Migration detection method of organic antimicrobial agent isothiazolinone

Abstract

The present invention discloses a migration detection method for an organic antimicrobial agent isothiazolinone, wherein the target detection substance of the migration detection method is an isothiazolinone antimicrobial agent, and the migration detection method comprises the following steps: preparing a migration solution: preparing a migration solution for dissolving the target detection substance; preparing a solution to be tested: immersing the test substance in the migration solution, filtering to obtain a solution to be tested; and liquid chromatography-mass spectrometry analysis: using a liquid chromatography-mass spectrometry analysis method to analyze and detect the solution to be tested, and obtaining a chromatogram-mass spectrum. The present invention provides a novel migration detection method, which can help correctly evaluate the chemical safety of consumer products, so as to effectively reduce production and inspection costs.

Description

Technical Field

[0001] The present application relates to the field of chemical detection, and in particular to a migration detection method of isothiazolinone, an organic antimicrobial agent. Background Art

[0002] Isothiazolinone derivatives are a general term for a class of compounds containing thiazolinone rings. Due to their strong antibacterial ability, small application dosage, good solubility, low toxicity, low residue, and easy degradation, this class of compounds has become the mainstream of current research and development of preservatives and fungicides, and is widely used in antifouling coatings, industrial water, leather, cosmetics, papermaking, textiles and other fields. In recent years, there have been increasing reports on the sensitization problem of isothiazolinone, which has attracted widespread attention from the society. The European Union has organized risk assessment work many times, and some countries and regions have also adjusted their regulations on the requirements and restrictions on its use in cosmetics and toys.

[0003] But the content of isothiazolinone chemical substances in different types of consumer goods (such as food packaging paper, cosmetics, textiles, toys, leather, etc.) is currently measured by measuring its total content. However, in the daily use of some consumer goods, the chemical substances added in its production process will not all come into contact with the skin, and will only partially migrate to the skin surface or skin sweat to cause harm to the human body. And the blind setting of total content or total addition of chemical substances such as isothiazolinone will cause the restriction standards to be too strict, and increase the technology and production costs of related industries. Therefore, it is necessary to develop a new test method to help correctly evaluate the chemical safety of consumer goods. Summary of the invention

[0004] The main purpose of the invention is to provide a migration detection method for organic antimicrobial agent isothiazolinone to help correctly evaluate the chemical safety of consumer products.

[0005] The present invention provides a migration detection method for an organic antimicrobial agent isothiazolinone, wherein the target detection substance of the migration detection method is an isothiazolinone antimicrobial agent, and the migration detection method comprises the following steps:

[0006] Preparing migration solution: preparing migration solution for dissolving the target detection substance;

[0007] Prepare the test solution: immerse the test sample in the migration solution and oscillate, filter the filtrate to obtain the test solution, wherein the oscillation frequency is 30-150r / min;

[0008] Liquid chromatography-mass spectrometry analysis: The solution to be tested is analyzed and detected by liquid chromatography-mass spectrometry analysis to obtain a chromatogram-mass spectrogram. The detection conditions of the liquid chromatography-mass spectrometry include:

[0009] Mobile phase A was water, mobile phase B was methanol,

[0010] Gradient elution program: 0min, 90% A-10% B; in the time period of 0-2min, the proportion of mobile phase A is 20%-40%, and the proportion of mobile phase B is 60%-80%; in the time period of 2-3min, the proportion of mobile phase A is 0%-5%, and the proportion of mobile phase B is 95%-100%; in the time period of 3-9min, the proportion of mobile phase A is 0%-5%, and the proportion of mobile phase B is 95%-100%; in the time period of 9-10min, the proportion of mobile phase A is 90%, and the proportion of mobile phase B is 10%;

[0011] Wherein, at any time point in the elution procedure, the sum of the proportion of mobile phase A and the proportion of mobile phase B is 100%.

[0012] Further preferably, in the above gradient elution program: within a time period of 2-3 minutes, the ratio change of mobile phase A and mobile phase B is set to be a linear change.

