KR100613400B1 - METHOD FOR DETERMINATION OF PAHs AND PCBs IN SAMPLE BY GC/MS WITH SPME - Google Patents

Abstract

The present invention provides a PAHs in a water sample or a biological sample by attaching a SPME (Solid Phase Microextraction) fiber coated with a stationary phase capable of adsorption of analyte to a headspace (HS) on the surface of a sample. adsorbing polyaromatic hydrocarbons and polychlorinated biphenyls (PCBs), desorbing the PAHs and PCBs adsorbed fibers at a hot gas chromatography inlet, and then analyzing them in a gas chromatography / mass spectrometer. The present invention relates to a method for simultaneously detecting trace amounts of PAHs and PCBs. The detection method of the present invention is applicable to biological samples including more complicated matrices such as urine or blood in addition to general water samples, and is environmentally and economically beneficial because it has excellent sensitivity, does not use an organic solvent, and does not require pretreatment. Do.

Description

Method of Simultaneous Detection of Residual PHAs and PCSs in Samples Using Solid Phase Trace Extraction / Gas Chromatographs / Mass Spectrometry {METHOD FOR DETERMINATION OF PAHs AND PCBs IN SAMPLE BY GC / MS WITH SPME}

1 is a schematic diagram of a solid-phase microextraction (SPME) device used in the present invention.

* Explanation of symbols in the drawings

(1) plunger (2) plunger holding screw

(3) Adjustable needle guide / depth gauge

(4) Septum-through Needle (5) Septum

(6) Needle with fiber (7) Fiber

(8) Samples (9) Stirring Bars

10 vials 11 aluminum block burner

Figure 2a and 2b is a graph showing the adsorption efficiency of PAHs and PCBs according to the SPME fiber type for water samples in the present invention.

Figure 3a and 3b is a graph showing the adsorption efficiency of PAHs and PCBs according to the SPME adsorption temperature for water samples in the present invention.

Figure 4a and 4b is a graph showing the adsorption efficiency of PAHs and PCBs according to the SPME adsorption time for the water sample in the present invention.

5a and 5b is a graph showing the adsorption efficiency of PAHs and PCBs according to the salting effect in the water sample in the present invention.

6a and 6b are graphs showing the area of each PAHs and PCBs according to the desorption time of the adsorptive fiber to the water sample in the present invention.

7a and 7b are graphs showing the area of each PAHs and PCBs according to the desorption temperature when the adsorption fibers for water samples in the present invention is desorbed at the inlet of gas chromatography.

8a and 8b are graphs showing the adsorption efficiency of PAHs and PCBs according to the salting effect in urine samples in the present invention.

9a and 9b are graphs showing the adsorption efficiency of PAHs and PCBs according to the SPME adsorption temperature for urine samples in the present invention.

10a and 10b are graphs showing the adsorption efficiency of PAHs and PCBs according to pH conditions for urine samples in the present invention.

Figure 11a and 11b is a graph showing the adsorption efficiency of PAHs and PCBs according to the salting effect in the blood sample in the present invention.

12a and 12b are graphs showing the adsorption efficiency of PAHs and PCBs according to the SPME adsorption temperature for blood samples in the present invention.

13a and 13b are graphs showing the adsorption efficiency of PAHs and PCBs according to pH conditions for blood samples in the present invention.

14A and 14B are chromatograms of 18 PAHs and 14 PCBs analyzed by a gas chromatography / mass spectrometer.

The present invention relates to a detection method for the simultaneous analysis of polyaromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) contained in a water sample or a biological sample. More specifically, by using solid phase microextraction (SPME), a sample pretreatment technique that extracts analytes without using organic solvents, 18 PAHs and 14 PCBs are simultaneously extracted from the sample, followed by gas Analysis using chromatography or gas chromatography / mass spectrometry.

 PAHs (polyaromatic hydrocarbons) are a group of compounds in which two or more aromatic rings are fused together and are produced by incomplete combustion when organic substances containing carbon and hydrogen are pyrolyzed at temperatures above 700 ° C. PAHs are released into the environment through a variety of pathways, including thermal power plants, home and industrial heating, incineration and tobacco smoke, and are exposed to humans through ingestion of contaminated air, workplaces or food. The International Cancer Research Council (IARC) reports benzo (a) pyrene; More than 10 PAHs, including B (a) P], have been reported to be carcinogenic in animals, and several epidemiologic studies have reported a significant increase in cancer incidence in workers exposed to PAHs. PAHs are mainly accumulated in the kidneys, liver, fat, spleen, thyroid and ovaries, and are known to show effects of cataracts, jaundice, kidney and liver disorders, dermatitis, erythrocyte destruction, and cancer.

In addition, polychlorinated biphenyls (PCBs) are organic chlorine compounds produced and used worldwide in the United States from the 1930s to the 1970s since they were first produced in the United States in 1929. It is widely used as insulating oil and heat medium of a capacitor. PCB congener is fat-soluble, and as the degree of substitution of chlorine increases, fat-soluble increases, and the accumulation tendency for organisms and soil increases. As the toxicity and bioresistance of this substance are known, its use has been banned since the 1970s, but it is still detected in biological and environmental media due to its high persistence and fat solubility. PCBs move easily through air, water, or soil, are exposed to the human body through contaminated air, water, food, and skin contact, and when they enter the body, they accumulate in adipose tissue, liver, breast milk, etc. Cases of detection in urine, blood, breast milk and the like have been reported. The International Cancer Research Council (IARC) and the US Environmental Protection Agency (EPA) define PCBs as carcinogens that can cause cancer, such as causing anemia of the PCBs, disorders of the liver, stomach, thyroid and reproductive function, and cancer. The adverse effects of the imagination have been reported.

