KR101490179B1 - Method for simultaneous analysis of phthalate components using hplc and mass spectrometry - Google Patents

Abstract

The present invention provides a method for detecting whether phthalates are contained in various kinds of edible salt. The method comprises: an extraction step; Concentration step; And detecting and quantifying the phthalates by introducing the measurement solution into a liquid chromatography and mass spectrometer, and at the same time, qualitative and quantitative analysis of various kinds of phthalates can be performed. The above method can simultaneously analyze various types of phthalate components by one analysis, thereby reducing the use of harmful extraction solvents, thereby achieving economic and environment-friendly effects, and increasing the sensitivity and accuracy of detection using a mass spectrometer .

Description

TECHNICAL FIELD The present invention relates to a method for simultaneously analyzing various kinds of phthalate components contained in a salt by using a liquid chromatography and a mass spectrometer,

The present invention relates to a method for simultaneously analyzing phthalate components of various species, that is, 12 kinds of phthalates, using liquid chromatography and mass spectrometry. The phthalate components, which are endocrine disrupting substances contained in edible salt, The present invention relates to a method for simultaneously measuring and estimating the presence or amount of harmful substances in salt and simultaneously measuring international environmentally regulated substances that may be contained in seawater.

Phthalate grades have been used worldwide to provide flexibility and resilience to plastic products from 2.0 to 4.3 tonnes per year worldwide due to convenience and price effectiveness (Balure et al. 1999; Peijnenburg and Struijs 2006) . Since phthalate components, which are abundant in plastics, do not form covalent bonds with plastics, phthalate components are naturally leached extensively into the environment during the production and use of plastics (Kondyli et al., 1992; Wittassek et al. 2007).

Phthalates classification is evaluated as a substance that disturbs major contaminants and endocrine systems in the US Environmental Protection Agency (US-EPA 2009) or in international environmental pollutant regulatory agencies (EFSA 2004, ATSDR 2002, CSTEE 2004, and ECB 2004) et al. 2005; Carrillo et al. 2007; Wilkinson and Lamb 1999).

2006, Heudorf et al. 2007; Api, 2001), human beings are exposed to phthalate ingredients through building materials, clothing, cosmetics, perfumes, food packaging containers, toys and various plastic materials.

Diethyl phthalate, a kind of phthalates, is known to cause changes in hormone concentration and sexual development. Dibutyl phthalate is known as a reproductive toxin, and diisobutyl phthalate is known to be involved in male developmental changes in particular. In addition, diisodecyl phthalate, diisononyl phthalate, and the like are potentially prohibited from being used in products and child care products that can come into contact with children's mouths at the US Consumer Product Safety Commission. The phthalates mentioned above have been monitored periodically to provide data on human exposure assessment (Heudorf et al. 2007).

This large amount of phthalate components is in continuous use and will eventually reach the marine environment as the environmental circulation releases to the environment (Teil et al. 2006; Xie et al. 2005). Therefore, the salt produced from sea water is very likely to be contaminated by the phthalate components released in large quantities to the global environment, and there is a potential source of contamination to the phthalate components in the process of producing and manufacturing edible salt. Will be. Natural salt produced from sea water by evaporation in the wind and sun contains a large proportion of physical and chemical impurities.

Salt from sea water will improve the quality and shape of salt through cleaning, filtration and heating and will be used in industries for food processing, textile, detergent, dye and chemical applications. The smelted salt is used as salt for table salt and diet by adding food additives such as sodium silicoaluminate and potassium iodide. In addition, some edible salt is produced by baking at 800 ° C or higher or repeatedly melting. The degree of contamination of the phthalate components in the salt may vary depending on the manufacturing process of the salt.

Analysis of phthalate components in various types of salt can be misleading due to contamination of experimental equipment, plastic containers, air pollution in laboratory, etc. Therefore, it is difficult to analyze, and due to high noise level, (Fankhauser-Noti and Grob 2007; Prokupkova et al. 2002.) Liquid-liquid extraction methods, clean-up methods, and GC-TOF / MS techniques have been used to analyze phthalate components.

In the following description, it is intended to provide a method for confirming the contamination level of phthalate in salt and accurately qualitatively and quantitatively analyzing whether various kinds of phthalates are contained in the sample. The above method is intended to provide a quantitative analysis method capable of simultaneously analyzing a large number of, for example, 12 phthalate components simultaneously, quickly, easily and accurately.