[0013] In some embodiments, the detection conditions of the liquid chromatography-mass spectrometry further include:

[0014] The chromatographic column selected was a reverse phase liquid chromatography column C18;

[0015] Flow rate: 0.20~0.50mL / min;

[0016] Injection volume: 10-30 μL;

[0017] The mass spectrometer adopts electrospray ion source positive ion mode analysis, data acquisition adopts multiple reaction monitoring mode, the ion source temperature range is set at 200-700°C, the curtain gas pressure range is set at 10-50psi, the collision gas pressure range is set at 2-12psi, the nebulizer gas pressure range is set at 20-80psi, the auxiliary gas pressure range is set at 20-80psi, and the capillary voltage range in positive ion mode is set at 3500-5500V.

[0018] Further preferably, the flow rate range is selected between: 0.25 and 0.35 mL / min;

[0019] More preferably, the injection volume is selected in the range of 10 to 20 μL.

[0020] More preferably, a reverse phase liquid chromatography column C18 with specification parameters of (2.1-4.6) mm*(50-100) mm*(2.6-5.0) μm is selected.

[0021] In some embodiments, the detection conditions of the liquid chromatography-mass spectrometry also include: the column temperature is set at 20-35°C.

[0022] In some embodiments, the mass spectrometer is analyzed in positive ion mode of an electrospray ion source, and data is collected in multiple reaction monitoring mode, with an ion source temperature of 600°C, a curtain gas pressure of 20psi, a collision gas pressure of 12psi, a nebulizer gas pressure of 55psi, an auxiliary gas pressure of 45psi, and a positive ion mode capillary voltage of 5500V.

[0023] In some embodiments, the isothiazolinone antibacterial agent specifically includes 2-methyl-4-isothiazolin-3-one (MIT), 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT), and 1,2-benzisothiazolin-3-one (BIT).

[0024] In some embodiments, the test object is a fabric; further, the fabric can be a textile such as clothing, sports protective gear, or a hat.

[0025] In some embodiments, when the test article is a fabric, the migration fluid is artificial sweat, and the components of the artificial sweat include: water and sodium chloride, wherein water is a solvent; the components of the migration fluid also include at least one of L-histidine hydrochloride monohydrate, sodium dihydrogen phosphate dihydrate, disodium hydrogen phosphate dodecahydrate, lactic acid, and urea.

[0026] In some embodiments, the migration solution comprises: water, sodium chloride, lactic acid and urea.

[0027] In some embodiments, in the migration solution, the concentration of sodium chloride is in the range of 3-7 g / mL, the concentration of lactic acid is in the range of 0.74-1.5 g / mL, and the concentration of urea is in the range of 0.74-1.5 g / mL.

[0028] In some embodiments, the pH of the migration solution is any value in the range of 3.0-7.0.

[0029] In some embodiments, the pH of the migration solution is any value in the range of 4.2 to 6.0, for example, the pH of the migration solution is 4.5, 5.0, 5.5, 6.0, etc. More preferably, the pH of the migration solution is 4.2, 4.8, 5.4 or 6.0.

[0030] In some embodiments, the pH of the migration solution is in the range of 4.4 to 5.6.

[0031] In some embodiments, the preparation of the test solution further comprises a migration step: immersing the test sample in the migration solution for a period of time to allow the target detection substance in the test sample to migrate into the migration solution.

[0032] In some embodiments, the migration step specifically includes: processing the test object into fragments, immersing the fragmented test object in the migration solution, and performing an oscillation operation at the same time; and obtaining the test solution after filtering.

[0033] In some embodiments, the oscillation frequency is set to 40-120 r / min, the oscillation time is 30-120 min, and the oscillation temperature is 20-40°C.

[0034] In some embodiments, the oscillation frequency is set to 85-95 r / min, the oscillation time is 60-120 min, and the oscillation temperature is 35-39°C.

[0035] Compared with the prior art, the technical solution of the present invention:

[0036] ① Using the migration value instead of the total content value can more accurately evaluate the chemical safety of the substance to be tested and fill the gap of the lack of migration detection method for isothiazolinone antimicrobial agents.

[0037] ② Using cheap and less toxic methanol-water as the mobile phase, the detection and separation effect is good and the chromatographic peaks are sharp.

[0038] ③ The migration detection conditions of liquid chromatography-mass spectrometry which are easy to apply and have excellent separation effect are proposed, especially a simple and feasible gradient elution procedure is proposed. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 The liquid chromatogram of the standard solution obtained by the migration detection method of the present invention (the concentration of MIT, CMIT, and BIT is 0.5 mg / L);

[0040] Figure 2 Comparison of the migration results of migration solution A and migration solution B on textile samples;

[0041] Figure 3 Comparison of the migration effects of different migration solution pH values ​​on textile samples;

[0042] Figure 4 Comparison of the migration effects of different oscillation frequencies on textile samples;

[0043] Figure 5 Comparison of the migration effects of different migration times on textile samples;

[0044] Figure 6 This is the liquid chromatogram of the real sample black sweatshirt;

[0045] Figure 7 This is the liquid chromatogram of the real sample black helmet strap.

