KR20030084841A - Assay methods of organic matters - Google Patents

Abstract

PURPOSE: An analysis method of an organic material is provided, to allow toxic carbanyl and benzopyrene(a) contained in drinking water to be analyzed simultaneously with a single detector. CONSTITUTION: The analysis method comprises the steps of pretreating a sample water; performing liquid chromatography of the pretreated sample water to separate the organic material contained in the sample water; and analyzing the separated organic material by using a UV detector. Preferably the liquid chromatography is HPLC. Preferably the organic material is carbanyl, benzopyrene(a) or their mixtures.

Description

Assay methods of organic matters

The present invention comprises the steps of pre-treatment inspection; A liquid chromatography step for separating organic matter present in the pretreated inspection; And it relates to an organic material analysis method comprising the step of passing the organic material separated by the liquid chromatography to the ultraviolet detector to measure and analyze the wavelength.

Organic material means a material composed of various elements including carbon. When organic substances are added to water, they are decomposed by microorganisms and consume oxygen in the water. Organic substances are generated in water as representative examples of food waste, manure, trash, pesticides or wastewater from barns.

Harmful organic substances present in water include carbanyl, benzopyrene (a), phenol, benzene, toluene and xylene. Among them, carbanyl and benzopyrene (a) are representative harmful organic substances. People who consumed water exceeding the standard of the two organic substances may develop serious diseases such as disorders of internal organs, difficulty in breathing, and cancer. .

Carbanyl is classified as a health hazard organic substance and it is stated that the standard value for drinking water is 0.07 ppm or less. Ingestion of carbanyl in large amounts can cause nausea, vomiting, diarrhea, decreased vision, dyslexia, muscle cramps, cyanosis and race (convulsions). have.

In addition, benzopyrene (a) is also a health hazard, the reference value that can be included in drinking water is 0.0007 ppm or less. Benzopyrene (a) is best known as a carcinogen and is one of the harmful organic substances found not only in water but also in the atmosphere.

In order to prepare a standard for analyzing the harmful organic substances, the Ministry of Environment has designated and enforced "Rules on Drinking Water Quality Standards and Inspection".

According to the rule, the carbonyl test method of the drinking water quality process test method can be tested by gas chromatography or fluorescence analysis by using liquid chromatography, benzopyrene (a) is a fluorescent substance by liquid chromatography It is described that fluorescence analysis can be performed. However, in the method of analyzing carbanyl, it is possible to analyze by gas chromatography or liquid chromatography, but especially in the case of fluorescence analysis using liquid chromatography, it is necessary to use a post column reaction. It is necessary to use a derivative reagent which is a carcinogenic substance such as mercaptoethanol. In addition, maintenance is difficult because the post-column reaction unit needs to be cleaned after the analysis.

On the other hand, it is not easy to simultaneously analyze the carbanyl and benchopyrene (a) through fluorescence analysis. This is because the excitation wavelength is 286 nm and the emission wavelength is 336 nm, whereas the analysis wavelength of benzopyrene (a) is different because the excitation wavelength is 296 nm and the emission wavelength is 406 nm.

Therefore, two pieces of equipment are required to examine both carbanyl and benzopyrene (a) by the methods presented above. That is, in order to analyze the harmful organic substances carbonyl and benzopyrene (a), expensive analysis equipment is required, and the analysis time and the number of testers also increase relatively.

Accordingly, the present inventors have developed a method capable of simultaneously analyzing organic substances such as carbanyl and benzopyrene (a) present in drinking water with a single analyzer and a detector.

Therefore, the present invention

Pretreatment of the inspection;

(Iii) liquid chromatography to separate the organics present in the inspection of step iv; And

And (b) passing the organic material separated in step iv through an ultraviolet detector to measure and analyze the wavelength, thereby providing an organic material analysis method.

1 is a graph showing ultraviolet spectra of carbanyl and benzopyrene (a) among organic substances present in drinking water.

Figure 2 shows a chromatogram of the simultaneous analysis of carbanyl and benzpyrene (a) by the organic material analysis method of the present invention.

Figure 3 shows a chromatogram of the analysis of carbonyl alone by the organic material analysis method of the present invention.

The present invention comprises the steps of pre-treatment inspection; A liquid chromatography step of separating the organic material present in the inspection by passing the pretreated inspection water through a column; And passing the organic material separated by the liquid chromatography through an ultraviolet detector to measure and analyze the wavelength, thereby providing an organic material analysis method.

In the present invention, the term "pretreatment" means a process of removing impurities and suspended substances present in the inspection in order to separate only the organic substances present in the water well. In the present invention, a filtration method using a filter may be used as a pretreatment method of inspection. The filter may use a filter having a pore size of 0.22 μm to 0.45 μm in consideration of particle sizes such as impurities and suspended substances present in the inspection, and filtration using the filter is performed at least once for accuracy of instrument analysis. Can be.

The organic material analysis method according to the present invention may be used for water containing harmful organic materials, but it is preferable to use drinking water.

