CN107589199B

CN107589199B - Fingerprint spectrum testing method for polycyclic aromatic hydrocarbon substances in indoor dust fall

Info

Publication number: CN107589199B
Application number: CN201710581357.7A
Authority: CN
Inventors: 阳琴
Original assignee: Guizhou University
Current assignee: Guizhou University
Priority date: 2017-07-17
Filing date: 2017-07-17
Publication date: 2020-06-16
Anticipated expiration: 2037-07-17
Also published as: CN107589199A

Abstract

The invention discloses a fingerprint spectrum testing method of polycyclic aromatic hydrocarbon substances in indoor dust fall, which comprises the following steps: the method comprises the following steps: (1) preparing a test solution; (2) chromatographic conditions are as follows: ZORBAX Eclipse PAHs C₁₈Chromatography column (2.1X 100mm, 1.8 μm, Agilent, USA) with flow rate of 0.8-1.2 mL/min^‑1Detection wavelength of 195-225nm, column temperature of 33-37 ℃, mobile phase: acid acetonitrile (A) -tetrahydrofuran water solution (B) with the volume ratio of 0.01-0.03 percent, and gradient elution is carried out; (3) the determination method comprises the following steps: sucking 1-20 μ l of test solution, injecting into liquid chromatograph, and measuring under the above chromatographic conditions. The method can realize the simultaneous and rapid identification of dozens of PAHs pollutants, even hundreds of PAHs pollutants, further comprehensively represent the pollution level and the characteristics of the PAHs pollutants, and can rapidly find the problem of environmental pollution and take treatment measures in time. The invention reduces the detection cost, improves the treatment efficiency and can greatly reduce the total treatment cost of pollution.

Description

Fingerprint spectrum testing method for polycyclic aromatic hydrocarbon substances in indoor dust fall

Technical Field

The invention relates to a method for quickly identifying persistent organic pollutants in indoor dust fall, in particular to a method for quickly identifying polycyclic aromatic hydrocarbon substances in indoor dust fall.

Background

Polycyclic Aromatic Hydrocarbons (PAHs) are volatile Hydrocarbons generated when organic matters such as coal, petroleum, wood, tobacco, organic high molecular compounds and the like are incompletely combusted, and are important environmental and food pollutants.

For the PAHs monitoring species, a gradual transition from early monitoring of 8 PAHs compounds to USEPA recommendation of optimal control of 16 PAHs compounds was made. In 1979, the U.S. Environmental Protection Agency (EPA) issued 129 priority pollutants, of which 16 are PAHs. In 8 months in 2014, the german technical equipment and consumer products committee (ATAV) published the PAHs control requirements of GS certified products, the new standard not only maintains the total amount limit of 18 kinds of PAHs and the limit of benzo (a) pyrene, but also increases the limit of 10 kinds of PAHs such as benzo (e) pyrene, benzo (a) anthracene, benzo (b) fluoranthene and naphthalene, and regulates the total amount limit of 7 kinds of PAHs such as acenaphthylene, fluorene, phenanthrene, pyrene, anthracene and fluoranthene. In this context, 18 distinct PAHs compounds are of wide interest in various fields including environmental studies. However, in the field of environmental research, as many as over 200 kinds of PAHs and derivatives thereof have been discovered so far, and a considerable part of PAHs pollutants with stronger carcinogenicity and obvious toxicity are far more than 18 kinds of PAHs which are widely concerned, and toxic pollutants of PAHs other than 18 kinds of PAHs are far from receiving due attention. PAHs pollutants are various in types and complex in composition, and dozens of or even hundreds of PAHs pollutants are difficult to be simultaneously and rapidly identified by the existing technology and method in the environmental field, so that the pollution level and the characteristics of the PAHs pollutants are comprehensively represented.

Therefore, in the prior art, tens of PAHs pollutants and even hundreds of PAHs pollutants are difficult to be identified simultaneously, the pollution level and the characteristics of the PAHs pollutants cannot be represented comprehensively, the problem of environmental pollution cannot be found quickly, and the environment cannot be treated by taking treatment measures in time. High detection cost, low treatment efficiency and high total pollution treatment cost.

