Establishment of a Solid Phase Microextraction Method for Rapid Detection of Trace Polar Glucocorticoid Drugs in Environmental Water

Abstract

The present invention relates to the field of analytical chemistry, and particularly to a solid-phase microextraction probe based on metal-organic framework SU101, its preparation method, and its application in the trace detection of pharmaceuticals and personal care products in water environment. The solid-phase microextraction probe provided by the present invention has environmentally friendly materials and significantly stronger extraction ability than commercial probes, and has the advantages of low detection limit and quantification limit, wide linear range, good reproducibility, etc. Therefore, it can be used to detect extremely trace amounts of polar drugs in environmental water, especially glucocorticoid drugs.

Description

Technical Field

[0001] The present invention relates to the field of analytical chemistry, and particularly to a solid-phase microextraction probe based on SU101, its preparation method and application. Background Art

[0002] Pharmaceutical and personal care products (PPCPs) are emerging pollutants that have been found to enter surface water and groundwater through domestic sewage, aquaculture wastewater, agricultural wastewater, and pharmaceutical industrial wastewater. Due to their wide use and bioaccumulation, the environmental risks and potential toxic effects of PPCPs have attracted increasing attention. Glucocorticoid drugs (GCs) are one of the important categories of PPCPs and have various effects on livestock and humans, such as immunosuppression, anti-inflammatory, and anti-allergic effects. Medicinal GCs are particularly stable and difficult to metabolize, and can remain in the environment for a long time. Relevant studies have shown that various GCs can be detected in treated wastewater and surface water, and they can have an adverse impact on the coagulation and reproduction of aquatic organisms, and even pose a threat to human health. Therefore, it is particularly important and urgent to establish a highly sensitive analytical method for trace GCs and other PPCPs in environmental water to study their behavior in environmental water.

[0003] Solid-phase microextraction (SPME) technology is a new type of sample pretreatment technology that integrates sampling, separation, enrichment, and injection, and has been applied in the fields of environment, biology, pharmacy, food, and clinical medicine. Existing studies have used SPME to detect PPCPs. However, SPME is still in the early stage of development in drug analysis, especially for the extraction of highly polar drugs, and there is little research. Summary of the Invention

[0004] The purpose of the present invention is to provide the application of the Bio-MOFs material SU101 in solid-phase microextraction and its application in detecting PPCPs in the environment. Another purpose of the present invention is to provide a solid-phase microextraction probe, its preparation method, and application. The probe can quickly detect extremely trace amounts of highly polar drugs, especially glucocorticoid drugs, in environmental water. The above purposes of the present invention are achieved through the following technical solutions:

[0005] On the one hand, the present invention provides the application of the Bio-MOFs material SU101 in solid-phase microextraction. Specifically, the bio-MOFs material SU101 is used to prepare the coating of the solid-phase microextraction probe.

[0006] SU101 (or SU-101) is an ellagic acid-based microporous metal-organic framework (MOFs), and its chemical formula is Bi2O(H2O)2(C 14H2O8)·nH2O. The specific structure of SU101 can be found in the reference Erik Svensson Grape, et al. A Robust and Biocompatible Bismuth Ellagate MOF Synthesized Under Green Ambient Conditions. J. Am. Chem. Soc., 2020 DOI: 10.1021 / jacs.0c07525.

[0007] On the other hand, the present invention also provides the application of the Bio-MOFs material SU101 in detecting pharmaceutical and personal care products (PPCPs) in the water environment.

[0008] The PPCPs include, but are not limited to, one or more of dehydroepiandrosterone, testosterone, androstenedione, erythromycin, trimethoprim, ofloxacin, levofloxacin, norfloxacin, ciprofloxacin, and carbamazepine.

[0009] In a preferred embodiment, the PPCPs are selected from glucocorticoid drugs, including, but not limited to, one or more of corticosterone, cortisone, fluorometholone, hydrocortisone, methylprednisolone, prednisolone, triamcinolone acetonide, and triamcinolone.

