KR20170023506A - Kit for diagnosis obesity using di-2-ethylhexyl phthalate metabolic ratio in biological material - Google Patents

Kit for diagnosis obesity using di-2-ethylhexyl phthalate metabolic ratio in biological material Download PDF

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KR20170023506A
KR20170023506A KR1020150118744A KR20150118744A KR20170023506A KR 20170023506 A KR20170023506 A KR 20170023506A KR 1020150118744 A KR1020150118744 A KR 1020150118744A KR 20150118744 A KR20150118744 A KR 20150118744A KR 20170023506 A KR20170023506 A KR 20170023506A
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표희수
이정애
온지원
김승기
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한국과학기술연구원
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    • G—PHYSICS
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    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract

The present invention relates to a kit for conveniently diagnosing obesity using a mole concentration ratio of di-2-ethylhexyl phthalate metabolic ratio in a biological material and a method for providing basic information for diagnosis.

Description

[0001] The present invention relates to a kit for diagnosing obesity using di-2-ethylhexyl phthalate metabolite ratio in a biological sample,

The present disclosure relates to a method for determining obesity using di-2-ethylhexyl phthalate metabolite ratio in a biological sample.

Obesity is an excess of fat tissue in the body, and over-consumption of nutrients over a long period of time over a long period of time leads to esterification of fatty acids and glucose, which are introduced into fat cells from plasma by energy imbalance, And is accumulated.

Conventionally, as a criterion of obesity, a method of defining obesity has been proposed when the body mass index (body mass index: body weight (kg) divided by the square of height (m)) is 25 to 30 or more. However, according to the above-mentioned body obesity index, since a lot of muscles other than fatty tissue are present and a person having a lot of weight is judged to be obese, there is a problem that accurate diagnosis is difficult.

Pediatric obesity refers to the case where the body weight is 20% or more of the standard weight per kidney in the age range from median to puberty. The factors of childhood obesity include excessive calorie intake, lack of exercise, genetic factors, environmental factors, Sexual obesity (congenital disorders, side effects of neurological and endocrinal diseases, etc.).

In adult obesity, the number of adipocytes is increased while the number of adipocytes is increased, while the number of adipocytes is increased in the case of childhood obesity. Because of the higher number of adipocytes in adults than in normal weight adults, 75-80% of childhood obesity is converted to adult obesity, is difficult to treat, and has a high likelihood of recurrence. In addition, growth hormone secretion can be inhibited and growth can be inhibited. If fat and cholesterol levels in the blood increase, hypertension, diabetes, fatty liver and cardiovascular diseases and complications resulting therefrom can develop early

Over the past 30 years, the rate of obesity in children and adolescents in the United States has increased dramatically from two to four times. In addition, the worldwide population of obese children and adolescents is rapidly increasing, It is necessary to develop the technology.

Assessing human exposure to phthalates using monoesters and their oxidized metabolites as biomarkers, Environmental Health Perspectives, 2003, vol 11, p1148-1151

In one aspect, an object of the present invention is to provide a kit which can easily diagnose obesity through a formula using molar ratio of di-2-ethylhexyl phthalate metabolite in phthalate.

In another aspect, an object of the present invention is to provide a method for providing basic information for diagnosing obesity using molar ratio of di-2-ethylhexyl phthalate metabolite in phthalate .

According to an aspect of the present invention,

Monoethylhydroxyhexyl phthalate and monoethylhexyl phthalate;

Monoethyloxohexyl phthalate and monoethyloxohexyl phthalate;

Monoethyl 6-hydroxyhexyl phthalate and monoethyl hexyl phthalate; And

Monoethyl carboxypentyl phthalate and monoethyl 6-hydroxyhexyl phthalate;

2-ethylhexyl phthalate group selected from the group consisting of di-2-ethylhexyl phthalate and di-2-ethylhexyl phthalate. Lt; / RTI >

In addition, an embodiment of the present invention is characterized in that,

Monoethylhydroxyhexyl phthalate and monoethylhexyl phthalate;

Monoethyloxohexyl phthalate and monoethyloxohexyl phthalate;

