Superparamagnetic lysozyme surface-imprinted polymer prepared by atom transfer radical polymerization and its application for protein separation

Molecular imprinting as a promising and facile separation technique has received much attention because of their high selectivity for target molecules. In this study, the superparamagnetic lysozyme surface-imprinted polymer was prepared by a novel fabricating protocol, the grafting of the imprinted polymer on magnetic particles in aqueous media was done by atom transfer radical polymerization (ATRP), and the properties of the imprinted polymer were characterized in detail. Its high selective adsorption and recognition to lysozyme demonstrated the separation ability of the magnetic imprinted material to template molecule, and it has been used for quick and direct separation of lysozyme from the mixture of standard proteins and real egg white samples under an external magnetic field. Furthermore, the elution of lysozyme from the imprinted material was achieved by PEG/sulphate aqueous two-phase system, which caused lysozyme not only desorption from the imprinted materials but also redistribution in the top and bottom phase of aqueous two-phase system. The aqueous two-phase system exhibited some of the extraction and enrichment effect to desorbed lysozyme. Our results showed that ATRP is a promising method for the protein molecularly imprinted polymer preparation.     

Synthesis, characterization and application of modified Pd nanoparticles as preconcentration probes for selective enrichment/analysis of proteins via hydrophobic interactions from real-world samples using nanoparticle-liquid-liquid microextraction coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

We introduce a novel preconcentrating technique by using surface modification of palladium nanoparticles (Pd-NPs) with octadecane thiol (ODT) prepared in toluene for selective and sensitive extraction of proteins (insulin, ubiquitin, lysozyme) from a variety of real-world samples including pancreas, mushroom, soybean and milk using nanoparticle-liquid-liquid microextraction (NP-LLME) coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS). The limit of detection (LOD) values obtained for gramicidin D and insulin in water and urine are between 17-37 nM (17-37 fmol) (with RSDs ranging from 5.3-7.2%) which are 10-20-fold enhancement in detection sensitivity compared with conventional MALDI-MS. The optimal sample pH for highest extraction efficiency of insulin, ubiquitin and lysozyme from biological samples was observed at sample pH ～ pI which could be due to the enhancement of hydrophobic interactions between proteins with the hydrophobic ligands of Pd-ODT NPs. In addition, we also found that with the addition of 1?M NaCl, signals could be significantly enhanced by using the current approach. It is an efficient, straightforward, sensitive and selective nanoprobe which can be widely applied for separation, enrichment and preconcentration of peptides or proteins from complex biological samples in proteome research.     

Suitability of selective pressurized liquid extraction combined with gas chromatography-ion-trap tandem mass spectrometry for the analysis of polybrominated diphenyl ethers

A one-step extraction and clean-up method using pressurized liquid extraction (PLE) (selective PLE) combined with gas chromatography-ion-trap tandem mass spectrometry (GC-ITMS-MS) was evaluated for the analysis of polybrominated diphenyl ethers (from tri- to hepta-PBDEs) at low concentrations in fish and shellfish samples. To this end, the performance of an on-line PLE extraction/clean-up method and of a classical Soxhlet extraction and clean-up method using a multi-layer modified silica column were compared. The two sample treatment methods provided similar results, although an important reduction in the sample treatment time (40 min per sample) was achieved using the selective PLE method. In addition, the suitability of the PLE combined with GC-ITMS-MS method was evaluated by comparing the results obtained in the analysis of fish samples with those obtained by gas chromatography-high resolution mass spectrometry (GC-HRMS). Good agreement between both techniques was obtained with differences between the mean values of less than 16%. The selective PLE method coupled to GC-ITMS-MS produced accurate results for PBDE determination with low limits of detection (1.0-16.8 pg g⁻¹ wet weight) and quantification (3.1-51 pg g⁻¹ wet weight) as well as good precision (RSD < 16%). This method has been applied to the analysis of PBDEs in fish and shellfish samples collected at fish markets in Catalonia (NE Spain).     

