KR20230122812A - Simultaneous determination method of phthalate in foods using gas chromatography and mass spectrometry - Google Patents

Abstract

The present invention relates to a method for simultaneous analysis of multi-component phthalates in food, and more specifically, the steps of extracting an analysis target and a first extraction solvent by stirring and allowing them to stand, forming a first extraction solvent layer after stationary; Separating the first extraction solvent layer and concentrating with nitrogen; preparing a measurement solution by adding a second extraction solvent to the nitrogen-enriched residue; and measuring and analyzing phthalates by introducing the measurement solution into a gas chromatography and mass spectrometer. A method for simultaneous analysis of multi-component phthalates in food, wherein the first extraction solvent includes a polar solvent, a non-polar solvent, or both, and the second extraction solvent includes a non-polar solvent. will be. In addition, it is further related to a simultaneous analysis system of multi-component phthalates in food using the above simultaneous analysis method.

Description

Simultaneous analysis method of multi-component phthalate contained in food using gas chromatography and mass spectrometry

The present invention relates to a method for simultaneous analysis of multi-component phthalates contained in food.

Phthalate is a plasticizer for giving flexibility and elasticity to plastics, and is widely used in various products such as household goods, industrial goods, medical supplies, food utensils, containers and packaging materials. However, since phthalates are known to cause endocrine disruptors (disorders) and carcinogenesis, as well as reproductive toxicity and hepatotoxicity, their use is regulated in some products. Exposure to the human body occurs through various routes, such as contact with products containing phthalate in daily life or inhalation of air. There is a need for a method capable of detecting the degree of exposure efficiently and with high sensitivity.

As a general method for analyzing components contained in current formulations, chromatography is used. Chromatography uses various solids or liquids as a stationary phase, and gas (eg gas chromatography) graph) or liquid as a mobile phase, and a sample is introduced into the mobile phase while allowing the mobile phase to pass through the stationary phase, and the difference in adsorption or distribution coefficient of the sample between the mobile phase and the stationary phase is used to separate the sample into components refers to the method

Since chromatography is fundamentally based on separation using the physical properties of the sample to be analyzed, that is, the partition coefficient between the stationary and mobile phases, molecular weight, and ionicity, the optimal separation and analysis conditions of the desired sample are uniformly defined or cannot be determined Depending on the target sample, various analysis factors, such as the selection of stationary and mobile phases, the flow rate of the mobile phase, and the selection of a detector that detects the separated and eluted sample, must be determined through interaction. In other words, since the determination of elements in this analysis method is important that can influence the analysis result, the optimal analysis method by chromatography must be selected individually for each substance, and the items according to analysis are limited, so it is not suitable for food mixed with various matrices. It is difficult to apply the sample pretreatment process including the extraction solvent. Accordingly, there is a need for a rapid and accurate analysis method capable of optimizing conditions for analyzing multi-component phthalate and increasing detection sensitivity and accuracy through an analyzer such as gas chromatography.

The present invention, in order to solve the above-mentioned problems, to analyze through the pretreatment method of samples of various formulations (e.g., liquid samples, high-moisture solid samples, low-moisture solid samples, fatty samples, non-lipid samples) It is to provide a method for simultaneous analysis of multi-component phthalates in food, which can increase analytical sensitivity, enable accurate, high-sensitivity detection, and reduce analysis time by removing and optimizing the interfering factors of substances.

The present invention, by using the method for simultaneous analysis of multi-component phthalates in food according to the present invention, increases analytical sensitivity, enables accurate and highly sensitive detection, and simultaneously analyzes multi-component phthalates in food, which can reduce analysis time. to provide the system.

However, the problem to be solved by the present invention is not limited to those mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

According to one embodiment of the present invention, the analysis target and the first extraction solvent are stirred and left to form a first extraction solvent layer, forming a first extraction solvent layer after stationary; Separating the first extraction solvent layer and concentrating with nitrogen; preparing a measurement solution by adding a second extraction solvent to the nitrogen-enriched residue; and measuring and analyzing phthalates by introducing the measurement solution into a gas chromatography and mass spectrometer, wherein the first extraction solvent includes a polar solvent, a non-polar solvent, or both, and the second extraction The solvent relates to a method for simultaneous analysis of multi-component phthalates in food, including a non-polar solvent.

According to an embodiment of the present invention, the first extraction solvent and the second extraction solvent include at least one selected from the group consisting of methanol, acetonitrile, n-hexane-saturated acetonitrile, and n-hexane, respectively, An extraction process may be performed by stirring the analyte and the first extraction solvent.