Monoethyl 6-hydroxyhexyl phthalate and monoethyl hexyl phthalate; And

Monoethyl carboxypentyl phthalate and monoethyl 6-hydroxyhexyl phthalate;

Measuring the molarity of the di-2-ethylhexyl phthalate metabolite contained in the di-2-ethylhexyl phthalate group selected from the group consisting of the di-2-ethylhexyl phthalate metabolite in the biological sample;

The molar concentration of the di-2-ethylhexyl phthalate metabolite measured from the molar concentration measurement unit in the biological sample is substituted into one or more mathematical formulas selected from the group consisting of the following equations (1) to (8) Calculating an ethylhexyl phthalate metabolite ratio; And

Diagnosing whether or not obesity is caused by comparing the metabolite ratio in the biological sample and the metabolic rate of the normal control;

A method for providing information for diagnosing obesity, comprising the steps of:

[Equation 1]

RMR A1 = molar concentration of monoethylhydroxyhexyl phthalate in biological sample / molar concentration of monoethylhexyl phthalate in biological sample

&Quot; (2) "

RMR A2 = molar concentration of monoethyloxohexyl phthalate in biological sample / molar concentration of monoethylhydroxyhexyl phthalate in biological sample

&Quot; (3) "

RMR AR = RMR A1 / RMR A2

&Quot; (4) "

RMR AT = RMR A1 * RMR A2

&Quot; (5) "

RMR B1 = molar concentration of monoethyl 6-hydroxyhexyl phthalate in the biological sample / molar concentration of monoethylhexyl phthalate in the biological sample

&Quot; (6) "

RMR B2 = molar concentration of monoethyl carboxypentyl phthalate in biological sample / molar concentration of monoethyl 6-hydroxyhexyl phthalate in biological sample

&Quot; (7) "

RMR BR = RMR B1 / RMR B2

&Quot; (8) "

RMR BT = RMR B1 * RMR B2

The present invention relates to the use of a relative metabolite ratio (RMR) obtained by applying a molar concentration in a biological sample to a metabolite of di-2-ethylhexyl phthalate having the largest amount of phthalate produced and consumed, The obesity degree of the subject can be easily and easily judged. The present invention solves the problematic accuracy in the determination of obesity by the existing body index (BMI, waist circumference, etc.) since the accurate analysis of the obesity degree is possible by directly analyzing metabolites in the biological sample.

Further, the present invention is useful for early diagnosis of the obesity or obesity possibility of pediatric obesity patients by using a metabolite of di-2-ethylhexyl phthalate, which shows a higher concentration of phthalates as the age is lower.

The present invention can determine the therapeutic effect of obesity or the success of disease prevention by a simple examination, and prevent the risk of developing a cardiovascular disease or diabetes due to obesity such as hypertension and high cholesterol.

Figure 1 shows the metabolic pathway of di-2-ethylhexyl phthalate in vivo.
Fig. 2 shows the result of GC-MS treatment of the sample obtained in Example 1. Fig.
(1: MEP, 1 ': MEP-d 4, 2: MiBP, 2': MiBP-d 4, 3: MnBP, 3 ': MnBP-d 4, 4: MEHP, 4': MEHP-d 4, 5 : MiNP, 5 ': MiNP- 13 C 4, 6: MBzP, 6': MBzP-d 4, 7: MEOHP, 7 ': MEOHP- 13 C 4, 8: MEHHP, 8': MEHHP- 13 C 4)

Hereinafter, the present invention will be described in detail.

Phthalates are plasticizers that give flexibility and elasticity to plastics and are widely used in a variety of products, such as household goods, industrial products, medical supplies, and baby products. Excessive exposure to phthalates in the human body is known to cause endocrine disruption and cancer, and reproductive toxicity and liver toxicity.

Accordingly, an embodiment of the present invention provides an obesity diagnosis kit comprising a measurement unit for measuring an excreted concentration of a metabolite of di-2-ethylhexyl phthalate having the highest production and consumption amount of the phthalate from a living body discharged from a human body And an information providing method for diagnosing obesity.