[0010] On the other hand, the present invention also provides a solid-phase microextraction probe (SU101 probe), which contains the following structure: (1) a stainless-steel wire carrier; and (2) a bio-MOFs material SU101 coating.

[0011] In a preferred embodiment, the solid-phase microextraction probe (SU101 probe) further contains (3) a neutral silicone sealant for attaching the SU101 coating to the stainless-steel carrier.

[0012] In a preferred embodiment, the length of the stainless-steel wire is 3 - 6 cm; more preferably 4 cm.

[0013] In a preferred embodiment, the length of the SU101 coating is 1 - 2 cm; more preferably 1 cm.

[0014] In a preferred embodiment, the thickness of the SU101 coating is 40 - 60 μm; more preferably 50 μm.

[0015] After use, the probe can be reused after soaking and drying in a polar solvent such as methanol.

[0016] On the other hand, the present invention also provides a preparation method of the solid-phase microextraction probe, and the method includes the following steps:

[0017] (1) Preparation of bio-MOFs material SU101: Ellagic acid and bismuth acetate with a molar ratio of 1:2 - 1:3 are reacted in an acetic acid aqueous solution with a volume ratio of 5 - 7%. The precipitate is taken, washed and dried to obtain the bio-MOFs material SU101.

[0018] (2) Preparation of the adhesive solution: Neutral silicone sealant and cyclohexane with a weight ratio of 1:4 - 1:6 are mixed to form a uniform adhesive solution, which is sealed for later use.

[0019] (3) Coating attachment: The front end of a clean stainless steel wire is immersed in the adhesive solution in step (2). After taking it out, it is immediately brought into full contact with the bio-MOFs material SU101 powder in step (1) to form a uniform coating, and then dried to remove cyclohexane.

[0020] Among them, there is no requirement for the sequence of steps S1 and S2.

[0021] In a preferred embodiment, in step (1), the prepared bio-MOFs material also needs to be ground.

[0022] In a preferred embodiment, in step (1), the precipitate is washed 3 times successively with deionized water and ethanol.

[0023] In a preferred embodiment, in step (1), the drying temperature is 75 - 100 °C.

[0024] In a preferred embodiment, in step (2), the mixing method is ultrasonic mixing.

[0025] In a preferred embodiment, in step (3), the drying temperature is 80 - 100 °C, and the drying time is 10 - 30 min.

[0026] In a preferred embodiment, in step (3), the clean stainless steel wire is ultrasonically cleaned successively with water, methanol and ethanol.

[0027] In a preferred embodiment, step (3) can be repeated multiple times.

[0028] In a preferred embodiment, in step (3), the finally prepared coating thickness is 1 - 2 cm. More preferably, before the first use of the probe, the coating is soaked in an organic solvent such as methanol for 0.5 - 2 h and then dried for later use.

[0029] On the other hand, the present invention provides a method for detecting pharmaceuticals and personal care products (PPCPs) in an aqueous environment. The method includes the following steps;

[0030] S1. Enrichment of the target analyte: Completely immerse the above-mentioned solid-phase microextraction probe in the water environmental sample solution, and extract the target analyte PPCPs at the selected extraction time and oscillation rate.

[0031] S2. Desorption of the target analyte: Immerse the solid-phase microextraction probe enriched with the target analyte in the desorption solvent, and desorb the target analyte at the selected oscillation rate and time to obtain the desorption solution.

[0032] S3. Use an analytical instrument to perform quantitative and / or qualitative analysis on the desorption solution.

[0033] In a preferred embodiment, the target analyte PPCPs are glucocorticoid drugs (GCs). The glucocorticoid drugs (GCs) include, but are not limited to, one or more of corticosterone, cortisone, fluorometholone, hydrocortisone, methylprednisolone, prednisolone, triamcinolone acetonide, and triamcinolone.

[0034] In a preferred embodiment, the target analyte PPCPs include, but are not limited to, one or more of dehydroepiandrosterone, testosterone, androstenedione, erythromycin, trimethoprim, ofloxacin, levofloxacin, norfloxacin, ciprofloxacin, and carbamazepine.