In order to accomplish the above object, a method for detecting phthalates according to an embodiment of the present invention comprises stirring a sample containing an analyte, a polar solvent, an internal standard material, and an extraction solvent and allowing to stand to form an aqueous solution layer and an extraction solvent layer ; Concentrating the extraction solvent layer separated from the aqueous solution layer to prepare a measurement solution; And introducing the measurement solution into a liquid chromatography and mass spectrometer to detect and quantify phthalates.

The phthalates may be selected from the group consisting of diethyl phthalate (DEP), dipropyl phthalate (DPrP), di-n-butyl phthalate (DBP), diisobutyl phthalate Di-n-pentyl phthalate (DPP), dicyclohexyl phthalate (DCHP), di-n-hexyl phthalate (DHP), benzylbutyl phthalate At least one selected from the group consisting of diethylhexyl phthalate (DEHP), diisononyl phthalate (DINP), diisodecyl phthalate (DIDP) and diethylhexyl adipate (DEHA) And may be analyzing them simultaneously.

The object to be analyzed may be one containing salt.

The sample in the extraction step may further comprise an ionic strength increasing agent.

The ionic strength enhancer may include any one selected from the group consisting of sodium chloride, sodium carbonate, sodium hydroxide, hydrochloric acid, and combinations thereof.

The extraction solvent may be any one selected from the group consisting of hexane, dichloromethane, n-butanol, cyclohexane, chloroform, ether, and combinations thereof.

The liquid chromatography may further comprise an ion activator in the mobile phase solvent.

The ion activating agent may be any one selected from the group consisting of sodium acetate (soidum acetate), ammonium formate, ammonium acetate, formic acid, acetic acid, trifluoroacetic acid, and combinations thereof .

The concentration of the extraction solvent layer in the concentration step may include filtration with anhydrous sodium sulfate.

The internal standard is diethylhexyl phthalate -d _4, di-butyl phthalate -d _4, benzyl butyl phthalate -d _4, and diethylhexyl adipate may be composed of any one sulfate selected from the group consisting of -d _8.

The mobile phase solvent may be any one selected from the group consisting of acetonitrile, distilled water, methanol, isopropanol, ammonium formate, ammonium acetate, formic acid, acetic acid, trifluoroacetic acid, .

The subject to be analyzed may be a coarse salt, a refined salt, a roasted salt, or a seasoning containing any one selected from the above.

The mobile phase solvent may be 0.1% or less, or 0.01-20 mM, based on the whole mobile phase solvent.

Hereinafter, the present invention will be described in more detail.

Among the various types of salt sold for food, the degree of contamination of the phthalate components may vary depending on the type of salt and the manufacturing process. It provides LC / MS analytical methods for identifying the phthalate contamination levels in edible salt and quantifying and quantifying which phthalates are contained in the salt to be analyzed among various phthalates.

In the analysis method according to an embodiment of the present invention, the phthalates to be detected include phthalates and adiposes. The phthalate has a basic structure represented by the following formula (1), and the adipate has a structure represented by the following formula (2).

[Chemical Formula 1]

(2)

For example, R and R 'each independently represent a substituted or unsubstituted alkyl group, a cycloalkyl group, and an aryl group, which may be the same or different, May be any one selected from the group consisting of The phthalates include compounds represented by the above formula (1) and isomers thereof. These are a kind of additives mainly used in molding of plastics, and their functions are different depending on kinds of functional groups or esters, and their products are also different. There are currently more than 20 species known, and the species is expected to continue to increase in the future, and is known to have a deleterious effect on human body even in trace amounts.

Specifically, the phthalates which can be detected by the above-mentioned analysis method include diethyl phthalate (DEP), dipropyl phthalate (DPrP), di-n-butyl phthalate (DBP) Diisobutyl phthalate (DIBP), di-n-pentyl phthalate (DPP), dicyclohexyl phthalate (DCHP), di-n-hexyl phthalate Diethylhexyl phthalate (DEHP), diisononyl phthalate (DINP), diisodecyl phthalate (DIDP), and diethylhexyl adipate , DEHA).

The phthalate-based detection method described below provides a method capable of simultaneously detecting various types of phthalates listed above in a single detection and determination process, and quantifying the phthalates as effective analysis data.

