Molecularly Imprinted Solid-phase Extraction Research Articles

Guiding Molecularly Imprinted Polymer Design by Pharmacophore Modeling.

Molecularly imprinted polymers (MIP) combine the selectivity of immunoaffinity chromatography with the robustness of common solid-phase extraction in what is referred to as molecularly imprinted solid-phase extraction (MISPE). This contribution shows how MIP design may be guided by pharmacophore modeling for the example of citrinin, which is an emerging mycotoxin from cereals. The obtained pharmacophore model allowed searching public databases for a set of citrinin-mimicking molecular surrogates. Imprinted and non-imprinted polymers were subsequently obtained through bulk and core-shell polymerization in the presence of these surrogates. Evaluation of their binding ability for citrinin and structurally related ochratoxin A revealed a promising MIP derived from rhodizonic acid. A protocol for MISPE of citrinin from cereals was subsequently developed and compared to immunoaffinity chromatography with respect to clean-up efficiency and recovery.

Optimization of a saccharin molecularly imprinted solid-phase extraction procedure and evaluation by MIR hyperspectral imaging for analysis of diet tea by HPLC

Saccharin was determined based on a new molecularly imprinted solid-phase extraction (MISPE) procedure. The polymer was synthesized with a hybrid monomer of metacrylic acid and 3-amino propril tetraethoxysilane and saccharin as template. After the synthesis, the saccharin removal from the MIP was verified by the UV analysis of the solutions used in the template removal procedure, as well as by the direct MIP analysis using FTIR hyperspectral image and chemometrics. The residual saccharin concentrations observed in the image analysis revealed a narrow concentration distribution consistent with a homogenous material. The MISPE was performed with homemade cartridges containing 200 mg of the MIP. The results obtained with standards and diet tea samples confirmed high affinity, adsorption capacity and selectivity of the MIP. The MISPE cartridge exhibited recoveries of 100 ± 3% in six extraction cycles. The diet tea analysis showed a significant reduction of the interferences, which can considerable simplifies the HPLC-UV analysis.

Computational modeling, fabrication, and characterization of the deep eutectic solvent-based green molecular cage for selective metronidazole extraction from plasma followed by UHPLC with diode array detector determination.

Four ternary deep eutectic solvents were computationally designed and synthesized, being used as candidate functional monomers in metronidazole molecular imprinting polymer synthesis, allowing selective extraction and determination by ultra high performance liquid chromatography with diode array detection. In terms of metronidazole selective extraction, the best results were obtained by (deep eutectic solvent)2 :(ethylene glycol dimethacrylate)11 , in which deep eutectic solvent is the functional monomer constructed by combining three components in 6:6:2 ratios of choline chloride:ethylene glycol:methacrylic acid. The effects of different parameters on molecular imprinted solid-phase extraction of metronidazole were thoroughly explored through screening design and response surface methodology. The adsorption mechanism findings show that the adsorption data are primarily fitted on the Freundlich model based on higher correlation coefficient. Kinetic experiments have shown that the mechanism of adsorption fits the pseudo-second-order model. The best extraction recovery (96.5%) was obtained in 25-min elution time, desorption temperature of 40°C, and 1.0mL ACN as eluent. Metronidazole was measured by a validated ultra high performance liquid chromatography with diode array detection method. The calibration of the method was linear in the range of 0.1-10 μg/mL with limits of detection and quantification of 0.03 and 0.1 μg/mL, respectively. The method was successfully applied for the determination of metronidazole in human plasma.

Computational design and synthesis of molecular imprinted polymers for selective solid phase extraction of sulfonylurea herbicides

A high-efficiency approach for the synthesis of molecularly imprinted polymers has been developed and further for the solid-phase extraction of sulfonylurea herbicides in food samples. Molecular simulation approach combined chemometric selected metsulfuron-methyl (MSM) and 2-trifluoromethyl acrylic acid (TFMAA) as the template and the monomer to synthesize the molecularly imprinted polymers (MIPs). Experimental validation confirmed that the MSM-imprinted polymers showed a higher selectivity and affinity to sulfonylurea herbicides. The optimized molecularly imprinted solid-phase extraction (MISPE) conditions, including loading, washing, and eluting conditions, were established. The developed MISPE technology combined HPLC-MSMS was successfully used for the determination of sulfonylurea herbicides in foods. Compared with commercial SPE columns, MISPE showed high affinity, excellent selectivity and low matrix effect. The recoveries of sulfonylurea herbicides spiked in four matrices were between 86.4% and 100.2%, with the relative standard deviations (RSD) in the range of 0.9%–10.5%.