Specifically, in one embodiment of the present invention,

Monoethylhydroxyhexyl phthalate and monoethylhexyl phthalate;

Monoethyloxohexyl phthalate and monoethyloxohexyl phthalate;

Monoethyl 6-hydroxyhexyl phthalate and monoethyl hexyl phthalate; And

Monoethyl carboxypentyl phthalate and monoethyl 6-hydroxyhexyl phthalate;

2-ethylhexyl phthalate group selected from the group consisting of di-2-ethylhexyl phthalate and di-2-ethylhexyl phthalate. Lt; / RTI >

In one embodiment, the molar concentration measuring unit in the biological sample of the di-2-ethylhexyl phthalate metabolism may include a substance or detection device capable of detecting the di-2-ethylhexyl phthalate metabolism from a biological sample have. A mass spectrometer such as a gas chromatography-mass spectrometry or a liquid chromatography-mass spectrometry may be used. However, it is preferable to use a mass spectrometer capable of measuring the molar concentration of the metabolite in the sample But is not limited thereto.

In one embodiment, the obesity diagnostic kit may further include an instruction sheet including an obesity diagnosis method, wherein the instruction sheet indicates a molar concentration of the di-2-ethylhexyl phthalate metabolite measured from the molar concentration measurement unit in the biological sample, Comprising the steps of: analyzing di-2-ethylhexyl phthalate metabolite ratio in a biological sample by substituting the formula into one or more mathematical formulas selected from the group consisting of the following formulas (1) to (8) An obesity diagnosis kit can be provided.

[Equation 1]

RMR A1 = molar concentration of monoethylhydroxyhexyl phthalate in biological sample / molar concentration of monoethylhexyl phthalate in biological sample

&Quot; (2) "

RMR A2 = molar concentration of monoethyloxohexyl phthalate in biological sample / molar concentration of monoethylhydroxyhexyl phthalate in biological sample

&Quot; (3) "

RMR AR = RMR A1 / RMR A2

&Quot; (4) "

RMR AT = RMR A1 * RMR A2

&Quot; (5) "

RMR B1 = molar concentration of monoethyl 6-hydroxyhexyl phthalate in the biological sample / molar concentration of monoethylhexyl phthalate in the biological sample

&Quot; (6) "

RMR B2 = molar concentration of monoethyl carboxypentyl phthalate in biological sample / molar concentration of monoethyl 6-hydroxyhexyl phthalate in biological sample

&Quot; (7) "

RMR BR = RMR B1 / RMR B2

&Quot; (8) "

RMR BT = RMR B1 * RMR B2

The step of diagnosing whether the observer is obese according to an embodiment of the present invention may include measuring the metabolic rate of di-2-ethylhexyl phthalate in the biological sample calculated by substituting the formula into one or more mathematical formulas selected from the group consisting of the mathematical formulas If the chevy differs from the di-2-ethylhexyl phthalate metabolite ratio of the normal control, it may be diagnosed as obesity.

In addition, an embodiment of the present invention is characterized in that,

Monoethylhydroxyhexyl phthalate and monoethylhexyl phthalate;

Monoethyloxohexyl phthalate and monoethyloxohexyl phthalate;

Monoethyl 6-hydroxyhexyl phthalate and monoethyl hexyl phthalate; And

Monoethyl carboxypentyl phthalate and monoethyl 6-hydroxyhexyl phthalate;

Measuring the molarity of the di-2-ethylhexyl phthalate metabolite contained in the di-2-ethylhexyl phthalate group selected from the group consisting of the di-2-ethylhexyl phthalate metabolite in the biological sample;

The molar concentration of the di-2-ethylhexyl phthalate metabolite measured from the molar concentration measuring unit in the biological sample is substituted into one or more mathematical formulas selected from the group consisting of the equations (1) to (8) Calculating an ethylhexyl phthalate metabolite ratio; And

Diagnosing whether or not obesity is caused by comparing the metabolite ratio in the biological sample and the metabolic rate of the normal control;

A method for providing information for diagnosing obesity, comprising the steps of:

The step of diagnosing obesity by comparing the ratio of the metabolite in the biological sample and the metabolic rate of the normal control group according to an embodiment of the present invention may be performed by substituting one or more mathematical expressions selected from the group consisting of the expressions 1 to 8 And judging that the di-2-ethylhexyl phthalate metabolite ratio in the calculated bio sample is different from the di-2-ethylhexyl phthalate metabolite ratio of the normal control group.

