CN113702529A

CN113702529A - Rapid high-throughput detection method for antibiotics in soil sample and sediment sample

Info

Publication number: CN113702529A
Application number: CN202110991925.7A
Authority: CN
Inventors: 吕敏; 宋乐慧; 陈令新; 郭晓彤; 丁晶
Original assignee: Yantai Institute of Coastal Zone Research of CAS
Current assignee: Yantai Institute of Coastal Zone Research of CAS
Priority date: 2021-08-27
Filing date: 2021-08-27
Publication date: 2021-11-26
Also published as: CN113899836A

Abstract

The invention discloses a method for quickly detecting antibiotics in a soil sample and a sediment sample in a high-throughput manner, which comprises the following steps: drawing a standard curve, pretreating a soil sample and a sediment sample, extracting (CS-acetonitrile extracting solution), salting out (anhydrous magnesium sulfate and trisodium citrate 4; 1 mixing), nitrogen-blowing, concentrating and fixing volume, measuring by high performance liquid chromatography-tandem mass spectrometry and quantifying by an external standard method. According to the detection method provided by the invention, by optimizing the QuEChERS pretreatment method, the extracting solution and the salting-out agent which can improve the solubility of the target antibiotic are used, and high performance liquid chromatography tandem mass spectrometry detection is combined, 25 antibiotics in the soil sample and the sediment sample can be extracted and detected simultaneously, the operation is simple and rapid, the detection limit is low, the stability is good, the detection efficiency is high, and the requirement of simultaneously detecting multiple antibiotics in the soil and the sediment can be basically met.

Description

Rapid high-throughput detection method for antibiotics in soil sample and sediment sample

Technical Field

The invention relates to a method for detecting antibiotics, in particular to a method for quickly detecting the antibiotics in a soil sample and a sediment sample in a high-throughput manner, and belongs to the technical field of environmental detection.

Background

As a medicament for treating infectious diseases, antibiotics are known as one of the most important discoveries in the 20 th century and are widely used in the aspects of treatment and prevention of diseases of human beings and animals and poultry and livestock breeding additives. In recent years, the problem of environmental contamination with antibiotics has received much attention due to their widespread use and their continuous release into the environment. Antibiotics have been reported to be detected to varying degrees in environments such as sewage, surface water, drinking water, soil and sediments, and have potential effects on the ecological environment and human health. Offshore sediments are an important trend for adsorbing and accumulating antibiotics, but the content detection difficulty of the antibiotics is high due to the relatively low content level of the antibiotics in the offshore sediments and the complex matrix. Therefore, the development of efficient, rapid and reliable pretreatment technology and detection and analysis method is the key to develop many research topics of antibiotics in the environment. At present, the application range of a sample pretreatment method and an instrumental analysis method for detecting and analyzing antibiotics in soil and sediments is narrow, and the treatment procedure is usually complicated and time-consuming, so that it is important to establish a method capable of quickly and simultaneously detecting multiple antibiotics in soil samples and sediment samples.

Disclosure of Invention

The invention aims to provide a method for quickly detecting antibiotics in soil samples and sediment samples at high flux.

In order to achieve the above object, the present invention adopts the following technical solutions:

the method for rapidly detecting the antibiotics in the soil sample and the sediment sample in high flux is characterized by comprising the following steps:

step 1, drawing a standard curve

Respectively preparing an antibiotic single standard solution and antibiotic mixed standard solutions with different mass concentrations by taking methanol as a solvent, and drawing a standard curve by taking a peak area as a vertical coordinate and the mass concentration of a target object as a horizontal coordinate;

step 2, pretreatment of soil sample and sediment sample

Freeze-drying a soil sample or a sediment sample, grinding and sieving, and accurately weighing 2g of the soil sample or the sediment sample for later use;

step 3, extraction

Adding 20mL of CS-acetonitrile extracting solution and 100 mu L of acetic acid solution with the volume concentration of 1% into the weighed soil sample or sediment sample of 2g, and performing ultrasonic extraction for 30 min;