Synthesis of molecularly imprinted polymers based on boronate affinity for diol-containing macrolide antibiotics with hydrophobicity-balanced and pH-responsive cavities

In this research, in order to separate and purify diol-containing macrolide antibiotics, like tylosin, from complex biological samples, molecularly imprinted polymer (MIP) based on boronate affinity for tylosin was synthesized by using precipitation polymerization method with 4-vinylphenylboronic acid (VPBA) and dimethyl aminoethyl methacrylate (DMAEMA) as pH-responsive functional monomers, and N,N'-methylene bisacrylamide (MBAA)/ ethylene glycol dimethacrylate (EGDMA) as the co-crosslinkers that balance the hydrophobicity of the MIP. The synthesized tylosin-MIP had the advantages of high adsorption capacity (120 mg/g), fast pH-responsiveness responsible for the accessibility of imprinted cavities, and high selectivity coefficient towards tylosin versus its analogues (2.8 versus spiramycin, 7.3 versus desmycosin) in an aqueous environment. The mechanism of boronate affinity between tylosin and VPBA in the form of charged hydrogen bonding was analyzed via density functional theory (DFT). MIPs were used to successfully separate diol-containing macrolides through molecularly imprinted solid phase extraction (MISPE). The results show that MIPs prepared in this method have a good application prospect in the separation and purification of the diol-containing macrolide antibiotics.

Selective solid-phase extraction of four phenylarsonic compounds from feeds, edible chicken and pork with tailoring imprinted polymer

The novel molecularly imprinted microspheres for four phenylarsonic compounds have been firstly prepared with the reversible addition-fragmentation chain transfer polymerization in a suspension system. The resulting polymeric microspheres were characterized by infrared spectrum, scanning electron microscope and differential scanning calorimetry. With serial adsorption experiments, the polymeric microspheres showed highly specific molecular recognition, fast mass transfer rate and robust adsorption of the substrates. Then, the imprinted polymer was used as the solid-phase extraction adsorbent to extract the phenylarsonic compounds from the feeds, edible chicken and pork. The cartridge was washed with 2 mL ethyl acetate and eluted with 3 mL of methanol- acetic acid (90:10, v/v). The recoveries of the molecularly imprinted solid-phase extraction (MISPE) column ranged from 83.4% to 95.1%. This work provided a versatile approach for the specific extraction of the organoarsenic compounds from complicated matrices and exhibited a bright future for the application of MISPE column.

Selective Solid-Phase Extraction Using Molecularly Imprinted Polymers for the Analysis of Target Pesticides in Cannabis Bud

ABSTRACT A molecularly imprinted solid-phase extraction (MISPE) procedure was developed for the GC-MS analysis of four high priority pesticides (atrazine, terbuthylazine, acetochlor and alachlor) in a cannabis bud sample matrix. The study demonstrated that the synthesised polymer had a high affinity and good selectivity for either chloroacetamide or triazine classes of pesticide used as a template molecule during the molecularly imprinted polymerisation reaction. The MISPE procedure was optimised in terms of loading, washing and elution fractions utilising a range of aqueous methanol solutions for optimal recovery and minimal matrix interferences. The optimal wash fraction was 20% (v/v) methanol in an aqueous solution, whilst 70% (v/v) was used for the elution fraction. The selectivity, accuracy and recovery of the MISPEs were verified using a synthesised non-imprinted polymer and a commercial C18 cartridge as reference sorbents in comparative experiments. Approximately 3 g of the cannabis bud sample was spiked at a 0.05 mg/kg maximum residue limit (MRL) concentration. The recovery of the four selected pesticides extracted from the spiked samples ranged between 76.4-85.0% when utilising the optimised MISPE methods, compared to 91.6-96.9% for the C18 SPE. However, the use of the MISPE resulted in enhanced selectivity, as evidenced by GC-MS analysis, due to the extraction of less matrix interferences. Therefore, it can be concluded that the MISPE is a viable pre-treatment method for selective pesticide analysis in cannabis flowers using GC-MS when selectivity is valued for the extraction of target pesticides from a complex sample matrix. Keywords: molecularly imprinted polymer; solid-phase extraction; Cannabis; pesticides; atrazine; terbuthylazine; acetochlor; alachlor

Molecularly Imprinted Polymers Coupled with Surface-Enhanced Raman Spectroscopy to Detect Chemical Hazards in Foods.