As such, PAHs and PCBs are emerging as global environmental issues due to their high human toxicity, wide distribution in the environment, ease of movement through air or water, environmental and in vivo stability and persistence, The process of monitoring and confirming the detection of these substances has become very important.

Since PAHs and PCBs are present in trace amounts in very complex matrices of environment and biomass, the extraction and concentration of PAHs and PCBs contained in a sample from sample matrix is essential for analyzing trace amounts more accurately. Liquid-liquid extraction and solid phase extraction are widely used for this analysis, but in these cases, due to the complicated process of sample pretreatment and the use of organic solvents, the human body and the environment are consumed with excessive time, cost and labor consumption. There is a problem that is not beneficial.

Therefore, there is a need for the development and application of PAHs and PCBs detection methods that are simple, economical, and have high selectivity and sensitivity, while minimizing the disturbing effects of highly complex matrices of environment and biomaterials.

According to the present invention, the PAHs and PCBs which are present in trace amounts in the environment and the living body are more easily and effectively extracted using solid phase microextraction (SPME), and then excellent gas selectivity and high sensitivity in gas chromatography or gas chromatography / mass spectrometer By providing analytical methods that can be used to analyze PSAs, PAHs and PCBs can be detected to meet the requirements of shorter analysis time, economics, and sensitivity, and to be applied not only to general water samples but also to biological samples such as urine and blood. For the purpose of

The present invention relates to a detection method for simultaneous analysis of PAHs and PCBs contained in water, urine and blood samples. More specifically, the present invention utilizes solid phase microextraction (SPME), which is one of the pretreatment techniques for sample extraction for analyte extraction, without the use of organic solvents. And co-extraction of PCBs, followed by analysis using gas chromatography or gas chromatography / mass spectrometry. The present invention exposes the SPME fiber, which has a solid phase having high interaction with the analyte on the outer wall of the fused silica core, to the headspace to adsorb the analyte in the sample solution, and then desorbs at a high temperature gas chromatography inlet. And separation and analysis directly on a gas chromatography / mass spectrometer.

After the simultaneous extraction and desorption of PAHs (polyaromatic hydrocarbons) and PCBs (polychlorinated biphenyls) in water samples using SPME fibers, the inventors used gas chromatography or gas chromatography / mass spectrometry. As a result of the analysis, it was found that PAHs and PCBs can be analyzed to trace concentrations quickly and economically. Based on experiments on these water samples, the extraction rate of PAHs and PCBs for urine and blood samples under several SPME adsorption conditions (adsorption temperature, salting effect, pH conditions) was investigated. It has been found that the present invention has been achieved.

More specifically, the present invention provides a sample to which a substance selected from the group consisting of NaCl, CaCl ₂ , MgSO ₄ , Na ₂ SO ₄ , (NH ₄ ) ₂ SO ₄ and KCl is added for salting effect; SPME fibers coated with a stationary phase capable of adsorption of the analyte were exposed to a headspace (HS) on the sample surface to adsorb PAHs and PCBs in the sample at 50 to 90 ° C. for at least 20 minutes; PAHs present in trace amounts in the sample, comprising the step of desorbing the fiber to which the PAHs and PCBs are adsorbed at a gas chromatography inlet at 220 to 310 ° C. for 1 to 10 minutes, followed by analysis on a gas chromatography / mass analyzer A method for simultaneous detection of PCBs.

Since the present invention does not use an organic solvent, it is very economical and environmentally advantageous, and since the extraction and concentration process are simultaneously performed on the SPME fiber exposed to the headspace, no special pretreatment and purification process is required. Along with the savings in time and labor, there is the advantage of preventing the loss of analytes and secondary contamination during the multiple steps of sample preparation. The detection method of the present invention can be usefully used for monitoring PAHs and PCBs remaining in trace amounts in biological samples such as urine, blood, sweat, breast milk, saliva and other body fluids, feces and hair, as well as general water samples.

In order to extract an analyte from a sample, the present invention uses a solid phase trace extraction method (SPME). Extracting and concentrating PAHs and PCBs from the sample matrix from the sample matrix is essential for more accurate trace analysis. To this end, conventional liquid-liquid extraction and solid phase extraction have been widely used, but these all require several steps of sample pretreatment and impurity removal, and they pose a problem of using highly toxic organic solvents. In the present invention, to solve this problem, the adsorption of the analyte using the SPME fiber, the SPME fiber is directly exposed to the headspace of the sample solution to adsorb the analyte, and then continuously at a high temperature gas chromatography inlet Desorption is performed directly on a gas chromatography / mass spectrometer. In this way, the SPME fibers are directly exposed to the headspace of the sample solution to adsorb the analyte, thereby preventing the SPME fibers from being rapidly contaminated by the sample matrix during SPME adsorption.

Like extraction with solvents, solid phase microextraction (SPME) is also important for selecting the appropriate fiber type, controlling the adsorption temperature and adsorption time, adding the appropriate salt and adjusting the pH to achieve optimal extraction efficiency. . The present inventors have completed the present invention by determining various adsorption and desorption conditions affecting the efficiency of the SPME, evaluating the analytical efficiency while changing them, and obtaining an optimum range for the SPME.