Disclosure of Invention

The invention aims to provide a fingerprint spectrum testing method for polycyclic aromatic hydrocarbon substances in indoor dust fall. The method can realize the simultaneous and rapid identification of dozens of PAHs pollutants, even hundreds of PAHs pollutants, further comprehensively represent the pollution level and the characteristics of the PAHs pollutants, and can rapidly find the problem of environmental pollution and take treatment measures in time. The invention reduces the detection cost, improves the treatment efficiency and can greatly reduce the total treatment cost of pollution.

The invention is realized by adopting the following technical scheme: a fingerprint spectrum testing method for polycyclic aromatic hydrocarbon substances in indoor dust fall comprises the following steps:

(1) preparing a test solution: taking 0.2g of indoor dust fall sample, adding 0.01-0.05 time weight of dehydrogenase and 15-25 times weight of dichloromethane solution containing 20-30% acetonitrile, performing ultrasonic extraction, performing microwave extraction, filtering, concentrating the filtrate to 0.8-1.2mL, transferring the concentrated solution to a chromatographic column, pre-washing the chromatographic column by using 8-12mL of n-hexane-diethyl ether solvent, eluting by using 8-12mL of n-hexane-dichloromethane, performing nitrogen blowing concentration to dryness, dissolving the residue by using 0.8-1.2mL of methanol, filtering by using a 0.45 mu m microfiltration membrane, and collecting the filtrate;

(2) chromatographic conditions are as follows: ZORBAX Eclipse PAHs C₁₈Chromatography column (2.1X 100mm, 1.8 μm, Agilent, USA) with flow rate of 0.8-1.2 mL/min^-1Detection wavelength of 195-225nm, column temperature of 33-37 ℃, mobile phase: acid acetonitrile (A) -tetrahydrofuran water solution (B) with the volume ratio of 0.01-0.03 percent, and gradient elution is carried out;

(3) the test method comprises the following steps: sucking 1-20 μ l of test solution, injecting into liquid chromatograph, testing under the above chromatographic conditions, and recording chromatogram.

In the fingerprint spectrum testing method of polycyclic aromatic hydrocarbon substances in indoor dust fall, ultrasonic extraction is carried out; ultrasonic extracting at 28-32 KHz for 10-14 min.

In the fingerprint spectrum testing method of polycyclic aromatic hydrocarbon substances in indoor dust fall, the microwave is extracted; microwave extraction is carried out for 4-6min under 680-720W.

In the fingerprint spectrum testing method of polycyclic aromatic hydrocarbon substances in indoor dust fall, the chromatographic column is arranged in the sample chamber; the filler in the chromatographic column is anhydrous sodium sulfate, silicon dioxide and copper powder.

In the fingerprint spectrum testing method for polycyclic aromatic hydrocarbon substances in indoor dust fall, the fillers in the chromatographic column are anhydrous sodium sulfate, silicon dioxide and copper powder; the mass ratio of anhydrous sodium sulfate, silicon dioxide and copper powder is 1: 1.5-2.5: 0.1-1.0.

In the fingerprint spectrum test method of polycyclic aromatic hydrocarbon substances in indoor dust fall, the n-hexane-ether solvent is adopted; the volume ratio of n-hexane to diethyl ether is 2.5-3.5: 1.

In the fingerprint spectrum testing method of polycyclic aromatic hydrocarbon substances in indoor dust fall, the n-hexane-dichloromethane; the volume ratio of n-hexane to dichloromethane is 0.8-1.2: 1.

In the fingerprint spectrum testing method of polycyclic aromatic hydrocarbon substances in indoor dust fall, a gradient elution program is carried out; the elution procedure was: 0-4.5min, 0% (A) -100% (B); 4.5-15.0min, 8% (A) -92% (B); 15.0-22.0min, 52% (A) -48% (B); 22.0-35.0, 70% (A) -30% (B); 35-50min, 95% (A) -5% (B); 50min, 95% (A) to 5% (B).

The inventors have conducted extensive testing of the method of the invention for a long period of time, in part as follows:

firstly, the optimized PAHs fingerprint spectrum test method in the indoor dust fall sample is as follows:

1. preparation of test solution

0.2g of the sample was collected, 0.03 times by weight of dehydrogenase and 20 times by weight of a dichloromethane solution containing 25% acetonitrile were added to the sample, and the mixture was subjected to ultrasonic extraction (30 kHz) for 12 minutes, followed by microwave extraction (700W) for 5 minutes, filtration, concentration of the filtrate to about 1mL, transfer of the concentrate to a column (1g of anhydrous sodium sulfate +2g of silica +0.5g of copper powder), pre-washing of the column with 10mL of n-hexane-ether solvent (volume ratio: 3+1), elution with 10mL of n-hexane-dichloromethane (1+1), nitrogen-blown concentration to dryness, dissolution of the residue in 1mL of methanol, filtration through a 0.45 μm microporous membrane, and analysis of the filtrate by HPLC.