[0046] The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0047] The scheme in the embodiment of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiment of the present invention. Obviously, the described embodiment is only a part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0048] It should be noted that the technical solutions between the various embodiments of the present invention can be combined with each other, but it must be based on the fact that ordinary technicians in the field can implement it. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0049] The present invention provides a migration detection method for an organic antimicrobial agent isothiazolinone, which comprises the following steps:

[0050] (I) Preparation of migration solution:

[0051] In some embodiments, sodium chloride, lactic acid, and urea are prepared into an initial migration solution; and then the pH of the initial migration solution is adjusted to 3-7 with dilute ammonia water to obtain a migration solution.

[0052] In some embodiments, L-histidine hydrochloride monohydrate, sodium chloride, and sodium dihydrogen phosphate dihydrate are prepared into an initial migration solution; and then the pH of the initial migration solution is adjusted to 3-7 with dilute ammonia water to obtain a migration solution.

[0053] In some embodiments, L-histidine hydrochloride monohydrate, sodium chloride, and disodium hydrogen phosphate dodecahydrate are prepared into an initial migration solution; and then the pH of the initial migration solution is adjusted to 3-7 with dilute ammonia water to obtain a migration solution.

[0054] Typically, ammonia water is used as the pH regulator of the migration solution. This is because the human body excretes nitrogen-containing wastes such as urea during exercise. In order to avoid introducing other heteroatoms, dilute ammonia water is preferred. When using a migration solution containing lactic acid, sodium hydroxide is not recommended as a pH regulator because sodium hydroxide reacts with lactic acid, destroying the composition of the migration solution and possibly affecting the accuracy of the test results.

[0055] (ii) Migration:

[0056] In some embodiments, a migration step is proposed to simulate the actual migration process between the fabric and the human body.

[0057] Take the textile to be tested and cut it into pieces, place the obtained pieces in a container, and add migration liquid to soak the textile pieces to be tested.

[0058] The container soaked with the textile fragments to be tested is placed in a constant temperature oscillator, the oscillation frequency is set in the range of 30 to 150 r / min, and the oscillation migration extraction is carried out in the temperature range of 20 to 40° C. for 30 to 120 minutes to obtain an extract.

[0059] The extract is filtered through a membrane filter, and the obtained filtrate is the solution to be tested.

[0060] In some embodiments, the relevant parameters of the migration step may also be: the oscillation frequency is set to 85-95 r / min, the oscillation time is 60-120 min, and the oscillation temperature is 35-39° C.

[0061] The present application adds an oscillation operation to the migration step, which can more realistically simulate the actual migration process between the fabric and the human body. That is, the migration step of the present application fully considers the movement characteristics in the actual application environment. Therefore, the migration effect is closer to reality and has more reference value for the evaluation of the chemical safety of consumer products.

[0062] (III) High performance liquid chromatography-mass spectrometry analysis:

[0063] Since isothiazolinone antimicrobial agents are highly polar compounds and are easily soluble in water, they cannot be extracted into organic solvents after migrating into the migration solution. Therefore, a high performance liquid chromatography-mass spectrometry method directly entering the aqueous phase is used for determination.

[0064] Through experiments, it was found that the C18 chromatographic column with special bonding and end-capping technology has an excellent retention effect on isothiazolinone antibacterial agents with strong polarity. Therefore, the C18 chromatographic column is preferably used in the embodiments of the present application.

[0065] Further preferred are chromatographic columns having specification parameters within the range of (2.1-4.6) mm*(50-100) mm*(2.6-5.0) μm.

[0066] At the same time, it was found through experiments that when a chromatographic column with a thick column diameter is used, especially 4.6mm×50mm×2.7μm or equivalent, the chromatographic column has an excellent separation effect on the target detection substance. This is because the composition of the migration liquid in the embodiment of the present application is not a pure solvent. The use of a chromatographic column with a thick column diameter can effectively reduce the adverse effects of multiple ions in the migration liquid, such as unsatisfactory peak shape, salt ions blocking the chromatographic column causing increased column pressure, affecting the chromatographic column and even the life of the chromatographic instrument.

