KR20140027735A - Method for simultaneous determination of phenol compound in biological material - Google Patents

Abstract

The present invention relates to a method for the simultaneous analysis of a phenol compound in a biological material, and more specifically, to a method for the simultaneous analysis of a phenol compound in a biological material, wherein the method includes a step of adjusting the pH of a living body sample and a step of extracting a phenolic compound from the living body sample in a liquid phase. According to one embodiment of the present invention, the pH of the living body sample is adjusted using potassium carbonate (K2CO3).

Description

TECHNICAL FIELD The present invention relates to a method for simultaneous determination of a phenol compound in a biological sample,

The present invention relates to a method for simultaneously analyzing a phenolic compound in a biological sample.

Phenolic compounds are endocrine disruptors that interfere with the normal function of the endocrine system. When they enter the body, they cause a decrease in reproductive capacity, hormone imbalance, or abnormalities of the cranial nervous system and the immune system. do. Phenol compounds have been widely used as a component of nonionic surfactants and have been used as antioxidants and stabilizers in the fields of petrochemical, steel and foods. They are not only included in various daily products such as plastics and glass bottles It is also contained in pesticides, antibiotics, and waste incineration, and many people are easily exposed to phenolic compounds.

When a phenolic compound enters the body, it is excreted mainly into the urine after metabolism, and some of it is also detected in adipose tissue and blood. Biological samples contain phenol compounds such as alkylphenol and chlorophenol which are difficult to analyze at the same time because they have different chemical properties. Therefore, it is necessary to analyze only one phenol compound selected from alkyl phenol or chlorophenol, And phenolic compounds were analyzed. Thus, there is a demand for a method for analyzing phenol compounds in a biological sample more efficiently and easily because the conventional method is complicated and time consuming and costly.

Biomed Environ Sci, XIAO Jing et al., 24 (2011), pp. 40-46

The present invention provides a method for simultaneously analyzing a plurality of phenolic compounds in a biological sample at a relatively simple and low cost with excellent sensitivity and accuracy, and a kit capable of performing the method.

According to an aspect of the present invention, there is provided a method for preparing a biological sample, comprising: adjusting a pH of a biological sample to 6 to 8; And a step of extracting a phenol compound in a biological sample, the pH of which is adjusted in the step, in a liquid phase.

According to another aspect of the present invention, there is provided a biosensor comprising: a pH adjuster for adjusting a pH of a biological sample to 6 to 8; And an extracting unit for extracting a phenol compound of the biological sample in a liquid phase. The present invention also provides a kit for analyzing a phenolic compound in a biological sample.

The phenolic compound assay method and assay kit according to the present invention can analyze phenolic compounds in biological samples with considerable simplicity and low cost with excellent sensitivity and accuracy, and can simultaneously analyze various phenolic compounds with a single treatment. As described above, the method and the kit for analyzing a phenol compound according to the present invention can quickly detect the presence or the concentration of a phenol compound in a large number of biological samples at a high sensitivity, So that the user can quickly respond to the request. Furthermore, the method and the kit for analyzing a phenol compound according to the present invention can be used as an index for determining the influence of phenol compound exposure in a body by analyzing the concentration of the phenol compound.

1 is a total ion chromatogram of 15 phenolic compounds: 1. 2,4-dichlorophenol (2,4-DCP), 2. t-butylphenol (t-BP), 3. 2 , 5-dichlorophenol (2,5-DCP), 4. 2,4-DCP-d ₃ , 5. n-butylphenol (n-BP), 6. 2,4,6-trichlorophenol (2, 4,6-TCP), 7. 2,4,5-trichlorophenol (2,4,5-TCP), 8. pentylphenol (n-PP), 9. n-hexylphenol (n-HX), 10. t-octylphenol (t-OP), 11.n-heptylphenol (n-HP), 12. nonylphenol (NP), 13. t-octylphenol (t-OP), 14. benzophenone-3 (BP-3), 15. Triclosan (TCS), 16. Bisphenol A (BPA), 17. BPA-d ₈

As used herein, the term "biological sample" is intended to encompass all samples derived from an organism, and specifically includes urine, blood or skin tissue. When the concentration of the phenolic compound in the biological sample increases, it is considered that the phenolic compound has been introduced into the body or is being introduced into the body. Therefore, by analyzing phenolic compounds in biological samples, it is possible to evaluate whether the subject is exposed to phenolic compounds. Since the phenol compound is excreted into the urine mainly through metabolism after metabolism, one aspect of the present invention can detect the phenol compound of the urine among the biological samples and analyze the degree of exposure to the phthalate of the subject. As used herein, an object includes an animal, including a human, that can be affected by exposure of the phenolic compound.