step 4, salting out

Adding 4g of anhydrous magnesium sulfate and 1g of trisodium citrate into the solution subjected to ultrasonic extraction, performing salting-out centrifugation for layering, and finally taking 5mL of supernatant into a brown glass bottle;

step 5, nitrogen blowing concentration constant volume

Blowing the supernatant to be nearly dry by nitrogen, dissolving the supernatant by using a proper amount of methanol, filtering the supernatant by using a filter membrane, transferring the supernatant to a sample injection bottle, blowing the supernatant to be nearly dry again by nitrogen, adding methanol to dissolve the supernatant to a constant volume of 1ml, and obtaining a soil sample solution or a sediment sample solution;

step 6, high performance liquid chromatography tandem mass spectrometry

Measuring the chromatographic peak area of the target substance by using a high performance liquid chromatography tandem mass spectrum;

step 7, external standard method quantification

And (3) calculating the concentration of the antibiotics in the soil sample solution or the sediment sample solution through the standard curve drawn in the step (1) based on the chromatographic peak area of the target object detected by the high performance liquid chromatography-tandem mass spectrometry.

The method for rapidly detecting the antibiotics in the soil sample and the sediment sample at high flux is characterized in that in the step 1, the antibiotics comprise: aureomycin, demeclocycline, doxycycline, oxytetracycline, tetracycline, sulfaclozine, sulfadiazine, sulfadimethoxine, sulfadimidine, sulfamethoxydiazine, sulfamethazine, sulfamethoxazole, trimethoprim, ciprofloxacin, lomefloxacin, norfloxacin, azithromycin, lincomycin, roxithromycin, spiramycin, tylosin, amoxicillin, penicillin G, penicillin V and chloramphenicol, for a total of 25.

The method for rapidly detecting the antibiotics in the soil sample and the sediment sample in the high throughput manner is characterized in that in the step 2, the soil sample and the sediment sample are ground and then pass through a 100-mesh sieve.

The method for rapidly detecting the antibiotics in the soil sample and the sediment sample in high flux is characterized in that in the step 3, the CS-acetonitrile extracting solution is prepared by adopting the following method:

(1) preparing a magnesium nitrate solution with the mass concentration of 50% and an ammonia water solution with the volume concentration of 2.5%, and mixing the magnesium nitrate solution and the ammonia water solution according to the volume ratio of 96:4 to obtain a magnesium nitrate-ammonia water mixed solution;

(2) dissolving 10.56g of sodium dihydrogen phosphate and 0.82mL of phosphoric acid in water, and diluting to 1L to obtain a phosphate solution; dissolving 80.0g of disodium ethylene diamine tetraacetate into 1L of phosphate solution to obtain disodium ethylene diamine tetraacetate-phosphate mixed solution; mixing the disodium ethylene diamine tetraacetate-phosphate mixed solution with acetonitrile according to the volume ratio of 1:1 to obtain acetonitrile mixed solution;

(3) and mixing the acetonitrile mixed solution with a magnesium nitrate-ammonia water mixed solution according to the volume ratio of 3:1 to obtain a CS-acetonitrile extracting solution.

The method for rapidly detecting the antibiotics in the soil sample and the sediment sample at high flux is characterized in that in the step 4, the specific operation of salting-out centrifugal layering is as follows:

adding anhydrous magnesium sulfate and trisodium citrate into the solution after ultrasonic extraction, immediately shaking vigorously for 3min, then performing ultrasonic treatment for 10min, then centrifuging at the rotating speed of 5000r/min for 10min, and finally taking the supernatant.