The term " salt " in the present invention is used to mean a chemical salt such as salt extracted from raw salt of salt water or salt of salt, and salt which is chemically synthesized. Means a salt containing various salts having different thicknesses, properties, flavors, aromas and colors, and the food additives containing them may also be a salt to be analyzed according to the present invention.

The method for detecting phthalates includes an extraction step, a concentration step, and a detection quantitative step.

In the extracting step, a sample containing an analyte, a polar solvent, an internal standard substance, and an extraction solvent is stirred and allowed to stand to form an aqueous solution layer and an extraction solvent layer.

The analysis object includes commercially available salt or an edible product containing the salt, and includes a coarse salt, a refined salt, a roasted salt, or a seasoning containing any one selected from the above, wherein the phthalates are included in the analysis target If it is necessary to detect whether or not it exists, it is sufficient.

The polar solvent may be any solvent capable of dissolving the analyte. For example, distilled water, deionized water, methanol, formamide, formic acid, glycerin, propylene glycol, and the like may be used.

The internal standard substance is a substance which is used as a standard for quantitative analysis after analyzing a certain amount of the sample, for example, diethylhexyl phthalate-d ₄ , dibutyl phthalate-d ₄ , benzyl butyl phthalate-d ₄ And diethylhexyl adipate-d ₈ may be used, but the present invention is not limited thereto.

For example, the extraction solvent may be selected from the group consisting of hexane, dichloromethane, n-butanol, cyclohexane, chloroform, ether, and the like. The extracting solvent may be any solvent that can extract the phthalates, Or a combination thereof.

The sample containing the analyte, the polar solvent, the internal standard substance, and the extraction solvent may further include an ionic strength increasing agent. The ionic strength enhancer can increase the strength of ions in the sample to facilitate layer separation in the extraction step.

The ionic strength enhancer may be any one selected from the group consisting of sodium chloride, salt (organic substances removed), sodium carbonate, sodium hydroxide, tripro acetic acid, phosphoric acid, formic acid, boric acid, acetic acid, And preferably sodium chloride can be applied. The salt from which the organics have been removed may be salt baked at a temperature of 600 ° C or more for 8 hours or more.

The sample is vigorously stirred and then allowed to stand to form an aqueous solution layer and an extraction solvent layer. The aqueous solution layer is removed, and the extraction solvent layer is separated to carry out the concentration step.

The extraction solvent layer may be concentrated to prepare a measurement solution. The concentration may be applied as long as the extraction solvent layer can be concentrated. For example, the extraction solvent layer may be filtrated with anhydrous sodium sulfate. .

The detection quantification step is a step of introducing the measurement solution into a liquid chromatography and mass spectrometer to detect and quantify phthalates.

The liquid chromatography and mass spectrometer are not limited to devices of a specific model, and apparatuses capable of continuously performing phase separation and mass analysis by liquid gromatograpin can be applied.

The mobile phase solvent used in the liquid chromatography may be any one selected from the group consisting of acetonitrile, formic acid, distilled water, methanol, acetic acid, and combinations thereof. The mobile phase solvent may further comprise an ion activator have. In the course of detecting the measurement solution, the ion activator inhibits [M + H] ⁺ ions and strongly promotes the formation of [M + Na] ⁺ ions. For example, sodium acetate , Ammonium formate or ammonium acetate, formic acid, acetic acid, trifluoroacetic acid, and combinations thereof, and preferably sodium acetate may be used.

The content of the ion activator contained in the mobile phase solvent may be 0.1% or less based on the whole mobile phase solvent, and may be in the range of 0.01-20 mM. When the mobile phase solvent is used in such a range of contents, the effect of [M + H] ⁺ ions is suppressed and that of [M + Na] ⁺ ions is maximized.

In this way, it is possible to measure whether or not the phthalates are contained in the analysis target, and the type and content thereof, if any, and it is possible to simultaneously detect various kinds of phthalates. Particularly, it is preferable to use one or more of diethyl phthalate (DEP), dipropyl phthalate (DPrP), di-n-butyl phthalate (DBP), diisobutyl phthalate (DIBP) di-n-pentyl phthalate (DPP), dicyclohexyl phthalate (DCHP), di-n-hexyl phthalate (DHP), benzylbutyl phthalate (BBP), diethylhexyl phthalate it is possible to simultaneously detect various phthalates ranging from diethylhexyl phthalate (DEHP), diisononyl phthalate (DINP), diisodecyl phthalate (DIDP) and diethylhexyl adipate (DEHA) Thus, the detection result can be obtained quickly, simply and accurately with a single analysis.