In the present specification, the term "biological sample" is a concept covering all samples derived from living organisms, and the living organism may include humans or animals other than humans. Specifically, the biological sample may include urine, blood or skin tissue derived from an organism. As a more specific example, the biological sample may be urine. The urine is a urine sample discharged from the human body. Specifically, the urine collected directly from the subject can be used. When a living body absorbs di-2-ethylhexyl phthalate and absorbs it, it is metabolized and excreted into the body in the form of a metabolite, which is mainly contained in urine.

The term "di-2-ethylhexyl phthalate metabolite" as used herein includes all metabolites in which di-2-ethylhexyl phthalate is absorbed and converted in vivo, It gradually transforms into a less toxic substance through metabolic processes, including but not limited to these substances. In one embodiment, the di-2-ethylhexyl phthalate metabolite is selected from the group consisting of monoethylhexyl phthalate, monoethylhexyl phthalate glucuronide, monoethylhydroxyhexyl phthalate, monoethylhydroxyhexyl phthalate glucuronide, monoethyl Monoethyl 6-hydroxyhexyl phthalate, monoethyl 6-hydroxyhexyl phthalate glucuronide, monoethyl carboxypentyl phthalate, and monoethyl carboxypentyl phthalate glucuronide But are not limited thereto.

As used herein, the term " mono-ethylhexyl phthalate (MEHP) "refers to a di-2-ethylhexyl phthalate metabolite, which is a compound of formula 1 below or a glucuronide form conjugated in vivo Lt; / RTI >

Figure pat00001

As used herein, the term " mono-ethyl-hydroxyhexyl phthalate (MEHHP) "refers to a di-2-ethylhexyl phthalate metabolite, wherein the compound of formula 2 below or the compound of formula 2 is conjugated in vivo And may be in the form of a croonide.

Figure pat00002

The term " mono-ethyl-oxohexyl phthalate (MEOHP) "as used herein also refers to a di-2-ethylhexyl phthalate metabolite, which may be a compound of formula 3 below or a compound of formula 3 conjugated in vivo It may be in the form of a Ronid.

Figure pat00003

As used herein, the term " mono-ethyl-6-hydroxyhexyl phthalate "also refers to a di-2-ethylhexyl phthalate metabolite, Gt; glucuronide < / RTI > form.

Figure pat00004

As used herein, the term "mono-ethyl-carboxypentyl phthalate (MECPP)" also refers to di-2-ethylhexyl phthalate as a metabolite of the compound of formula 5 below or the compound of formula 5 conjugated in vivo It may be in the form of a Ronid.

Figure pat00005

The compounds of formulas 1 to 5 and their glucuronide forms may be metabolites produced in the course of metabolism of di-2-ethylhexyl phthalate in vivo, and such metabolism in vivo is disclosed in FIG. 1 . 1, it can be seen that each phthalate metabolite is converted into a conjugated glucuronide form in the body. Thus, each "di-2-ethylhexyl phthalate phthalate" as used herein refers to both pure di-2-ethylhexyl phthalate metabolites and di-2-ethylhexyl phthalate metabolites conjugated in vivo in glucuronide form .

The above formula of the present invention may use the molar concentration of each metabolite, but may be calculated using the mole fraction. The "mole fraction" may mean the percentage by which each compound constitutes when the molar amount of di-2-ethylhexyl phthalate is 100. Therefore, the term "mole fraction of the di-2-ethylhexyl phthalate metabolite" as used herein includes the "mole fraction of the enzyme hydrolyzate of the di-2-ethylhexyl phthalate metabolite" present in the biological sample.