According to an embodiment of the present invention, the first extraction solvent includes methanol and n-hexane, and the nitrogen-concentrating step is to separate the n-hexane layer of the first extraction solvent layer and concentrate it with nitrogen can

According to an embodiment of the present invention, the first extraction solvent includes n-hexane and n-hexane-saturated acetonitrile, and the nitrogen-concentrating step includes n-hexane-saturated acetonitrile in the first extraction solvent layer. It may be nitrogen enrichment by separating the layers.

According to an embodiment of the present invention, the first extraction solvent may include n-hexane, and the nitrogen-concentrating step may include separating the n-hexane layer of the first extraction solvent layer and enriching it with nitrogen.

According to one embodiment of the present invention, the phthalates are dibutyl phthalate (DBP: di-n-butyl phthalate), butylbenzyl phthalate (BBP: benzyl-n-butyl phthalate), diethylhexyl phthalate (DEHP: di -(2-ethylhexyl) phthalate), di-n-octyl phthalate (DNOP: di-n-octyl phthalate), diethylhexyl adipate (DEHA: di-(2-ethylhexyl) adipate), diisononyl phthalate (DINP : diisononyl phthalate) and diisodecyl phthalate (DIDP: diisodecyl phthalate).

According to one embodiment of the present invention, during the step of measuring the components of the phthalates, the transport rate of the sample in the gas chromatography in the measuring device equipped with gas chromatography and mass spectrometry is 0.5 mL / min or more, The temperature of the sample injection unit is 250 ° C to 330 ° C, and the oven temperature is heated from 100 ° C or higher to 250 ° C to 330 ° C at a heating rate of 5 ° C to 10 ° C / min, and the sample is separated within 30 to 40 minutes In the mass spectrometer, the separated sample may be ionized by passing through an interface of 250 ° C to 330 ° C.

According to one embodiment of the present invention, the quantitative limit of the phthalates may be 0.1 ug/mL or less.

According to one embodiment of the present invention, the preparation of the measurement sample; An analysis unit of a measurement sample including a gas chromatography and a mass spectrometer; and a data processing unit processing the data acquired by the analysis unit. It relates to a simultaneous analysis system for multi-component phthalates in food, using the simultaneous analysis method according to the present invention.

According to one embodiment of the present invention, the simultaneous analysis system may perform qualitative and quantitative analysis of multi-component phthalates in food.

The present invention provides a simultaneous analysis method for multi-component phthalates (e.g., 7 types of phthalates) contained in food by one-time analysis by gas chromatography and mass spectrometry. And, the simultaneous analysis method can be quickly and conveniently analyzed, and can provide environmental, economic and time benefits by reducing the use of harmful extraction solvents.

The present invention can provide a method for quickly and accurately monitoring the residual amount of phthalate in food being distributed by establishing a method for analyzing phthalate mixed in food, rather than a test method used for existing utensils, containers and packaging.

The present invention can provide a detection and quantitative analysis method capable of simultaneously detecting and quantifying multi-component phthalates (eg, 7 phthalate components) present in food quickly and conveniently with one analysis, and an analysis system using the same. .

1 shows the results of simultaneous analysis of multi-component phthalates by gas chromatography and mass spectrometry of measurement solution 1 prepared in Example of the present invention according to an embodiment of the present invention.
Figure 2 shows the results of simultaneous analysis of multi-component phthalates by gas chromatography and mass spectrometry in Measurement Solution 1 prepared in Example of the present invention according to another embodiment of the present invention.
Figure 3 shows the results of simultaneous analysis of multi-component phthalates by gas chromatography and mass spectrometry in Measurement Solution 1 prepared in Example of the present invention according to another embodiment of the present invention.
4a to 4g show calibration curves obtained from standard materials of 7 phthalates in an embodiment of the present invention, according to an embodiment of the present invention. That is, FIG. 4a is a calibration curve for di-n-butyl phthalate (DBP), FIG. 4b is a calibration curve for benzyl-n-butyl phthalate (BBP), and FIG. 4c is a calibration curve for diethylhexyl A calibration curve of adipate (DEHA: di- (2-ethylhexyl) adipate), Figure 4d is a calibration curve of diethylhexyl phthalate (DEHP: di- (2-ethylhexyl) phthalate), Figure 4e is a di-n- It is a calibration curve of di-n-octyl phthalate (DNOP), Figure 4f is a calibration curve of diisodecyl phthalate (DIDP), Figure 4g is a calibration curve of diisononyl phthalate (DINP: diisononyl phthalate) it's a curve
FIG. 4h shows the results of simultaneous analysis of seven phthalates detected by gas chromatography and mass spectrometry in a mixture of standard materials of seven phthalates according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. In describing the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the subject matter of the present invention, the detailed description will be omitted. In addition, the terms used in this specification are terms used to appropriately express preferred embodiments of the present invention, which may vary according to the intention of a user or operator or customs in the field to which the present invention belongs. Therefore, definitions of these terms will have to be made based on the content throughout this specification. Like reference numerals in each figure indicate like elements.