[0067] The reference working conditions for HPLC are as follows:

[0068] a) Chromatographic column: C18 chromatographic column (2.1~4.6)mm*(50~100)mm*(2.6~5.0)μm;

[0069] b) Column temperature 20~35℃;

[0070] c) Flow rate: 0.20-0.50 mL / min;

[0071] d) Injection volume: 10-30 μL;

[0072] e) mobile phase A is ultrapure water, mobile phase B is methanol; the gradient elution program is: 0 min, 90% A-10% B; in the 0-2 min time period, the proportion of mobile phase A is 20%-40%, and the proportion of mobile phase B is 60%-80%; in the 2-3 min time period, the proportion of mobile phase A is 0%-5%, and the proportion of mobile phase B is 95%-100%; in the 3-9 min time period, the proportion of mobile phase A is 0%-5%, and the proportion of mobile phase B is 95%-100%; in the 9-10 min time period, the proportion of mobile phase A is 90%, and the proportion of mobile phase B is 10%;

[0073] Wherein, at any time point in the elution procedure, the sum of the proportion of mobile phase A and the proportion of mobile phase B is 100%.

[0074] The mass spectrometer was analyzed in electrospray ion source positive ion mode, and data was acquired in multiple reaction monitoring mode. The ion source temperature range was set at 200-700°C, the curtain gas pressure range was set at 10-50psi, the collision gas pressure range was set at 2-12psi, the nebulizer gas pressure range was set at 20-80psi, the auxiliary gas pressure range was set at 20-80psi, and the capillary voltage range in positive ion mode was set at 3500-5500V.

[0075] The reference working conditions of the high performance liquid chromatography are further preferred as follows:

[0076] a) Chromatographic column: C18 chromatographic column (2.1~4.6)mm*(50~100)mm*(2.6-5.0)μm;

[0077] b) Column temperature 25~35℃;

[0078] c) Flow rate: 0.25-0.35 mL / min;

[0079] d) Injection volume: 10-20 μL;

[0080] e) Mobile phase A was ultrapure water, and mobile phase B was methanol; the gradient elution program was: 0 min, 90% A-10% B; 0-2 min, 40% A-60% B; 2-3 min, 0% A-100% B; 3-9 min, 0% A-100% B; 9-9.1 min, 90% A-10% B; 9.1-10 min, 90% A-10% B.

[0081] Among them, one or more 90% A and 10% B elution programs can be set within the 9-10 min time period, just to ensure the normal switching of the mobile phase ratio. For example: In some embodiments, the gradient elution program can also be: 0 min, 90% A-10% B; 0-2 min, 40% A-60% B; 2-3 min, 0% A-100% B; 3-9 min, 0% A-100% B; 9-10 min, 90% A-10% B.

[0082] It is further preferred that the mass spectrometer adopts electrospray ion source positive ion mode analysis, data acquisition adopts multiple reaction monitoring mode, ion source temperature is 600°C, air curtain gas pressure is 20psi, collision gas pressure is 12psi, nebulizer gas pressure is 55psi, auxiliary gas pressure is 45psi, and positive ion mode capillary voltage is 5500V. The mass spectrometer voltage conditions are shown in the following table:

[0083] Table 1. Compound monitoring ion pairs and related parameter settings

[0084]

[0085] (1) Matrix effect verification experiment:

[0086] The external standard method was used for quantification, therefore, the matrix effect was verified.

[0087] It should be noted that in the detection industry, the matrix refers to the components in the sample other than the analyte. Since the matrix may significantly interfere with the detection process of the target detection substance and affect the accuracy of the detection results, the industry calls these effects and interferences matrix effects. The most commonly used method to remove the matrix effect is to establish a calibration curve through a standard sample with a known analyte concentration while keeping the matrix in the sample as unchanged as possible. However, when the percentage value of the matrix effect is between 90% and 110%, it proves that the matrix effect of the matrix on the target detection substance is weak and can be ignored, and no special treatment of the matrix is ​​required.

[0088] Preparation of migration solution: 5 g of sodium chloride, 1 g of lactic acid, and 1 g of urea were prepared into 900 mL of initial migration solution; then the pH of the initial migration solution was adjusted to 6.0 with dilute ammonia water, and the volume was fixed to 1000 mL to obtain the migration solution.