Synthesis,characterization, and application of griseofulvin surface molecularly imprinted polymers as the selective solid phase extraction sorbent in rat plasma samples

In present study, an investigation was carried out to develop and validate an analytical method for the selective extraction and determination of griseofulvin (GSF) from plasma samples. For this purpose, a rational approach was made to synthesize and characterize the surface molecularly imprinted polymers (SMIPs). The SMIPs were utilized as solid phase extraction (SPE) sorbents. The SMIPs were prepared by using GSF as template molecule on the surface of modified silica particles through a non-covalent technique. The particles demonstrated high adsorption capacity (119.1 µg/mL), fast adsorption equilibrium time (30 min) and good recognition selectivity for the template drug. The scanning electron microscopy and infrared spectroscopy were used to explain the structural and morphological characteristics of the SMIPs and surface non-imprinted polymers. The SPE method was combined with HPLC for plasma analysis. The method validation results demonstrated that the established method possessed good linearity for GSF ranging from 0.1 to 50 µg/mL (R² = 0.997). The limit of detection for this method was 0.02 µg/mL for rat plasma samples. The recoveries of GSF from spiked plasma samples were (90.7–97.7%) and relative standard deviations were (0.9–4.5%). Moreover, the SMIPs as selective SPE sorbent can be reused more than 8 times which is a clear advantage over commercial SPE sorbents. Finally, the usefulness of the proposed strategy was assessed by extraction and detection of GSF in real rat plasma samples.     

SPE/RIA vs LC/MS for measurement of low levels of budesonide in plasma

A radioimmunoassay is described that measures budesonide in plasma after solid-phase extraction (SPE/RIA) of the analyte. The performance of the assay was compared with that of a selective LC/MS method. The limit of quantitation of budesonide determined for the LC/MS and SPE/RIA assay was 50 pg/mL and 120 pg/mL, respectively. Based on quality control samples, a higher variability was observed for the SPE/RIA (CV between 4.5 and 23.0%) than for the LC/MS method (CV between 7.5 and 12.5%). Plasma samples obtained from healthy volunteers after administration of budesonide rectal foam were assayed by both methods. In a subset of samples, these results were compared with those measured by direct RIA to evaluate the selectivity of two assays. About two times higher budesonide levels were measured with the direct RIA (lacking the extraction step), presumably because of cross-reactivity with budesonide metabolites, indicating that the extraction step in SPE/RIA is necessary for selectivity. Both SPE/RIA and LC/MS methods were found to be selective, sensitive and suitable for pharmacokinetic studies. Results obtained from the two methods were compared with a number of statistical methods. Ratios of results obtained for the clinical samples were close to 1 (ratio LC-MS/ SPE/RIA = 0.98 +/- 0.27). Linear regression indicated a slope of 1.17 +/- 0.0378. The concordance correlation (r = 0.91) indicated that the agreement between both methods was fair while the Bland-Altman plot indicated that the agreement was less pronounced at higher concentrations (1-3 ng/mL). In summary, the results confirm that the SPE/RIA is an alternative to HPLC/MS and that among the statistical methods tested the concordance correlation analysis was judged to be the most informative test to assess the comparability of two methods.     

CdSe量子点探针共振光散射法检测溶菌酶

基于CdSe量子点与溶菌酶之间的相互作用, 采用共振光散射法建立了简单快速检测溶菌酶的新方法. 在优化的实验条件下, 当溶菌酶浓度在0.01~0.8 μmol/L范围内变化时, 散射光强度与溶菌酶浓度间呈现良好的线性关系, 相关系数为0.9960. 此方法对溶菌酶的检出限为5.2 nmol/L, 对0.09 μmol/L溶菌酶5次平行测定的相对标准偏差为2.1%, 此方法选择性较好, 常见离子、蛋白质及常见氨基酸对溶菌酶检测干扰较小, 这种新方法已被用于合成样品中溶菌酶的检测, 并取得较好结果. 为进一步考察CdSe量子点与溶菌酶之间的相互作用, 进行了圆二色光谱、透射电子显微镜和荧光寿命表征研究, 结果表明, CdSe量子点与溶菌酶的结合不仅使溶菌酶的构象发生变化, 也使CdSe量子点的分散状态和荧光寿命发生了改变.     