The SPME fiber means that a stationary phase for adsorption and extraction of an analyte of interest is coated on the outer wall of the fused silica core, and various types and coating thicknesses may be selected and used according to the type of analyte of interest. Can be. In consideration of the polarity, volatility, and molecular size of the analyte of interest, it is important to select an appropriate fiber for the proper utilization of the SPME. In such a fiber selection, the fixed phase coated on the fiber is the most important criterion. After a certain period of time with the fiber exposed to the sample, the analyte is equilibrated and partitioned into the stationary phase. At this time, the time to reach the adsorption equilibrium depends on the size of the analyte or the thickness of the fixed phase coated on the fiber, and the thicker the coating thickness of the fiber, the longer the time to reach the adsorption equilibrium. The coating thickness of the fiber varies depending on the type of fiber used, but it is preferably about 7 to 100 µm in consideration of the adsorptive capacity, analysis efficiency, adsorption equilibrium time, and the like.

Considering the physical and chemical properties (polar and volatility) of the coated stationary phase and PAHs and PCBs, the analytes, the available SPME fibers are shown in Table 1 below. As can be seen in Table 1, in the present invention, as the SPME fiber, polydimethylsiloxane (PDMS) fiber, polydimethylsiloxane-divinylbenzene (PDMS-DVB) fiber, polyacrylate (PA) fiber, One selected from the group consisting of carboxyl-PDMS fibers, and DVB-PDMS-carboxyl fibers can be used, of which polyacrylate (PA) fibers are most suitable.

Fiber along stationary phase Cover thickness (㎛) polarity Cloth stability Temperature (℃) Application method Desirable application Polydimethylsiloxane (PDMS) 100 Nonpolar Unbound 280 GC / HPLC volatile 30 Nonpolar Unbound 280 GC / HPLC Nonpolar Semivolatiles 7 Nonpolar Combined 340 GC / HPLC Medium to nonpolar semivolatiles PDMS-DVB (StableFlex fiber) 65 Bipolar Bridged 270 GC Polar volatiles 60 Bipolar Bridged 270 HPLC Prevailing 65 Bipolar Bridged 270 GC Polar volatiles Polyacrylate (PA) 85 polarity Bridged 320 GC / HPLC Polar semivolatiles (phenols) Carboxen-PDMS (StableFlex fiber) 75 Bipolar Bridged 320 GC Trace volatiles 85 Bipolar Bridged 320 GC Trace volatiles Carbowax / DVB (StableFlex fiber) 65 polarity Bridged 265 GC Polar analytes (alcohols) 70 polarity Bridged 265 GC Polar analytes (alcohols) DVB-PDMS-Carboxen 50/30 Bipolar Bridged 270 GC Aromatic substance

  * StableFlex fiber: 2 cm fiber assembly without spring

If the adsorption temperature is lower than 50 ℃, the extraction rate is very low, the extraction rate is increased above 50 ℃, above 90 ℃, because the sample boils and the fiber is affected by moisture, the extraction rate is reduced, so the adsorption temperature is 50 to 90 ℃ is suitable. It is preferable that the adsorption time is 20 minutes or more, and if the adsorption time is 20 minutes or more, the extraction rate is steadily increased, but in consideration of the molecular size, polarity of the analyte, the thickness of the fiber, and the economics with respect to the experiment time, the present embodiment Was fixed at 50 minutes.

In addition, for the salting effect, CaCl ₂ , MgSO ₄ , Na ₂ SO ₄ , (NH ₄ ) ₂ SO ₄ , KCL and NaCl and the like can be used, in the embodiment of the present invention, in the case of water samples Sample 2 0.1 to 0.6 g of NaCl per ml, 0.1 to 0.4 g of NaCl per 2 ml of sample for urine samples and 0.1 to 0.4 g of NaCl per ml of sample for blood samples were added. It is based on the water solubility (≥10 g / 100 μl) for NaCl and the description of Legal Medicine 1999; 1: 231-7, Journal of Chromatography A, 902 (2000) 267-287; It is the value obtained from the efficiency test by determining the addition range. Therefore, the addition amount of a substance other than NaCl can be appropriately determined in consideration of self-solubility and equivalent weight having ordinary skill in the art.

In addition, the SPME used in the present invention can be applied to biological samples of various matrices such as urine, blood, sweat, breast milk, saliva and other body fluids, feces and hair, and when applied to such biological samples, trace amounts present in the sample In order to increase the adsorption rate of the analyte, it is necessary to minimize the interference effect on various polymers such as proteins and fats present in the biological sample, so that the polymer material is hydrolyzed under a strong acid of pH 3 or lower or a strong base of pH 11 or higher. It is then preferred to adsorb the material to be analyzed. In particular, in the case of urine samples, all strong acid conditions of pH 3 or less or strong base conditions of pH 11 or more are suitable, and in the case of blood samples, strong base conditions of pH 11 or more are suitable.

In addition, the desorption temperature of the analyte adsorbed on the fiber at the high temperature gas chromatography inlet is 220 to 310 in consideration of the physicochemical properties (molecular weight, boiling point, freezing point and molecular form, etc.) of the PAHs and PCBs as the analyte. ℃ is preferred. Desorption time is suitable for 1 to 10 minutes, and if the desorption time is shorter than this, the adsorbed material is not completely desorbed, if it is longer than this is not preferable because the life of the fiber is shortened under high temperature.