Preparation of molecularly imprinted polymers coupled with surface-enhanced Raman spectroscopy (MIP-SERS) sensor and its application in detecting chemical hazards in food matrices are described. Sample cleaning is achieved by molecularly imprinted solid-phase extraction (MISPE), and target molecules are detected by SERS. Procedures of MIP synthesis, MISPE preparation, SERS substrate preparation, spectral collection, data analysis, and food analysis application are described.

Molecularly Imprinted Solid-Phase Extraction Sorbents for the Selective Extraction of Drugs from Human Urine.

Development of molecularly imprinted solid-phase extraction (MISPE) sorbents for the removal of drugs from human urine is described. MISPE sorbents are synthesized through free radical polymerization mechanism and non-covalent imprinting approach. After the completion of the polymerization, bulk polymer is ground using an analytical mill and then wet-sieved. Later on, MISPE sorbent has been dry-packed into solid-phase extraction cartridges after the extraction of template drug molecule. Eluates from the cartridges are analyzed using a spectrophotometer or spectrofluorimeter for the determination of target analyte by referring to the calibration curves.

Bisphenol A migration to alcoholic and non-alcoholic beverages – An improved molecular imprinted solid phase extraction method prior to detection with HPLC-DAD

The great concern about food contamination and human exposure to bisphenol A (BPA) necessitated the development of a green and highly selective method for the efficient monitoring of BPA migration from the packaging materials to alcoholic and non-alcoholic beverages. Molecularly imprinted solid phase extraction (MISPE) coupled to high pressure liquid chromatography-diode array detection (HPLC-DAD) using an imprinted sol–gel silica-based hybrid inorganic–organic polymeric sorbent (sol–gel MIP) was applied in the determination of BPA in trace levels. The sorbent mass was optimized. The highest absolute recovery (RE = 84.1%) was obtained with the use of 50 mg of mass sorbent. The extraction protocol involved 0.5 mL sample loading, 15 min of holding time and elution with 2 mL ACN. The imprinting factor (IF) was equal to 3.63 ± 0.11 (n = 3). The developed MISPE-HPLC-DAD method was validated and demonstrated excellent linearity (0.997), high recovery (RE: 99.5% ± 5.7, n = 3), intra-assay repeatability over the range 92.4–98.1%, n = 3; and inter-assay precision over the range 81.2–102.3%, n = 3. Overall, 36 alcoholic and non-alcoholic beverages available in the Greek market were tested for BPA and the quantification results were below the regulation limit for all the samples over the range 0.011–0.041ng mL−1.

Synthesis and characterization of α-mangostin imprinted polymers and its application for solid phase extraction

α-mangostin imprinted polymers have been synthesized by a non-covalent imprinting approach with α-mangostin as a template molecule. The α-mangostin molecularly imprinted polymers (MIPs) prepared by radical polymerization using methacrylic acid, ethlylene glycol dimethacrylate, benzoyl peroxide, and acetonitrile, as a monomer, crosslinker, initiator, and porogen, respectively. The template was removed by using methanol:acetic acid 90:10 (v/v). The physical characteristics of the polymers were investigated by Fourier Transform Infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and thermogravimetric analysis (TGA). The rebinding studies were carried out by batch methods. The results exhibited that the MIPs was able to adsorb the α-mangostin at pH 2 and the contact time of 180 min. The kinetic adsorption data of α-mangostin performed the pseudo-second order model and followed the Langmuir isotherm model with the adsorption capacity of 16.19 mg.g-1. MIPs applied as a sorbent material in solid-phase extraction, namely molecularly imprinted solid-phase extraction (MISPE) and it shows the ability for enrichment and clean-up of α-mangostin from the complex matrix in medicinal herbal product and crude extract of mangosteen (Garcinia mangostana L.) pericarp. Both samples, respectively, which were spiked with α-mangostin gives recovery more than 90% after through by MISPE in all concentration ranges.