2. Chromatographic conditions

ZORBAX Eclipse PAHs C₁₈Column chromatography (2.1X 100mm, 1.8 μm, Agilent, USA), flow rate 1mL min^-1The detection wavelength is 220nm, the column temperature is 35 ℃, and the sample injection amount is 10 mu l.

3. Gradient elution procedure

The change of A, B phase is shown in the following table 1, using acidic acetonitrile (containing acetic acid 0.05 vol.%) as phase a and aqueous solution containing tetrahydrofuran 0.02 vol.% (as phase B), with different analysis time nodes:

table 1 mobile phase gradient elution procedure

Second, detailed research procedure

1. Sample preparation method

(1) Extraction of PAHs from samples

The extraction is carried out by adopting the following method: and taking 0.2g of indoor dust fall sample, respectively adding 0.03 time weight of dehydrogenase and 20 times weight of dichloromethane solution containing 25% acetonitrile, ultrasonically extracting (30 kilohertz) for 12 minutes, then performing microwave extraction (700W) for 5 minutes, and filtering to obtain the product.

(2) Purification research of PAHs in extracting solution

1) Evaluation index

① PAHs recovery rate, 2-fluorobiphenyl was added to the extract solution prepared under the above item (1) at the beginning of purification to average the effect of different purification methods, and the recovery rate was calculated as follows:

② time of purification process the time required for the different purification processes was recorded by timing from the start of the purification process.

2) Different purification treatment methods

① selection of packing combinations in the column, the type and volume of elution solvent was fixed, and different packing combinations were investigated under these conditions, and the results are shown in Table 2.

TABLE 2 purification Effect of different Filler combinations

As can be seen from table 2, from the point of view of recovery: the recovery of the combination of "1 g anhydrous sodium sulfate +2g silica +0.5g copper powder" was 93%, which is significantly better than the other combinations. From the time-consuming point of view: in each combination, the time consumed after copper powder addition was significantly less than without copper powder, indicating that the addition of copper powder to the filler combination significantly reduced the time required for purification.

② optimization of solvent elution systems based on the physical and chemical properties of PAHs, different solvent elution systems were optimized using a combination of "1 g anhydrous sodium sulfate +2g silica +0.5g copper powder" as the filler, with the results shown in Table 3:

TABLE 3 purification effectiveness of different solvent elution systems

As can be seen from table 3, from the recovery rate perspective: the recovery rate of pre-washing the chromatographic column by 10mL of n-hexane-diethyl ether solvent (volume ratio of 3+1) and then eluting by 10mL of n-hexane-dichloromethane (1+1) is basically equivalent to that of pre-washing the chromatographic column by 10mL of n-hexane-diethyl ether solvent (volume ratio of 3+1) and then eluting by 10mL of n-hexane-trichloromethane (1+1), and the recovery rate is obviously superior to that of other solvent systems; from the time-consuming point of view: the time required for the "pre-washing of the column with 10mL of n-hexane-diethyl ether solvent (volume ratio 3+1) and then elution with 10mL of n-hexane-dichloromethane (1+ 1)" is substantially the same as the time required for the "pre-washing of the column with 10mL of n-hexane-diethyl ether solvent (volume ratio 3+1) and then elution with 10mL of n-hexane-chloroform (1+ 1)". It can be seen that the elution ability and effect of "pre-washing the chromatographic column with 10mL of n-hexane-ether solvent (volume ratio 3+1) and then eluting with 10mL of n-hexane-dichloromethane (1+ 1)" and "pre-washing the chromatographic column with 10mL of n-hexane-ether solvent (volume ratio 3+1) and then eluting with 10mL of n-hexane-trichloromethane (1+ 1)" are consistent, but considering that the toxicity of trichloromethane is significantly higher than that of dichloromethane, the finally selected solvent elution system is: the column was pre-washed with 10mL of n-hexane-ether solvent (volume ratio 3+1) and then eluted with 10mL of n-hexane-dichloromethane (1+ 1).