1 is a schematic diagram of an SPME apparatus used in one embodiment of the present invention. As can be seen in Figure 1, in the present invention by placing the SPME fibers in the headspace to adsorb the analysis material, it is possible to prevent rapid contamination of the SPME fibers from the sample matrix.

As such, the analyte adsorbed on the fiber is desorbed at the hot gas chromatography inlet and injected into the column and analyzed directly at the gas chromatography / mass spectrometer. In the present invention, all gases generally used in gas chromatography are possible as gases which can be used as mobile phases in the analysis, and gases which are inert to oxidation, that is, nitrogen, helium and hydrogen, are particularly preferable. In addition, in the case of a column that can be used according to the present invention, there is no particular limitation on the length and the inner diameter of the column, but only a moderate polarity.

Hereinafter, the present invention will be described in more detail with reference to the following examples, but the present invention is not limited to the following examples.

The reagents used in the following examples are as follows:

- 16 kinds of PAHs standard mixture: n- hexane 1 each reference material 1 ㎍ per ㎖, Accustandard社[naphthalene (Naphthalene), acenaphthylene (Acenaphthylene), acenaphthene (Acenaphthene), fluorene (Fluorene), phenanthrene (Phenanthrene), Anthracene, Fluoranthene, Pyrene, Benzo (a) anthracene, Chrysene, Benzo (b) Fluoranthene (b) fluoranthene, benzo (k) fluorane thene, benzo (a) pyrene, indeno (1,2,3-cd) pyrene ( Indeno (1,2,3-cd) pyrene), Dibenz (a, h) anthracene, Benzo (g, h, i) perylene perylene)]

Perylene standard solution: 1 μg per 1 mL of n-hexane, Fluka

-Benzo (e) pyrene standard solution: 1 μg per 1 mL of n-hexane, Sigma

14 PCBs standard solution: 1 μg of each standard per ml of n-hexane, Accustandard [3,4,4 ', 5-tetrachlorobiphenyl, 3,3', 4,4 '-Tetrachlorobiphenyl, 2', 3,4,4 ', 5-pentachloro biphenyl, 2,3', 4,4 ', 5-pentachlorobiphenyl , 2,3,4,4 ', 5-pentachlorobiphenyl, 2,3,3', 4,4'-Pentachloro biphenyl, 3,3 ', 4,4' , 5-pentachlorobiphenyl, 2,3 ', 4,4', 5,5'-hexaxabibiphenyl, 2,3,3 ', 4,4', 5-hexachloro Hexachloro biphenyl, 2,3,3 ', 4,4', 5'-Hexachlorobiphenyl, 2,2 ', 3,4,4', 5,5'-heptachlorobi Heptachlorobiphenyl, 3,3 ', 4,4', 5,5'-Hexa chlorobiphenyl, 2,2 ', 3,3', 4,4 ', 5-heptachlorobiphenyl (Heptachlorobiphenyl), 2,3,3 ', 4,4', 5,5'-heptachlorobiphenyl]

Phenanthrene-d ₁₀ solution (internal standard for PAHs, ISTD1): 5 μg per 1 mL of n-hexane, Accustandard

Pyrene-d ₁₀ solution (internal standard for PCBs, ISTD2): 5 μg per ml of n-hexane, Accustandard

Example 1: Adsorption efficiency test of PAHs and PCBs according to SPME fiber type in water sample

Prepare 5 4 ml vials, add 2 ml of blank (third distilled water) to each vial, and add 20 µl of a mixture of PAHs and PCBs standard, and add phenanthrene-d10, an internal standard (ISTD). After adding 1 µl of a pyrene-d10 mixed solution, 0.5 g of NaCl was added. The fiber holders with PDMS 100 μm, PDMS 30 μm, PDMS 7 μm, PDMS / DVB 65 μm and PA 85 μm fibers, respectively, were fixed in the headspace above the water surface in five vials, respectively, and then at the needle of the fiber holder The fibers were removed and exposed at about 1 cm above the water surface. At this time, the temperature of a heater was adjusted to 90 degreeC, the vial containing a sample was heated, and the adsorption time of a fiber was 30 minutes.

In order to confirm the adsorption degree of the fiber adsorbed with PAHs and PCBs in this process, the fiber adsorbed with the PAHs and PCBs was immediately injected into a gas chromatography inlet at 310 ° C. for 5 minutes. The PAHs and PCBs were separated by raising the oven temperature of the gas chromatography and detected by a mass spectrometer to compare the degree of adsorption according to the size of selected ions of each material shown in Table 2 below. The obtained result is shown in FIG. As can be seen from Figure 2, among the five fibers, the most suitable degree of adsorption was shown overall when the adsorption using PA fibers.

Example 2: Test of Adsorption Efficiency of PAHs and PCBs on SPME Adsorption Temperature in Water Samples

Prepare 6 4 ml vials, add 2 ml of the blank sample to each vial, add 20 µl of the PAHs and PCBs standard solution mixture, and 1 µl of the internal standard phenanthrene-d10 and pyrene-d10 solution. After the addition, 0.5 g of NaCl was added. After fixing the fiber holder with PA 85 um fiber to the headspace above the water of the solution in the vial, the fiber was removed from the needle of the fiber holder and exposed at a position about 1 cm above the water. At this time, the temperature of a heater was adjusted to 30 degreeC, 40 degreeC, 50 degreeC, 70 degreeC, 90 degreeC, and 100 degreeC, respectively, and the vial containing a sample was heated and the adsorption time of a fiber was 30 minutes.