Development and validation of an SPE HG-AAS method for determination of inorganic arsenic in samples of marine origin

The present paper describes a novel method for the quantitative determination of inorganic arsenic (iAs) in food and feed of marine origin. The samples were subjected to microwave-assisted extraction using diluted hydrochloric acid and hydrogen peroxide, which solubilised the analytes and oxidised arsenite (As(III)) to arsenate (As(V)). Subsequently, a pH buffering of the sample extract at pH 6 enabled selective elution of As(V) from a strong anion exchange solid-phase extraction (SPE) cartridge. Hydride generation atomic absorption spectrometry (HG-AAS) was applied to quantify the concentration of iAs (sum of As(III) and As(V)) as the total arsenic (As) in the SPE eluate. The results of the in-house validation showed that mean recoveries of 101-104% were achieved for samples spiked with iAs at 0.5, 1.0 and 1.5 mg·kg(-1), respectively. The limit of detection was 0.08 mg kg(-1), and the repeatability (RSD(r)) and intra-laboratory reproducibility (RSD(IR)) were less than 8% and 13%, respectively, for samples containing 0.2 to 1.5 mg kg(-1) iAs. The trueness of the SPE HG-AAS method was verified by confirming results obtained by parallel analysis using high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry. It was demonstrated that the two sets of results were not significantly different (P < 0.05). The SPE HG-AAS method was applied to 20 marine food and feed samples, and concentrations of up to 0.14 mg kg(-1) of iAs were detected.     

C_(18)膜萃取/液相色谱-质谱联用法测定极地水体中有机磷酸酯

建立了C_(18)膜萃取/液相色谱-质谱联用技术测定极地水体中10种有机磷酸酯(OPEs)的方法。根据优化后的样品前处理及仪器方法,利用C_(18)膜富集4 L水体中的10种OPEs,经二氯甲烷超声提取,在电喷雾正离子模式下,采用选择反应监测(SRM)模式进行分析,线性相关系数为0.994 4~0.999 9。10种OPEs的加标回收率为64.1%~115%,方法检出限为0.08~0.55 ng/L。该方法适用于极地水体中10种OPEs的分析,利用该方法对北极水体样品中的10种OPEs进行检测,测得冰川融水中∑OPEs的质量浓度为0.64~6.64 ng/L,海水中∑OPEs的质量浓度为0.09~2.03 ng/L。     

Molecularly imprinted polymer for solid-phase extraction of rutin in complicated traditional Chinese medicines

In the present work, an improved and direct approach for the preparation of molecularly imprinted polymers (MIPs) was proposed. The MIPs were prepared based on bulk polymerization by water-bath heating and ultrasonic elution of the template, using rutin as the template, acrylamide (AM) as the functional monomer and 2,2'-azobisisobutyronitrile (AIBN) as the cross linker. Molecularly imprinted polymers prepared by other elution methods, including microwave-assisted extraction and conventional Soxhlet extraction, were used for comparison and the results showed that the ultrasonic elution method is the best. The synthesized MIPs were characterized by Fourier transform infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM). High performance liquid chromatography (HPLC) was used to evaluate the adsorption properties and recognition mechanism of the MIPs. Structurally similar compounds including quercetin and genistein were utilized for verifying the molecular selectivity and characterizing the recognition capability of the MIPs. The MIPs were used as a sorbent for the solid phase extraction of rutin, and the resultant cartridge showed a good extraction performance. Thus, a molecularly imprinted solid-phase extraction (MISPE) procedure for selective pre-concentration of rutin from complicated traditional Chinese medicine (TCM) samples was proposed. Various elution parameters that affect the adsorption capacity of the polymer were evaluated to optimize the selective pre-concentration of rutin. The characteristics of the MISPE method were validated by HPLC. The recoveries ranged from 85% to 91% for TCMs, which demonstrated that this MISPE-HPLC method could be applied to pre-concentrate and determinate rutin directly from complicated TCM samples in the presence of other interfering substances.     

Polarographic determination of cyanide as nickelcyano complex in blood plasma after selective extraction in a methylene blue impregnated polyethylene column

A method for the determination of cyanide in blood plasma by differential pulse polarography (DPP) is described without a drastic acidification of the sample. Cyanide was determined as tetracyanonickelate(II)-anion complex after a microwave-acid assisted cleanup and a selective complex extraction in a polyethylene methylene blue (PE-MB) impregnated column. The cyano complex was eluted from the column with water/acetonitrile and determined by pulse-polarography at –380 mV (Ag/AgCl). The linear range of calibration was obtained from 1.2 to 9.6 μg of cyanide with r = 0.99 and RSD = 9% of 1.2 μg of cyanide. A detection limit of 40 μg L^–1 was calculated and the recoveries of cyanide from spiked samples were about 80%. This method was compared with the classical pyridine-pyrazolone method.     