The method for rapidly detecting the antibiotics in the soil sample and the sediment sample in the high throughput is characterized in that in the step 6, the high performance liquid chromatography is carried out under the following conditions:

the chromatographic column is Waters-ACQUITY UPLC BEH C18 with specification of 100 × 2.1mm and 1.7 μm; the column temperature was 35 ℃; the mobile phase A is an aqueous solution containing 0.01 percent of formic acid, and the mobile phase B is a 100 percent acetonitrile solution; the sample injection volume is 5 mu L, and the flow rate of the mobile phase is 0.5 mL/min; the detection time for each sample was 15min, and the mobile phase elution procedure is shown in the following table:

time (min)	Flow rate (mL/min)	A(％)	B(％)
				0	0.5	95	5
0.5	0.5	95	5
				5	0.5	90	10
10	0.5	40	60
				12	0.5	5	95
13	0.5	5	95
				13.1	0.5	95	5
15	0.5	95	5

The method for rapidly detecting the antibiotics in the soil sample and the sediment sample at high flux is characterized in that in step 6, the tandem mass spectrometry is carried out under the following conditions:

selecting an electrospray ion source, and simultaneously carrying out scanning detection in a positive ion mode and a negative ion mode; ion spray voltage: 5500V in positive ion mode, and-4500V in negative ion mode; the air pressure of the air curtain is 40 psi; the spray gas pressure was 55 psi; the auxiliary heating air pressure was 55psi and the mass spectrometric detection parameters for the 25 antibiotics are given in the following table:

the invention has the advantages that: according to the detection method provided by the invention, by optimizing the QuEChERS pretreatment method, the extracting solution and the salting-out agent which can improve the solubility of the target antibiotic are used, and high performance liquid chromatography tandem mass spectrometry detection is combined, 25 antibiotics in the soil sample and the sediment sample can be extracted and detected simultaneously, the operation is simple and rapid, the detection limit is low, the stability is good, the detection efficiency is high, and the requirement of simultaneously detecting multiple antibiotics in the soil and the sediment can be basically met.

Drawings

FIG. 1 is a total ion chromatogram of a mixed standard solution (100. mu.g/L) of 25 antibiotics shown in Table 1;

FIG. 2 is a graph showing the calculated recovery rates of the 25 antibiotics shown in Table 1 under different extraction conditions;

FIG. 3 is a graph showing the results of calculation of the recovery rates of 25 antibiotics shown in Table 1 under different salting-out conditions.

Detailed Description

The detection method provided by the invention can be used for simultaneously and rapidly detecting 25 antibiotics in the soil sample and the sediment sample (the information of the 25 antibiotics is shown in table 1), and specifically comprises the following steps: drawing a standard curve, pretreating a soil sample and a sediment sample, extracting, salting out, blowing nitrogen, concentrating and fixing volume, measuring by high performance liquid chromatography-tandem mass spectrometry and quantifying by an external standard method.

Information on Table 125 antibiotics

The invention is described in detail below with reference to the figures and the embodiments.

First, the concentration of 25 antibiotics in the sample of offshore surface sediment in Bay of Liaodong is detected

Step 1, drawing a standard curve

Methanol is used as a solvent, antibiotic single standard solutions with the mass concentration of 1000mg/L are respectively prepared and stored at the temperature of minus 20 ℃.

Using methanol as a solvent, preparing an antibiotic mixed standard solution with the mass concentration of 1mg/L, and storing at the temperature of minus 20 ℃.

The antibiotic mixed standard solution with the mass concentration of 1mg/L is diluted by methanol to prepare antibiotic mixed standard solutions with the mass concentrations of 0.1 mu g/L, 0.5 mu g/L, 1 mu g/L, 2 mu g/L, 5 mu g/L, 10 mu g/L, 20 mu g/L, 40 mu g/L and 100 mu g/L respectively.

The peak area is plotted as the ordinate (y) and the mass concentration of the target substance is plotted as the abscissa (x) to obtain a standard curve.

Step 2, sediment sample pretreatment

And 8, 29 months in 2018, collecting a surface sediment sample in the offshore region of the Bay in Liaodong, and freezing and storing the collected sediment sample at the temperature of minus 20 ℃ in a dark place.

Freeze-drying the sediment sample stored in a dark place, grinding the sediment sample through a 100-mesh sieve, and accurately weighing 2g of the sediment sample for later use.