[0035] In a preferred embodiment, in step S1, the extraction time is 20 - 60 min; the oscillation rate is 400 - 700 rpm; more preferably, the extraction time is 30 - 50 min; the oscillation rate is 500 - 700 rpm.

[0036] In a preferred embodiment, in step S2, the desorption time is 10 - 40 min; the oscillation rate is 400 - 700 rpm; more preferably, the desorption time is 10 - 30 min; the oscillation rate is 500 - 700 rpm.

[0037] In a preferred embodiment, the desorption solvent is selected from any one or more of the following: methanol, acetonitrile, methanol containing 0.1% formic acid by volume, acetonitrile containing 0.1% formic acid by volume, and a mixed solvent of methanol and acetonitrile (volume ratio of methanol to acetonitrile is 2:3). More preferably, the desorption solvent is a mixed solvent of methanol and acetonitrile (volume ratio of methanol to acetonitrile is 2:3).

[0038] In a preferred embodiment, the analytical instrument is selected from HPLC / MS / MS. More preferably, the analytical conditions of the HPLC / MS / MS are: For GCs drugs, use Express Separation was carried out using an F5 column (4.6 mm×100 mm, 2.7 μm, Supelco, USA), and the other 10 polar drugs were separated using a ZORBAX SB-C18 column (2.1 mm×150 mm, 3.5 μm, Agilent, USA). The column temperature was 25 - 35 °C. An aqueous solution containing 0.1% formic acid (A) and a mixed solution of methanol and acetonitrile (B) were used as the mobile phase. The temperature of the mass spectrometry ion source was 400 - 600 °C, and the injection volume was 5 - 10 μL.

[0039] Advantageous technical effects of the present invention:

[0040] (1) The present invention utilizes an environmentally friendly bio-MOFs to establish a green, economical, and environmentally friendly solid-phase microextraction probe, which is particularly suitable for extracting polar drugs, especially glucocorticoid drugs.

[0041] (2) The extraction ability of the solid-phase microextraction probe of the present invention is strong, and the extraction ability for the target analyte is 7.7 - 3669 times that of commercial PDMS, PDMS / DVB, PA, CAR / PDMS, and DVB / CAR / PDMS probes. Therefore, it can be used to detect extremely trace amounts of drugs in environmental water.

[0042] (3) The detection method for pharmaceuticals and personal care products (PPCPs) in the water environment of the present invention has the advantages of low detection limit and quantification limit, wide linear range, good reproducibility, etc., and can meet the requirements of trace detection in real environmental water samples. Description of the Drawings

[0043] Figure 1 Shows the morphology of the solid-phase microextraction probe of the present invention, where (a) SU101, (b) bare steel wire, and (c) the self-made SU101 probe are the morphologies under a scanning probe microscope;

[0044] Figure 2 Shows the optimized conditions for using the solid-phase microextraction probe of the present invention, where (a) desorption solvent, (b) extraction time, (c) desorption time, and (d) oscillation rate are the optimized curve graphs;

[0045] Figure 3 Shows the comparison of the extraction results of the SU101 probe and commercial PDMS, PDMS / DVB, PA, CAR / PDMS, and DVB / CAR / PDMS probes for 8 polar glucocorticoid drugs under the same conditions;

[0046] Figure 4The extraction results of SU101 probe and commercial PDMS, PDMS / DVB, PA, CAR / PDMS and DVB / CAR / PDMS probes for other 10 polar drugs under the same conditions were compared. DETAILED DESCRIPTION

[0047] The technical solution of the present invention is further described below by specific embodiments, which do not limit the protection scope of the present invention. Some non-essential modifications and adjustments made by others based on the concept of the present invention still fall within the protection scope of the present invention.