Throughout the specification, when a member is said to be located “on” another member, this includes not only a case where a member is in contact with another member, but also a case where another member exists between the two members.

Throughout the specification, when a certain component is said to "include", it means that it may further include other components rather than excluding other components.

Hereinafter, a method for simultaneous analysis of multi-component phthalates in food of the present invention and its utilization will be described in detail with reference to examples and drawings. However, the present invention is not limited to these examples and drawings.

The present invention relates to a method for simultaneous analysis of multi-component phthalates in food, and according to an embodiment of the present invention, preparing a measurement sample; And it may include the step of analyzing. According to an embodiment of the present invention, the step of preparing the measurement sample is performed by applying an extraction solvent to the analysis target (eg, food sample) and performing extraction and nitrogen concentration one or more times or more repeatedly to perform gas chromatography and It may be to prepare a measurement sample capable of analyzing multi-component phthalates using a mass spectrometer.

According to an embodiment of the present invention, the extraction solvent may include a polar solvent (eg, a water-soluble solvent), a non-polar solvent (eg, a non-aqueous aliphatic hydrocarbon-based solvent), or both, and the type of measurement sample and the following According to the preparation process mentioned above, it can be appropriately selected for carrying out the simultaneous analysis method of the present invention.

According to an embodiment of the present invention, the extraction solvent includes at least one solvent selected from the group consisting of methanol, acetonitrile, n-hexane saturated acetonitrile, and n-hexane, and by applying the extraction solvent, In the process of pre-treatment and analysis of samples for multi-component phthalates analysis, the amount of solvent used and time required can be reduced. preferably n-hexane; Alternatively, a mixture of n-hexane and at least one solvent selected from n-hexane-saturated acetonitrile, acetonitrile, and methanol is used as an extraction solvent, and after extraction, the n-hexane layer is separated, and after nitrogen concentration, it is dissolved in n-hexane and measured It can be prepared as a sample.

As an example of the present invention, the mixing ratio (v/v) of n-hexane to the rest of the solvent is 1:0.1 to 1:1; 1:0.5 to 1:1; or 1:0.8 to 1:1. When included within the above range, accuracy, reproducibility and sensitivity can be improved in the simultaneous analysis process of multi-component phthalates through the extraction process of the analyte including various matrices, formulations, and the like.

According to one embodiment of the present invention, the step of preparing the measurement sample, the analysis target and the first extraction solvent to stir and stand still to extract; Separating the first extraction solvent layer and concentrating with nitrogen; and preparing a measurement solution by adding a second extraction solvent to the nitrogen-enriched residue.

As an example of the present invention, in the extraction step, the analysis target is food, and the food can be applied without limitation as long as it can be consumed by humans and animals, for example, solids, liquids, gels, emulsions, It may be in the form of a slurry, and the like, and may be, for example, tissues and/or extracts of plants and animals, edible compounds, etc., but is not limited thereto. For example, it may be a liquid sample, a water-containing sample (eg, a water-containing solid sample), an oily sample, a non-fatty sample, and the like. For example, the analyte may be pulverized material collected from food.

As an example of the present invention, in the extraction step, the first extraction solvent may include at least one solvent selected from the group consisting of methanol, acetonitrile, n-hexane-saturated acetonitrile, and n-hexane. That is, the extraction process may be performed by stirring the first extraction solvent and the analyte and leaving them still.

As an example of the present invention, after stirring and standing in the extraction step, the layers may be separated according to the first extraction solvent (eg, water-soluble, water-insoluble), moisture contained in the analyte, solids, and the like. For example, a first extraction solvent layer is formed, and layers may be separated according to the first extraction solvent within the first extraction solvent layer.

As an example of the present invention, the step of separating the first extraction solvent layer and concentrating with nitrogen is to separate the first extraction solvent layer from the mixture of the first extraction solvent and the analyte and concentrate it with nitrogen after the extraction. , When the first extraction solvent layer forms two or more solvent layers, the desired layer may be separated.