Step 3, extraction

1. Optimized extract

Selecting two buffers of CS and EDTA-McIlvaine and acetonitrile: two extracting agents of methanol/3: 1, combining the buffer solution and the extracting agents two by two to obtain 4 combination schemes:

(1) CS buffer-acetonitrile combination protocol;

(2) CS buffer-acetonitrile: a methanol/3: 1 combination scheme;

(3) EDTA-McIlvaine buffer-acetonitrile combination protocol;

(4) EDTA-McIlvaine buffer-acetonitrile: methanol/3: 1 combination scheme.

For the CS buffer-acetonitrile combination protocol: preparing CS-acetonitrile extracting solution, adding 20mL of CS-acetonitrile extracting solution and 100 mu L of acetic acid solution with the volume concentration of 1% into a weighed sediment sample of 2g, and performing ultrasonic extraction for 30 min.

For CS buffer-acetonitrile: methanol/3: 1 combination scheme: preparing CS-acetonitrile/methanol extracting solution, adding 20mL of CS-acetonitrile/methanol extracting solution and 100 mu L of acetic acid solution with the volume concentration of 1% into a weighed sediment sample of 2g, and performing ultrasonic extraction for 30 min.

For the EDTA-McIlvaine buffer-acetonitrile combination protocol: preparing EDTA-McIlvaine buffer solution, adding 10mL of EDTA-McIlvaine buffer solution into a weighed sediment sample of 2g, vortexing for 1min, adding 10mL of acetonitrile, vortexing for 1min, finally adding 100 mu L of acetic acid solution with the volume concentration of 1%, and ultrasonically extracting for 30 min.

For EDTA-McIlvaine buffer-acetonitrile: methanol/3: 1 combination scheme: preparing EDTA-McIlvaine buffer solution, adding 10mL of EDTA-McIlvaine buffer solution into a weighed 2g sediment sample, vortexing for 1min, and then adding 10mL of acetonitrile: methanol/3: 1, vortex for 1min, finally add 100 μ L of 1% acetic acid solution, ultrasonic extract for 30 min.

Wherein, the CS-acetonitrile extracting solution is prepared by the following method:

(i) preparing a magnesium nitrate solution with the mass concentration of 50% and an ammonia water solution with the volume concentration of 2.5%, and mixing the magnesium nitrate solution with the mass concentration of 50% and the ammonia water solution with the volume concentration of 2.5% according to the volume ratio of 96:4 to obtain a magnesium nitrate-ammonia water mixed solution;

(ii) dissolving 10.56g of sodium dihydrogen phosphate and 0.82mL of phosphoric acid in water, and diluting to 1L to obtain phosphate Solution (SPB); dissolving 80.0g of disodium ethylene diamine tetraacetate into 1L of SPB to obtain disodium ethylene diamine tetraacetate-phosphate mixed solution; mixing the disodium ethylene diamine tetraacetate-phosphate mixed solution with acetonitrile according to the volume ratio of 1:1 to obtain acetonitrile mixed solution;

(iii) and mixing the acetonitrile mixed solution with a magnesium nitrate-ammonia water mixed solution according to the volume ratio of 3:1 to obtain a CS-acetonitrile extracting solution.

The CS-acetonitrile/methanol extract is prepared by the following method:

(ii) dissolving 10.56g of sodium dihydrogen phosphate and 0.82mL of phosphoric acid in water, and diluting to 1L to obtain phosphate Solution (SPB); dissolving 80.0g of disodium ethylene diamine tetraacetate into 1L of SPB to obtain disodium ethylene diamine tetraacetate-phosphate mixed solution; mixing acetonitrile and methanol according to a volume ratio of 3:1, and then mixing the mixture with an ethylene diamine tetraacetic acid-phosphate mixed solution according to a volume ratio of 1:1 to obtain an acetonitrile/methanol mixed solution;

(iii) and mixing the acetonitrile/methanol mixed solution with a magnesium nitrate-ammonia water mixed solution according to the volume ratio of 3:1 to obtain a CS-acetonitrile/methanol extracting solution.