[0089] Prepare mixed standard solution based on blank matrix: Take MIT, CMIT, BIT negative blank textile samples (9 in total) of three common sports protective gear lining materials: cotton, linen, and nylon. Based on the above migration method, use the migration solution to impregnate the three negative blank textile samples, and prepare a mixed standard solution A1 with a concentration of MIT, CMIT, and BIT of 0.05 mg / L and another mixed standard solution A2 with a concentration of MIT, CMIT, and BIT of 0.5 mg / L.

[0090] Preparation of standard solution: directly add liquid MIT, CMIT and BIT into the migration solution to prepare a standard solution B1 with a concentration of MIT, CMIT and BIT of 0.05 mg / L and another standard solution B2 with a concentration of MIT, CMIT and BIT of 0.5 mg / L.

[0091] The above mixed standard solution A1, mixed standard solution A2, standard solution B1 and standard solution B2 were tested on the machine, and the matrix effect results obtained are shown in Table 2 and Table 3, among which the chromatographic result of standard solution B2 is shown in Figure 1 shown.

[0092] Table 2. Matrix effects at 0.5 mg / L concentration

[0093]

[0094]

[0095] Table 3. Matrix effects at 0.05 mg / L concentration

[0096]

[0097] From the data in Tables 2 and 3, we can see that the matrix effects of the blank matrices of the three negative textile materials on the target detection substances are all between 90% and 110%, and the matrix effect of the textile materials is weak and can be ignored. The above data show that in actual daily testing, the migration solution can be used to prepare a mixed standard solution, which is convenient for quantitative calculation.

[0098] Among them, relative standard deviation (RSD) is usually used to express the precision of analytical test results.

[0099] The calculation formula for relative standard deviation is:

[0100]

[0101] Where S is the standard deviation (also expressed as SD), It is the average value of the peak area of ​​the target detection substance.

[0102] The standard deviation refers to the average of the distances that each data deviates from the mean. This value can reflect the degree of dispersion of a data set. Its calculation formula is:

[0103]

[0104] In this embodiment, x refers to the peak area of ​​the target detection substance.

[0105] (2) Detection limit and linear correlation experiment of the migration detection method of the present invention:

[0106] The above migration solution was used to prepare a mixed standard working solution of three isothiazolinones (MIT, CMIT, BIT) with a concentration level of 0.005-0.5 mg / L by stepwise dilution. Specifically, blank samples of three textile materials were first impregnated with the migration solution to perform migration, and then the obtained liquid was used to prepare a mixed standard working solution of isothiazolinones.

[0107] The mixed standard solution was then tested on a test machine, and the test results were listed in Table 4.

[0108] The results in Table 4 show that within the concentration range of 0.005 to 0.5 mg / L, the correlation coefficient R 2 The method is greater than or equal to 0.999; at the same time, the method calculates the instrument detection limit and quantitative limit of the three isothiazolinone substances by three times the signal-to-noise ratio (S / N=3) of the standard solution, and calculates and determines the method detection limit and method quantitative limit by comparing the matrix effect. The migration detection method proposed by the present invention has quantitative limits of 3μg / L, 0.7μg / L, and 0.7μg / L for MIT, CMIT, and BIT, respectively, and the quantitative limit is lower than 1 / 10 of the migration limit, and the accuracy and sensitivity meet the requirements.

[0109] Table 4. Linearity and detection limits of migration assays

[0110]

[0111] (3) Accuracy verification of the migration detection method of the present invention:

[0112] A certain amount of target detection substance is added to the blank matrix and used as the sample to be tested on the machine. The ratio of the actual detection result of the target detection substance to the actual amount of the added target detection substance is called the spike recovery rate, or the recovery rate of a certain target detection substance. Theoretically, when there is no error, the amount of target detection substance added can be measured, that is, the recovery rate should be 100%, but in practice, due to the existence of systematic errors and accidental errors, the recovery rate is not necessarily 100%. Therefore, the recovery rate is a statistic for evaluating the error size of the detection method.

[0113] In order to verify the accuracy of the high performance liquid chromatography-mass spectrometry analysis method adopted in the present invention, the recovery rate and relative standard deviation working curves of three different textiles were tested, including low (the concentrations of MIT, CMIT, and BIT were all 10 μg / L), medium (the concentrations of MIT, CMIT, and BIT were all 60 μg / L), and high (the concentrations of MIT, CMIT, and BIT were all 200 μg / L). The experimental results are shown in Tables 5 to 7 below.