Facilitating serum determination of neuron specific enolase at clinically relevant levels by coupling on-line molecularly imprinted solid-phase extraction to LC-MS/MS

The identification and quantification of biomarkers is essential for the diagnosis, treatment, and long-term monitoring of many human diseases. In the present work, macromolecular synthetic receptors with pre-determined affinity and selectivity for the signature peptide of a prognostically significant small cell lung cancer (SCLC) biomarker - neuron-specific enolase (NSE) – were prepared in a porous polymer microsphere format using a template-directed synthesis strategy performed under precipitation polymerization conditions. The polymer microspheres were packed into short trap columns and then exploited as molecularly selective sorbents in a fully automated, on-line molecularly imprinted solid-phase extraction (MISPE) protocol. The on-line MISPE protocol was optimised with respect to the composition of the loading mobile phase, the flow rate, and the extraction time. The molecularly imprinted polymers (MIPs) showed high affinity and useful selectivity for the peptide target - the hexapeptide ELPLYR - compared to non-imprinted control polymers. The MIPs were able to retain the biomarker on-column for extraction times of up to 20 min, and the on-line MISPE method enabled complete recovery of the biomarker over the linear range 10–100 ng mL−1 when the biomarker was present in spiked ammonium bicarbonate solution (R2 = 0.994). For extractions of ELPLYR from very complex biological matrices, the recoveries of ELPLYR from reversed-phase SPE (RP-SPE)-treated and untreated digested human serum were 100.8 ± 6.2% and 61.6 ± 1.9%, respectively. Extractions of ELPLYR from spiked untreated digested serum were linear in the range of 7.5–375 ng mL−1 (R2 = 0.99). The limit of detection (LOD) and limit of quantification (LOQ) for the biomarker in digested serum were estimated to be 1.8 ng mL−1 and 6.0 ng mL−1, respectively, which is below the median reference level of NSE in humans (8.6 ng mL−1). This work sets in place the basis for a new diagnostic tool for SCLC that is sensitive, robust, automated, and antibody-free, and which works very well with complex human plasma samples.

Determination of fenthion in urine samples using molecularly imprinted nanoparticles: modelling and optimisation by response surface methodology

ABSTRACT In the present study, a molecularly imprinted solid-phase extraction (MISPE) technique coupled with high-performance liquid chromatography was applied for selective extraction and recognition of fenthion insecticide in the biological sample. The nanoparticles of fenthion imprinted polymer were prepared by precipitation polymerisation using methacrylic acid (MAA) as functional monomer, ethylene glycol dimethacrylate (EGDMA) as cross-linker, and 70 mL of chloroform. Morphological and structural characterisation was achieved by scanning electron microscopy and infrared spectrometry. The optimisation of parameters being effective on extraction yield was carried out by employing the central composite design (CCD). The empirical models were developed to link the output response and input variables and the proposed models were tested for significance through analysis of variance (ANOVA). The optimum conditions leading to the maximum efficiency of MISPE were sorbent mass of 8.61 mg, sample pH 4.93, sample flow rate of 2.07 mLmin−1, as well as 1.94 mL of methanol/acetic acid (95.7:4.3, v/v) at the flow rate of 1 mLmin−1 as elution solvent. Under the optimised conditions, a linear range of 20–120 μgL−1 was obtained and the limit of detection was found to be 4.58 μgL−1. Analysis of spiked urine samples demonstrated the satisfactory efficiency of the designed protocol ranging from 92.69 to 95.64% and the relative standard deviation (RSD) was calculated to be lower than 3.75%.

Design and performance of novel molecularly imprinted biomimetic adsorbent for preconcentration of prostate cancer biomarker coupled to electrochemical determination by using multi-walled carbon nanotubes/Nafion®/Ni(OH)2-modified screen-printed electrode