Water-compatible molecularly imprinted polymer for the selective recognition of fluoroquinolone antibiotics in biological samples

A novel water-compatible molecularly imprinted polymer (MIP), prepared with enrofloxacin (ENR) as the template, has been optimised for the selective extraction of fluoroquinolone antibiotics in aqueous media. The results of a morphological characterisation and selectivity tests of the polymer material for ENR and related derivatives are reported. High affinity for the piperazine-based fluoroquinolones marbofloxacin, ciprofloxacin, norfloxacin and ofloxacin was observed, whereas no retention was found for nonrelated antibiotics. Various parameters affecting the extraction efficiency of the polymer have been optimised to achieve selective extraction of the antibiotics from real samples and to reduce nonspecific interactions. These findings resulted in a MISPE/HPLC-FLD method allowing direct extraction of the analytes from aqueous samples with a selective wash using just 50% (v/v) organic solvent. The method showed excellent recoveries and precision when buffered urine samples fortified at five concentration levels (25–250 ng mL⁻¹ each) of marbofloxacin, ciprofloxacin, norfloxacin, enrofloxacin and sarafloxacin were tested (53–88%, RSD 1–10%, n = 3). Moreover, the biological matrix of the aqueous samples did not influence the preconcentration efficiency of the fluoroquinolones on the MIP cartridges; no significant differences were observed between the recovery rates of the antibiotics in buffer and urine samples. The detection limits of the whole process range between 1.9 and 34 ng mL^–1 when 5-mL urine samples are processed. The developed method has been successfully applied to preconcentration of norfloxacin in urine samples of a medicated patient, demonstrating the ability of the novel MIP for selective extraction of fluoroquinolones in urine samples.     

Determination of triazine herbicides in environmental water samples by high-performance liquid chromatography using graphene-coated magnetic nanoparticles as adsorbent

In this paper, a graphene-based Fe(3)O(4) magnetic nanoparticles (G-Fe(3)O(4) MNPs) was used as the adsorbent for the magnetic solid-phase extraction of some triazine herbicides (atrazine, prometon, propazine and prometryn) in environmental water samples followed by high performance liquid chromatography-diode array detection (HPLC-DAD). After the extraction, the adsorbent can be conveniently separated from the aqueous samples by an external magnet. The main factors influencing the extraction efficiency including the amount of the MNPs, the extraction time, the pH of sample solution, and desorption conditions were studied and optimized. Under the optimized experimental conditions, a good linearity was observed in the range of 0.1-50.0 ng mL(-1) for all the analytes, with the correlation coefficients (r) ranging from 0.9996 to 0.9999. The limits of detection of the method ranged between 0.025 and 0.040 ng mL(-1). Good reproducibility was obtained with the relative standard deviations below 5.2%. The developed method was applied to the analysis of the triazine herbicides in different water samples (lake, river and reservoir). The recoveries of the method were in the range between 89.0% and 96.2%.     

Rapid determination of pesticide residues in herbs using selective pressurized liquid extraction and fast gas chromatography coupled with mass spectrometry

A selective pressurized liquid extraction and gas chromatography coupled with triple quadrupole mass spectrometer method was developed for simultaneous determination of 52 pesticide residues in medicine and food dual-purpose herbs. The developed extraction method integrated extraction and cleanup processes for sample preparation. The sorbents, 5 g Florisil and 100 mg graphitized carbon black, were placed inside the extraction cell to remove matrix interferences. Optimized conditions of selective pressurized liquid extraction were ethyl acetate as extraction solvent, 120°C of extraction temperature, 6 min of static extraction time, 50% of flush volume extracted for two cycles. An ultra inert capillary GC-MS HP-5 UI column (20 m × 0.18 mm id, 0.18 μm) and column backflush system were used for the analysis. Multiple-reaction monitoring was employed for the quantitative analysis with electron ionization mode. All calibration curves showed good linearity (r(2) > 0.995) within the test ranges. The average recoveries of most pesticides were from 81 to 118%. The validated method was successfully applied for the determination of pesticide residues in four herbs. The results indicate that selective pressurized liquid extraction and GC-MS/MS is a sensitive and reliable analytical method for the simultaneous determination of multiple pesticide residues in herbs.     