As an example of the present invention, in the step of preparing a measurement solution by adding the second extraction solvent, the second extraction solvent is at least one selected from the group consisting of methanol, acetonitrile, n-hexane-saturated acetonitrile, and n-hexane The above solvent may be included, preferably n-hexane. For example, the residue in the measurement solution is 90 wt% or less; 80 wt% or less; 50 wt% or less; 30 wt% or less; or 0,1 wt% to 20 wt%.

According to an embodiment of the present invention, the step of preparing the measurement sample is the step of extracting by stirring and standing still the analyte and the first extraction solvent containing methanol and n-hexane, the first extraction in the extraction step A solvent layer (methanol layer and n-hexane layer) can be formed. Separating the n-hexane layer of the first extraction solvent layer and concentrating with nitrogen; and preparing a measurement solution by adding n-hexane to the nitrogen-enriched residue. For example, the analyte may be a liquid sample and a solid sample having a lot of moisture.

According to an embodiment of the present invention, the step of preparing the measurement sample is the step of extracting by stirring and standing still the analyte and the first extraction solvent containing n-hexane and n-hexane saturated acetonitrile, the extraction In the step, a first extraction solvent layer (n-hexane layer and n-hexane-saturated acetonitrile layer) may be formed. Separating the n-hexane-saturated acetonitrile layer of the first extraction solvent layer and concentrating with nitrogen; and preparing a measurement solution by adding n-hexane to the nitrogen-enriched residue. For example, the analyte may be a fatty sample.

According to an embodiment of the present invention, the step of preparing the measurement sample is the step of stirring and stationary extraction of the first extraction solvent containing the analyte and n-hexane, and the first extraction solvent layer in the extraction step. (Example: n-hexane layer) can be formed. Separating the n-hexane layer of the first extraction solvent layer and concentrating with nitrogen; and preparing a measurement solution by adding n-hexane to the nitrogen-enriched residue. For example, the analyte may be a water-free solid sample and a water-free non-fatty solid sample.

According to an embodiment of the present invention, the step of measuring and analyzing the phthalates is a detection quantification for detecting and quantifying the phthalates by introducing the measurement sample (eg, a measurement solution) into a chromatography and mass spectrometer in the analysis device. analysis can be made.

As an example of the present invention, the temperature of the sample injection part of the analysis device is 250 ℃ to 330 ℃; or 280 °C to 300 °C.

As an example of the present invention, the transport rate of the sample in the gas chromatography of the analyzer is 0.5 Ml/min or more; 0.8 or higher; 1 or more, and an inert gas can be used as a carrier gas.

As an example of the present invention, the oven temperature of the analyzer may be heated at a heating rate of 5 °C to 10 °C/min at 100 °C or higher, and the sample may be separated within 30 to 40 minutes. For example, the oven may correspond to a chromatography area. for example, 250 °C to 330 °C; 280 ° C to 320 ° C; Alternatively, it may be heated to 280 °C to 300 °C and maintained for 10 to 20 minutes. In this way, the sample can be separated and analyzed within 30 to 40 minutes.

As an example of the present invention, the separated sample in the mass spectrometer region of the analyzer is 250 ℃ to 330 ℃; 280 ° C to 320 ° C; Alternatively, it may be ionized by passing through the interface at 280 °C to 300 °C.

As an example of the present invention, the analysis device may use a device known in the art capable of chromatography and mass spectrometry, which is not specifically mentioned in this document.

According to one embodiment of the present invention, the phthalates are dibutyl phthalate (DBP: di-n-butyl phthalate), butylbenzyl phthalate (BBP: benzyl-n-butyl phthalate), diethylhexyl phthalate (DEHP: di -(2-ethylhexyl) phthalate), di-n-octyl phthalate (DNOP: di-n-octyl phthalate), diethylhexyl adipate (DEHA: di-(2-ethylhexyl) adipate), diisononyl phthalate (DINP : at least two selected from the group consisting of diisononyl phthalate) and diisodecyl phthalate (DIDP: diisodecyl phthalate); 3 or more species; Alternatively, five or more kinds may be included, and preferably all of the above-mentioned phthalates may be simultaneously analyzed.

According to an embodiment of the present invention, in the step of measuring the components of the phthalates, multi-component phthalates included in food can be simultaneously detected and quantitatively analyzed.

According to one embodiment of the present invention, the quantitative analysis may be performed using a calibration curve obtained from a standard solution of phthalates to be analyzed.