The present paper describes the development of a new electroanalytical method for prostate cancer biomarker (sarcosine) by coupling molecularly imprinted solid-phase extraction (MISPE) with electrochemical determination at a MWCNT/Nafion®/Ni(OH)2-modified screen-printed electrode using a flow injection analysis (FIA) system. The synthesized MIP prepared by using dual functional monomers 2-acrylamido-2-methyl-1-propanesulfonic acid and 4-vinylpyridine was characterized by Fourier Transform Infrared (FT-IR) spectroscopy, Scanning Electron Microscopy (SEM) and nitrogen adsorption/desorption measurements. The optimized pH for adsorption of sarcosine was 5.0 and the elution was completed with MeOH:HAc (90:10, v/v), whilst the electrochemical determination was performed in FIA system using a sample volume of 121.0 μL, 0.05 mol L−1 NaOH carrier solution, a flow rate of 2.0 mL min−1 and applied potential of 0.7 V vs Ag/AgCl. The proposed method provided enrichment factor of 50.6-fold, linear analytical curve from 3.2 to 25.0 μmol L−1 (R2 = 0.995), and limit of detection of 0.96 μmol L−1. The precision was evaluated in terms of intra (n = 6) and interday (n = 2) for concentrations of 10.0, 15.0, and 20.0 μmol L−1, giving rise to low percentages of relative standard deviations (RSD %), ranging from 2.2 to 4.3%. A highly improved response for sarcosine using MIP when compared to NIP (non-imprinted polymer) was obtained. The method also exhibited higher selectivity toward sarcosine compared to glycine, alanine, proline, and urea. Analysis of synthetic urine samples yielded recoveries in the range of 95.0–108.0%, thereby indicating the absence of matrix effect and the feasibility of the method for sarcosine determination.

Molecularly imprinted polymers for selective extraction of rosmarinic acid from Rosmarinus officinalis L.

This study provides a robust and reproducible approach for selective extraction of rosmarinic acid (RA) using molecularly imprinted polymers (MIPs). Computational modeling and UV spectroscopic analysis were performed to optimize MIP synthesis. Consequently, six different bulk and surface imprinted polymers were generated using RA as the template. Binding performance of the imprinted polymers was evaluated using static equilibrium and complementary dynamic rebinding experiments. Despite the high selectivity of thus generated surface imprinted polymers, the corresponding bulk polymers exhibited better binding performance when serving as sorbents during solid phase extraction (SPE). An optimized molecularly imprinted solid phase extraction (MISPE) protocol was developed in respect to loaded amount of RA, composition of the loading solution, washing solvent, and elution volume. Thereby, a remarkably selective extraction of RA from real-world Rosmarinus officinalis L. extract with a recovery rate and purity of 81.96 ± 6.33% and 80.59 ± 0.30%, respectively, was achieved.

Development of selective preconcentration/clean-up method for imidazolinone herbicides determination in natural water and rice samples by HPLC-PAD using an imazethapyr imprinted poly(vinylimidazole-TRIM)

The development of a novel molecularly imprinted solid-phase extraction (MISPE) method for simultaneous preconcentration of imazapyr (IMP), imazapic (IMZ) and imazethapyr (IMT) with determination by HPLC-PAD (High performance liquid chromatography – photodiode-array detector) is proposed. The polymer synthesis was performed using imazethapyr as template molecule and 1-vinylimidazole as functional monomer. The method is based on preconcentration of 100.0 mL of sample through 200.0 mg of molecularly imprinted poly(vinylimidazole-TRIM) (MIP-1VN) at pH 4.0, followed by elution with 2.0 mL of MeOH:CH2Cl2:HAc (34:62:4, v/v). The range of analytical curve (0.29–200.0, 0.21–200.0 and 0.15–200.0 µg L−1), limits of detection (0.09, 0.06 and 0.04 µg L−1) and preconcentration factors (92, 96 and 98) determined for the herbicides, IMP, IMZ and IMT, respectively, were greatly superior when compared with those ones obtained with commercial adsorbents. The analytical method was successfully applied to spiked surface water and rice samples with good results of recovery values (86–107%).

Application of Molecularly Imprinted Polymers (MIP) and Flowing Atmospheric-Pressure Afterglow Mass Spectrometry (FAPA-MS) to Analysis of Nonsteroidal Anti-Inflammatory Drugs (NSAIDs)

In recent years, the rapid development of the pharmaceutical industry and the extensive and illicit use of painkillers have led to increased levels of nonsteroidal anti-inflammatory drugs (NSAIDs) in the environment. In view of the significant impact of NSAIDs on living organisms, including humans, their presence in the environment needs to be continuously monitored at trace levels. For this purpose, a combination of molecularly imprinted solid-phase extraction (MISPE) and HPLC-MS analysis is commonly used. MISPE has been utilized in direct, fast, and ecological analysis of drugs using a flowing atmospheric-pressure afterglow ion source for mass spectrometry (FAPA-MS). The new method was applied herein in the determination of naproxen, diclofenac, and ibuprofen. The linear dependence of the intensity of analytical signals on the amount of drugs is in the range of 0.2 μg to 1 g and the method detection limit (MDL) for all drugs is 0.2 μg in environmental samples. The new method also decreased the number of analytical stages, the time and cost of analysis, and the organic solvent consumption, besides being environmentally friendly.