In the present specification, a phenol compound is a concept including all compounds containing a phenol group. In one aspect of the invention, the phenolic compound comprises at least one of an alkyl phenol and a chlorophenol. As used herein, an alkylphenol comprises a phenolic compound comprising a C ₁ to C ₁₅ , specifically a C ₃ to C ₁₀ alkyl group, wherein the chlorophenol is a phenol containing from 1 to 6, in particular from 2 to 4, chloro groups ≪ / RTI >

In another aspect of the present invention, the phenolic compound is selected from the group consisting of t-butylphenol (t-BP, Formula 1), n-butylphenol (n-BP, Formula 2), n-pentylphenol (n-HX, Formula 4), n-heptylphenol (n-HP, Formula 5), t-octylphenol (t-OP, Formula 6), n-octylphenol (2, 3, or 4), at least one of nonylphenol (NP, Formula 8), bisphenol A (BPA, Formula 9), and benzophenone- 4-DCP, Formula 11), 2,5-dichlorophenol (2,5-DCP, Formula 12), 2,4,6-trichlorophenol , 5-trichlorophenol (2,4,5-TCP, Formula 14), and triclosan (TCS, Formula 15).

According to one aspect of the present invention, a method for analyzing a phenolic compound in a biological sample can simultaneously analyze an alkylphenol compound and a chlorophenol compound contained in a biological sample after performing a single treatment on a biological sample. Specifically, the phenol compounds of the above-mentioned formulas (1) to (15) can be simultaneously analyzed. Previously, it was difficult to simultaneously analyze the phenolic compounds contained in biological samples having different chemical characteristics. Therefore, only one to four small phenolic compounds were selectively analyzed for one biological sample. On the other hand, according to one aspect of the present invention, there is provided a method for analyzing a phenol in a biological sample, which comprises performing a single treatment with respect to one biological sample and then analyzing various phenolic compounds having different chemical properties, specifically 5 to 20 species, , More specifically 10 to 15 kinds of phenol compounds can be analyzed at the same time, so that the time and cost required for the analysis can be reduced, so that the analysis can be performed easily and efficiently. In addition, phenol compounds can be detected accurately and precisely.

According to an aspect of the present invention, there is provided a method for analyzing a phenolic compound in a biological sample, which comprises preparing a plurality of phenol compounds, specifically, 10 to 15, more preferably 10 to 15, Since phenol compounds of species can be analyzed at the same time, it is convenient because a large amount of sample is not required.

The method for analyzing a phenolic compound in a biological sample according to an aspect of the present invention may include a step of hydrolyzing a biological sample with an enzyme. Since phenol compounds such as alkylphenols and chlorophenols are released into urine in the form of sulfuric acid or glucuronic acid, a hydrolysis step is required to analyze phenol compounds that are not associated with them. . Unlike the hydrolysis method using an acid or base generally used, the enzyme hydrolysis method is suitable because it can secure an excellent recovery rate of the phenol compound while reducing the background matrix of the phenolic compound analysis. In another aspect of the present invention, the enzyme used in the hydrolysis includes an enzyme capable of hydrolyzing sulfuric acid or hydrochloric acid, specifically,? -Glucuronidase and arylsulfatase arylsulfatase). In another aspect of the present invention, the step of hydrolyzing the enzyme may be carried out at a weakly acidic condition of 4 to 6, and an acetic acid buffer solution may be used in setting the biological sample to a slightly acidic condition.