(3) Optimized resulting sample preparation method

The optimal sample preparation method obtained by the above-mentioned preferences and studies of "(1) and (2)" is: 0.2g of the sample was collected, 0.03 times by weight of dehydrogenase and 20 times by weight of a dichloromethane solution containing 25% acetonitrile were added to the sample, and the mixture was subjected to ultrasonic extraction (30 kHz) for 12 minutes, followed by microwave extraction (700W) for 5 minutes, filtration, concentration of the filtrate to about 1mL, transfer of the concentrate to a column (1g of anhydrous sodium sulfate +2g of silica +0.5g of copper powder), pre-washing of the column with 10mL of n-hexane-ether solvent (volume ratio: 3+1), elution with 10mL of n-hexane-dichloromethane (1+1), nitrogen-blown concentration to dryness, dissolution of the residue in 1mL of methanol, filtration through a 0.45 μm microporous membrane, and analysis of the filtrate by HPLC.

2. Optimization and screening of chromatographic conditions

(1) Mobile phase composition optimization and screening

Methanol-water and acetonitrile-water systems are the first choice mobile phase systems of reversed phase liquid chromatography, and the two systems mainly have differences in the aspects of absorbance, chromatographic pressure, elution capacity, separation selectivity and the like. In the ultraviolet absorption spectra of HPLC grade acetonitrile and methanol, the absorption of acetonitrile is small, especially in the short wavelength band. HPLC grade methanol is mainly used for eliminating impurities with ultraviolet absorption, so that the absorbance value at a specific wavelength is controlled within a specification value. HPLC grade acetonitrile is suitable for high sensitivity analysis of samples in short wavelength bands. In addition, HPLC grade acetonitrile produced fewer ghost peaks when subjected to gradient elution. The pressures of the mobile phase elution with different organic solvent types and different mixing ratios in the chromatographic column are greatly different. The ratio of the methanol-water and acetonitrile-water mixing systems is related to the chromatographic pressure, and the same composition ratio of the methanol-water system has a higher chromatographic pressure than the acetonitrile-water system. The same ratio of methanol-water and acetonitrile-water, usually acetonitrile-water, has a stronger elution capacity, especially at lower organic phase ratios, this trend is more pronounced. Methanol is protic and acetonitrile is aprotic due to the large difference in chemical properties between methanol and acetonitrile, so if good separation cannot be obtained using acetonitrile, methanol can be used.

Based on the above analysis, the present invention pairs CH sequentially₃CN-Water (group 1), CH₃OH-Water (group 2), CH₃CN-0.01H₃PO₄(group 3) and CH₃OH-0.03 H₃PO₄(group 4) four mobile phase systems were intensively studied.

The results of comparing the performance of the four mobile phase systems are shown in FIG. 1. As shown in fig. 1, from the viewpoint of the separation degree, the numbers of the groups 1 to 4 with the separation degrees of more than 1.5 are respectively 150, 113, 108 and 113, and the group 1 has a better separation effect; for the number of chromatographic peaks, the numbers of chromatographic peaks in groups 1 to 4 are respectively 160, 143, 137 and 141, and the number of chromatographic peaks in group 1 is large; peak area, with reference to group 4, the peak area of groups 1-3 was 1.3, 1.1 and 0.96 times that of group 4, indicating that the peak area of the chromatogram was the largest in group 1. According to three assessment indexes of comprehensive separation degree, chromatographic peak number and peak area, the group 1 is CH₃The CN-water system has better separation and elution capacity, so that CH is selected₃The CN-water system serves as a mobile phase system for chromatographic separation.

(2) Detection wavelength

In order to obtain as much chromatographic information as possible, four wavelengths, 220nm, 254nm, 310nm and 340nm, were selected for simultaneous measurement, while a full wavelength scan study was performed using a diode array detector, the 3D plot of which is shown in fig. 2. The contrast research shows that the chromatographic peak appears most under the wavelength of 220nm, and the absorption intensity of most chromatographic peaks is the maximum at the wavelength of 220m, so that the wavelength of 220nm is finally taken as the detection wavelength for collecting the PAHs fingerprint in the indoor dust fall sample.