In order to confirm the adsorption degree of the fiber adsorbed PAHs and PCBs in the above process, the fiber adsorbed PAHs and PCBs was immediately injected into the gas chromatography inlet of 310 ℃ desorption for 5 minutes. Each PAHs and PCBs were separated while raising the oven temperature of the gas chromatography, detected by mass spectrometry, and the adsorption degree according to the size of selected ions of each material shown in Table 1 of Example 1 was compared. The results are shown in FIG. As can be seen in Figure 3, the adsorption efficiency was excellent when the adsorption temperature is 50 ℃ or more, in particular, showed the most excellent adsorption efficiency when the adsorption temperature is 90 ℃.

Example 3: Adsorption efficiency test of PAHs and PCBs according to SPME adsorption time in water samples

Prepare 6 4 ml vials, add 2 ml of blank sample to each vial, add 20 µl of PAHs and PCBs standard solution, and add 1 µl of the internal standard phenanthrene-d10 and pyrene-d10 solution. After the addition, 0.5 g of NaCl was added. The fiber holder with PA 85 μm fiber was fixed in the headspace above the water of the solution in the vial, and then the fiber was removed from the needle of the fiber holder and exposed at about 1 cm above the water surface. At this time, the temperature of the heater was adjusted to 90 ° C., and the vial containing the sample was heated, and the adsorption time of the fiber was adsorbed for 10 minutes, 20 minutes, 30 minutes, 50 minutes, 70 minutes, and 90 minutes, respectively.

In order to confirm the adsorption degree of the fiber adsorbed by the PAHs and PCBs by the above process, the fiber adsorbed on the PAHs and PCBs was immediately injected into the gas chromatography inlet at 310 ° C. for 5 minutes. The PAHs and PCBs were separated and detected by mass spectrometry while raising the oven temperature of the gas chromatography to compare the degree of adsorption according to the size of the selected ions of each material shown in Table 1 of Example 1 above. The results are shown in FIG. As can be seen in Figure 4, both the PAHs and PCBs showed an increase in the adsorption rate in more than 20 minutes, the adsorption increased after 50 minutes for PCBs, but the analysis time and the simultaneous extraction of PAHs and PCBs When 50 minutes was found to be the optimum adsorption time.

Example 4 Adsorption Efficiency Test of PAHs and PCBs with Salting Effect in Water Samples

Prepare 6 4 ml vials, add 2 ml of the blank sample to each vial, add 20 µl of the PAHs and PCBs standard solution mixture, and mix the internal standard (ISTD) phenanthrene-d10 with pyrene-d10. 1 μl of solution was added. Add 0.1 g, 0.1 g, 0.2 g, 0.4 g, 0.6 g, 1 g of NaCl to each vial, fix the fiber holder with PA 85 μm fiber to the headspace above the water surface of the solution in the vial, and then The fibers were removed from the needle of and exposed at a position about 1 cm above the water surface. At this time, the temperature of the heater was adjusted to 90 ° C. to heat the vial containing the sample, and the adsorption time of the fiber was 50 minutes.

In order to confirm the adsorption degree of the fiber adsorbed by the PAHs and PCBs by the above process, the fiber adsorbed on the PAHs and PCBs was immediately injected into the gas chromatography inlet at 310 ° C. for 5 minutes. The PAHs and PCBs were separated and detected by mass spectrometry while increasing the oven temperature of the gas chromatography, and the adsorption degree according to the size of the selected ions of each material shown in Table 1 of Example 1 was compared. The results are shown in FIG. As can be seen in FIG. 5, when NaCl was added in an amount of 0.1 to 0.6 g with respect to 2 ml of the water sample, overall stability and high adsorption were shown.

Example 5 Area Test of Each PAHs and PCBs with Desorption Time of Adsorbent Fiber

Prepare 6 4 ml vials, add 2 ml of blank sample to each vial, add 20 µl of PAHs and PCBs standard solution, and add 1 µl of an internal standard phenanthrene-d10 and pyrene-d10 solution. After the addition, 0.1 g of NaCl was added. The fiber holder with PA 85 μm fiber was fixed in the headspace above the water of the solution in the vial, and then the fiber was removed from the needle of the fiber holder and exposed at about 1 cm above the water surface. At this time, the temperature of a heater was adjusted to 90 degreeC, the vial containing a sample was heated, and the adsorption time of the fiber was 50 minutes.

In order to confirm the adsorption degree of the fiber adsorbed by the PAHs and PCBs by the above process, the fiber adsorbed by the PAHs and PCBs was immediately injected into the gas chromatography inlet at 310 ° C. for 0.5 minutes, 1 minute, 2 minutes, and 5 minutes, respectively. Desorption for 10 minutes, 20 minutes. Each PAHs and PCBs were separated and raised with a mass spectrometer while raising the oven temperature of the gas chromatography, and the area according to the size of selected ions of each material shown in Table 1 of Example 1 was compared. The results are shown in FIG. As can be seen in Figure 6, when the desorption time is 1 to 10 minutes, in particular, showed a beneficial effect when 5 minutes.

Example 6 Area Test of Each PAHs and PCBs According to Desorption Temperature During Adsorption Fiber Desorption at Inlet of Gas Chromatography

Prepare 6 4 ml vials, add 2 ml of the blank sample to each vial, add 20 µl of the PAHs and PCBs standard solution mixture, and add 1 µl of the internal standard phenanthrene-d10 and pyrene-d10 solution. After the addition, 0.1 g of NaCl was added. The fiber holder equipped with PA 85 μm fiber was fixed in the headspace above the water of the solution in the vial, and then the fiber was removed from the needle of the fiber holder and exposed at about 1 cm above the water surface. At this time, the temperature of a heater was adjusted to 90 degreeC, the vial containing a sample was heated, and the adsorption time of the fiber was 50 minutes.