[0114] Table 5. 10μg / L spike recovery

[0115]

[0116] Table 6. 60μg / L spike recovery

[0117]

[0118]

[0119] Table 7. 200μg / L spike recovery

[0120]

[0121] The validation method standard of chemical detection method is that when the recovery rate is between 80-120% and the relative standard deviation is within 10%, the detection method is reasonable. The above data show that the recovery rate and relative standard deviation of the target detection substances (MIT, CMIT, BIT in this example) meet the requirements under different textile materials and different spiked concentrations.

[0122] Among them, the recovery rates of MIT in the three materials were 87.2%-102.6%, with RSDs of 3.0%-8.8%; the recovery rates of CMIT in the three materials were 88.2%-113.1%, with RSDs of 2.6%-9.4%; and the recovery rates of BIT in the three materials were 86.7%-114.8%, with RSDs of 4.2%-8.3%.

[0123] The following are specific embodiments.

[0124] Example 1

[0125] Preparation of migration solution A: 900 mL of initial migration solution was prepared by mixing 5 g of sodium chloride, 1 g of L-histidine hydrochloride monohydrate, and 1 g of sodium dihydrogen phosphate dihydrate; the pH of the initial migration solution was then adjusted to 6.0 with dilute ammonia water, and the volume was fixed to 1000 mL to obtain the migration solution.

[0126] The measuring area is 10cm 2 The textile to be tested is cut into 1 cm2 Place the fragments in a stoppered glass bottle and add 10 mL of migration solution.

[0127] The stoppered glass bottle was placed in a constant temperature oscillator, the oscillation frequency was set to 90 r / min, and the oscillation migration extraction was performed at 37° C. for 60 min to obtain an extract.

[0128] The extract is filtered through a membrane filter, and the obtained filtrate is the solution to be tested.

[0129] Put the solution to be tested on the machine for testing. The reference working conditions of HPLC are as follows:

[0130] a) Chromatographic column: C18 chromatographic column (4.6mm×50mm×2.7μm) or equivalent;

[0131] b) Column temperature 30°C;

[0132] c) Flow rate: 0.3 mL / min;

[0133] d) Injection volume: 15 μL;

[0134] e) Mobile phase A was ultrapure water, and mobile phase B was methanol; the gradient elution program was: 0 min, 90% A-10% B; 0-2 min, 40% A-60% B; 2-3 min, 0% A-100% B; 3-9 min, 0% A-100% B; 9-9.1 min, 90% A-10% B; 9.1-10 min, 90% A-10% B.

[0135] The mass spectrometer voltage conditions are shown in Table 8 below:

[0136] Table 8. Compound monitoring ion pairs and related parameter settings

[0137]

[0138] Example 2

[0139] The only difference between Example 2 and Example 1 is that the preparation method of the migration solution B in Example 2 is: 5g sodium chloride, 1g lactic acid, and 1g urea are prepared into 900mL of the initial migration solution; then the pH of the initial migration solution is adjusted to 6.0 with dilute ammonia water, and the volume is fixed to 1000mL to obtain the migration solution B.

[0140] Example 3

[0141] The only difference between Example 3 and Example 1 is that the migration solution C in Example 3 is composed of water, sodium chloride, L-histidine hydrochloride monohydrate and disodium hydrogen phosphate dodecahydrate.

[0142] The textiles to be tested in Examples 1-3 are positive samples of three textile materials, cotton, linen and acrylic, containing MIT, CMIT and BIT. Each group of examples is repeated three times, and the average value is taken to calculate the corresponding migration amount. Among them, the composition of migration solution A and migration solution C is basically the same, so, Figure 2 The migration results of migration solution A and migration solution B are listed in FIG.

[0143] By analyzing Figure 2 From the results, we can see that the migration rules of the three textile materials are consistent. Among them, MIT has a better migration effect in migration solution A than in migration solution B, but CMIT and BIT have better migration effects in migration solution B than in migration solution A. Therefore, if the migration of three target detection substances is to be considered at the same time, migration solution B would be a better choice, and the composition of migration solution B is very close to the composition of real human sweat.

[0144] Embodiment 4-7

[0145] Based on the same method and parameters as in Example 2, Examples 4-7 respectively selected four pH values ​​of 4.2, 4.8, 5.4, and 6.0 for the migration solution to perform verification and compare the effects of different pH values ​​on the migration efficiency.

[0146] Similarly, positive samples of cotton, linen and acrylic textile materials containing MIT, CMIT and BIT were weighed in Examples 4-7, where each group of examples was repeated three times and the average value was taken for calculation. The corresponding migration results are shown in Figure 3 .

[0147] By analyzing Figure 3 The results show that the migration patterns of the three textile materials are consistent. Among them, the migration amount of MIT increases with the increase of the pH of the migration solution, reaching the maximum at pH = 5.4, and tends to be flat or even slightly decreased at pH = 6.0. The migration amount of CMIT and BIT increases with the increase of pH, and reaches the maximum at pH = 6.0.

[0148] Embodiment 8-11

[0149] Based on the same method and parameters as in Example 2, four oscillation frequencies of 40 r / min, 60 r / min, 90 r / min, and 120 r / min were selected in Examples 8-11, respectively.

[0150] Similarly, positive samples of cotton, linen and acrylic textile materials containing MIT, CMIT and BIT were weighed in Examples 8-11, where each group of examples was repeated three times and the average value was taken for calculation. The corresponding migration results are shown in Figure 4 .

[0151] By analyzing Figure 4 The results show that the migration rules of the three textile materials are consistent. Among them, the migration of MIT, CMIT, and BIT increases with the increase of frequency. At 40r / min to 60r / min, the increase of migration is the fastest and the slope is the highest; at 60r / min to 90r / min, the increase decreases and the slope decreases. At the same time, the migration data basically reaches the highest at 90r / min, and even if the oscillation frequency is increased, the migration amount remains basically stable.

[0152] Examples 12-15

[0153] Based on the substantially same method and parameters as in Example 2, four migration times of 30 min, 60 min, 90 min and 120 min were selected in Examples 12-15, respectively.

[0154] Similarly, positive samples of cotton, linen and acrylic textile materials containing MIT, CMIT and BIT were weighed in Examples 12-15, respectively. Each group of examples was repeated three times, and the average value was taken for calculation. The corresponding migration results are shown in Figure 5 .

[0155] By analyzing Figure 5 The results show that the migration patterns of the three textile materials are consistent. Among them, the migration of MIT, CMIT, and BIT increases with time. The increase is fastest and the slope is highest at 30min to 60min. The migration of most samples no longer increases at 60min to 120min. At the same time, if the migration time is too long, it will affect the detection efficiency and manufacturing production cost of daily large-scale samples.

[0156] Theoretically, the longer the migration time, the more conducive it is for substances in the sample to migrate into the migration solution. However, too long a migration time will bring unnecessary costs. Therefore, in order to control the detection cost, it is not advisable to set too long a migration time (in this embodiment, it refers to the oscillation time).

[0157] Example 16-Example 85

[0158] Based on the same method and parameters as in Example 2, the migration of MIT, CMIT and BIT was tested on 70 commercially available textile samples. Among them, 4 textile samples migrated out of the target detection substances, but none of them exceeded the specified limit, and the target detection substances were not detected in the remaining textile samples. This shows that the 70 commercially available textile samples surveyed all meet the requirements and are of good quality.

[0159] The migration results of the above four textile samples are listed in the table. The liquid chromatogram of the real sample black sweater can be found in Figure 6 , the liquid chromatography of the real sample black helmet band can be found in Figure 7 , combined with Figure 1 It can be seen from the standard spectrum in that the retention time of each target detection substance (referring to the three substances MIT, CMIT, and BIT) is basically consistent, which further proves that the migration detection method of the present invention is stable and consistent in the detection of the three substances MIT, CMIT, and BIT.

[0160] Table 9. Migration values ​​of target test substances of commercially available samples (mg / L)

[0161]

[0162]

[0163] In summary, the present invention provides a new migration detection method for organic antimicrobial agents, which uses migration value detection instead of total content value detection, can more accurately evaluate the chemical safety of the substance to be tested, and fills the gap in the migration test method of isothiazolinone antimicrobial agents. By detecting the migration amount of isothiazolinone antimicrobial agents in consumer products, combined with the critical sensitization amount of isothiazolinone antimicrobial agents causing human allergies, it can help countries or regions establish a safe use level assessment mechanism for isothiazolinone substances, effectively reduce product quality and safety risks, improve standard compliance, and reduce production and inspection costs.