[0048] Example 1 Preparation method of solid phase microextraction probe (SU101 probe)

[0049] S1. Stir ellagic acid and bismuth acetate in a molar ratio of 1:2 in a 6% by volume acetic acid aqueous solution at room temperature for 2 days. Take the precipitate, wash it three times with deionized water and ethanol in sequence, and finally dry it in a vacuum drying oven overnight to obtain a bio-MOFs material SU101. This product is a yellow powder, which is ground evenly for standby use.

[0050] S2. Ultrasonic mix neutral silicone sealant and cyclohexane in a weight ratio of about 1:5 to form a uniform viscous solution, and seal it for later use.

[0051] S3. Cut the stainless steel wire into about 4 cm lengths, clean it with water, methanol and ethanol ultrasonically, and dry it for later use. Immerse the front end of the cleaned stainless steel wire in the sticky solution in step S2, and immediately contact it with the pre-ground powder in step S1 to form a uniform coating. Place it in an 80°C oven and dry it for 20 minutes to remove cyclohexane. Repeat the above operation several times to finally obtain an SPME probe with a uniform SU101 coating. Figure 1 In (c), the coating thickness is between 40-60 μm, preferably 50 μm. Before the first use, the coating of the homemade probe is scraped to 1 cm with a knife, of course, it can be longer, such as 2 cm, and soaked in methanol for 1 hour, and dried for later use.

[0052] Example 2 Optimization of solid phase microextraction probe conditions

[0053] S1. Enrichment of target analytes: The solid phase microextraction probe in Example 1 was completely immersed in a 10 mL glass vial containing 10 mL of sample solution, and the target analytes were extracted at a selected extraction time and oscillation rate.

[0054] S2. Desorption of the target analyte: Place an inner cannula containing 100 μL of desorption solvent in a 2 mL vial, completely immerse the probe in step S1 therein, and desorb the target analyte at the selected oscillation rate and time to obtain a desorbed solution. After a certain desorption time, take out the solid-phase microextraction probe and immerse it in methanol. It can be reused after drying. The eluate is stored in a -20 °C refrigerator until quantitative analysis of the analyte by coupling with an HPLC-MS / MS instrument, or it can be directly analyzed on the machine.

[0055] S3. HPLC-MS / MS analysis: Quantitative analysis of the eluate containing the analyte is performed by HPLC-MS / MS. The parameters of HPLC-MS / MS are as follows: For GCs drugs, Express separation is carried out using an Express F5 column (4.6 mm × 100 mm, 2.7 μm, Supelco, USA), and for the other 10 polar drugs, separation is carried out using a ZORBAX SB-C18 column (2.1 mm × 150 mm, 3.5 μm, Agilent, USA). The column temperature is 30 °C. An aqueous solution containing 0.1% formic acid (A) and a mixed solution of methanol and acetonitrile (v / v = 2:3, B) are used as the mobile phase. The temperature of the mass spectrometry ion source is 400 °C, and the injection volume is 10 μL.

[0056] Table 1 Gradient elution program

[0057]

[0058] Table 2 MS / MS parameters of the target analyte

[0059]

[0060] Using the method of controlling variables, the four factors of desorption solvent, extraction time, desorption time, and oscillation rate in the solid-phase microextraction step are optimized. The experiment is optimized among 5 desorption solvents: methanol, acetonitrile, methanol containing 0.1% (v / v) formic acid, acetonitrile containing 0.1% (v / v) formic acid, and a mixed solvent of methanol and acetonitrile (volume ratio of methanol to acetonitrile is 2:3); the extraction time is optimized within 0 - 60 min; the desorption time is optimized within 0 - 40 min; the oscillation rate is optimized within 0 - 700 rpm.

[0061] The results are shown in Figure 2 , according to the experimental results, the final optimized conditions are:

[0062] The desorption solvent is selected as a mixed solvent of methanol and acetonitrile (volume ratio of methanol to acetonitrile is 2:3);

[0063] The extraction time is 20 - 60 min; the oscillation rate is 400 - 700 rpm; more preferably, the extraction time is 30 - 50 min; the oscillation rate is 500 - 700 rpm; even more preferably, the extraction time is selected as 40 min and the oscillation rate is selected as 600 rpm.

