Dynamic in situ growth of bonded-phase silica nanospheres on silica capillary inner walls for open-tubular liquid chromatography.

Silica nanospheres (SNS) were grown on the inner walls of silica capillaries through a dynamic in situ nucleation process to prepare a highly porous and large accessible surface area substrate. The SNS were then functionalized with octadecyl (C18), 3-aminopropyltriethoxysilane (APTES), beta-cyclodextrin (ß-CD), and amino groups to develop robust and efficient chromatographic stationary phases. The modified silica capillaries were exploited for open-tubular liquid chromatography (OT-LC) and open-tubular capillary electrochromatography (OT-CEC) applications. The prepared stationary phases were compared to conventional capillaries in terms of separation performance. The synthesis process was optimized, and the bonded-phase stationary phases were characterized by the electron microscopy technique. The effects of different solvents, additives, and functional groups on the geometry and chromatographic resolving power of the SNS were envisaged. The capillaries modified with octadecyl groups were evaluated for the separation of non-steroidal anti-inflammatory drugs, phenones, alkenylbenzenes, and enantiomers of chlorophenoxy herbicides. As an application instance, an SNS-C18-coated capillary was utilized for the separation of alkenylbenzenes from clove extract and protein digest medium, through OT-LC and OT-CEC techniques, respectively. The ß-CD functionalized capillary was applied for the OT-CEC separation of a dichlorprop racemic mixture.

In-line sample concentration in capillary electrophoresis by cyclodextrin to admicelle microextraction.

Cyclodextrins (CDs) as a pseudophase in pseudophase-to-pseudophase microextraction (P2ME) in capillary zone electrophoresis (CZE) are proposed. In this P2ME mode called CD to admicelle ME, a long plug of dilute analyte solution prepared in cetyltrimethylammonium bromide (CTAB) at the critical micellar concentration was injected into the capillary. This formed CTAB admicelles at the interface between the solution and the negatively charged capillary surface, where the analytes were trapped. The injection of CD solution released the admicelles and the analytes from the capillary surface due to the formation of stable CD/CTAB inclusion complexes. The analytes are concentrated at the CD front during injection and voltage separation. Various neutral CDs were found to be effective for CD to admicelle ME. To implement this in-line sample concentration technique in CZE, CD concentration, sample injection time, and sample:CD solution injection ratio were optimized. The optimized conditions for five model anionic analytes, namely, 4-bromophenol, sulindac, sulfamethizole, 4-vinylbenzoic acid, and succinylsulfathiazole, were 20 mM α-CD in 20 mM sodium tetraborate (pH 9.2) solution, sample injection time of 370 s, and CD:sample injection ratio of 1:2. The sensitivity enhancement factors (SEFs) were between 112 and 168. The SEFs of sulindac and sulfamethizole in particular were similar to previously published off-line microextraction techniques, which are typically time-consuming. The calculated values of LOQ, intra-/inter-day (n = 6/n = 10, 3 days) repeatability, and linearity (R2) of CD to admicelle ME were 0.0125-0.05 µg/mL, 1.5-4.6%, 1.8-4.8%, and ≥0.999, respectively. Finally, the potential of CD to admicelle ME to the analysis of artificial urine samples was demonstrated.

Pseudophase-to-solvent microextraction for in-line sample concentration of anionic analytes in capillary zone electrophoresis.

We present pseudophase-to-solvent microextraction (PSME) as a simple pressure-driven in-line sample concentration technique in capillary zone electrophoresis (CZE) for small organic anions. The developed technique is like solid-phase extraction; however, the chromatographic phase is a pseudophase, i.e., in-situ cetyltrimethylammonium bromide (CTAB) interfacial micelles. A large volume of sample (e.g., > 12 capillary volumes) prepared in [CTAB] ∼critical micelle concentration was injected into the capillary. The elution and enrichment of analytes trapped in the CTAB coating was facilitated by the high concentration of methanol in the background solution, which was introduced from the outlet end. Co-electroosmotic flow CZE was conducted after the concentrated analytes reached the tip of the inlet. Parameters such as sample loading time and elution time were optimised. Under optimised conditions, the SEFs of 187-573 achieved for the model anions (4-nitrophenol, 4-vinylbenzoic acid, mecoprop, indoprofen, sulfamethizole and sulindac) were comparable to previously reported off-line microextraction techniques. The calculated LOD (S/N = 3), LOQ (S/N = 10), intra-/inter- (n = 6/n = 9, 3 days) day repeatability and linearity (R2s) values in PSME-CZE were 4.2-20.1 ng/mL, 13.8-67.1 ng/mL, 5%, 10% and > 0.990, respectively. The PSME-CZE of fortified urine samples showed % recovery values of 93-108% with %RSDs (n = 3) of 4-10% for the model analytes.

Chiral separation using cyclodextrins as mobile phase additives in open-tubular liquid chromatography with a pseudophase coating.