The EDTA-McIlvaine buffer solution is prepared by the following method:

12.9g of citric acid, 27.5g of disodium hydrogenphosphate and 37.2g of disodium ethylenediaminetetraacetate were dissolved in water and the volume was set to 1L, to obtain EDTA-McIlvaine buffer (pH 4.0).

The results of the standard recovery calculations for the 25 antibiotics for the 4 combination schemes are shown in FIG. 2.

As can be seen from fig. 2, for most antibiotics, the combination scheme using acetonitrile as extractant compares acetonitrile: the combined methanol/3: 1 extraction reagent scheme has better recovery of antibiotics. When the pH value of the extracting solution is close to the pKa of the antibiotic, the antibiotic is in an isoelectric state and has high solubility in the extracting solution, compared with an EDTA-McIlvaine buffer solution, the pH value of a CS buffer solution is relatively lower and is close to the pKa of most of the antibiotics, the CS buffer solution can improve the solubility of the antibiotic and prevent the antibiotic from being hydrolyzed, and the macrolide antibiotic has high pKa compared with other antibiotics, so that the recovery rate of most of other antibiotics in the CS buffer solution is high except the macrolide antibiotic.

The recovery rates of the antibiotics were combined to finally determine that the CS buffer-acetonitrile combination scheme is the most preferable extraction scheme of the present invention.

2. Extracting antibiotic with CS-acetonitrile extract (CS buffer-acetonitrile combination scheme)

To a weighed 2g sample of the deposit, 20mL of CS-acetonitrile extract and 100. mu.L of 1% by volume acetic acid solution were added and extracted with ultrasound for 30 min.

Step 4, salting out

1. Optimized salting-out agent

The water absorbent and salting-out agent are added to carry out salting-out centrifugal layering, so that the separation of a water phase and an organic phase can be promoted, and meanwhile, the pH value of a buffer system can be adjusted to obtain a better recovery rate.

Anhydrous magnesium sulfate is selected as a water absorbent, three salts of sodium chloride, sodium acetate and trisodium citrate which can respectively lead the pH value of a buffer system to be relatively low, relatively high and relatively moderate are selected as salting-out agents, the water absorbent and the salting-out agents are combined according to the mass ratio of 4:1 to carry out salting-out centrifugal layering on the solution after ultrasonic extraction, and the influence results of the three salting-out agents on the recovery rates of 25 antibiotics are shown in figure 3.

As can be seen from FIG. 3, the recovery rates of 25 antibiotics were generally better when trisodium citrate was selected as the salting-out agent.

2. Salting out of the solution with trisodium citrate

Adding 4g of anhydrous magnesium sulfate and 1g of trisodium citrate into the solution after ultrasonic extraction, immediately shaking vigorously for 3min, then carrying out ultrasonic treatment for 10min, then centrifuging at the rotating speed of 5000r/min for 10min, and finally taking 5mL of supernatant into a brown glass bottle.

Step 5, nitrogen blowing concentration constant volume

And (3) blowing nitrogen to the supernatant to be dry, dissolving the supernatant by using a proper amount of methanol, filtering the supernatant by using a 0.22-micron filter membrane, transferring the supernatant into a 1.5ml sample injection bottle, blowing nitrogen to be dry again, adding methanol to dissolve the supernatant until the volume is reduced to 1ml, obtaining a sample solution, and storing the sample solution at the temperature of minus 20 ℃ for detection.