In addition, the organic material analysis method according to the present invention can simultaneously detect the harmful substances such as carbonyl, benzopyrene (a), phenol, benzene, toluene and xylene included in the inspection, but preferably has the greatest adverse effect on the human body It may be used to detect carbanyl or benzopyrene (a).

In addition, the organic material analysis method according to the invention is characterized in that it comprises the step of separating the organic materials present in the inspection by using a liquid chromatography analysis method.

The liquid chromatography is liquid-liquid chromatography (Liquid-Liquid Chromatography, LLC) and liquid-solid chromatography that separates organic matter present in the inspection by passing a liquid or solid column and a liquid as a mobile phase as the fixed phase. (Liquid-Solid Chromatography, LSC), etc., and the type of chromatography can be classified into adsorption chromatography (Adsorption Chromatography (AC), partition chromatography (Partition Chromatography (PC), ions) It can be further divided into exchange chromatography (Ion-exchange Chromatography (IC)) and size exclusion chromatography (Size-exclusion Chromatography, SEC). On the other hand, the liquid chromatography can be divided into two types according to the fixed phase column and the mobile phase. One is reverse phase chromatography, and the fixed phase of the column is nonpolar, the mobile phase is polar, and the analyte is polar. The other is normal phase chromatography, which can be used to analyze materials that are well soluble in mobile phases where the fixed phase of the column is polar, the mobile phase is nonpolar, and the analyte is nonpolar.

Liquid chromatography used in the present invention belongs to liquid-solid chromatography, and the classification according to column and mobile phase belongs to reverse phase chromatography. That is, the above-listed chromatography shows that the mobile phase is liquid and polar, the stationary phase is solid and nonpolar, and the organic materials to be analyzed in the present invention, preferably carbanyl and benzopyrene (a), are well dissolved and nonpolar in the polar mobile phase. Since it adsorbs well on the stationary phase, the above-listed chromatography can be used.

In the present invention, for analyzing the organic material present in the inspection by using liquid chromatography, an analyzer including 1) a mobile phase 2) a pump and 3) a fixed phase (hereinafter referred to as a 'column') may be used. As an analyzer, a High Performance Liquid Chromatograph System (HPLC) can be used.

The mobile phase used in the analyzer may be used by mixing two solvents as a liquid. In general, a mobile phase may be prepared by mixing water and an organic solvent in a predetermined ratio and determining a ratio of water and an organic solvent according to the time when the analyte is separated. Organic solvents used in the present invention include, but are not limited to, acetonitrile; Methanol; Tetrahydrofuran; Isopropanol; Any one of the group consisting of ethanol may be used, preferably acetonitrile may be used.

In addition, the mobile phase used in the present invention can be used with a ratio of water and an organic solvent of about 15: 85. This is because the polarity of the organic material present in the inspection, preferably carbanyl and benzopyrene (a) is different. That is, the polarity of the carbanyl is relatively high, so that it is separated first and benzopyrene (a) is later separated. On the other hand, when the amount of the organic solvent is increased, the separation time of the analyte is shortened, and the separation ability may be reduced.

The pump used in the analyzer is to serve to push the mobile phase at a constant flow rate and pressure, it is preferable to use that there is no chemical interaction with the solvent of the mobile phase inside the pump.

The column used in the analyzer is preferably an inner diameter of 3 to 4.6 mm, and a length of 10 to 30 cm. On the other hand, the filler filled in the column may be used, such as silica (silica), alumina (alumina), molecular sieve, and porous polymer, but preferably octadecylsilane (denoted ODS or C-18) on the silica stationary phase. It is preferable to use a substance combined with these.

On the other hand, the organic material analysis method according to the invention is characterized in that it comprises the step of measuring the wavelength of the organic material present in the inspection. In particular, the present invention is characterized by analyzing the wavelength of carbanyl or benzopyrene (a) present in the test.

Carbanyl and benzopyrene (a) has an absorbance value in the ultraviolet region, the maximum absorption wavelength can be seen by examining the ultraviolet spectrum (see Figure 1). As shown in Figure 1, the maximum absorption wavelength of the carbonyl is 220nm, 275nm, the maximum absorption wavelength is 220nm. Benzopyrene (a) has a maximum absorption wavelength of 260 nm, 280 nm, and 296 nm, and a maximum absorption wavelength of 296 nm.

Therefore, the carbonyl or benzopyrene (a) present in the separated test using liquid chromatography is passed through an ultraviolet detector to detect two wavelengths, the maximum absorption wavelength of carbanyl 220 nm and the maximum absorption wavelength of benzopyrene (a). Selecting and analyzing nm can determine whether carbanyl or benzopyrene (a) is present in the test (see FIGS. 2 and 3).

Finally, the organic material analysis method according to the present invention is to separate the organic material present in the inspection by using liquid chromatography, and then pass it through an ultraviolet detector to measure the wavelength, which is analyzed by a data processing system using a computer It can be determined that there is an organic substance present in the test, that is, carbanyl or benzopyrene (a).