Molecularly imprinted polymer solid phase extraction coupled with liquid chromatography-high resolution mass spectrometry for the detection of gonyautoxins 2&3 in seawater

Paralytic shellfish toxins (PSTs) cause risks to human health through food chains. Understanding the change of PSTs in seawater is critical for predicting the safety of seafood. Most reported methods for the detection of PSTs in microalgae or shellfish are not applicable in seawater because of extremely low concentration and matrix interferences. High resolution mass spectrometry (HRMS), quadrupole exactive orbitrap detects molecular ions accurately, and molecularly imprinted solid-phase extraction (MISPE) is recognized effective to reduce the matrix interference. GTXs 2&3 are two of common marine toxins in PSTs. In this study, a sensitive method consisting MISPE and liquid chromatography LC-HRMS was developed for the detection of GTXs 2&3 with a limit of detection (LOD) of 47.4 ng/L in seawater. With this method, samples obtained from the estuaries of the Shuangtaizi and Daliao Rivers were analyzed, and the results indicated the concentrations were lower than LOD in the area under investigation.

Direct extraction and detection of malachite green from marine sediments by magnetic nano-sized imprinted polymer coupled with spectrophotometric analysis

This research describes the application of magnetic molecularly imprinted nano-sized polymers (MMIPs) for the selective extraction and fast detection of malachite green (MG) from marine sediment samples followed by UV–Vis spectrophotometry. The novel material was prepared by surface imprinting using methacrylic acid as the functional monomer for fixing the template molecules. The polymers obtained at each step were thoroughly studied by transmission electron microscopy, FTIR spectroscopy and thermogravimetric analysis. Simultaneously, the adsorption performances of the resulting nanoparticles were analysed in detail and an excellent affinity with the MG was revealed. Further, the main parameters of magnetic molecular imprinted solid-phase extraction (MMIP-SPE) were screened via multivariate optimization methods. The magnetic nanoparticles were used as special adsorbents to directly extract MG from crude marine sediment extracts. The developed method exhibits satisfactory recoveries from the spiked samples, ranging from 80.40 to 92.96% with an RSD of less than 5.18% (n = 3).

Synthesis and characterization of a molecularly imprinted polymer (MIP) for solid-phase extraction of the antidiabetic gliclazide from human plasma.

Gliclazide is a sulfonylurea frequently prescribed for the management of type 2 diabetes mellitus in elderly patients and for patients with chronic renal or hepatic diseases. Even though it is considered a safer alternative, the drug can provoke side effects in some patients, especially hypoglycemia, due to the high interindividual variability. Therefore, the quantification of gliclazide in biological samples is usually recommended in order to assure efficacy and safety of the pharmacotherapy. However, due to the complexity of biological matrices, therapeutic monitoring can be very challenging, especially in the sample preparation step. For that reason, the synthesis and characterization of a novel and selective molecularly imprinted polymer (MIP) was proposed to be employed as sorbent for the extraction of gliclazide from human plasma samples by a molecularly imprinted solid-phase extraction (MISPE) procedure. Synthesis conditions were optimized (monomer, crosslinker and porogen) and the polymer was characterized for its morphological, physicochemical and stability properties. The influence of drug concentration, solvent composition and pH on the coefficient of distribution (Kd) and imprinting factor (IF) were studied, as well as repeatability between batches and selectivity. A bioanalytical method was developed applying the developed MIP as sorbent in solid phase extraction and liquid chromatography using a Poroshell 120 C18 (100×4.6mm, 4μm) column, acetonitrile and 10mM potassium phosphate buffer pH3.0 (50:50) at a flow-rate of 1.2mL/min as mobile phase, temperature of 30°C, injection volume of 40μL and detection at 230nm. The best reaction yield, extraction capacity, and selectivity was obtained using 2-hydroxyethyl methacrylate (2-HEMA), ethyleneglycol dimethacrylate (EGDMA) and acetonitrile. The optimized MIP showed coefficient of distribution (Kd) of 59.85μg/g, imprinting factor (IF) of 1.60, and selectivity for gliclazide and other sulfonylureas compared to possible concurrent drugs. The developed method by MISPE-HPLC-UV showed to be appropriate to determine gliclazide in human plasma samples.