全二维气相色谱-飞行时间质谱测定地下水中低环多环芳烃及其衍生物

建立了地下水中低环多环芳烃及其衍生物的全二维气相色谱-飞行时间质谱(GC×GC-TOF MS)检测方法。对比研究了液液萃取(LLE)和固相萃取(SPE)对地下水中低环多环芳烃及其衍生物的提取效率,优选液液萃取为前处理方法。在优化条件下,除1,2,3,4-四氢萘(r=0.987 2)和联苯(r=0.989 9)外,其它目标物在0.1~1 000μg/L范围内具有良好的线性关系,相关系数(r)均大于0.99。地下水的平均加标回收率为63.3%~111%,除喹啉的相对标准偏差(RSD,n=6)为24.9%外,其余目标物的RSD均小于9.5%,方法检出限在1.63~14.7 ng/L之间。该方法用于河北地区6个地下水样中低环多环芳烃及其衍生物的检测,4个样品有检出,最高浓度达353 ng/L。     

Molecularly imprinted solid-phase extraction and high-performance liquid chromatography with ultraviolet detection for the determination of urinary trans,trans-muconic acid: a comparison with ionic exchange extraction

A new molecularly imprinted polymer (MIP) has been synthesized for the selective extraction of trans,trans-muconic acid (ttMA) from urine samples, followed by high-performance liquid chromatography analysis with ultraviolet detection. The synthesis was based on non-covalent interactions, and 4-vinylpyridine was used as a functional monomer. The analytical calibration curve was prepared using a pool of five urine samples of non-smokers spiked with ttMA standards with concentrations that ranged from 0.3 to 10 mg L(-1) (r(2) = 0.999). The limit of quantification was 0.3 mg L(-1) (lower than the biological exposure limits suggested by the ACGIH). The within-day and between-day precision and accuracy presented relative standard deviations and relative errors of less than 15%. The analytical frequency was 4 h(-1) (considering extraction and separation/quantification steps), and the same MIP cartridge was efficiently used for approximately 100 cycles. All figures of merit were similar or better than those obtained by the procedure based on ionic exchange extraction. The proposed method could be an interesting alternative for the routine analysis of ttMA in urine for biological monitoring procedures of human exposure to benzene.     

分散固相萃取-在线凝胶色谱-气相色谱-质谱联用法快速检测紫菜中的农药多残留

建立了紫菜中农药多残留的在线凝胶色谱-气相色谱-质谱联用（GPC-GC/MS）检测方法。以有机氯、有机磷、三嗪类和菊酯类的19种农药为目标物,对比了丙酮、丙酮/二氯甲烷（1:1,v/v）和乙腈3种有机溶剂的提取效果,通过石墨化炭黑粉（GCB）和N-丙基乙二胺粉（PSA）分散固相萃取净化和GPC在线净化,气相色谱-质谱联用法分析,外标法定量。结果表明,此方法实现了在线净化与分析检测的自动化,缩短了分析时间。分析物在10~1000 μg/L范围内线性关系良好,相关系数r>0.995;采取GPC大体积进样和气相色谱进样口的程序升温方式提高了检测灵敏度,检出限为0.005~0.03 mg/kg。方法的平均添加回收率在70%~120%之间,相对标准偏差（RSD）均小于15%。该方法简单、快速、具有良好的回收率和重复性,适用于紫菜样品中农药多残留的快速灵敏检测。     

Selective pressurized liquid extraction of polychlorinated dibenzo-p-dioxins, dibenzofurans and dioxin-like polychlorinated biphenyls from food and feed samples