The phthalate detection method is a very valid analysis method in terms of the degree of contamination of the phthalate component from the viewpoint of the US Environmental Protection Agency (EPA) recommended food standard (US-EPA 2009), and the pollution degree of the sea water It can be used as a scale.

The technical problems and the detection method to be achieved by the present invention are not limited to the phthalate component but can be applied to other materials having different molecular structure, molecular weight, solubility and polarity.

The phthalates detection method of the present invention can simultaneously analyze 12 phthalate components in one analysis, and can perform analysis quickly and easily. It is possible to obtain economical and environment-friendly effects by reducing the use of harmful extraction solvent, The analyzer can be used to increase detection sensitivity and accuracy.

1 is a graph showing changes in the detected molecular weights of [M + Na] ⁺ in a condition containing 0.05 mM sodium acetate (NaOAc, ion activator) under the condition of mobile phase solvent of LC / MS according to Example 1 of the present invention FIG.
2 is a graph showing changes in the detected molecular weights of [M + Na] ⁺ in the absence of 0.05 mM sodium acetate (NaOAc, ion activator) under LC / MS mobile phase solvent conditions according to Example 2 of the present invention It is a graph showing.
3 is a graph showing the chromatogram as a result of phthalate detection of mineral water used as a reference value in Examples 3 to 6 of the present invention.
4 is a graph showing a chromatogram as a result of phthalate detection of natural salt (coarse salt) according to Example 3 of the present invention.
FIG. 5 is a graph showing a chromatogram as a result of phthalate detection of purified salt according to Example 4 of the present invention. FIG.
6 is a graph showing a chromatogram as a result of phthalate detection of a salt containing a food additive according to Example 5 of the present invention.
7 is a graph showing a chromatogram as a result of phthalate detection of baked salt according to Example 6 of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Of the 36 commercially available edible salts sold in the Korean market, 12 phthalate components (diethyl phthalate (DEP), dipropyl phthalate (DPrP), di-n-butyl phthalate , Diisobutyl phthalate (DIBP), di-n-pentyl phthalate (DPP), dicyclohexyl phthalate (DCHP), di-n-hexyl phthalate , DHP, benzylbutyl phthalate (BBP), diethylhexyl phthalate (DEHP), diisononyl phthalate (DINP), diisodecyl phthalate (DIDP) and diethylhexyl Diethylhexyl adipate (DEHA)} was measured.

In the following measurement, liquid chromatography-mass spectrometry (LC-TOF / MS) was performed using liquid chromatograph, 1200 serise model of Agilent, USA, mass spectrometer of LECO of USA, Unique®HT TOFMS model. The analysis conditions are shown in Table 1 below.

LC , model name Agilent ^®  1200 series Column, Column 5 cm ZORBAX ^® EclipseXDB-C18column (4.6 mmi.d.and 1.8 μm particle diameter, Agilent Technologies, USA Flow rate 0.8 ml / min Injection volume 5 μl Mobile phase A, mobile phase A 0.1% formic acid Mobile phase B, mobile phase B acetonitrile in 0.05mM sodium acetate Isocratic run time 20 min TOF / MASS , model name LECO Unique ^® HT TOFMS Source, Source High Flow ESI source Voltage, Voltage (-) 2700 v Nebulizer Pressure 400 kPa Desolvation Temp. 200 ℃ Nebulizer Pressure 400 kPa Interface temp. 100 ℃ Acquisition rate 3.13 spectra / second parameter , variable Desolvation 7000 cc / min Desolvation temp. 400 ° C Electrospray -4500 v Nozzle -160 v skimmer -80 v Quad RF -260 v Einzel vertical deflect 0.6 v

Columns for liquid chromatography were a 5 cm ZORBAX® Eclipse XDB-C18 column (4.6 mm id and 1.8 μm particle diameter, Agilent Technologies, USA). The mobile phases were 90% acetone nitrile and 10% formic acid. The flow rate of the mobile phase was maintained at 0.8 ml / min for 20 min and the injection volume was 5 μL. The analytes separated from the detection and quantification step are set to be introduced into the mass spectrometer. At this time, the detected ions of the phthalate components are subjected to qualitative and quantitative analysis by selecting the characteristic ion [M + Na] ⁺ of the mass spectrum.