In order to confirm the adsorption degree of the fibers adsorbed by the PAHs and PCBs by the above process, the fibers adsorbed by the PAHs and PCBs are respectively adjusted to a temperature of 180 ° C., 200 ° C., 220 ° C., 250 ° C., 280 ° C., and 310 ° C. in advance. It was injected into the prepared gas chromatography inlet and desorbed for 5 minutes. Each PAHs and PCBs were separated and raised with a mass spectrometer while raising the oven temperature of the gas chromatography, and the area according to the size of selected ions of each material shown in Table 1 of Example 1 was compared. The results are shown in FIG. As can be seen in FIG. 7, when the desorption temperature is 220 to 310 ° C., the overall stable and high area is shown.

Example 7: Adsorption efficiency test of PAHs and PCBs according to salting effect in urine samples

Prepare 4 4 vials, add 2 ml of urine sample to each vial, add 20 μl of PAHs and PCBs mixed standard solution, and add 1 μl of the internal standard phenanthrene-d10 and pyrene-d10 mixed solution. Put in. NaCl was added to each vial at 0 g, 0.1 g, 0.2 g and 0.4 g, respectively, and the fiber holder with PA 85 μm fiber was fixed in the headspace above the water surface of the solution in the vial, and then the fiber at the needle of the fiber holder Was removed and exposed at a position of about 1 cm above the water surface. Based on the experiment on the water sample, the temperature of the heater was adjusted to 90 ° C., and the vial containing the sample was heated, and the adsorption time of the fiber was 50 minutes.

In order to confirm the adsorption degree of the fiber adsorbed by the PAHs and PCBs by the above process, the fiber adsorbed on the PAHs and PCBs was immediately injected into the gas chromatography inlet at 310 ° C. for 5 minutes. The PAHs and PCBs were separated and detected by mass spectrometry while increasing the oven temperature of the gas chromatography, and the adsorption degree according to the size of the selected ions of each material shown in Table 1 of Example 1 was compared. The results are shown in FIG. As can be seen in FIG. 8, when 0.1 to 0.4 g, especially 0.2 g of NaCl was added to 2 ml of urine sample, the overall degree of adsorption was improved.

Example 8: Adsorption efficiency test of PAHs and PCBs with SPME adsorption temperature in urine samples

Prepare three 4 ml vials, add 2 ml of urine sample to each vial, add 20 µl of the PAHs and PCBs standard solution mixture and mix the internal standard (ISTD) with phenanthrene-d10 and pyrene-d10. After adding 1 μl of the solution, 0.2 g of NaCl was added. The fiber holder with PA 85 μm fiber was fixed in the headspace above the water of the solution in the vial, and then the fiber was removed from the needle of the fiber holder and exposed at about 1 cm above the water surface. At this time, the temperature of a heater was adjusted to 70 degreeC, 80 degreeC, and 90 degreeC, respectively, and the vial containing a sample was heated and the adsorption time of the fiber was 50 minutes.

In order to confirm the adsorption degree of the fiber adsorbed by the PAHs and PCBs by the above process, the fiber adsorbed on the PAHs and PCBs was immediately injected into the gas chromatography inlet at 310 ° C. for 5 minutes. The PAHs and PCBs were separated and detected by mass spectrometry while increasing the oven temperature of the gas chromatography, and the adsorption degree according to the size of the selected ions of each material shown in Table 1 of Example 1 was compared. The results are shown in FIG. As can be seen in Figure 9, when the adsorption temperature is 90 ℃, it showed the highest adsorption efficiency.

Example 9: Adsorption efficiency test of PAHs and PCBs according to pH conditions in urine samples

Prepare 5 4 ml vials, add 2 ml of urine sample to each vial, add 20 µl of the PAHs and PCBs standard solution mixture, and mix the internal standard phenanthrene-d10 and pyrene-d10 solution 1 After adding μL, 0.2 g of NaCl was added. Adjust the pH of the sample under conditions of no pH adjust, pH <2 (6M HCl), pH ≒ 3 (citric acid), pH ≒ 11 (K ₂ CO ₃ ), pH ≒ 13 (KOH) After adjustment, the fiber holder with the PA 85 μm fiber was fixed in the headspace above the water of the solution in the vial, and then the fiber was removed from the needle of the fiber holder and exposed at about 1 cm above the water surface. At this time, the temperature of a heater was adjusted to 90 degreeC, the vial containing a sample was heated, and the adsorption time of the fiber was 50 minutes.

In order to confirm the adsorption degree of the fiber adsorbed by the PAHs and PCBs by the above process, the fiber adsorbed on the PAHs and PCBs was immediately injected into the gas chromatography inlet at 310 ° C. for 5 minutes. The PAHs and PCBs were separated and detected by mass spectrometry while increasing the oven temperature of the gas chromatography, and the adsorption degree according to the size of the selected ions of each material shown in Table 1 of Example 1 was compared. The results are shown in FIG. As can be seen in FIG. 10, the highest adsorption degree was shown in conditions of pH 3 or below or pH 11 and above, especially pH 13.