Selective pressurized liquid extraction (PLE) of polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) from various food and feed samples was performed with a selective PLE method previously developed for bulk PCBs. The method utilizes sulfuric acid impregnated silica inside the extraction cell to oxidize coextracted fat. Extractions were performed at 100 degrees C with n-heptane for 5 min in two cycles. Data obtained by selective PLE combined with gas chromatography/high-resolution mass spectrometry (GC-HRMS) were compared to concentrations derived from reference laboratories applying conventional sample preparation and GC-HRMS. Experiments performed on spiked vegetable oil, naturally contaminated crude fish oil and oil containing compound feed samples showed good results for these relatively simple matrices. The accuracy was generally +/-20% as compared to spiked levels or to values obtained by the reference laboratories. The precision, measured as the relative standard deviation (RSD) for 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalency values (TEQs), was below 10% in all cases. The method was also tested on naturally contaminated herring tissue, chicken tissue, pork tissue and sepiolitic clay, which all caused some trouble. It was observed that sufficient amounts of sodium sulfate should be used for dehydration of tissue samples and additionally, the cells should not be packed too dense in order to avoid suppressed extraction efficiency. Once this was attended to, satisfactory data could be obtained, except for sepiolithic clay. This study demonstrates that selective PLE can be applied with success to a number of food and feed matrices in analysis of PCDD/Fs and dl-PCBs. Since the fat removal step is on-line, the selective PLE method will reduce time and solvent consumption for sample preparation as compared to traditional clean-up.     

多壁碳纳米管固相萃取-在线提取蛋清中的溶菌酶

建立了以羧基功能化的多壁碳纳米管作为吸附剂在线提取蛋清中溶菌酶的新方法. 将多壁碳纳米管经氧化和提纯后装入固相萃取柱, 在顺序注射系统中实现对蛋清中溶菌酶的在线选择性萃取, 以碳酸盐缓冲溶液定量洗脱, 进样量为2 mL时, 富集倍率为12, 吸附率和洗脱率均为100%, 采样速率为10 h-1, 精密度为3.0%. 实验结果表明, 该方法所用试剂量少, 提取速度快, 每100 mL蛋清可提取溶菌酶0.4 g, 且溶菌酶纯度较高.     

Sensitive gas chromatographic-mass spectrometric method for the determination of gacyclidine in rat plasma and spinal cord dialyzates

A sensitive gas chromatographic-mass spectrometric (GC-MS) procedure is described for the selective determination of gacyclidine (a non-competitive N-methyl-D-aspartate antagonist) in rat plasma and spinal cord dialyzates. It involves a single-step liquid-liquid extraction of plasma samples and dialyzates with hexane (pH 8.0) and the use of phencyclidine as an internal standard. The compounds were separated on a GC capillary column and specifically detected by MS in the selected-ion monitoring mode. Gacyclidine and its internal standard were monitored by using the fragment ions at m/z 206 and 200, respectively. The method was accurate and reproducible (intra- and inter-day reproducibility < 12%) with a limit of quantification of 1.6 ng ml-1 using 100 microliters plasma of dialyzate samples. The calibration curves for rat plasma and Ringer's solution were linear (r2 > 0.996) over a range from 1.6 to 200 ng ml-1. The extraction efficiency was close to 100%. This simple and rapid assay (total run time < 10 min) was validated for a pilot pharmacokinetic study in healthy rats after intravenous injection of a bolus dose of gacyclidine (2.5 mg kg-1).     

Protein recognition via ion-coordinated molecularly imprinted supermacroporous cryogels

Molecular imprinting is a method for making selective binding sites in synthetic polymers using a molecular template. The aim of this study is to prepare lysozyme-imprinted supermacroporous cryogels which can be used for the purification of lysozyme (Lyz) from egg white. N-Methacryloyl-(L)-histidinemethylester (MAH) was chosen as the metal-coordinating monomer. In the first step, Cu2+ was complexed with MAH and the lysozyme-imprinted poly(HEMA-MAH) [Lyz-MIP] cryogel were produced by free radical polymerization initiated by N,N,N',N'-tetramethylene diamine (TEMED) in an ice bath. After that, the template (i.e., lysozyme) was removed using 0.05 M phosphate buffer containing 1M NaCl (pH 8.0). The maximum lysozyme adsorption capacity was 22.9 mg/g polymer. The relative selectivity coefficients of Lyz-MIP cryogel for lysozyme/bovine serum albumin and lysozyme/cytochrome c were 4.6 and 3.2 times greater than non-imprinted poly(HEMA-MAH) (NIP) cryogel, respectively. Purification of lysozyme from egg white was also monitored by determining the lysozyme activity using Micrococcus lysodeikticus as substrate. The purity of the desorbed lysozyme was about 94% with recovery about 86%. The Lyz-MIP cryogel could be used many times without decreasing the adsorption capacity significantly.