(3) Chromatographic column

The chromatographic column is a carrier for chromatographic separation, each compound in a sample enters a detector after being separated on the chromatographic column, a characteristic signal of the compound is captured by the detector and then information (chromatographic peak) on a chromatogram is converted, and qualitative and quantitative analysis is carried out according to retention time and peak area of the chromatographic peak. Therefore, the proper chromatographic column is very important for collecting PAHs fingerprint in the dustfall sample. In the research process of the invention, the chemical characteristics of PAHs are fully considered, and reversed-phase liquid chromatography is selected for separation, so that a reversed-phase chromatographic column needs to be optimized. The invention selects a conventional silica gel bonded chromatographic column Eclipse XDB-C8(4.6 multiplied by 250mm, 5 mu m), ZORBAXSB-C18(4.6 multiplied by 250mm, 5 mu m) and a special chromatographic column ZORBAXeclipse PAHs C for PAHs₁₈(4.6X 250mm, 5 μm) for comparison. The results showed that conventional columns Eclipse XDB-C8 (4.6X 250mm, 5 μm) and ZORBAXSB-C18 (4.6X 250mm, 5 μm) although capable of separating PAHs, were significantly less effective than ZORBAX Eclipse PAHs C in analysis in terms of peak profile and resolution of each spectral peak₁₈(4.6X 250mm, 5 μm) chromatography column. ZORBAX Eclipse PAHs C₁₈The (4.6 multiplied by 250mm, 5 mu m) can completely separate each PAHs in the dustfall sample, and has better separation degree and sharp chromatographic peak type, thereby meeting the requirement of chromatographic analysis and evaluation. Therefore, ZORBAXeclipse PAHs C was selected₁₈(4.6X 250mm, 5 μm) chromatography column was used for subsequent studies.

(4) Optimization of gradient elution procedure

Different gradient elution procedures have decisive influence on the separation effect of each PAHs component in the sample, so that in the research process of the invention, the following different elution procedures are researched by taking the peak type, the mathematical theory and the separation degree of chromatographic peaks as evaluation indexes:

gradient elution procedure 1:

time (min)	Phase A (%)	Phase B (%)
			0	0	100
4.5	8	92
			15.0	52	48
22.0	70	30
			35.0	95	5
50.0	95	5

Gradient elution procedure 2:

gradient elution procedure 3:

time (min)	Phase A (%)	Phase B (%)
			0	0	100
6	6	94
			14.0	45	55
26.0	65	35
			40.0	95	5
50.0	95	5

Gradient elution procedure 4:

time (min)	Phase A (%)	Phase B (%)
			0	0	100
6	10	90
			17.0	40	60
28.0	80	20
			36.0	95	5
50.0	95	5

Gradient elution procedure 5:

time (min)	Phase A (%)	Phase B (%)
			0	0	100
7	7	93
			12.0	59	41
24.0	75	26
			37.0	92	8
50.0	95	5

Gradient elution procedure 6:

time (min)	Phase A (%)	Phase B (%)
			0	0	100
7.5	9	91
			16.0	54	46
23.0	73	27
			38.0	91	9
50.0	95	5

Gradient elution procedure 7:

time (min)	Phase A (%)	Phase B (%)
			0	0	100
6.5	5	95
			15.5	53	47
25.0	77	23
			38.5	93	7
50.0	95	5

The results show that the gradient elution procedure 1 has the best effect in terms of the peak shape, mathematical and resolution of each chromatographic peak, and the gradient elution procedure has the best resolution, sharp chromatographic peak shape and the largest number of chromatographic peaks, and meets the requirements of chromatographic analysis and evaluation. Therefore, gradient elution procedure 1 was chosen as the final analysis condition.

(5) Quality assurance and quality control

1) Precision experiment

Precision experiments are commonly used to assess the accuracy and reliability of a newly created qualitative or quantitative analytical method, by simultaneously measuring the same test sample using defined measurement conditions, recording at least 6 parallel measurements, calculating the relative mean standard deviation (RSD) values between the different measurements, and considering the established analytical method to have good precision if the RSD value is less than 3%.