According to an embodiment of the present invention, the quantitative analysis process includes preparing a standard stock solution; Preparing a mixed standard solution; Preparing a standard solution for preparing a calibration curve; and obtaining a calibration curve by analyzing a standard solution for preparing a calibration curve using a gas chromatograph and a mass spectrometer. and performing quantitative analysis using the calibration curve. can include

As an example of the present invention, the step of preparing the standard stock solution is a step of preparing individual standard stock solutions of phthalate components set to be analyzed, for example, dibutyl phthalate (DBP: di-n-butyl phthalate), butyl benzyl Phthalate (BBP:benzyl-n-butyl phthalate), diethylhexylphthalate (DEHP:di-(2-ethylhexyl) phthalate), di-n-octylphthalate (DNOP:di-n-octyl phthalate), diethylhexyladi At least two phthalate components are selected from the group consisting of di-(2-ethylhexyl) adipate (DEHA), diisononyl phthalate (DINP), and diisodecyl phthalate (DIDP), and the selected Individual standard stock solutions may be prepared by dissolving individual standard forms of phthalate components in the second extraction solvent (eg, non-polar solvent) (eg, n-hexane). Two or more standard stock solutions may be prepared according to the number of selected phthalate components.

As an example of the present invention, in the step of preparing the mixed standard solution, a second extraction solvent (eg, a non-polar solvent) (eg, n-hexane) is added to each of the prepared standard stock solutions, and the individual mixed standard solutions are prepared can do.

As an example of the present invention, in the step of preparing the standard solution for preparing the calibration curve, a second extraction solvent (eg, non-polar solvent) (eg, n-hexane) is regularly applied to each of the prepared mixed standard solutions for individual calibration curves. A standard solution can be prepared. The concentration (ug/mL) of the standard solution for preparing the calibration curve is 0.01 to 1; 0.05 to 0.5; It may be 0.1 to 0.5.

As an example of the present invention, in the quantitative analysis using the calibration curve, each of the prepared standard solutions for preparing the calibration curve is injected into an analysis equipment and analyzed with a gas chromatograph and a mass spectrometer to obtain a calibration curve for each phthalate component. . For example, the calibration curve for each component may be obtained according to at least one of the measurement conditions (temperature, time, etc.), the concentration of the standard solution for preparing the calibration curve, and the solvent.

According to one embodiment of the present invention, the simultaneous analysis method of the multi-component phthalates, 0.1 ug / mL or less; 0.09 ug/mL or less; 0.07 ug/mL or less; or 0.02 ug/mL to 0.09 ug/mL; Or it may have a detection limit (eg, quantitative detection limit) of 0.07 ug/mL to 0.09 ug/mL. For example, DBP, BBP, DEHP, DNOP and DEHA are each 0.01 ug/mL to 0.05 ug/mL ug/mL; or 0.02 ug/mL to 0.03 ug/mL, for example, the detection limits of DINP and DIDP are, respectively, 0.06 to 0.09 ug/mL; or a limit of detection of 0.07 to 0.09 ug/mL.

The present invention relates to a system for simultaneous analysis of multi-component phthalates in food using the method for simultaneous analysis of multi-component phthalates in food according to the present invention. For example, the above-mentioned measurement solution may be applied to the simultaneous analysis system.

According to one embodiment of the present invention, the system for simultaneous analysis of multi-component phthalates in food includes a measurement sample preparation unit; An analysis unit of a measurement sample including a gas chromatography and a mass spectrometer; and a data processing unit that processes the data acquired by the sample analysis unit.

According to one embodiment of the present invention, the measurement sample preparation unit may prepare a measurement solution for analysis by taking a food sample to be analyzed and applying an extraction process and a concentration process using an extraction solvent according to the present invention.

According to an embodiment of the present invention, the analysis unit of the measurement sample introduces the prepared measurement solution into an analysis device and distributes it to a gas chromatography and mass spectrometer through a separation tube so that measurement can be made.

As an example of the present invention, the conveyance rate of the sample in the chromatography of the analysis device is 0.5 mL/min or more, the temperature of the sample injection part is 250 ° C to 330 ° C, and the oven temperature is 100 ° C or more to 250 ° C to 250 ° C warming up to 330 °C at a heating rate of 5 °C to 10 °C/min, for example, 250 °C to 330 °C; 280 ° C to 320 ° C; Alternatively, it may be heated to 280 °C to 300 °C and maintained for 10 to 20 minutes. In this way, the sample can be separated within 30 to 40 minutes. The mass spectrometer, the separated sample is 250 ℃ to 330 ℃; 280 ° C to 320 ° C; Alternatively, it may be ionized by passing through the interface at 280 °C to 300 °C.