The method for analyzing a phenolic compound in a biological sample according to an aspect of the present invention includes a step of hydrolyzing a biological sample with an enzyme and then adjusting the pH of the biological sample to 6 to 8, specifically 6.5 to 7.5. By including the step of neutralizing the pH of the biological sample in this manner, the method according to one aspect of the present invention can simultaneously analyze phenolic compounds having different chemical properties, specifically alkylphenol and chlorophenol, with excellent recovery and accuracy .

In one aspect of the invention, pH of the biological sample is sodium carbonate as a carbonate, in particular (Na ₂ CO _3), are exemplified potassium carbonate (K ₂ CO _3), calcium carbonate (CaCO ₃₎ or magnesium carbonate (MgCO ₃₎ Lt; RTI ID = 0.0 > alkaline < / RTI > In another aspect of the present invention, the pH of the biological sample can be adjusted with an aqueous carbonate solution of 1 to 10%, specifically 3 to 7%.

The method for analyzing a phenolic compound in a biological sample according to an aspect of the present invention includes a step of controlling the pH of the biological sample and then extracting a liquid phase of the phenolic compound in the biological sample. In one aspect of the present invention, the liquid phase extraction of a biological sample is carried out by one or more extraction solvents selected from among methyl-tert-butyl ether, ethyl acetate and ether, Lt; RTI ID = 0.0 > methyl-tert-butyl < / RTI > The method according to one aspect of the present invention can simultaneously analyze phenol compounds having different chemical properties such as alkylphenol and chlorophenol by a single treatment by performing liquid phase extraction instead of solid phase extraction.

In one aspect of the present invention, the extraction solvent may be used in a volume of 0.9 to 2 times, in particular 1 to 1.5 times, the volume of the biological sample. In another aspect of the present invention, the extraction solvent may be extracted several times by adding 1 to 5 times, specifically 2 to 3 times.

The method for analyzing a phenolic compound in a biological sample according to an aspect of the present invention may further include a step of quantitatively analyzing a phenol compound by a gas chromatography-mass spectrometer after extracting a liquid phase of the phenolic compound. This step can be used to analyze the amount and concentration of phenolic compounds in biological samples.

In one aspect of the present invention, the internal standard method can be used for quantitative analysis. In the internal standard method, first, standard substances are added to samples in various concentration ratios, and a calibration curve is made using the ratio of the peak height (width) and the concentration ratio. Means a quantitative analysis method in which an accurate amount of an internal standard substance is put into an unknown sample to measure the ratio of the peak height between the internal standard substance and the sample and then the concentration ratio of the unknown sample and the internal standard substance is determined on the resulting calibration curve. In one aspect of the invention, the internal standard is as, that is, as an internal standard for the alkyl phenol is bisphenol-A in -d ₈ (BPA-d _8, Formula 16), an internal standard of 2-chlorophenol, 4-Dichlorophenol-d ₃ (2,4-DCP-d ₃ , Formula 17) can be used.

In one aspect of the present invention, the quantitative analysis can be performed in a selective ion monitoring (SIM) mode of the GC-MS. The SIM mode monitors the mass to charge ratio (m / z) for only a few selected ions, thereby improving the precision and accuracy of quantitative analysis. The SIM mode also allows time programming of ions by elution time for analysis of several analytes.

The method for analyzing a phenolic compound in a biological sample according to an aspect of the present invention includes a method for detecting a phenolic compound in a biological sample and a method for analyzing the amount or concentration of a phenolic compound in the biological sample.

According to an aspect of the present invention, there is provided a biosensor comprising: a pH controller for adjusting a pH of a biological sample to 6 to 8; And an extracting unit for extracting a phenol compound of the biological sample in a liquid phase. The present invention also provides a kit for analyzing a phenolic compound in a biological sample. The kit for analyzing a phenolic compound in a biological sample according to an aspect of the present invention may enable a method of analyzing a phenolic compound in a biological sample according to another aspect of the present invention.

In the kit according to one aspect of the present invention, the pH adjuster may adjust the pH of the phenolic compound in the biological sample to 6 to 8, specifically, 6.5 to 7.5, and may include sodium carbonate (Na ₂ CO ₃ ), potassium carbonate (K ₂ CO ₃ ) , Calcium carbonate (CaCO ₃ ), or magnesium carbonate (MgCO ₃ ) can be used to adjust the pH of the biological sample.