In the research process of the invention, the established PAHs fingerprint spectrum test method in the dustfall sample is subjected to systematic precision test. The method comprises the following specific steps: and taking a proper amount of the dust fall sample, continuously injecting samples for 6 times according to an optimal sample solution preparation method and chromatographic conditions, and recording the atlas. Taking a large peak of about 34min as a reference, respectively recording the retention time and the chromatographic peak area of a common chromatographic peak in a chromatogram, and calculating respective RSD values, wherein the result shows that the peak area (A) and the retention time (T) are_R) RSD values are 0.47% and 2.46% respectively, and the precision test requirement is met. Therefore, the method established by the invention is considered to have good precision and can be used for correlation analysis and evaluation.

2) Repeatability test

The repeatability test is that the same sample is used as a research object, the established analysis method is subjected to parallel test research, under the condition that operators, instruments, equipment, places and the like are not changed, the same concentration of a sample is taken in a specified range, at least 6 parts of results are evaluated, and if the relative mean standard deviation (RSD) value of the measured results is less than 3%, the repeatability of the method is considered to be good. In the research process of the invention, 0.5g of dust fall sample powder, 6 parts in total, are taken, the test solution is prepared according to the established method, and the chromatogram is measured and recorded. And taking a large peak of about 34min as a reference, recording the relative peak area and the relative retention time of the common peak, calculating the RSD value of the common peak, and observing the reproducibility of the experiment. The results are shown in FIG. 3 and indicate the peak area (A) and retention time (T) between the common chromatographic peaks shown in the chromatograms in 6 samples_R) RSD values are 0.50% and 2.51% respectively, and the requirement of a repeatability test is met, so that the established PAHs fingerprint spectrum acquisition method can repeatedly reproduce a test result and can be used for correlation analysis and evaluation.

3) Stability test

The stability test is usually used for evaluating the stability, no decomposition and no deterioration of a target substance in a test solution in a time range, so that the established analysis test method is guided to perform analysis test in a reasonable time range, and the accuracy and reliability of a test result are ensured. Stability ofThe test usually requires the same sample solution to be tested and analyzed at different time nodes, and the relative standard deviation between the test results is calculated. In the research process of the invention, the dedusting test sample solution is taken and measured at 6 different times of 0h, 2h, 4h, 8h, 12h and 24h according to the optimized chromatographic conditions. The results show that the peak-to-peak area (A) and the retention time (T) are shared_R) RSD values of 0.49% and 2.51% respectively indicate that the test solution has good stability within 24 h.

(6) Sample testing

Taking the indoor dust fall sample of the house in the Guiyang area as an example, the fingerprint spectrum of the indoor sample of a certain house is measured by adopting the established measuring method, and the measuring result is shown in the table 4 and is as follows:

from fig. 4, it can be seen visually that the indoor dustfall contains about 38 chromatographic peaks, namely: the dust fall sample contains 38 PAHs substances. Through the test of the fingerprint, the PAHs substances in the dust fall sample are quickly identified.

The above-mentioned rapid identification is only the identification of PAHs pollutants from the qualitative point of view of substances, and in order to further characterize the content level of PAHs substances in the sample, each specific PAHs was quantitatively tested, and the results are shown in Table 10.

TABLE 10 PAHs measurement results in a dustfall sample

Therefore, the method can realize the simultaneous and rapid identification of dozens of PAHs pollutants, even hundreds of PAHs pollutants, further comprehensively represent the pollution level and the characteristics of the PAHs pollutants, quickly find the problem of environmental pollution and take treatment measures in time. The invention reduces the detection cost and improves the treatment efficiency, so the invention can greatly reduce the total treatment cost of pollution.

Drawings

FIG. 1 is a graph of performance of different mobile phase systems;

FIG. 2 is a full wavelength scanning 3D plot of an indoor dustfall sample;

FIG. 3 is a chromatogram of a repetitive experimental study;

FIG. 4 is a fingerprint of PAHs substances in an indoor dustfall sample of a certain household.

Example 1.