According to an embodiment of the present invention, the data processing unit may process the data acquired by the sample analysis unit to output results of detection and quantitative analysis of multi-component phthalates in food.

Although described with reference to preferred embodiments of the present invention, the present invention is not limited thereto, and within the scope not departing from the spirit and scope of the present invention described in the following claims, detailed description of the invention and accompanying drawings Various modifications and variations of the present invention can be made.

Example

1. Sample pretreatment

Samples were taken from food from 3 types of remaining 7 types of phthalates and pretreated as follows to prepare a measurement solution for gas chromatography and mass spectrometry.

1) Measurement solution 1: Treatment of liquid samples and solid samples with a lot of moisture

Precisely take 1 g of sample in a 20 mL screw cap vial, add 5 mL of methanol, mix, add 5 mL of n-hexane, shake vigorously, and let stand. Then, the supernatant (n-hexane layer) was transferred to a tube for nitrogen concentration, and 5 mL of n-hexane was added to the lower layer (sample + methanol layer), vigorously shaken, and allowed to stand. Water was dissolved in 0.5 mL of n-hexane to prepare a test solution.

2) Measurement solution 2: Treatment of fatty samples

1 g of sample was precisely taken in a 20 mL screw cap vial, 5 mL of n-hexane was added, vigorously shaken, and then allowed to stand. The n-hexane layer was transferred to a separate 20 mL screw cap vial, and 5 mL of n-hexane saturated acetonitrile was added thereto, vigorously shaken, and allowed to stand. Transfer the lower layer (n-hexane-saturated acetonitrile) to a tube for nitrogen concentration, add 5 mL of n-hexane-saturated acetonitrile to the upper layer (n-hexane), shake it, and leave it to stand. Combine the lower layer with a nitrogen concentration tube and concentrate with nitrogen. was dissolved in 0.5 mL of n-hexane to make a test solution.

3) Measurement solution 3: treatment of solid sample (non-fat) without water

1 g of sample was precisely taken in a 20 mL screw cap vial, 5 mL of n-hexane was added, vigorously shaken, and then allowed to stand. Transfer the n-hexane layer to a tube for nitrogen concentration, add 5 mL of n-hexane to the residue, shake vigorously, and stand still. Combine the n-hexane layer with a tube for nitrogen concentration and concentrate the residue with 0.5 mL of n-hexane. It was dissolved and used as a test solution.

2. Gas chromatography and mass spectrometry analysis

2-1)

Measuring solution 1, measuring solution 2 and measuring solution 3 prepared in the above sample pretreatment using an Agilent 7693A series injector for gas chromatography and mass spectrometry (Agilent 5977B mass selective detector directly connected to an Agilent 8890 gas chromatograph from Agilent Technologies). Each was aliquoted and 1 μl of the sample was injected. HB-5 mass (cross-connected 5% phenylmethylsilicon, length 30 m, inner diameter 0.25 mm, film thickness 0.25 um) was used as the separation tube, and ultra-high purity (99.999%) helium at a flow rate of 1 mL/min was used as the carrier gas. was used. The temperature of the inlet was 260 °C, and a splitless injection method was used. Oven conditions were set to warm from 120 °C to 280 °C. The electron energy used for ionization was 70 eV, and a method (SIM mode) was used to select and detect only a few specific ions among the analyte materials. GC-MS operating conditions are summarized in Table 1 below.

1 shows the detection results of multi-component phthalates in Measurement Solution 1.

Gas chromatograph Column HB-5MS UI (30 m × 0.25 mm × 0.25 um) Flow rate 1 mL/min, He Injection vol. 1ul, splitless Injection temp. 260℃ Oven temp. 120 ℃ (2 min) → 10 ℃/min → 280 ℃ (15 min) Mass spectrometer Interface temp. 280℃ Ionization voltage 70eV Analyzer Quadrupole Selective Ion Monitoring mode Compound Selected ion DBP 149, 223, 205 BBP 149, 206, 91 DEHA 129, 147, 112 DEHP 149, 167, 279 DNOP 149, 279, 261 DINP 149, 293, 127 DIDP 149, 307, 167

2-2)

Divi-1701 (cross-linked 14% cyanopropylphenyl-86% methylpolysiloxane, length 30 m, inner diameter 0.25 mm, film thickness 0.25 um) was used as the separation tube for gas chromatography and mass spectrometry. Ultra-pure (99.999%) helium at a flow rate of mL/min was used. The temperature of the inlet was 240 °C, and a splitless injection method was used. The oven temperature was set at 33 minutes by raising the temperature from 120 ° C (2 min) to 280 ° C by 10 ° C / min. The results are shown in FIG. 2 as the detection results of multi-component phthalates in Measurement Solution 2.