Quantitative analysis was performed by combining the areas of the peaks in the chromatograms, and then calculating the calibration curve using the internal standard method. Quantitative curves used concentrations of each phthalate in the range of 0.5-100 ng / g.

The quantitation ion, correlation coefficient, limit of detection, limit of quantitation, and relative standard deviation of analytes for qualitative and quantitative analysis are shown in Table 2 below .

No Phthalates
Phthalates ^A Quantitation ion
(m / z) Correlation coefficient ^B
( R ² ) LOD
( ng  g ^-One ) ^C LOQ
( ng  g ^-One ) ^C Recovery
(%) ^D RSD
(%) ^D One DEP 245 0.999 1.9 5.7 68.1 3.9 2 DPrP 273 0.997 2 6 85.9 6 3 BBP 335 0.999 2.1 6.3 108.4 4.5 4 DBP / DIBP 301 0.996 1.2 3.6 103.5 4.4 5 DPP 329 0.997 2.8 8.6 96.3 4.4 6 DCHP 353 0.997 2.7 8.1 97.4 4.7 7 DHP 357 0.998 2.7 8.2 104.4 4.3 8 DEHA 393 0.999 2 5.9 103.6 3 9 DEHP 413 0.999 1.5 4.5 104.6 2.7 10 DINP 441 0.999 1.4 4.3 101 4.2 11 DIDP 469 0.999 1.8 5.5 91.9 4.6

In Table 2, the abbreviations indicating the phthalates represented by A means the following substances.

1) DEP (Diethyl phthalate, diethyl phthalate)

2) DPrP (Dipropyl phthalate, dipropyl phthalate)

3) BBP (benzylbutyl phthalate, benzylbutyl phthalate)

4) DBP (Dibutyl phthalate, Dibutyl phthalate), DIBP (Diisobutyl phthalate, Diisobutyl phthalate)

5) DPP (dipentyl phthalate)

6) DCHP (Dicyclohexyl phthalate, dicyclopenic phthalate)

7) DHP (Dihexyl phthalate, dihexyl phthalate)

8) DEHA (diethylhexyl adipate, diethylhexyl adipate)

9) DEHP (Diethylhexyl phthalate, diethylhexyl phthalate)

10) DINP (diisononyl phthalate, diisononyl phthalate)

11) DIDP (Diisodecyl phthalate, diisodecyl phthalate)

In Table 2, the correlation coefficient indicated by B was obtained using a 5-point calibration curve prepared in the range of 0.5-100 ng / g, and the LOD / LOQ of C was calculated using the ICH guideline Q2R1. ≪ / RTI > In addition, experiments in D were repeated 7 times a day, 5 times a day (inter-day), resulting in average recovery and relative standard deviation (RSD).

All of the reagents used in the experiments were HPLC grade reagents.

Example  One

After the addition of the distilled water 200 ml edible salt 40 g, the internal standard (diethyl phthalate -d _4, DEHP-d _4, below using the same internal standard), was added a hexane 10 ml, and sodium chloride ( To remove the organic matter, 40 g of salt (baked at 800 ° C for 8 hours, ionic strength increasing agent) was added and the mixture was shaken for 20 minutes (extraction step).

The mixture thus obtained was separated into a hexane layer and an aqueous solution layer. Only the hexane layer was filtered through anhydrous sodium sulfate and concentrated to 500 μl with nitrogen gas to prepare a measurement solution (concentration step).

The measurement solution was introduced into a liquid chromatography / mass spectrometer (LC-TOF / MS) to be detected and the content thereof was determined (detection / quantification step). Specifically, variation in the detected molecular weight of [M + Na] ⁺ was measured by containing 0.05 mM sodium acetate (soidum acetate, NaOAc, ion activator) in the mobile phase solvent condition of LC / MS, .

Example  2 and background test

Example 2 was carried out in the same manner as in Example 1 except that the detection of [M + Na] ⁺ was carried out in the absence of 0.05 mM sodium acetate (soidum acetate, NaOAc, ion activator) And the variation of the molecular weight is measured. The spectrum of the LC / MS measurement result of Example 2 is shown in Fig.