As used herein, the terms mono-ethylhexyl phthalate (MEHP), mono-ethyl-hydroxyhexyl phthalate (MEHHP), mono-ethyl-oxohexyl phthalate (MEOHP) In order to determine the molar concentration of monoethyl-6-hydroxyhexyl phthalate and monoethyl-carboxypentyl phthalate (MECPP), the amount of the compound contained in the biological sample It is necessary to quantify. Any method known in the art can be used without limitation, but the method described in Applicant's patent application No. 10-2012-0060835 can be used. That is, the biological sample is treated with an enzyme such as? -Glucuronidase to enzymatically decompose the metabolite of the phthalate, and the resulting solution is treated with a C 5 to C 6 saturated hydrocarbon solvent and ether (N, O-bis- (trimethylsilyl) -trifluoroacetamide (BSTFA)) and trimethyl chlorosilane (hereinafter, referred to as " trimethylsilyl trifluoroacetamide " silane, TMCS), and finally, the phthalate metabolite can be quantitatively analyzed by isotope dilution method.

In the present specification, "RMR" in the above Equations 1 to 8 may mean a relative metabolite ratio. That is, RMR A1 may be obtained by dividing the molar concentration of monoethylhydroxyhexyl phthalate in the biological sample by the molar concentration of monoethylhexyl phthalate in the same biological sample. In addition, RMR A2 may be obtained by dividing the molar concentration of monoethyloxohexyl phthalate by the molar concentration of monoethylhydroxyhexyl phthalate in the same biological sample. In addition, RMR AR may be one obtained by dividing RMR A1 by RMR A2 , and RMR AT may be obtained by multiplying RMR A1 by RMR A2 .

In addition, RMR B1 may be obtained by dividing the molar concentration of monoethyl 6-hydroxyhexyl phthalate in the biological sample by the molar concentration of monoethylhexyl phthalate in the same biological sample. In addition, RMR B2 may be obtained by dividing the molar concentration of monoethyl carboxypentyl phthalate by the molar concentration of monoethyl 6-hydroxyhexyl phthalate in the same biological sample. In addition, RMR BR may be obtained by dividing RMR B1 by RMR B2 , and RMR BT may be obtained by multiplying RMR B1 by RMR B2 .

The present invention newly defines a formula using the molar concentration of a phthalate metabolite as described above, and compared the molarity ratio of the di-2-ethylhexyl phthalate metabolite contained in the biological sample discharged from the human body to the control, It is easy to judge whether or not it is possible.

Hereinafter, the present invention will be described more specifically with reference to the following examples. However, the following examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

[Example] Quantification of di-2-ethylhexyl phthalate metabolite

1. Treatment of urine samples

A total of 245 men and women between the ages of 7 and 14 years, normal, overweight, and obesity were enrolled in this study.

To 1 mL of each urine sample were added 4 kinds of internal standard substances such as monoethylhexyl phthalate-d 4 , monoethylhydroxyhexyl phthalate- 13 C 4 , monoethyloxohexyl phthalate- 13 C 4 and monoethyl carboxypentyl phthalate- 13 C 4 25 mu l of the mixed solution (1 mu g / mL) was added, and 700 mu l of 1M ammonium acetate buffer solution (pH 7.0) was added thereto so as to have a pH of 6.0 to 6.5. The mixture was homogenized using a vortexer, and 50 μl of β-glucuronide was added thereto, followed by decomposition at 37 ° C. for 2 hours.

1 mL of 2M acetic acid was added to the decomposed sample so that the pKa (3.4-3.6) of the monophthalate ester was taken into consideration, and 0.5 g of anhydrous sodium sulfate was added thereto as a salting agent. Then, hexane and ethyl ether were added at a ratio of 8: And extracted by shaking for 20 minutes with 3 mL of the extracted solvent. After the extraction, the urine layer and the solvent layer were separated by rotating at 2800 rpm in a centrifuge, and the urine layer was frozen in a freezer at -21 캜 to separate it from the solvent layer. The frozen urine sample was dissolved at room temperature, and 3 mL of a mixed solvent of fresh hexane and ethyl ether was added, and the extraction procedure substantially the same as above was repeated.

Then, the extraction solvent was evaporated with a nitrogen concentrator, and then dried in a vacuum dryer using P 2 O 5 / KOH / silica gel for 30 minutes. After drying, 99: 1 (v / v) of N, O-bistrimethylsilyl-trifluoroacetamide (BSTFA) and trimethylchlorosilane (TMCS) ) Mixed solution (Sigma-Aldrich, Steinheim, Germany) was added, and the mixture was allowed to react at 65 ° C for 30 minutes to effect trimethylsilylation.

2. Gas Chromatograph Mass Spectrometer (GC-MS) Analysis

Agilent 7693 A Series injector was used in a GC-MS instrument (Agilent 5975 C mass selective detector directly connected to Agilent 7890 A gas chromatograph from Agilent Technologies, Santa Clara, Calif., USA) Lt; / RTI >

The separation tube used was ultra-2 (cross-linked 5% phenylmethyl silicone, length 25 m, internal diameter 0.2 mm, film thickness 0.33 μm) and carrier gas was ultrapure (99.9999%) helium at a flow rate of 1 mL / Respectively. The temperature of the inlet was 280 ° C and a 1: 5 split injection mode was used. The oven conditions were set to warm from 100 占 폚 to 310 占 폚. The electron energy used for ionization was 70 eV, and selected ion monitoring (SIM mode) was used. GC-MS operating conditions are shown in Table 1 below.

Condition Separator Ultra-2
(Cross-linked 5% phenylmethyl silicone, 25 m x 0.2 mm ID, 0.33 um film thickness) Carrier gas A flow rate of 1.0 mL / min of helium Injection mode Split mode (5: 1) Dose 2 μl Acquisition mode SIM mode in GC-MSD Ionization EI at 70 eV Solvent delay time 3 minutes Total time 25 minutes Temperature programming Initial 100 ° C # Heating rate (° C / min) Temperature (℃) Time (minutes) One 15 200 5 2 10 270 0 3 30 310 5

The molar concentrations of the di-2-ethylhexyl phthalate metabolites in the urine obtained through the urine sample treatment and GC-MS analysis are shown in Table 2 below.

group gender Molar concentration in terms of creatinine (pmol / gcr) MEHP MEHHP MEOHP MECPP normal Girl Number of samples 76 76 76 76 Standard Deviation 43.6 128.0 95.6 353.5 Geometric mean 50.8 143.2 116.0 339.4 stay Number of samples 63 63 63 63 Standard Deviation 40.1 76.8 66.0 208.9 Geometric mean 37.7 79.9 78.3 189.5 Total Number of samples 139 139 139 139 Standard Deviation 42.4 113.5 86.1 309.4 Geometric mean 44.4 109.9 97.1 260.6 Overweight Girl Number of samples 21 21 21 21 Standard Deviation 27.7 56.7 43.1 136.8 Geometric mean 43.3 109.9 86.7 244.7 stay Number of samples 15 15 15 15 Standard Deviation 14.4 41.0 30.3 76.8 Geometric mean 26.9 71.2 60.2 148.7 Total Number of samples 36 36 36 36 Standard Deviation 24.7 53.2 40.0 125.1 Geometric mean 35.5 91.7 74.5 198.8 obesity Girl Number of samples 46 46 46 46 Standard Deviation 44.0 269.7 143.3 507.2 Geometric mean 47.1 138.3 103.6 302.2 stay Number of samples 24 24 24 24 Standard Deviation 26.7 66.6 53.0 164.8 Geometric mean 31.4 103.9 83.8 217.0 Total Number of samples 70 70 70 70 Standard Deviation 40.0 224.3 121.5 430.0 Geometric mean 41.0 125.4 96.3 269.8

3. Relative to obesity Metabolism ratio  ( RMR ) Calculation

The RMRs were calculated by Equations 1 to 8 using the data obtained from Table 2 of Example 1, and the results according to the degree of obesity according to each group are shown in Table 3.