[0164] At the same time, the migration detection method uses inexpensive and less toxic methanol-water as the mobile phase, and has good detection and separation effects. It can be seen from the chromatogram in the attached figure that the chromatographic peaks of the target detection substance are sharp and clear; in addition, the migration detection method proposes liquid chromatography-mass spectrometry detection conditions that are suitable for actual detection industry applications and have excellent separation effects, especially a simple and feasible gradient elution procedure.

[0165] The above description is only a partial or preferred embodiment of the present invention. Neither the text nor the drawings can limit the scope of protection of the present invention. All equivalent operation transformations made by using the contents of the present invention specification and drawings under the overall concept of the present invention, or direct / indirect application in other related technical fields are included in the scope of protection of the present invention.

Claims

1. A migration detection method for an organic antimicrobial agent isothiazolinone, characterized in that: The target detection substance of the detection method is an isothiazolinone antibacterial agent, and the detection method comprises the steps of: Preparing migration solution: preparing migration solution for dissolving the target detection substance; Prepare the test solution: immerse the test sample in the migration solution and oscillate, and obtain the test solution after filtering, wherein the oscillation frequency is 30-150r / min; Liquid chromatography-mass spectrometry analysis: The solution to be tested is analyzed and detected by liquid chromatography-mass spectrometry analysis to obtain a chromatogram-mass spectrogram. The detection conditions of the liquid chromatography-mass spectrometry include: Mobile phase A was water, mobile phase B was methanol, Gradient elution program: 0min, 90% A-10% B; in the time period of 0-2min, the proportion of mobile phase A is 20%-40%, and the proportion of mobile phase B is 60%-80%; in the time period of 2-3min, the proportion of mobile phase A is 0%-5%, and the proportion of mobile phase B is 95%-100%; in the time period of 3-9min, the proportion of mobile phase A is 0%-5%, and the proportion of mobile phase B is 95%-100%; in the time period of 9-10min, the proportion of mobile phase A is 90%, and the proportion of mobile phase B is 10%; wherein, at any time during the elution procedure, the sum of the proportion of mobile phase A and the proportion of mobile phase B is 100%; Chromatographic column: C18 chromatographic column, the specification parameters of C18 chromatographic column are (2.1~4.6)mm*(50~100)mm*(2.6~5.0)μm; The product to be tested is a textile, and the textile includes cotton, linen or acrylic; The isothiazolinone antibacterial agents include 2-methyl-4-isothiazoline-3-one, 5-chloro-2-methyl-4-isothiazoline-3-one and 1,2-benzisothiazolin-3-one; The components of the migration solution include: water, sodium chloride, lactic acid and urea; The pH of the migration solution is 3.0-7.

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2. The migration detection method according to claim 1, characterized in that The detection conditions of the liquid chromatography-mass spectrometry also include: Flow rate: 0.20-0.50mL / min; Injection volume: 10-30 μL; The mass spectrometer adopts electrospray ion source positive ion mode analysis, data acquisition adopts multiple reaction monitoring mode, the ion source temperature range is set at 200-700°C, the curtain gas pressure range is set at 10-50psi, the collision gas pressure range is set at 2-12psi, the nebulizer gas pressure range is set at 20-80psi, the auxiliary gas pressure range is set at 20-80psi, and the capillary voltage range in positive ion mode is set at 3500-5500V.

3. The migration detection method according to claim 1, characterized in that In the migration solution, the concentration range of sodium chloride is 3-7 g / mL, the concentration range of lactic acid is 0.74-1.5 g / mL, and the concentration range of urea is 0.74-1.5 g / mL.

4. The migration detection method according to claim 1 or 2, characterized in that: The preparation of the test solution also includes a migration step: immersing the test sample in the migration solution for 30 to 120 minutes and shaking the sample to allow the target detection substance in the test sample to migrate into the migration solution.

5. The migration detection method according to claim 4, characterized in that: The migration step includes: processing the sample to be tested into fragments, immersing the fragmented sample to be tested in the migration solution and shaking; and filtering to obtain the solution to be tested.

6. The migration detection method according to claim 1 or 2, characterized in that: The frequency of the oscillation is set to 40-120 r / min, and the temperature is set to 20-40°C.

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