2-3)

HB-5 mass (cross-connected 5% phenylmethylsilicone, length 30 m, inner diameter 0.25 mm, film thickness 0.25 um) was used as a separation tube for gas chromatography and mass spectrometry. As a carrier gas, a flow rate of 1 mL/min was used. Ultra-high purity (99.999%) helium was used. The temperature of the inlet was 300 °C, and a splitless injection method was used. The oven conditions were set at 100 ° C to rise at 15 ° C per minute to reach 200 ° C, heated again at 5 ° C per minute to 270 ° C, and maintained for 15 minutes so that the analyte components could be separated in 38 minutes. As a result, it is shown in FIG. 3 as the detection result of multi-component phthalates in Measurement Solution 3.

3. Creation of calibration curve

A calibration curve was prepared by selecting concentrations of 0.1, 0.5, 1, 2, and 3 ug/mL of the mixed solution of seven standard substances of phthalate. Quantitative analysis was performed using a dilution method without an extraction process, and all calibration curves showed linearity (R ² > 0.99 or more) within the quantitative range. The results are shown in Figures 4a to 4g. In FIG. 4h, it can be confirmed that all 7 types of phthalates were simultaneously detected in the mixed solution of 7 types of standard substances.

The detection and quantification limit of the analysis method is the standard deviation of the y-intercept multiplied by 3.3 times the average value of the slope divided by the average value of the slope using a calibration curve obtained by analyzing the standard solution three times. The divided value was set as the limit of quantification. The limit of quantification of each analyte of DBP, BBP, DEHP, DNOP, and DEHA was 0.02 ug/mL, and the limit of quantification of DINP and DIDP, which appeared in a broad peak shape when analyzed as a standard material, was 0.07 ~ 0.09 ug/mL. mL level was shown.

4. Verification of validity

The recovery rate was evaluated by selecting concentrations of 0.05 ug/mL and 0.3 ug/mL for the 7 kinds of phthalate standard mixture solutions. In addition, in the case of precision, analysis was performed by different analysts and analysis dates for analysis reproducibility test. The recovery rate and precision evaluation results are shown in Table 2.

division 0.05ug/mL 0.3ug/mL LOD
(ug/mL) LOQ
(ug/mL) Recovery rate (%) RSD (%) Recovery rate (%) RSD (%) DBP 77.0 6.64 87.1 7.04 0.009 0.02 BBP 90.4 0.94 90.3 6.97 0.012 0.03 DEHP 77.0 7.94 99.7 6.78 0.010 0.03 DNOP 73.6 7.33 102.9 6.73 0.010 0.02 DEHA 110.5 8.99 89.2 6.44 0.013 0.03 DINP 100.4 0.94 90.3 8.10 0.032 0.09 DIDP 87.1 4.28 102.9 7.67 0.025 0.08

In Table 2, the recovery rate is to evaluate the degree of recovery by measuring the ratio of the result obtained through the extraction process to the result obtained without the extraction process. ~ 91.4%, DEHP recovery 70.0 ~ 81.2%, DNOP recovery 70.0 ~ 79.8%, DEHA recovery 100 ~ 119.8%, DINP recovery 99.8 ~ 101.5%, and DIDP recovery 84.8 ~ 91.4%. In addition, when 0.3 ug/mL was added, the recovery rate of DBP was 80.1 ~ 91.1 %, the recovery rate of BBP was 83.0 ~ 94.4 %, the recovery rate of DEHP was 91.9 ~ 104.2 %, the recovery rate of DNOP was 94.9 ~ 107.4 %, the recovery rate of DEHA was 83.0 ~ 94.4 %, DINP The recovery rate was 83.0 ~ 97.7% and the DIDP recovery rate was 94.9 ~ 110.7%.

In the case of precision in Table 2, the analysis was performed by changing the analyzer and the analysis date for the reproducibility test, and when 0.1 ppm was added, DBP was 5.28 to 5.48%, BBP was 2.72 to 7.64%, and DEHA was 2.98 to 6.25%. , DEHP ranged from 3.28 to 7.65%, DNOP ranged from 3.35 to 6.51%, DINP ranged from 3.75 to 6.36%, and DIDP ranged from 3.53 to 4.88.