Step 6, high performance liquid chromatography tandem mass spectrometry

The high performance liquid chromatography tandem mass spectrometry is carried out under the following conditions:

(1) the column was Waters-ACQUITY UPLC BEH C18 (100X 2.1mm, 1.7 μm); the column temperature was 35 ℃; the mobile phase A is an aqueous solution containing 0.01 percent of formic acid, and the mobile phase B is a 100 percent acetonitrile solution; the sample injection volume is 5 mu L, and the flow rate of the mobile phase is 0.5 mL/min; the detection time of each sample is 15min, and the mobile phase elution program is shown in table 2;

(2) selecting an electrospray ionization (ESI) source by tandem mass spectrometry, and simultaneously carrying out scanning detection in a positive ion mode and a negative ion mode; ion spray voltage: 5500V (positive ion mode), -4500V (negative ion mode); air curtain gas (CUR) pressure was 40 psi; the pressure of the spray gas (ion source gas 1, GS1) was 55 psi; the supplemental heating gas (GS 2) pressure was 55 psi. The mass spectrometric detection parameters for the 25 antibiotics are shown in table 3.

Table 2 mobile phase elution procedure

Table 325 Mass Spectrometry detection parameters for antibiotics

Step 7, external standard method quantification

And (3) calculating the concentration of the antibiotics in the sample solution through the standard curve drawn in the step (1) based on the chromatographic peak area of the target substance detected by the high performance liquid chromatography tandem mass spectrometry. The calculated concentrations of the 25 antibiotics in the samples of the superficial sediments in the gulf of Liaodong are shown in Table 4.

TABLE 42018 concentration of 25 antibiotics in samples of superficial sediments in Bay of Liaodong province (. mu.g/kg)

nd represents not detected.

As can be seen from table 4, the detection rates of 14 antibiotics (CTC, DMCTC, DOC, TC, SCZ, SFT, SMX, CIP, LFX, NOF, ATM, RXM, AMC, CPC) in the samples of superficial deposits in the gulf of jeaodong were all over 30%.

Therefore, the detection method provided by the invention can be used for simultaneously detecting 25 antibiotics in the surface sediment sample of the Bay of Liaodong, has high detection efficiency, and can be used for simultaneously detecting the 25 antibiotics in the marine sediment.

Second, method verification

1. Detection limit and quantification limit

The detection limit of the instrument (LOD) is calculated by the signal-to-noise ratio S/N being 3 and the quantitative limit of the instrument (LOQ) being 10, and the detection limit of the Method (MDL) is calculated by the following formula:

MDL＝LOD·V/(R·m)

wherein V is the constant volume of the sample; r is the recovery rate; and m is the extracted sample mass.

The results of the calculations of the linear range, correlation coefficient, instrument detection limit, instrument quantitation limit, and method detection limit for the 25 antibiotics in the samples of surface deposits offshore in the gulf of jeaodong are shown in table 5.

Table 525 antibiotics linear range, correlation coefficient, instrument detection limit, instrument quantitation limit and method detection limit

2. Accuracy and precision of detection method

The sediment with extremely low concentration of the target substance is used as a control group, and the concentration of the sediment is used as a baseline value of blank control. 2 different concentrations of target, 5. mu.g/kg (low concentration) and 25. mu.g/kg (high concentration), were added, 3 replicates were set for each concentration, the concentrations of 25 antibiotics in the sediment were measured using the above-described test method, and the average spiked recovery and relative standard deviation of the 25 antibiotics were calculated.

The calculated average spiked recoveries and relative standard deviations for the 25 antibiotics are shown in table 6.

Average spiked recovery and relative standard deviation of 625 antibiotics in Table

As can be seen from Table 6, the average spiked recovery rates of 22 other antibiotics in the sediment sample except Sulfadiazine (SDZ), Ciprofloxacin (CIP) and Norfloxacin (NOF) were 40.35-142.92%, and the relative standard deviations of the 22 other antibiotics except Sulfadimethoxine (SFT), Sulfamethoxazole (SMX) and Spiramycin (SRY) were less than 20%.

That is to say, the detection method provided by the invention has the advantages of low detection limit, high recovery rate, good stability, simple and quick operation and high detection efficiency, can be used for simultaneously detecting 25 antibiotics in a sediment sample, and can basically meet the requirement of simultaneously detecting multiple antibiotics in sediment.