[0064] The desorption time is 10 - 40 min; the oscillation rate is 400 - 700 rpm; more preferably, the desorption time is 10 - 30 min; the oscillation rate is 500 - 700 rpm; even more preferably, the desorption time is selected as 20 min and the oscillation rate is selected as 600 rpm.

[0065] Comparison of extraction results of glucocorticoid drugs with different probes in Example 3

[0066] In order to take into account the optimal extraction and desorption conditions of other commercial probes, under the conditions of 40 min extraction time, 60 min desorption time, 500 rpm oscillation rate and methanol as the desorption solvent, the extraction performance of the solid-phase microextraction probe of the present invention for 8 glucocorticoid drugs (corticosterone, cortisone, fluorometholone, hydrocortisone, methylprednisolone, prednisolone, triamcinolone acetonide, triamcinolone) was evaluated by comparing with the experimental results of commercial PDMS, PDMS / DVB, PA, CAR / PDMS and DVB / CAR / PDMS probes. The remaining steps refer to Example 2.

[0067] The results are shown in Figure 3 , even under the non-optimal solid-phase microextraction conditions of the SU101 probe of the present invention, the extraction ability of the SU101 probe of the present invention for 8 glucocorticoid drugs is at least 7.7 times higher than that of commercial probes, and the highest increase is 3669 times, showing a very significant effect.

[0068] Comparison of extraction results of other polar drugs with different probes in Example 4

[0069] Meanwhile, in order to further verify the excellent extraction ability of SU101 for polar drugs, the effects were also compared with 5 commercial probes for another 10 polar drugs (dehydroepiandrosterone, testosterone, androstenedione, erythromycin, trimethoprim, ofloxacin, levofloxacin, norfloxacin, ciprofloxacin and carbamazepine) under the same conditions. The remaining steps and experimental conditions refer to Example 3.

[0070] The results are shown in Figure 4, Similarly, even under the optimal solid-phase microextraction conditions of the SU101 probe of the present invention, the extraction ability of the SU101 probe of the present invention for other 10 polar drugs is significantly better than that of 5 commercial probes. The extraction ability of the SU101 probe for ten polar drugs is 1.4 - 1311 times that of other probes. The SU101 probe of the present invention can not only be applied to the detection of glucocorticoid drugs, but also to the detection of other PPCPs, with a wide range of applications.

[0071] Example 5 Verification of Detection Method

[0072] Under the optimized conditions of a selected extraction time of 40 min, a desorption time of 20 min, an oscillation rate of 600 rpm, and a mixed solvent (a mixed solvent of methanol and acetonitrile with a volume ratio of 2:3) as the desorption solvent, a methodological investigation was carried out on the established solid-phase microextraction method for the rapid detection of trace polar glucocorticoid drugs in environmental water. The investigation contents included the detection limit, quantification limit, linear range, single-needle reproducibility, and multi-needle reproducibility.

[0073] The results are shown in Table 3. The results indicate that the established method is linear in the range of 5 - 10000 ng L -1 and has a satisfactory linear coefficient (R 2 ≥0.9974). The detection limit is as low as 0.070 - 1.5 ng L -1 , and the quantification limit can reach 0.24 - 4.9 ng L -1 . The reproducibility is expressed by the relative standard deviation (RSD) value. The RSD values of single-needle reproducibility (n = 3) and multi-needle reproducibility (n = 6) are both less than 9%. These excellent experimental performances prove that the probe and analytical method of the present invention have high sensitivity and stability and can meet the requirements of trace detection.

[0074] Table 3 Methodological Investigation of the Established SPME-HPLC Method

[0075]

[0076] Example 6 Detection Method for Glucocorticoid Drugs in Water Environment

[0077] Real water samples were collected from three different locations in the Pearl River of Guangzhou. After filtering with a 0.22 μm filter membrane, the samples were detected by the above-established method.

[0078] The specific method is as follows:

[0079] S1. Enrichment of target analytes: The solid-phase microextraction probe of the present invention was completely immersed in a 10 mL glass vial containing 10 mL of sample solution, and extracted for 40 min at an oscillation rate of 600 rpm.

[0080] S2. Elution of the target analyte: Then, place an inner cannula containing 100 μL of desorption solvent (a mixed solvent of methanol and acetonitrile with a volume ratio of 2:3) in a 2 mL vial, completely immerse the probe adsorbed with the target in it, desorb the analyte at an oscillation rate of 600 rpm for 20 min of elution, take out the solid-phase microextraction probe and immerse it in methanol, and it can be reused after drying. The eluate is stored in a -20 °C refrigerator for use when analyzing or can be used directly.

[0081] S3. HPLC-MS / MS analysis: Use HPLC-MS / MS to quantitatively analyze the eluate containing the analyte. The parameters of HPLC-MS / MS are referred to in Example 2.

[0082] Meanwhile, to verify the reliability of the method, before sample adsorption, 0.5 and 5 μg L -1 of the standard substances of 8 polar glucocorticoid drugs in the following table were added to the sample respectively, and the recovery rate was determined using the linear equation established in Step Example 5. The results are shown in Table 4, and the obtained recovery rate is 70.8% - 120%, verifying the practicability and feasibility of this method.

[0083] Table 4 Analysis of environmental water samples and determination of recovery rate

[0084]

[0085]

Claims

1. Application of Bio-MOFs material SU101 in solid-phase microextraction, characterized in that, The extraction targets include dehydroepiandrosterone, testosterone, androstenedione, erythromycin, trimethoprim, carbamazepine, corticosterone, cortisone, fluorometholone, hydrocortisone, methylprednisolone, prednisolone, triamcinolone acetonide, and / or triamcinolone.

2. The application according to claim 1, wherein the bio-MOFs material SU101 is used to prepare the coating of the solid-phase microextraction probe.

3. The application of the bio-MOFs material SU101 in detecting pharmaceuticals and personal care products (PPCPs) in the water environment; the PPCPs include dehydroepiandrosterone, testosterone, androstenedione, erythromycin, trimethoprim, carbamazepine, corticosterone, cortisone, fluorometholone, hydrocortisone, methylprednisolone, prednisolone, triamcinolone acetonide, and / or triamcinolone.

4. A detection method for pharmaceuticals and personal care products (PPCPs) in an aqueous environment, characterized in that, The PPCPs include dehydroepiandrosterone, testosterone, androstenedione, erythromycin, trimethoprim, carbamazepine, corticosterone, cortisone, fluorometholone, hydrocortisone, methylprednisolone, prednisolone, triamcinolone acetonide, and / or triamcinolone; The method comprises the following steps: S1. Enrichment of the target analyte: Completely immerse the solid-phase microextraction probe in the water environment sample solution and oscillate to extract the target analyte; S2. Desorption of the target analyte: Immerse the solid-phase microextraction probe enriched with the target analyte in the desorption solvent and desorb the target analyte under oscillation to obtain the desorbed solution; S3. Use an analytical instrument to perform quantitative and / or qualitative analysis on the desorbed solution; The solid-phase microextraction probe described in step S1 includes a stainless steel wire carrier and a SU101 coating.

5. The detection method according to claim 4, characterized in that, The pharmaceuticals and personal care products (PPCPs) are selected from glucocorticoid drugs (GCs); In step S1, the extraction time is 20 - 60 min; the oscillation rate is 400 - 700 rpm; In step S2, the desorption time is 10 - 40 min; the oscillation rate is 400 - 700 rpm; In step S2, the desorption solvent is selected from any one or more of the following: methanol, acetonitrile, methanol containing 0.1% formic acid by volume, acetonitrile containing 0.1% formic acid by volume, and a mixed solvent of methanol and acetonitrile; In step S3, the analytical instrument is selected from HPLC / MS / MS.

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