In the kit according to one aspect of the present invention, the extracting unit can be subjected to liquid-phase extraction of a biological sample using at least one extraction solvent selected from among methyl-tert-butyl ether, ethyl acetate and ether have.

The kit according to one aspect of the present invention may further comprise a hydrolyzing unit for enzymatic hydrolysis of a biological sample, and the enzyme may include at least one of? -Glucuronidase and arylsulfatase.

The kit according to one aspect of the present invention is characterized by using only a small amount of a biological sample of 0.01 to 5 ml, specifically 0.1 to 4 ml, more specifically, 1 to 3 ml, to convert the alkylphenol and chlorophenol in the biological sample, Butylphenol, n-butylphenol, n-pentylphenol, n-hexylphenol, n-heptylphenol, t-octylphenol, n-octylphenol, nonylphenol, bisphenol A, benzophenone-3,4-dichlorophenol , 2,5-dichlorophenol, 2,4,6-trichlorophenol, 2,4,5-trichlorophenol and triclosan can be simultaneously analyzed.

A kit according to one aspect of the present invention includes a kit for detecting a phenol compound in a biological sample or a kit for analyzing a phenol compound concentration in a biological sample.

The kit according to one aspect of the present invention may further include an instruction that discloses a method for analyzing or detecting a phenol compound in a biological sample.

Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to experimental examples. The following experimental examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, but the scope and scope of the present invention are not limited thereto.

[Experimental Example]

1. Treatment of urine samples

BPA-d ₈ as an internal standard substance To 2 mL of a urine sample to which 2,4-DCP-d ₃ (C / D / N isotope, Quebec, Canada) was added at concentrations of 1 μg / mL and 5 μg / mL, respectively, 1 mL of 0.2 M sodium acetate The pH was adjusted to 4.5 to 5 by adding a buffer solution (0.2 M sodium acetate buffer, pH 5.2). Then, 50 μl of β-glucuronidase / arylsulfatase was added and hydrolyzed at 55 ° C. for 1 hour. To the hydrolyzed urine sample, 50 to 100 μl of a 5% aqueous solution of potassium carbonate (K ₂ CO ₃ ) was added to adjust the pH to 6.5 to 7.5. Next, 2.5 mL of MTBE (methyl-tert-butyl ether) was added as an extraction solvent and extracted twice. The obtained extract was evaporated at 40 ° C using a Turbo Vap LV nitrogen evaporator and sufficiently dried in a vacuum desiccator for 30 minutes or longer using P ₂ O ₅ / KOH.

2. Gas Chromatography-Mass Spectrometry (GC-MS) Analysis

50 μl of BSTFA / TMCS 100: 1 (v / v) mixed solution (Sigma-Aldrich, Steinheim, Germany) was added to the sample in order to increase the volatility of the sample and to effectively separate it from other phenol compounds. The reaction was allowed to proceed for 15 minutes to be derivatized. 2 μl of the derivatized sample was injected into the GC-MS instrument. The GC-MS instrument used an Agilent 5975 mass selective detector connected directly to an Agilent 7890 Plus gas chromatograph from Agilent Technologies, Calif., USA. The sample was also injected into the GC using an Agilent 7683B series injector.

The separation tube used was Ultra-2 (crosslinked 5% phenylmethylpolysiloxane, length 25 m, ID 0.25 mm ID, film thickness 0.33 탆) and temperature programming for volatilization was carried out at an initial temperature of 120 ° C for 1 minute The temperature was raised to 130 占 폚 at 20 占 폚 / min, the temperature was further raised to 150 占 폚 at 5 占 폚 / min, the temperature was raised to 320 占 폚 at 20 占 폚 / min, The sample injection amount was 2 μl, and helium gas having a purity of 99.999% was flowed at a flow rate of 1.0 mL / min. The injection mode was set to a split mode (ratio 5: 1). The electron energy used for ionization was 70 eV, and selected ion monitoring (SIM mode) was used. GC-MS operating conditions are summarized in the table below.