Hereinafter, the present invention will be described by the following examples. The following examples illustrate the invention and should be construed as not limiting the scope thereof.

<Example>

Example 1

Determination of maximum absorption wavelength of carbanyl and benzopyrene (a)

Maximum absorption wavelengths of carbanyl and benzopyrene (a) were measured with an ultraviolet spectrophotometer (Beckman Du 7500, Beckman Instruments, U.S.A.), which measures the ultraviolet spectrum.

First, 10 mg of carbanyl and benzopyrene (a) were each taken into a 50 ml volumetric flask, and 25 ml of methanol was added to dissolve it. 0.1 N hydrochloric acid and methanol were mixed in a 1: 1 solution to fill the final solution to 50 ml (each concentration 200 ppm). 1 ml of the solution was taken into a 100 ml volumetric flask, and the final solution was filled with a solution of 0.1 N hydrochloric acid and methanol in a 1: 1 ratio (each concentration of 1 ppm). Again, 1 ml of the solution was added to a 100 ml volumetric flask, and 0.1 N hydrochloric acid and methanol were mixed 1: 1 to fill the final solution with 100 ml. The concentration of carbanyl and benzopyrene (a) was 10 ppb, respectively. Standard solution was used. The standard solution was placed in a quartz cell of 1 cm and scanned at an ultraviolet radiation field (200 nm to 340 nm) to analyze the spectrum.

The ultraviolet spectrums of the above-described carbanyl and benzopyrene (a) are shown in FIG. 1.

As shown in Fig. 1, the maximum absorption wavelength of carbanyl was 220 nm and 275 nm, and the maximum absorption wavelength was 220 nm. The maximum absorption wavelength of benzopyrene (a) was 260 nm, 280 nm, and 296 nm, and the maximum absorption wavelength was 296 nm.

The maximum absorption wavelength of the carbanyl and benzopyrene (a) was used as compared to the wavelength of the carbanyl and benzopyrene (a) present in the inspection analyzed in the following examples.

Example 2

Inspection and Pretreatment

The inspection to be measured was ground water or purified water. The sampling time of the inspection was 09:00 ~ 14:00, and the sampling amount was 4ℓ.

The pretreatment of the collected inspection was filtered with a filter having a pore size of 0.45 μm to remove impurities and suspended substances from the inspection.

Example 3

Analysis using high performance liquid chromatography system

The pretreatment test in Example 2 was analyzed for the carbonyl and benzopyrene (a) present in the test using a high performance liquid chromatography system (Thermo Seperation Products, U.S.A.). The analyzer consists of 1) mobile phase reservoir 2) degassing device 3) pump (Quaternary: supplying 4 different mobile phases with one pump) 4) sample auto-injector 5) column 6) UV field detector (trade name: PAD) and 7 Data processing system (software name: CROMQUST).

The analysis procedure is as follows. To the mobile phase reservoir the ratio of water to acetonitrile was added at 15:85 and the mobile phase was pumped. Thereafter, 20 µl of the pretreatment sample prepared in Example 2 was injected into the inlet through the sample autoinjector, and the inspection was passed through the column together with the mobile phase. The column was an ODS column in which octadecyl silane was bonded onto a silica stationary column, and an inner diameter of the column was 4.6 mm and a length of 25 cm. After the inspection passed the column, the separated analyte was passed through a UV field detector. Passed analytes were converted into electrical signals, sent to a data processing system, and displayed as chromatograms. This is shown in FIGS. 2 and 3.

As shown in Fig. 2, when the carbanyl and benzopyrene (a) separated by the liquid chromatography were passed through an ultraviolet wave detector, it was possible to simultaneously detect the carbanyl and benzopyrene (a) when the absorption wavelength was 296 nm. there was. Compared with the maximum absorption wavelength of carbanyl and benzopyrene (a) measured in Example 1, the maximum absorption wavelength of carbanyl was 220 nm and the maximum absorption wavelength of benzopyrene (a) was 296 nm. .

On the other hand, as shown in FIG. 3, when the organic material present in the inspection was analyzed by the ultraviolet wave field detector, when the absorption wavelength was 220 nm, carbanyl having a maximum absorption wavelength of 220 nm could be detected alone. This can be seen from the fact that the maximum absorption wavelength of carbanyl measured in Example 1 is 220 nm.

The present invention relates to a method for analyzing organic substances present in the inspection, and in particular, since the carbonyl or benzopyrene (a) can be analyzed by a single analyzer and a detector, the organic substance analysis method of the present invention is economical, efficient and analysis time. There is an advantage that can be shortened.

Claims (3)

Pretreatment of the inspection; (Iii) liquid chromatography to separate the organics present in the inspection of step iv; And (B) passing the organic material separated in step (v) through an ultraviolet detector to measure and analyze the wavelength. The method of claim 1, wherein the liquid chromatography is a high performance liquid chromatography system (HPLC). The method of claim 1, wherein the organic material is an individual or a mixture of carbanyl or benzopyrene (a).