Example 10: Adsorption efficiency test of PAHs and PCBs according to salting effect in blood samples

Prepare 4 4 vials, add 1 ml of blood sample and 1 ml of tertiary distilled water to each vial, add 20 μl of PAHs and PCBs standard solution mixture, and add Phenanthrene-d10, an internal standard (ISTD), and pie. 2 μl of the Len-d10 mixed solution was added. Add 0.1 g, 0.1 g, 0.2 g, 0.4 g of NaCl to each vial, fix the fiber holder with PA 85 μm fiber to the headspace above the water surface of the solution in the vial, and then remove the fiber from the needle of the fiber holder. It was removed and exposed at a position of about 1 cm above the water surface. Based on the experiment on the water sample, the temperature of the heater was adjusted to 90 ° C. to heat the vial containing the sample, and the adsorption time of the fiber was 50 minutes.

In order to confirm the adsorption degree of the fiber adsorbed by the PAHs and PCBs by the above process, the fiber adsorbed on the PAHs and PCBs was immediately injected into the gas chromatography inlet at 310 ° C. for 5 minutes. The PAHs and PCBs were separated and detected by mass spectrometry while increasing the oven temperature of the gas chromatography, and the adsorption degree according to the size of the selected ions of each material shown in Table 1 of Example 1 was compared. The results are shown in FIG. As can be seen in FIG. 11, the highest degree of adsorption was seen when NaCl was added in an amount of 0.1 to 0.4 g, especially 0.1 g, for 1 ml of blood sample.

Example 11 Adsorption Efficiency Test of PAHs and PCBs with SPME Adsorption Temperature in Blood Samples

Prepare three 4 ml vials, add 1 ml of blood sample and 1 ml of tertiary distilled water to each vial, and add 20 µl of a mixture of PAHs and PCBs standards, and add phenanthrene-d10, an internal standard (ISTD). 2 microliters of pyrene-d10 mixed solution was added, and 0.1 g of NaCl was added thereto. The fiber holder with PA 85 μm fiber was fixed in the headspace above the water of the solution in the vial, and then the fiber was removed from the needle of the fiber holder and exposed at about 1 cm above the water surface. At this time, the temperature of a heater was adjusted to 70 degreeC, 80 degreeC, and 90 degreeC, respectively, and the vial containing a sample was heated and the adsorption time of the fiber was 50 minutes.

In order to confirm the adsorption degree of the fiber adsorbed by the PAHs and PCBs by the above process, the fiber adsorbed by the PAHs and PCBs was immediately injected into the gas chromatography inlet at 310 ° C. and desorbed for 5 minutes. Each PAHs and PCBs were separated by raising the oven temperature of the gas chromatography and detected by mass spectrometry to compare the degree of adsorption according to the size of the selected ions of each material shown in Example 1 above. The results are shown in FIG. As can be seen in Figure 12, when the adsorption temperature is 90 ℃ showed the highest degree of adsorption.

Example 12: Adsorption efficiency test of PAHs and PCBs according to pH conditions in blood samples

Prepare 5 4 ml vials, add 1 ml of blood sample and 1 ml of tertiary distilled water to each vial, add 20 µl of the PAHs and PCBs standard solution mixture, and place the internal standard (ISTD) and phenanthrene-d10 and pie. 2 µl of the len-d10 mixed solution was added and 0.1 g of NaCl was added thereto. The pH of each vial was adjusted as follows: no pH adjust, pH <2 (6M HCl), pH ≒ 3 (citric acid), pH ≒ 11 (K ₂ CO ₃ ), pH ≒ 13 ( KOH). The fiber holder with PA 85 μm fiber was fixed in the headspace above the water of the solution in the vial, and then the fiber was removed from the needle of the fiber holder and exposed at about 1 cm above the water surface. At this time, the temperature of a heater was adjusted to 90 degreeC, the vial containing a sample was heated, and the adsorption time of the fiber was 50 minutes.

In order to confirm the adsorption degree of the fibers adsorbed by the PAHs and PCBs by the above process, the fibers adsorbed on the PAHs and PCBs were immediately injected into a gas chromatography inlet at 310 ° C. for 5 minutes. The PAHs and PCBs were separated and detected by mass spectrometry while increasing the oven temperature of the gas chromatography, and the adsorption degree according to the size of the selected ions of each material shown in Table 1 of Example 1 was compared. The results are shown in FIG. As can be seen in FIG. 13, the highest adsorption degree was shown at a pH of 11 or above, especially at pH = 11.

Example 13

According to the optimum conditions obtained in Examples 1 to 6, after adding an appropriate amount of NaCl for the salting effect to the sample, the PA fiber was installed in the headspace of the sample vial and the temperature of the heater was adjusted to 90 ° C. for 50 minutes. Adsorbed. The fiber to which the analyte was adsorbed was desorbed at a gas chromatography inlet at 310 ° C. for 5 minutes, and then injected into a gas chromatography / mass spectrometer, followed by 18 PAHs, 14 PCBs, and 2 types. A chromatogram of the internal standard of was obtained.