Comparing the results of FIG. 1 and FIG. 2, it was confirmed that the amount of [M + Na] ⁺ ion detected differs depending on the presence or absence of 0.05 mM sodium acetate in the HPLC mobile phase. This is because, when sodium acetate as an ion activator is further added to the mobile phase to detect and quantify the phthalates contained in the salt, the [M + H] ⁺ ion is inhibited and the [M + Na] ⁺ ion is strongly It is the result that shows that it was possible to do it.

In the following experiment, mineral water for sale was used as a blank test sample in the same manner as in Example 1, and the results are shown in FIG. This is to obtain the correlation between the presence of factors that may be contaminated in the experimental process, the validity of the test, and the data.

Example  3 to 6: Various salts contained in various salts Phthalate  Detection experiment

The commercially available salt was purchased and the following experiment was carried out according to each of the following types.

Example  3

After adding 200 ml of third distilled water to 40 g of natural salt (coarse salt), the mixture containing 10 ml of internal standard substance and hexane was shaken for 20 minutes (extraction step).

The mixture thus obtained was separated into a distilled water layer and an aqueous solution layer. Only the hexane layer was filtered through anhydrous sodium sulfate and concentrated to 500 μl with nitrogen gas to prepare a measurement solution (concentration step).

The measurement solution was introduced into a liquid chromatography / mass spectrometer (LC-TOF / MS) to be detected and the content thereof was determined (detection / quantification step). Specifically, the change in the detected molecular weight of [M + Na] ⁺ was measured by containing 0.05 mM sodium acetate (soidum acetate, NaOAc, ion activator) in the mobile phase solvent condition of LC / MS, . The quantitative analysis results are shown in Tables 3 and 4 (shown as sample "A").

Example  4

Experiments were carried out in the same manner as in Example 3 except that tablets were used in place of natural salt (coarse salt), and the results were shown in FIG. 5, Table 3 and Table 4 (indicated by sample "B").

Example  5

The experiment was carried out in the same manner as in Example 3 except that the salt contained in the food additive was used in place of natural salt (coarse salt), and the results were shown in FIG. 6, Table 3 and Table 4 .

Example  6

Experiments were carried out in the same manner as in Example 3 except that the salt was used instead of natural salt (coarse salt), and the results were shown in FIG. 7, Table 3 and Table 4 (shown as sample "D").

3 to 7, the chromatogram and mass spectrum of the background sample, the coarse salt, the thin salt, the food additive-containing salt and the baked salt are shown in FIG. 3-7, Method.

Diethylhexyl phthalate-d ₄ (DEHP-d ₄ ), an internal standard substance, is similar in properties to LC chromatograms and has a similar retention time and is not eluted well. However, mass spectrometry has different masses It is also confirmed that it is possible.

Quantitative analysis results and ranges for each sample are shown in Table 3 and Table 4.

No Phthalates Phthalate detection frequency  (%) Sample A B C D One DEP 52.6 66.7 50 0 2 DPrP 0 0 0 0 3 BBP 0 0 0 0 4 DBP / DIBP 63.2 33.3 10 0 5 DPP 0 0 0 0 6 DCHP 0 0 0 0 7 DHP 0 0 0 0 8 DEHA 0 0 0 0 9 DEHP 100 66.7 80 0 10 DINP 0 0 0 0 11 DIDP 0 0 0 0

No Phthalates Phthalate  ( ng  g ^-One ) Mean Range Median Sample A B C D A B C D A B C D One DEP 15.4 15.9 13.9 - ND-54.0 ND-25.1 ND-40.7 ND 17.5 22.8 10.4 - 2 DPrP - - - - ND ND ND ND - - - - 3 BBP - - - - ND ND ND ND - - - - 4 DBP
/ DIBP 63.9 13.5 7.2 - ND-293.7 ND-30.7 ND-23.2 ND 22.6 5.3 6.2 - 5 DPP - - - - ND ND ND ND - - - - 6 DCHP - - - - ND ND ND ND - - - - 7 DHP - - - - ND. ND ND ND - - - - 8 DEHA - - - - ND ND ND ND - - - - 9 DEHP 457.8 33.1 85.7 - 21.7-913.1 ND-63.5 ND-343.8 ND 501.8 35.7 40.2 - 10 DINP - - - - ND ND ND ND - - - - 11 DIDP - - - - ND ND ND ND - - - -

* The abbreviations of the respective phthalates are the same as those used in Table 2.