[Equation 1]

RMR A1 = molar concentration of monoethylhydroxyhexyl phthalate in biological sample / molar concentration of monoethylhexyl phthalate in biological sample

&Quot; (2) "

RMR A2 = molar concentration of monoethyloxohexyl phthalate in biological sample / molar concentration of monoethylhydroxyhexyl phthalate in biological sample

&Quot; (3) "

RMR AR = RMR A1 / RMR A2

&Quot; (4) "

RMR AT = RMR A1 * RMR A2

&Quot; (5) "

RMR B1 = molar concentration of monoethyl 6-hydroxyhexyl phthalate in the biological sample / molar concentration of monoethylhexyl phthalate in the biological sample

&Quot; (6) "

RMR B2 = molar concentration of monoethyl carboxypentyl phthalate in biological sample / molar concentration of monoethyl 6-hydroxyhexyl phthalate in biological sample

&Quot; (7) "

RMR BR = RMR B1 / RMR B2

&Quot; (8) "

RMR BT = RMR B1 * RMR B2

group gender RMR_A1 RMR_A2 RMR_AT RMR_AR RMR_BT normal Girl Number of samples 76 76 76 76 76 Standard Deviation 1.77 .10 1.31 2.62 5.04 Geometric mean 2.82 .81 2.28 3.48 6.68 stay Number of samples 63 63 63 63 63 Standard Deviation 1.65 .29 1.24 2.27 3.35 Geometric mean 2.12 .98 2.08 2.16 5.02 Total Number of samples 139 139 139 139 139 Standard Deviation 1.74 .23 1.28 2.52 4.45 Geometric mean 2.48 .88 2.19 2.80 5.87 Overweight Girl Number of samples 21 21 21 21 21 Standard Deviation 1.40 .10 1.06 2.06 3.34 Geometric mean 2.54 .79 2.00 3.21 5.65 stay Number of samples 15 15 15 15 15 Standard Deviation 1.40 .14 .84 2.38 3.33 Geometric mean 2.65 .85 2.24 3.13 5.53 Total Number of samples 36 36 36 36 36 Standard Deviation 1.38 .12 .97 2.17 3.29 Geometric mean 2.58 .81 2.10 3.18 5.60 obesity Girl Number of samples 46 46 46 46 46 Standard Deviation 2.51 .11 1.42 4.73 6.83 Geometric mean 2.93 .75 2.20 3.92 6.41 stay Number of samples 24 24 24 24 24 Standard Deviation 1.44 .13 .99 2.25 2.79 Geometric mean 3.31 .81 2.67 4.10 6.92 Total Number of samples 70 70 70 70 70 Standard Deviation 2.19 .12 1.29 4.04 5.75 Geometric mean 3.06 .77 2.35 3.98 6.58

As a result, RMR_A1 was increased, RMR_A2 was decreased, and RMR_AT, RMR_AR and RMR_BT were increased in the experimental group composed of male and female children with obesity. This means that the molar concentration of the di-2-ethylhexyl phthalate metabolite in the biological sample according to the present invention is measured and the obesity of the subject can be diagnosed by analyzing the molar concentration in the biological sample.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