A fingerprint spectrum testing method for polycyclic aromatic hydrocarbon substances in indoor dust fall comprises the following steps:

(1) preparing a test solution: taking 0.2g of indoor dust fall sample, adding 0.03 time weight of dehydrogenase and 20 times weight of dichloromethane solution containing 25% acetonitrile, performing ultrasonic extraction, performing microwave extraction again, filtering, concentrating the filtrate to 1.0mL, transferring the concentrated solution to a chromatographic column, pre-washing the chromatographic column by using 10mL of n-hexane-diethyl ether solvent, eluting by using 10mL of n-hexane-dichloromethane, performing nitrogen blowing concentration to dryness, dissolving the residue by using 1.0mL of methanol, passing through a 0.45 mu m microporous filter membrane, and collecting the filtrate;

(2) chromatographic conditions are as follows: ZORBAX Eclipse PAHs C₁₈Column chromatography (2.1X 100mm, 1.8 μm, Agilent, USA), flow rate 1.0mL min^-1Detection wavelength of 220nm, column temperature of 35 ℃, mobile phase: acid acetonitrile (A) -tetrahydrofuran water solution (B) with the volume ratio of 0.02 percent, and gradient elution is carried out;

(3) the determination method comprises the following steps: sucking 10 μ l of test solution, injecting into liquid chromatograph, and measuring under the above chromatographic conditions.

The ultrasonic extraction; is ultrasonic extracted at 30 Hz for 12 min.

The microwave extraction; microwave extracting at 700W for 5 min.

The chromatography column; filling materials in the chromatographic column are anhydrous sodium sulfate, silicon dioxide and copper powder; the mass ratio of the anhydrous sodium sulfate to the silicon dioxide to the copper powder is 1: 2.0: 0.8.

the n-hexane-ether solvent; the volume ratio of n-hexane to diethyl ether is 3.0: 1.

The normal hexane-dichloromethane; the volume ratio of n-hexane to dichloromethane was 1.0: 1.

Performing gradient elution; the elution procedure was: 0-4.5min, 0% (A) -100% (B); 4.5-15.0min, 8% (A) -92% (B); 15.0-22.0min, 52% (A) -48% (B); 22.0-35.0, 70% (A) -30% (B); 35-50min, 95% (A) -5% (B); 50min, 95% (A) to 5% (B).

Example 2.

(1) preparing a test solution: taking 0.2g of indoor dust fall sample, adding 0.05 times weight of dehydrogenase and 25 times weight of dichloromethane solution containing 30% acetonitrile, performing ultrasonic extraction, performing microwave extraction again, filtering, concentrating the filtrate to 1.2mL, transferring the concentrated solution to a chromatographic column, pre-washing the chromatographic column by using 12mL of n-hexane-diethyl ether solvent, eluting by using 12mL of n-hexane-dichloromethane, performing nitrogen blowing concentration to dryness, dissolving the residue by using 1.2mL of methanol, passing through a 0.45 mu m microporous filter membrane, and collecting the filtrate;

(2) chromatographic conditions are as follows: ZORBAX Eclipse PAHs C₁₈Column chromatography (2.1X 100mm, 1.8 μm, Agilent, USA), flow rate 1.2mL min^-1Detection wavelength 225nm, column temperature 37 ℃, mobile phase: acid acetonitrile (A) -tetrahydrofuran water solution (B) with the volume ratio of 0.03 percent, and gradient elution is carried out;

(3) the determination method comprises the following steps: sucking 20 μ l of test solution, injecting into liquid chromatograph, and measuring under the above chromatographic conditions.

The ultrasonic extraction; the ultrasonic extraction is carried out at 32 KHz for 14 min.

The microwave extraction; is microwave-extracted at 720W for 6 min.

The chromatography column; filling materials in the chromatographic column are anhydrous sodium sulfate, silicon dioxide and copper powder; the mass ratio of the anhydrous sodium sulfate to the silicon dioxide to the copper powder is 1: 1.5: 0.1.

the n-hexane-ether solvent; the volume ratio of n-hexane to diethyl ether is 3.5: 1.

The normal hexane-dichloromethane; the volume ratio of n-hexane to dichloromethane was 1.2: 1.

Performing gradient elution; the gradient elution procedure was: 0-4.5min, 0% (A) -100% (B); 4.5-15.0min, 8% (A) -92% (B); 15.0-22.0min, 52% (A) -48% (B); 22.0-35.0, 70% (A) -30% (B); 35-50min, 95% (A) -5% (B); 50min, 95% (A) to 5% (B).

Example 3.