Thirdly, detecting the concentration of 25 antibiotics in forest and farmland soil samples of 36 sampling points in Henan, Hunan, Yunnan, Zhejiang, Liaoning and Fujian

The soil with extremely low concentration of the target substance is used as a control group, and the concentration of the soil is used as a baseline value of blank control. Adding 25 mug/kg of target, setting 3 parallel samples, detecting the concentrations of 25 antibiotics in forest and farmland soil of 36 sampling points in six places of Henan, Hunan, Yunnan, Zhejiang, Liaoning and Fujian by using the method for detecting the concentrations of 25 antibiotics in the sample of the offshore surface sediment of the Liaodong Bay, and calculating the average standard addition recovery rate and the relative standard deviation of the 25 antibiotics.

The detection limit (MDL) of the method is calculated by the following formula:

MDL＝LOD·V/(R·m)

The average spiked recovery, relative standard deviation, and method detection limits for the 25 antibiotics in the soil samples were calculated as shown in table 7.

TABLE 7 mean spiked recovery (spiked concentration 25. mu.g/kg), relative standard deviation and method detection limit for 25 antibiotics in soil samples

As can be seen from Table 7, the average normalized recovery rate of the 23 antibiotics in the soil sample is 38.96-118.68% except for Oxytetracycline (OTC) and Spiramycin (SRY), and the relative standard deviation of the 23 antibiotics except for Spiramycin (SRY) and Tylosin (TYL) is less than 20%.

The method of the invention is used for detecting and analyzing 25 antibiotics in soil samples of forests and farmlands of 36 sampling points in six places of Henan, Hunan, Yunnan, Zhejiang, Liaoning and Fujian in 2020, and the results are shown in Table 8.

TABLE 82020 year 25 antibiotic concentrations (ug/kg) in soil samples of different regions of China

As can be seen from Table 8, the detection method provided by the present invention can be used to detect the concentrations of 25 antibiotics in marine sediment samples, and can also be used to detect the concentrations of 25 antibiotics in soil samples.

It should be noted that the above-mentioned embodiments do not limit the present invention in any way, and all technical solutions obtained by using equivalent alternatives or equivalent variations fall within the protection scope of the present invention.

Claims

1. The method for rapidly detecting the antibiotics in the soil sample and the sediment sample in high flux is characterized by comprising the following steps:

step 1, drawing a standard curve

step 2, pretreatment of soil sample and sediment sample

step 3, extraction

step 4, salting out

step 5, nitrogen blowing concentration constant volume

step 6, high performance liquid chromatography tandem mass spectrometry

step 7, external standard method quantification

2. The method for rapid high-throughput detection of antibiotics in soil and sediment samples according to claim 1, wherein in step 1, the antibiotics comprise: aureomycin, demeclocycline, doxycycline, oxytetracycline, tetracycline, sulfaclozine, sulfadiazine, sulfadimethoxine, sulfadimidine, sulfamethoxydiazine, sulfamethazine, sulfamethoxazole, trimethoprim, ciprofloxacin, lomefloxacin, norfloxacin, azithromycin, lincomycin, roxithromycin, spiramycin, tylosin, amoxicillin, penicillin G, penicillin V and chloramphenicol, for a total of 25.

3. The method for rapid high throughput detection of antibiotics in soil and sediment samples according to claim 1, wherein in step 2, the soil and sediment samples are ground and then screened through a 100 mesh screen.

4. The method for rapid high-throughput detection of antibiotics in soil and sediment samples according to claim 1, wherein in step 3, the CS-acetonitrile extracting solution is prepared by the following method:

5. The method for rapid high-throughput detection of antibiotics in soil and sediment samples according to claim 1, wherein in step 4, the specific operation of salting-out centrifugation layering is as follows:

6. The method for rapid high-throughput detection of antibiotics in soil and sediment samples according to claim 1, wherein in step 6, the high performance liquid chromatography is performed under the following conditions:

7. the method for rapid high-throughput detection of antibiotics in soil and sediment samples according to claim 1, wherein in step 6, tandem mass spectrometry is performed under the following conditions: