds 2 min.
Determination of heating time:
in the experiment, fat and precipitated protein are further separated by heating, and if the heating time is too short, the protein precipitator has incomplete action; the nitrite is easy to be oxidized and decomposed to reduce the concentration if the length is too long. Therefore, an experiment for an optimum heating time was conducted. After weighing and adding various reagents, heating on an alcohol lamp for 1.0, 1.5, 2.0, 3.0, 5.0 and 8.0min, and keeping other steps unchanged, and the colorimetric results are shown in the following table 6.
TABLE 6 relationship between heating time and absorbance
1.0 | 1.5 | 2.0 | 3.0 | 4.0 | 5.0 | 8.0 | |
1 | 0.240 | 0.261 | 0.261 | 0.252 | 0.250 | 0.549 | 0.226 |
2 | 0.241 | 0.260 | 0.260 | 0.250 | 0.250 | 0.248 | 0.221 |
3 | 0.009 | 0.012 | 0.011 | 0.009 | 0.008 | 0.009 | 0.006 |
When the data are plotted, it can be seen from FIG. 2 that the optimum heating time is 1.5-2.0min because the absorbance value is maximum at 1.5 min.
Conditions of acidity of the developer:
as can be seen from the formula of the color reaction based on the experimental principle, 2H is present in the diazotization+To participate in the reaction, i.e., to proceed under acidic conditions, in order to search for suitable acidic conditions, 2mL of nitrite standard solutions each having a concentration of 2. mu.g/mL were added to 7 10mL cuvettes previously charged with 0.02g of a chromogenic powder reagent, and hydrochloric acid (1+1) was added dropwise to the solutions at different pH values, thereby obtaining Table 7.
TABLE 7 relationship between pH and absorbance
Reference numerals | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
pH value | 0.5 | 1.0 | 1.5 | 2.5 | 3.5 | 4.0 | 7.0 |
Absorbance value | 0.167 | 0.165 | 0.170 | 0.210 | 0.206 | 0.173 | 0.068 |
The absorbance values were best plotted from the above data (FIG. 3) at pH 2.5.
When the nitrite content in the sample is less than 0.06g/kg, the acidity is controlled at pH2.5-3.0, which completely meets the experimental requirements, and if the nitrite content is more than 0.06g/kg, the sample can be diluted and then made.
Selection of the amount of buffer:
the buffer serves to stabilize acidity throughout the process, and therefore, its amount has some effect on the assay results. Weighing 10 parts of each 1.0g sample in a 15mL test tube, adding 5mL of water, adding 1mL of 0.5mol/L NaOH solution, shaking uniformly, mixing, adding 120g/L ZnSO4.7H2Mixing O1 mL, addingbuffer solution 0.00, 0.50, 1.00, 1.50, 2.00, 2.50, 3.50, 4.00 and 5.00mL, adding water to scale, and measuring absorbance value.
TABLE 10 correlation of buffer usage to absorbance values
Reference numerals | Amount of buffer (mL) | Absorbance (A) |
1 | 0.00 | 0.243 |
2 | 0.50 | 0.263 |
3 | 1.00 | 0.273 |
4 | 1.50 | 0.267 |
5 | 2.00 | 0.279 |
7 | 3.50 | 0.263 |
8 | 4.00 | 0.264 |
9 | 5.00 | 0.262 |
From the above graph (FIG. 4), it can be seen that the absorbance value is best when the amount of buffer is 2 mL. When the nitrite content in the sample is less than 0.06g/kg, the dosage of the buffer is 0.5-50 ml.
Example 3
The experimental conditions were the same as in example 2, the amount ofthe solid color developing agent added was 0.02g, and the liquid to be measured was visually evaluated by a spectrophotometer and a standard color gradation, which resulted in an absorbance value of 0.027, a content of 0.73. mu.g, and a light red-purple color gradation. The g/kg value can be obtained from the standard color scale.
Example 4
The conditions were the same as in example 3, the μ g value was 3.81 and the standard color scale was magenta.
Example 5
The conditions were the same as in example 3, the μ g value was 1.15 and the standard color scale was deep red-purple.
Compared with the prior national standard method, the invention has the following advantages:
the following are the data for the nitrite content measurements in the two samples.
Table 9 determination of nitrite content (μ g) in saloon sausages.
Reference numerals | 1 | 2 | 3 | 4 | 5 | 6 |
The invention | 9.2436 | 9.0894 | 9.0123 | 9.4340 | 9.4340 | 9.5378 |
National standard law | 9.2858 | 9.2315 | 9.4480 | 8.9610 | 9.8295 | 9.9868 |
TABLE 10 determination of nitrite content in Songjiang children's intestines (μ g)
Reference numerals | 1 | 2 | 3 | 4 | 5 | 6 |
The invention | 8.1636 | 8.2007 | 8.9309 | 8.3350 | 8.8550 | 8.8550 |
National standard law | 8.8663 | 8.4128 | 8.5765 | 8.7988 | 8.3118 | 9.1236 |
The data are statistically processed, and the two methods have no significant difference when the t test P is more than 0.05.
The invention has the advantages that:
the invention uses N-1-naphthyl ethylenediamine hydrochloride, sulfanilamide, PVC to make solid color developing agent, the standing time is long, and stable (as long as closed, dark preservation), the color developing agent of the existing national standard method is 0.4% sulfanilamide acid solution and 0.2% N- (1-naphthyl) -ethylenediamine solution, unstable, after preserving for a week, must reformulate, only suitable for laboratory test, and the time consuming is long, the required reagent amount is large, the tedious operation, the solid color developing agent that the invention provides is because the formulation is rational, the color developing speed is fast, the color developing range is very wide, the reagent is miniaturized, the sample volume is small, the time consuming is short, when the national standard method destroys the hydrolytic action of enzyme, heat 15min on the water bath, while this method only needs to heat 1.5min on the alcohol lamp, the whole testing process only needs 5min, is suitable for on-the spot food hygiene monitoring and detection of nitrite food poisoning, in the invention, the coupling reaction is carried out after the nitrite and the sulfanilamide are diazotized, so that the nitrite decomposition caused by the interference of certain components in a sample such as ascorbic acid and the like can be reduced, and therefore, the method has high sensitivity and high accuracy in determination.
Claims (9)
1. A solid color developing agent is characterized by mainly comprising the following components in parts by weight: the color reagent is composed of naphthyl ethylenediamine hydrochloride, sulfanilamide or sulfanilic acid, and PVC carrier, wherein the weight ratio of naphthyl ethylenediamine hydrochloride to sulfanilamide is 0.1-3: 10, and the weight ratio of PVC to the total weight of two color reagents is 1-15: 1.
2. The solid developer according to claim 1, wherein the dosage ratio of the naphthyl ethylenediamine hydrochloride to the sulfanilamide amide is 1: 10; the ratio of the PVC to the total amount of the two color developing agents is 10: 1.
3. The preparation method of the solid color developing agent is characterized by comprising the following steps of:
(1) weighing color-developing agent naphthyl ethylenediamine hydrochloride, sulfanilamide or sulfanilic acid at a ratio of 0.1-3: 10;
(2) adding a polar organic solvent which can dissolve the color developing agent but not the PVC, and completely dissolving the color developing agent;
(3) adding carrier PVC resin, and shaking uniformly, wherein the ratio of PVC to the total amount of the two color developing agents is 1-15: 1;
(4) evaporating, introducing nitrogen gas to volatilize the solvent to dryness, and obtaining the uniform solid color developing agent.
4. The method for preparing a solid color developer according to claim 3, wherein the organic solvent is acetonitrile.
5. A method for preparing a solid developer according to claim 3, characterized in that: the dosage ratio of the naphthyl ethylenediamine hydrochloride to the sulfanilamide is 1: 10; the ratio of the total weight of the PVC and the two color developing agents is 10: 1.