To determine the concentration of these PAHs and PCBs, the analysis was performed using a Polaris-Q ion-trap mass analyzer (thermo-Finnigan) and gas chromatography connected thereto. Electron impact method (Electron impact, EI) was used as the ionization method, the temperature of the ion source was 220 ℃. Selected ion monitoring (SIM) method was used for quantification, and the selected ions are shown in Table 1 above. The column used in the analysis was an ultra 2 column with a length of 25 m, an inner diameter of 0.2 mm and a film thickness of 0.33 μm, helium as the mobile phase gas, and the flow rate was 1 ml / min for the first 12 minutes. The next 11 minutes were programmed to 0.8 ml / min and the remaining 17 minutes back to 1 ml / min. The temperature of the column was initially elevated to 230 ° C. at 15 ° C./min at 80 ° C., then maintained at 230 ° C. for 2 minutes, and again at 245 ° C. at 5 ° C./min, and then at 245 ° C. for 6 minutes. After holding, the temperature was raised to 300 ° C at a rate of 10 ° C / min, and maintained at 300 ° C for 15 minutes. The temperature of the sample inlet was 310 ° C., and the temperature of the gas chromatograph and the mass spectrometer connection device was 300 ° C.

Example 14

After the PAHs and PCBs standard mixed solution was added to the actual water quality, urine, and blood samples, and the internal standard phenanthrene-d10 and pyrene-d10 mixed solutions were added, and according to the optimum conditions obtained in Examples 1 to 12, After adding NaCl for salting effect to the sample and adjusting the pH, PA fiber was installed in the headspace of the sample vial, and the temperature of the heater was adjusted to 90 ° C. for 50 minutes, followed by adsorbing fiber Was desorbed at a gas chromatography inlet at 310 ° C. for 5 minutes and then injected into a gas chromatography / mass spectrometer to obtain calibration curves for PAHs and PCBs for each matrix. Good linearity of at least 0.99 was achieved in the concentration range of 0.02 to 5 ng / ml for water samples, 0.02 to 5 ng / ml for urine samples and 0.05 to 20 ng / ml for blood samples.

As described above, the present invention more easily and effectively extract PAHs and PCBs present in trace amounts in biological samples such as water, urine, blood, and the like using gas phase chromatography or gas chromatography. Provides analytical methods for selective and superior sensitivity analysis in graphimetric / mass spectrometers.

Therefore, the method according to the present invention can satisfy the requirements of shortening the analysis time, economical efficiency, sensitivity, and the like, and can be widely applied to the analysis of biological samples such as body fluids, urine, blood and the like as well as general water samples. Can be.

Claims (9)

delete delete delete Urine, blood, sweat, saliva, breast milk and other body fluids, feces and hair For the salting effect, a substance selected from the group consisting of NaCl, CaCl ₂ , MgSO ₄ , Na ₂ SO ₄ , (NH ₄ ) ₂ SO ₄ and KCl is added, By adjusting the pH of the biological sample to pH 3 or less or pH 11 or more to hydrolyze the polymer material in the biological sample, to prepare a sample that minimizes the interference with the adsorption of the analyte, A solid-phase microextraction (SPME) fiber coated with a stationary phase capable of adsorption of the analyte was mounted in the headspace above the sample surface to adsorb PAHs and PCBs in the sample at 50 to 90 ° C. for at least 20 minutes. Desorbing the PAHs and PCBs adsorbed fibers in a gas chromatography inlet at 220 to 310 ℃ for 1 to 10 minutes, and then analyzing in a gas chromatography / mass spectrometer, Simultaneous detection of PAHs and PCBs present in trace amounts in biological samples. According to claim 4, For the salting effect, in the biological samples other than blood in the biological sample, NaCl is added in an amount of 0.1 to 0.4 g per 2 ml of the sample, in the case of blood, NaCl in 0.1 to 1 ml per Adsorption is carried out after addition in an amount of 0.4 g. delete The method according to claim 4 or 5, wherein the fixed phase coated on the SPME fiber has a thickness of 7 to 100 m. The method of claim 7, wherein the SPME fibers are polydimethylsiloxane (PDMS) fiber, polydimethylsiloxane-divinylbenzene (PDMS-DVB) fiber, polyacrylate (PA) fiber, carboxyl-PDMS fiber and DVB-PDMS- And a carboxyl fiber. The method according to claim 4 or 5, wherein the analysable PAHs are naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluorane Fluoranthene, Pyrene, Benzo (a) anthracene, Chrysene, Benzo (b) fluoranthene, Benzo (k) fluorine Lancene (Benzo (k) fluoranthene), Benzo (a) pyrene, Indeno (1,2,3-cd) pyrene (Indeno (1,2,3-cd) pyrene) , Dibenz (a, h) anthracene and benzo (g, h, i) perylene, and the analyzed PCBs include 3, 4,4 ', 5-Tetrachlorobiphenyl, 3,3', 4,4'-Tetrachlorobiphenyl, 2 ', 3,4,4', 5-pentachlorobiphenyl ( Pentachloro biphenyl), 2,3 ', 4,4', 5-pentachlorobiphenyl, 2,3,4,4 ', 5-pentachlorobiphenyl, 2,3,3', 4,4'-pentachloro Pentachloro biphenyl, 3,3 ', 4,4', 5-pentachlorobiphenyl, 2,3 ', 4,4', 5,5'-hexachlorobiphenyl, 2,3,3 ', 4,4', 5-Hexachloro biphenyl, 2,3,3 ', 4,4', 5'-Hexachlorobiphenyl, 2,2 ' , 3,4,4 ', 5,5'-heptachlorobiphenyl, 3,3', 4,4 ', 5,5'-hexaxachlorobiphenyl, 2,2', 3,3 ', 4,4', 5-heptachlorobiphenyl, and 2,3,3 ', 4,4', 5,5'-heptachlorobiphenyl Way.

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