* ND means not detected.

In Tables 3 and 4, A is natural salt (coarse salt), B is refined salt, C is salt contained in food additive, D is 800? It means salt baked above. DBP and DIBP were not separated on the chromatogram and were indicated together.

In the above experimental results, a typical chromatogram of the mineral water shown in FIG. 3, which is the result of the background test, is shown. The peak of DEHP-d ₄ used as an internal standard was 25 ng / g, which was relatively high in blank water compared to other phthalate components. DEP, DBP / DIBP and DEHP in the background samples show trace amounts, which are used as reference values for other samples.

FIG. 4 shows a typical chromatogram of natural salt (coarse salt) according to Example 3, wherein DEHP and DBP / DIBP were relatively higher than the internal standard, and as a result, Among the analyzed salt samples, DEHP was detected as 21.7-913.1, and DBP / DIBP was detected as ND-293.7.

FIG. 5 shows a typical chromatogram of purified salt according to Example 4, where DEHP, DEP, and DBP / DIBP were detected in smaller amounts than coarse salt. ND-63.5 was detected in DEHP and ND-30.7 was detected in DBP / DIBP among the analyzed salt samples as shown in Table 3 and Table 4.

Fig. 6 shows a typical chromatogram of the salt added with the food according to Example 5. Fig. DEHP was found to be higher than that of other salts, which seems to be derived from the food additives contained in the salt.

Fig. 7 shows that phthalate components are not contained in a typical chromatogram of baked salt according to Example 6. Fig.

3 to 7, Table 3 and Table 4, it was confirmed that the total content and the content of the twelve phthalates were contained in the samples of the respective examples. In particular, It is possible to detect it simultaneously.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, Of the right.

Claims (12)

An extraction step of stirring a sample containing an analyte, a polar solvent, an internal standard material and an extraction solvent and allowing the sample to stand to form an aqueous solution layer and an extraction solvent layer;
Concentrating the extraction solvent layer separated from the aqueous solution layer to prepare a measurement solution; And
And a detection quantitation step of introducing the measurement solution into a liquid chromatography and mass spectrometer to detect and quantify the phthalates, wherein the liquid chromatography further comprises an ion activator in the mobile phase solvent , And a method for detecting phthalates. The method according to claim 1,
The phthalates may be selected from the group consisting of diethyl phthalate (DEP), di-n-butyl phthalate (DBP), diisobutyl phthalate (DIBP) and diethylhexyl phthalate , Wherein the phthalates are at least one selected from the group consisting of the phthalates. The method according to claim 1,
Wherein the analysis object comprises salt, and the sample in the extraction step further comprises an ionic strength enhancer. The method of claim 3,
Wherein the ionic strength enhancer comprises any one selected from the group consisting of sodium chloride, sodium carbonate, sodium hydroxide, hydrochloric acid, and combinations thereof. The method according to claim 1,
Wherein the extraction solvent is any one selected from the group consisting of hexane, dichloromethane, n-butanol, cyclohexane, chloroform, ether, and combinations thereof. delete The method according to claim 1,
Wherein the ion activator is any one selected from the group consisting of sodium acetate, ammonium formate, ammonium acetate, formic acid, acetic acid, trifluoroacetic acid, and combinations thereof. Detection method of phthalates. The method according to claim 1,
Wherein the concentration of the extraction solvent layer in the concentration step comprises filtration with anhydrous sodium sulfate. The method according to claim 1,
The internal standard is diethylhexyl phthalate -d _4, di-butyl phthalate -d _4, benzyl butyl phthalate -d _4, and diethylhexyl adipate -d of, phthalates from the group consisting of ₈ to which the selected one Detection method. The method according to claim 1,
Wherein the mobile phase solvent is selected from the group consisting of acetonitrile, distilled water, methanol, isopropanol, ammonium formate, ammonium acetate, formic acid, acetic acid, trifluoroacetic acid, ≪ / RTI > The method according to claim 1,
Wherein the object to be analyzed is a seasoning containing coarse salt, refined salt, baked salt, or any one selected from the group consisting of the above. The method according to claim 1,
The mobile phase solvent may be 0.1% or less, or 0.01-20 mM, based on the whole mobile phase solvent.

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