Monoethylhydroxyhexyl phthalate and monoethylhexyl phthalate;
Monoethyloxohexyl phthalate and monoethyloxohexyl phthalate;
Monoethyl 6-hydroxyhexyl phthalate and monoethyl hexyl phthalate; And
Monoethyl carboxypentyl phthalate and monoethyl 6-hydroxyhexyl phthalate;
2-ethylhexyl phthalate group selected from the group consisting of di-2-ethylhexyl phthalate and di-2-ethylhexyl phthalate. . The obesity diagnostic kit according to claim 1, wherein the biological sample comprises at least one of urine, blood and skin tissue. 2. The method of claim 1, wherein the obesity diagnostic kit further comprises an instruction sheet including an obesity diagnostic method,
The molar concentration of the di-2-ethylhexyl phthalate metabolite measured from the molar concentration measuring unit in the biological sample is substituted into one or more mathematical formulas selected from the group consisting of the following equations (1) to (8) Analyzing the ethylhexyl phthalate metabolite ratio; And
Diagnosis of obesity from the analysis result;
And an obesity diagnostic kit.
[Equation 1]
RMR A1 = molar concentration of monoethylhydroxyhexyl phthalate in biological sample / molar concentration of monoethylhexyl phthalate in biological sample
&Quot; (2) "
RMR A2 = molar concentration of monoethyloxohexyl phthalate in biological sample / molar concentration of monoethylhydroxyhexyl phthalate in biological sample
&Quot; (3) "
RMR AR = RMR A1 / RMR A2
&Quot; (4) "
RMR AT = RMR A1 * RMR A2
&Quot; (5) "
RMR B1 = molar concentration of monoethyl 6-hydroxyhexyl phthalate in the biological sample / molar concentration of monoethylhexyl phthalate in the biological sample
&Quot; (6) "
RMR B2 = molar concentration of monoethyl carboxypentyl phthalate in biological sample / molar concentration of monoethyl 6-hydroxyhexyl phthalate in biological sample
&Quot; (7) "
RMR BR = RMR B1 / RMR B2
&Quot; (8) "
RMR BT = RMR B1 * RMR B2 4. The method according to claim 3, wherein the step of diagnosing obesity of the instruction sheet comprises the steps of diagnosing obesity if di-2-ethylhexyl phthalate metabolism ratio in a biological sample is different from di-2-ethylhexyl phthalate metabolism ratio of a normal control group And an obesity diagnostic kit. Monoethylhydroxyhexyl phthalate and monoethylhexyl phthalate;
Monoethyloxohexyl phthalate and monoethyloxohexyl phthalate;
Monoethyl 6-hydroxyhexyl phthalate and monoethyl hexyl phthalate; And
Monoethyl carboxypentyl phthalate and monoethyl 6-hydroxyhexyl phthalate;
Measuring the molarity of the di-2-ethylhexyl phthalate metabolite contained in the di-2-ethylhexyl phthalate group selected from the group consisting of the di-2-ethylhexyl phthalate metabolite in the biological sample;
The molar concentration of the di-2-ethylhexyl phthalate metabolite measured from the molar concentration measuring unit in the biological sample is substituted into one or more mathematical formulas selected from the group consisting of the following equations (1) to (8) Calculating an ethylhexyl phthalate metabolite ratio; And
Diagnosing whether or not obesity is caused by comparing the metabolite ratio in the biological sample and the metabolic rate of the normal control;
The method comprising the steps of:
[Equation 1]
RMR A1 = molar concentration of monoethylhydroxyhexyl phthalate in biological sample / molar concentration of monoethylhexyl phthalate in biological sample
&Quot; (2) "
RMR A2 = molar concentration of monoethyloxohexyl phthalate in biological sample / molar concentration of monoethylhydroxyhexyl phthalate in biological sample
&Quot; (3) "
RMR AR = RMR A1 / RMR A2
&Quot; (4) "
RMR AT = RMR A1 * RMR A2
&Quot; (5) "
RMR B1 = molar concentration of monoethyl 6-hydroxyhexyl phthalate in the biological sample / molar concentration of monoethylhexyl phthalate in the biological sample
&Quot; (6) "
RMR B2 = molar concentration of monoethyl carboxypentyl phthalate in biological sample / molar concentration of monoethyl 6-hydroxyhexyl phthalate in biological sample
&Quot; (7) "
RMR BR = RMR B1 / RMR B2
&Quot; (8) "
RMR BT = RMR B1 * RMR B2 6. The method of claim 5, wherein the biological sample comprises at least one of urine, blood and skin tissue. 6. The method according to claim 5, wherein the metabolic rate of the body sample is compared with the metabolic rate of a normal control,
When di-2-ethylhexyl phthalate metabolism ratio in the biological sample calculated by substituting into one or more mathematical formulas selected from the group consisting of the above-mentioned formulas (1) to (8) differs from the di-2-ethylhexyl phthalate metabolism ratio of the normal control group And determining that the obesity is obese.
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Assessing human exposure to phthalates using monoesters and their oxidized metabolites as biomarkers, Environmental Health Perspectives, 2003, vol 11, p1148-1151

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