parameter Condition Separator ultra-2 (cross-linked 5% phenylmethylpolysiloxane
25 m x 0.25 mm ID x 0.33 m film thickness) Carrier gas Helium (flow rate 1.0 mL / min) Injection mode Split mode (5: 1) Dose 2 μl Acquisition mode SIM mode in GC-MSD Ionization EI at 70 eV Solvent delay time 5 minutes Total time 16 minutes Temperature programming Initial 120 ° C # Heating rate (° C / min) Temperature (℃) Time (minutes) One 0 120 One 2 20 130 0.5 3 5 150 4 4 20 320 8.5 5 0 320 2

The retention times of the analytes were compared using the internal reference material and the reference material, and the peaks were identified by comparing the peak height ratios of the phenol compounds to be analyzed with respect to the internal standard material peaks. The total ion chromatogram (TIC) of 15 phenolic compounds is shown in FIG. Phenolic compounds that were not separated on chromatogram showed different m / z values and were sufficiently distinguished in SIM mode. The results of the identification of each peak in Figure 1 are as follows: 1. 2,4-Dichlorophenol (2,4-DCP), 2. t-butylphenol (t-BP), 3. 2,5-Dichlorophenol 2,5-DCP), 4. 2,4-DCP-d ₃ , 5. n-butyl phenol (n-BP), 6. 2,4,6- trichlorophenol , 7. 2,4,5-trichlorophenol (2,4,5-TCP), 8. pentyl phenol (n-PP), 9. n-hexyl phenol (n-HX), 10. t- (t-OP), 11. n-heptylphenol (n-HP), 12. nonylphenol (NP), 13. t-octylphenol (t-OP), 14. benzophenone- 15. Triclosan (TCS), 16. Bisphenol A (BPA), 17. BPA-d ₈

Specific ions on the mass spectrum for SIM mode analysis are shown in Table 2.

Phenol compound Retention time Selected ion t-BP 5.60 207 , 222, 151 n-BP 6.55 179 , 222, 207 n-PP 7.63 179 , 236, 221 n-HX 8.56 179 , 250, 235 n-HP 9.34 179 , 264. 249 t-OP 8.77 207 , 278, 263 n-OP 10.04 179 , 278, 263 NP 9.51 ~ 9.88 221 , 193, 249 2,4-DCP 5.57 219 , 234 2,5-DCP 5.83 219 , 234 2,4,6-TCP 7.37 255 , 270 2,4,5-TCP 7.63 255 , 270 BPA 12.31 357 , 372, 207 BP-3 11.72 285 , 242, 300 TCS 11.98 347 , 362, 200 BPA-d ₈ 12.27 365 , 380, 211 2,4-DCP-d ₃ 5.82 222 , 237

3. Creating a calibration curve

(N-BP), n-pentyl phenol (n-PP) and n-hexyl phenol (n-butyl phenol) Octylphenol (n-OP), nonylphenol (NP), bisphenol A (BPA), and benzophenone-3 (2,4-dichlorophenol), 2,5-dichlorophenol (2,5-DCP), 2,4,6-trichlorophenol (2,4- , 6-TCP), 2,4,5-trichlorophenol (2,4,5-TCP) and triclosan (TCS) were analyzed. The assay range of phenol compounds was determined to be 0.2 to 50 ng / mL for nonylphenol and bisphenol A, and 0.1 to 50 ng / mL for other phenol compounds, depending on the concentrations detected in human urine samples. The calibration curve concentration range was determined by selecting the concentration of 0.1, 0.2, 0.5, 1, 2, 5, 10, 20 and 50 ng / mL for the phenol standard substance mixture in the substantially same manner as the experimental example of the present invention Respectively. The detection limit was measured by the signal to noise (S / N) method and the concentration at which the signal-to-noise ratio was 10 or more was expressed as the quantitative limit (LOQ). The calibration range and linearity correlation coefficient of each compound are summarized in Table 3 below. As can be seen from the results below, the calibration curve for all phenolic compounds showed linearity (r ² > 0.996) within the quantitative range.

compound Black range
(ng / mL) inclination Intercept R ² Detection limits (ng / mL) Quantitation limits (ng / mL) t-BP 0.1 to 50 0.0461 00035 0.9994 0.05 0.1 n-BP 0.1 to 50 0.0651 -0.0099 0.9997 0.02 0.1 n-PP 0.1 to 50 0.0880 0.0066 0.9987 0.02 0.1 n-HX 0.1 to 50 0.0980 0.0185 0.9997 0.02 0.1 n-HP 0.1 to 50 0.1036 0.0050 0.9982 0.02 0.1 t-OP 0.1 to 50 0.1170 0.0851 0.9995 0.05 0.1 n-OP 0.1 to 50 0.1121 0.0018 0.9993 0.05 0.1 NP 0.2 to 50 0.0031 0.0314 0.9971 0.05 0.2 2,4-DCP 0.1 to 50 0.0439 0.0018 1.0000 0.05 0.1 2,5-DCP 0.1 to 50 0.0333 0.0025 1.0000 0.05 0.1 2,4,6-TCP 0.1 to 50 0.0549 0.0422 0.9967 0.05 0.1 2,4,5-TCP 0.1 to 50 0.1194 0.0029 0.9991 0.05 0.1 BPA 0.2 to 50 0.1298 0.0906 0.9990 0.05 0.2 BP-3 0.1 to 50 0.1012 -0.0140 0.9997 0.05 0.1 TCS 0.1 to 50 0.0164 0.0011 0.9998 0.05 0.1

4. Verification of validity

In addition, concentrations of 0.5, 1, and 5 ng / mL were selected for the accuracy and precision of 15 phenolic compound standards. (Recovery,%), Accuracy (%) and Accuracy (%) based on the results of 5 replicates and 5 replicates for 0.5, 1, and 5 ng / precision,%) were evaluated and shown in Table 4 (alkylphenol) and Table 5 (chlorophenol) below. As can be seen in Tables 4 and 5, the recovery rates of all phenolic compounds ranged from 82.1% to 110.5%, with precision ranging from 0.1% to 17.4% and accuracy ranging from 88.5% to 116.4%, with excellent recovery, accuracy and accuracy .

Alkylphenol density
(ng / mL) Repeat 5 times Repeat for 5 days Recovery rate Precision accuracy Recovery rate Precision accuracy t-BP 0.5 99.0 4.3 107.5 89.9 3.5 101.7 One 94.4 4.2 108.7 98.3 1.8 109.1 5 88.6 7.9 107.8 92.4 1.6 99.1 n-BP 0.5 102.3 10.9 92.7 83.6 4.0 100.2 One 88.0 11.7 104.7 93.7 2.9 109.3 5 87.8 3.8 93.9 89.3 0.2 103.3 n-PP 0.5 103.3 4.7 95.6 91.1 9.6 97.6 One 92.2 8.3 93.5 87.8 4.7 109.0 5 86.7 5.4 106.7 89.6 1.5 97.7 n-HX 0.5 87.9 17.4 106.8 87.8 2.0 103.8 One 92.9 8.9 96.4 90.8 6.6 111.8 5 96.7 10.8 96.9 85.2 5.6 99.3 n-HP 0.5 97.5 7.9 90.4 89.5 7.8 97.7 One 97.5 11.6 99.6 84.8 11.0 104.4 5 107.1 6.1 98.7 82.8 7.6 107.7 t-OP 0.5 102.8 7.1 95.9 90.8 0.5 100.4 One 98.8 7.6 93.6 90.4 4.6 107.7 5 89.9 9.7 88.5 86.1 1.5 97.4 n-OP 0.5 92.7 6.9 96.5 91.5 6.7 104.8 One 104.4 14.8 98.7 96.9 6.5 97.5 5 85.2 4.5 93.3 88.4 2.1 101.9 NP 0.5 110.5 9.9 106.0 90.1 5.5 99.0 One 96.8 9.8 98.3 102.3 4.0 108.3 5 105.6 6.0 95.8 106.4 3.1 97.2 BPA 0.5 88.1 8.3 102.1 93.3 4.1 99.6 One 91.3 7.4 116.4 83.9 5.4 102.2 5 109.5 5.9 109.8 107.5 5.0 101.1 BP-3 0.5 97.5 5.6 105.7 105.5 1.6 106.6 One 107.9 6.2 113.2 92.3 9.7 106.4 5 101.8 4.9 96.4 83.5 6.5 98.1

Chlorophenol density
(ng / mL) Repeat 5 times Repeat for 5 days Recovery rate Precision accuracy Recovery rate Precision accuracy 2,4-
DCP 0.5 88.2 8.8 103.4 98.9 1.7 101.9 One 108.0 5.0 111.7 103.9 3.4 106.5 5 97.6 2.0 100.2 109.2 8.0 93.1 2,5-
DCP 0.5 91.2 6.1 100.7 82.1 8.7 102.2 One 107.3 5.4 113.2 93.7 4.5 94.0 5 106.0 3.3 106.2 103.0 5.3 91.3 2,4,6-TCP 0.5 92.6 6.5 101.2 83.3 2.8 104.4 One 87.3 4.6 103.6 97.4 3.8 109.4 5 104.0 6.1 101.3 101.1 9.4 96.3 2,4,5-TCP 0.5 89.7 8.2 100.8 83.5 0.1 101.8 One 106.7 10.5 110.7 85.9 6.2 108.0 5 103.6 3.5 97.6 106.7 3.2 91.3 TCS 0.5 85.0 2.8 98.6 101.8 9.9 104.1 One 109.5 7.4 111.3 100.6 6.7 110.6 5 105.8 3.6 97.6 106.7 3.2 91.6

As described above, the effectiveness of the method for analyzing phenolic compounds in a biological sample according to the present invention was verified. Therefore, it can be confirmed that the method according to the present invention can simultaneously detect and analyze 15 kinds of phenol compounds in a biological sample.

Claims (11)

Adjusting the pH of the biological sample to 6-8; And
Method for analyzing a phenolic compound in a biological sample comprising the step of liquid phase extraction of the phenolic compound in the biological sample adjusted to pH in the step.
The method of claim 1,
The biological sample comprises 0.01 ml to 5 ml of urine.
The method of claim 1,
Adjusting the pH of the biological sample is a method of adjusting the pH with potassium carbonate (K ₂ CO ₃ ).
The method of claim 1,
The liquid phase extraction may be performed using at least one extraction solvent selected from methyl-tert-butyl ether, ethyl acetate and ether.
The method of claim 1,
and prior to adjusting the pH, hydrolyzing the biological sample with an enzyme.
The method of claim 1,
Further comprising the step of analyzing the phenolic compound with a gas chromatography-mass spectrometer after the step of liquid phase extraction.
The method of claim 1,
Phenolic compounds include alkylphenols and chlorophenols.
The method of claim 1,
The method comprises t-butylphenol, n-butylphenol, n-pentylphenol, n-hexylphenol, n-heptylphenol, t-octylphenol, n-octylphenol, nonylphenol, bisphenol A, benzophenone-3 in biological samples , 2,4-dichlorophenol, 2,5-dichlorophenol, 2,4,6-trichlorophenol, 2,4,5-trichlorophenol and triclosan at the same time.
PH adjusting unit for adjusting the pH of the biological sample to 6 to 8; And
A kit for analyzing a phenolic compound in a biological sample, comprising an extraction unit for extracting a phenolic compound in a biological sample in a liquid phase.
The method of claim 9,
Wherein the extract comprises at least one extraction solvent selected from methyl-tert-butyl ether, ethyl acetate and ether.
The method of claim 9,
The kit includes t-butylphenol, n-butylphenol, n-pentylphenol, n-hexylphenol, n-heptylphenol, t-octylphenol, n-octylphenol, nonylphenol, bisphenol A, benzophenone-3 in biological samples. , Kit for simultaneously analyzing 2,4-dichlorophenol, 2,5-dichlorophenol, 2,4,6-trichlorophenol, 2,4,5-trichlorophenol and triclosan.

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