Gas chromatography and mass spectrometry, which are used for component analysis of cosmetics or industrial products such as existing utensils and packaging containers, have limited analysis items, and in the analysis of foods mixed with various matrices (or formulations), including extraction solvents, Although there is a difficult problem in applying the sample pretreatment process, the method for simultaneous analysis of multi-component phthalates according to the present invention is a sample of various formulations (e.g., liquid samples, high-moisture solid samples, low-moisture solid samples, fatty samples, Non-fat samples), etc., to remove and optimize the interfering factors of the substance to be analyzed, thereby increasing the analytical sensitivity in the analysis of multi-component phthalates contained in food and providing a detection method capable of qualitative and quantification with accurate and high sensitivity. can do.

The present invention, by preparing a measurement solution by the extraction and dilution process using the extraction solvent according to the present invention, in food that can solve the high deviation of the result value, low accuracy, etc. A simultaneous analysis method for multi-component phthalates can be provided.

As described above, although the embodiments have been described with limited examples and drawings, those skilled in the art can make various modifications and variations from the above description. For example, even if the described techniques are performed in a different order from the described method, and/or the described components are combined or combined in a different form than the described method, or substituted or replaced by other components or equivalents. Appropriate results can be achieved. Therefore, other implementations, other embodiments, and equivalents of the claims are within the scope of the following claims.

Claims (10)

Stirring and stilling the analysis target and the first extraction solvent to extract, forming a first extraction solvent layer after standing;
Separating the first extraction solvent layer and concentrating with nitrogen;
preparing a measurement solution by adding a second extraction solvent to the nitrogen-enriched residue; and
measuring and analyzing phthalates by introducing the measurement solution into a gas chromatography and mass spectrometer;
including,
The first extraction solvent includes a polar solvent, a non-polar solvent, or both,
Wherein the second extraction solvent comprises a non-polar solvent,
Method for simultaneous analysis of multi-component phthalates in food.
According to claim 1,
The first extraction solvent and the second extraction solvent,
Each containing at least one selected from the group consisting of methanol, acetonitrile, n-hexane saturated acetonitrile and n-hexane,
Method for simultaneous analysis of multi-component phthalates in food.
According to claim 1,
The first extraction solvent includes methanol and n-hexane,
The step of nitrogen enrichment is to separate the n-hexane layer of the first extraction solvent layer and concentrate it with nitrogen,
Method for simultaneous analysis of multi-component phthalates in food.
According to claim 1,
The first extraction solvent includes n-hexane and n-hexane saturated acetonitrile,
The step of nitrogen concentration is to separate the n-hexane-saturated acetonitrile layer of the first extraction solvent layer and concentrate it with nitrogen,
Method for simultaneous analysis of multi-component phthalates in food.
According to claim 1,
The first extraction solvent includes n-hexane,
The step of nitrogen enrichment is to separate the n-hexane layer of the first extraction solvent layer and concentrate it with nitrogen,
Method for simultaneous analysis of multi-component phthalates in food.
According to claim 1,
The phthalates,
Dibutyl phthalate (DBP: di-n-butyl phthalate), butylbenzyl phthalate (BBP: benzyl-n-butyl phthalate), diethylhexyl phthalate (DEHP: di-(2-ethylhexyl) phthalate),
di-n-octyl phthalate (DNOP), di-(2-ethylhexyl) adipate (DEHA), diisononyl phthalate (DINP) and diisodecyl phthalate (DIDP: diisodecyl phthalate) containing two or more selected from the group consisting of,
Method for simultaneous analysis of multi-component phthalates in food.
According to claim 1,
In the measuring device equipped with gas chromatography and mass spectrometry during the step of measuring the components of the phthalates
In the gas chromatography, the transport rate of the sample is 0.5 mL/min or more, the temperature of the sample injection part is 250 °C to 330 °C, and the oven temperature is 5 °C to 10 °C from 100 °C or more to 250 °C to 330 °C. /min, and the sample is separated within 30 to 40 minutes,
In the mass spectrometer, the separated sample is ionized by passing through an interface of 250 ° C to 330 ° C,
Method for simultaneous analysis of multi-component phthalates in food.
According to claim 1,
The limit of quantification of the phthalates is 0.1 ug / mL or less,
Method for simultaneous analysis of multi-component phthalates in food.
preparation of the measurement sample;
An analysis unit of a measurement sample including a gas chromatography and a mass spectrometer; and
a data processing unit that processes the data acquired by the analysis unit;
including,
Using the simultaneous analysis method of any one of claims 1 to 8,
Simultaneous analysis system for multi-component phthalates in food.
According to claim 9,
The simultaneous analysis system,
Qualitative and quantitative analysis of multi-component phthalates in food,
Simultaneous analysis system for multi-component phthalates in food.