6. A method for determining nitrite by using solid color developing agent is characterized in that; it comprises the following steps:
(1) weighing a sample, cutting, adding water, adding a NaOH solution, and fully oscillating and mixing;
(2) adding ZnSO47H2O, keeping boiling and oscillation during heating;
(3) addition of NH4Adding water into the AC buffer solution, and uniformly mixing;
(4) adding HCl into a centrifugal tube added with a solid color developing agent to adjust the pH value;
(5) filtering the mixed solution in the step (3), adding the filtered mixed solution into the centrifugal tube in the step (4), and closing a cover to shake;
(6) and (5) carrying out visual colorimetry on the result and the standard color gradation, and measuring the content.
7. The method for measuring nitrite using solid color developer according to claim 6, wherein: the amount of the added solid color developing agent is 0.01-0.04g, and the amount corresponds to that the content of nitrite is less than 0.06g/kg, and the color developing time is more than 2 min; the pH value range is 2.5-3.0.
8. The method for measuring nitrite using solid color developer according to claim 6, wherein the method comprisesIn the following steps: the concentration of NaOH added into the sample is 0.3-1.0mol/L, and the dosage is 1.5-0.5 mL; ZnSO4&7H2The concentration of O is 60-120g/L, the dosage is 2-1mL, and the heating time is 1.5-2 min.
9. The method for measuring nitrite using solid color developing reagent according to claim 6, wherein the buffer solution is ammonium acetate buffer solution, and the amount thereof is 0.5-5.0 mL.
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Cited By (5)
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CN1319711C (en) * | 2001-11-05 | 2007-06-06 | 日本碍子株式会社 | Honeycomb structural body forming ferrule, and method of manufacturing ferrule |
CN103234959A (en) * | 2013-04-17 | 2013-08-07 | 上海绿帝环保科技有限公司 | Water quality nitrate detection agent |
CN109632677A (en) * | 2019-02-20 | 2019-04-16 | 深圳锦秀大唐环保科技有限公司 | Absorbent and preparation method thereof for spectrophotometry measurement |
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CN113588569A (en) * | 2021-07-23 | 2021-11-02 | 湖南省药品检验研究院(湖南药用辅料检验检测中心) | Preparation method of colorimetric block for identifying dye intermediate and colorimetric method thereof |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
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JPS60233552A (en) * | 1984-05-02 | 1985-11-20 | Terumo Corp | Test piece |
JPS6130768A (en) * | 1984-07-23 | 1986-02-13 | Eiken Kagaku Kk | Composition for measuring nitrite |
JPH0623744B2 (en) * | 1986-06-20 | 1994-03-30 | テルモ株式会社 | Test piece for nitrite detection |
CN1082612A (en) * | 1992-07-09 | 1994-02-23 | 季岳军 | The measuring method of activity of super-oxide dismutase |
CN1040141C (en) * | 1993-04-29 | 1998-10-07 | 付广录 | Reagent paper for monitoring food sanitation and producing process thereof |
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CN1319711C (en) * | 2001-11-05 | 2007-06-06 | 日本碍子株式会社 | Honeycomb structural body forming ferrule, and method of manufacturing ferrule |
CN103234959A (en) * | 2013-04-17 | 2013-08-07 | 上海绿帝环保科技有限公司 | Water quality nitrate detection agent |
CN109632677A (en) * | 2019-02-20 | 2019-04-16 | 深圳锦秀大唐环保科技有限公司 | Absorbent and preparation method thereof for spectrophotometry measurement |
CN109632677B (en) * | 2019-02-20 | 2022-01-07 | 深圳锦秀大唐环保科技有限公司 | Absorbent for spectrophotometry determination and preparation method thereof |
CN112763068A (en) * | 2020-12-29 | 2021-05-07 | 广西贺州市科隆粉体有限公司 | Method for detecting chromaticity of calcium carbonate powder |
CN113588569A (en) * | 2021-07-23 | 2021-11-02 | 湖南省药品检验研究院(湖南药用辅料检验检测中心) | Preparation method of colorimetric block for identifying dye intermediate and colorimetric method thereof |
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