(1) preparing a test solution: taking 0.2g of indoor dust fall sample, adding 0.01-time weight of dehydrogenase and 15-time weight of dichloromethane solution containing 20% acetonitrile, performing ultrasonic extraction, performing microwave extraction, filtering, concentrating the filtrate to 0.8mL, transferring the concentrated solution to a chromatographic column, pre-washing the chromatographic column by using 8mL of n-hexane-diethyl ether solvent, eluting by using 8mL of n-hexane-dichloromethane, performing nitrogen blowing concentration to dryness, dissolving the residue by using 0.8mL of methanol, passing through a 0.45-micrometer microporous filter membrane, and collecting the filtrate;

(2) chromatographic conditions are as follows: ZORBAX Eclipse PAHs C₁₈Column (2.1X 100mm, 1.8 μm, Agilent, USA) at a flow rate of 0.8 mL/min^-1Detection wavelength of 195nm, column temperature of 33 ℃, mobile phase: acid acetonitrile (A) -tetrahydrofuran water solution (B) with the volume ratio of 0.01 percent, and gradient elution is carried out;

(3) the determination method comprises the following steps: sucking 1 μ l of test solution, injecting into liquid chromatograph, and measuring under the above chromatographic conditions.

The ultrasonic extraction; is ultrasonic extracted at 28 KHz for 10 min.

The microwave extraction; microwave extracting at 680W for 4 min.

The chromatography column; filling materials in the chromatographic column are anhydrous sodium sulfate, silicon dioxide and copper powder; the mass ratio of the anhydrous sodium sulfate to the silicon dioxide to the copper powder is 1: 2.5: 1.0.

the n-hexane-ether solvent; the volume ratio of n-hexane to diethyl ether is 2.5: 1.

The normal hexane-dichloromethane; the volume ratio of n-hexane to dichloromethane was 0.8: 1.

Performing gradient elution; the elution procedure is 0-4.5min, 0% (A) -100% (B); 4.5-15.0min, 8% (A) -92% (B); 15.0-22.0min, 52% (A) -48% (B); 22.0-35.0, 70% (A) -30% (B); 35-50min, 95% (A) -5% (B); 50min, 95% (A) to 5% (B).

Claims

1. A fingerprint test method for polycyclic aromatic hydrocarbon substances in indoor dust fall is characterized in that: the method comprises the following steps:

(2) chromatographic conditions are as follows: ZORBAXeclipsePHASC 18 column 2.1 × 100mm, 1.8 μm, Agilent, USA, flow rate 0.8-1.2mL min^-1Detection wavelength of 195-225nm, column temperature of 33-37 ℃, mobile phase: acid acetonitrile A-tetrahydrofuran water solution B with the volume percentage of 0.01-0.03 percent, and gradient elution is carried out, wherein the elution procedure is as follows: 0-4.5min, 0% A-100% B; 4.5-15.0min, 8% A-92% B; 15.0-22.0min, 52% A-48% B; 22.0-35.0, 70% A-30% B; 35-50min, 95% A-5% B; 50min, 95% A-5% B;

2. The fingerprint spectrum testing method for the polycyclic aromatic hydrocarbon substances in the indoor dust fall as claimed in claim 1, is characterized in that: the ultrasonic extraction is carried out at 28-32 KHz for 10-14 min.

3. The fingerprint spectrum testing method for the polycyclic aromatic hydrocarbon substances in the indoor dust fall as claimed in claim 1, is characterized in that: the microwave extraction is performed at 680-720W for 4-6 min.

4. The fingerprint spectrum testing method for the polycyclic aromatic hydrocarbon substances in the indoor dust fall as claimed in claim 1, is characterized in that: the filler in the chromatographic column is anhydrous sodium sulfate, silicon dioxide and copper powder.

5. The fingerprint spectrum testing method for the polycyclic aromatic hydrocarbon substances in the indoor dust fall, as claimed in claim 4, is characterized in that: the filling materials in the chromatographic column are anhydrous sodium sulfate, silicon dioxide and copper powder, and the mass ratio of the anhydrous sodium sulfate to the silicon dioxide to the copper powder is 1: 1.5-2.5: 0.1-1.0.

6. The fingerprint spectrum testing method for the polycyclic aromatic hydrocarbon substances in the indoor dust fall as claimed in claim 1, is characterized in that: the n-hexane-diethyl ether solvent is prepared by mixing n-hexane and diethyl ether at a volume ratio of 2.5-3.5: 1.

7. The fingerprint spectrum testing method for the polycyclic aromatic hydrocarbon substances in the indoor dust fall as claimed in claim 1, is characterized in that: the volume ratio of the n-hexane to the dichloromethane is 0.8-1.2: 1.