The chiral separation of various analytes (dichlorprop, mecoprop, ibuprofen, and ketoprofen) was demonstrated with different cyclodextrins as mobile phase additives in open-tubular liquid chromatography using a stationary pseudophase semipermanent coating. The stable coating was prepared by a successive multiple ionic layer approach using poly(diallyldimethylammonium chloride), polystyrene sulfonate, and didodecyldimethyl ammonium bromide. Increasing concentrations (0-0.2 mM) of various native and derivatized cyclodextrins in 25 mM sodium tetraborate (pH 9.2) were investigated. Chiral separation was achieved for the four test analytes using 0.05-0.1 mM ß-cyclodextrin (resolution between 1.11 and 1.34), γ-cyclodextrin (resolution between 0.78 and 1.27), carboxymethyl-ß-cyclodextrin (resolution between 1.64 and 2.59), and 2-hydroxypropyl-ß-cyclodextrin (resolution between 0.71 and 1.76) with the highest resolutions obtained with 0.1 mM carboxymethyl-ß-cyclodextrin. %RSD values were <10%. This is the first demonstration of chiral open-tubular liquid chromatography using achiral chromatographic coatings and cyclodextrins as mobile phase additives.

Micelle to cyclodextrin stacking in open-tubular liquid chromatography using capillaries coated with surfactant admicelles.

In-line sample concentration by micelle to cyclodextrin stacking (MCDS) in open-tubular liquid chromatography (OT-LC) with UV detection is described. OT-LC of two sets of analytes (small-molecule drugs and neutral alkenylbenzenes) was by the use of a fused-silica capillary that was coated with admicelles of didodecyldimethyl ammonium bromide (DDAB). These admicelles acted as a stationary chromatographic pseudophase. The mobile phase was 25 mM sodium tetraborate in 10% methanol, pH 9.2. MCDS was by long pressure injection of samples prepared in 10 mM hexadecyltrimethyl ammonium bromide (CTAB) in 25 mM sodium tetraborate, pH 9.2 (buffer), followed by injection of 50 mM α-CD in buffer (CD solution). Stacking was by application of voltage at -20 kV prior to pressure-driven OT-LC. The factors that influenced MCDS such as type and concentration of CD, concentration of CTAB in the sample, injection time ratio of the sample and the CD solution and stacking time were studied. Under optimised conditions, sensitivity enhancement factors (SEFs) were between 19 and 23, linear ranges were between 0.5 and 10 µg/mL with r2 > 0.99 and inter-day/intra-day repeatability in retention time and peak area were ≤5.6% for the model small-molecule drugs. Application to real samples was by the determination of potentially toxic alkenylbenzenes (SEFs = 10 to 12) in basil-leaf and whole-clove extracts. The assay results were comparable to those obtained from an in-house high-performance liquid chromatography-UV method.

Pseudophase microextraction for in-line sample concentration in micellar electrokinetic chromatography.

Pseudophase microextraction (PPME) as a simple in-line sample concentration technique in micellar electrokinetic chromatography (MEKC) is presented. In contrast to popular electric-field driven stacking techniques in MEKC such as sweeping, PPME is pressure-driven. The technique afforded up to 403-2968x improvements in peak heights for fenoprop, dichlorprop, 1- and 2-naphthol compared to typical injection. Under the same MEKC conditions, the improvements in PPME were up to 23-59x better compared to sweeping. Briefly in PPME, the entire capillary was loaded (up to 20 capillary volumes) with the sample prepared in a dilute solution of cetyltrimethylammonium bromide ([CTAB] > critical surface aggregation concentration). The CTAB formed aggregates at the inner capillary walls and these aggregates acted as a stationary chromatographic pseudophase. After clean-up via flushing the capillary with purified water, the MEKC background solution (BGS) with sodium dodecyl sulfate was then introduced by pressure from the outlet end to elute the retained analytes. The analytes concentrate at front of the BGS and the front was moved to the inlet end of the capillary prior to MEKC. Optimization strategies and current limitations in PPME-MEKC are described. The linear ranges using a 4 capillary volume sample load obtained for fenoprop, dichlorprop, 1- and 2-naphthol were between 1 and 160 ng/mL (r2s ≥ 0.996), LOQs = 1-2.5 ng/mL and repeatability %RSDs (n = 6) were ≤5% (intra-day) and ≤7% (inter-day) (using low analyte concentrations 1-5x LOQ). PPME-MEKC with simple dilution of fortified real samples (no off-line sample concentration) was also able to detect low levels of dichlorprop (10 ng/mL, limit set in Australia) and 1- and 2-naphthol (7.5-15 ng/mL) in a drinking water and natural water sample, respectively (% recovery = 84-108%). The concept of PPME may find use in other modes of capillary electrophoresis and other nano-microscale separations.

Electroosmotic flow assisted pseudophase to pseudophase microextraction for stacking in capillary zone electrophoresis.

A stacking technique is proposed to improve the poor detection sensitivity of capillary zone electrophoresis (CZE) with UV detection. A long injection (e.g., 12.4 cm plug) of model anionic analytes prepared in a dilute solution of hexadecyltrimethylammonium bromide (CTAB) was enriched 26-34-x (compared to a typical or 2.1 mm sample injection) via the injection of a micellar solution of sodium dodecyl sulfate (SDS) prior to CZE separation. During sample injection, the CTAB formed a stationary pseudophase coating, which trapped the analytes at the inner walls of a fused silica capillary. The SDS micelles then released the CTAB admicelles via the formation of solution CTAB-SDS catanionic micelles during SDS plug injection and voltage application. As the SDS micelles moved through the sample zone, the formation of the catanionic micelles then released and accumulated the analytes at the front of the injected SDS zone. The stacking technique is called electroosmotic flow (EOF) assisted pseudophase to pseudophase microextraction because the EOF was essential for the formation of CTAB-SDS catanionic micelles for microextraction. Also, the CTAB and SDS aggregates are both pseudophases, which were used to retain and release the analytes from the capillary wall, respectively.

Sweeping-micellar electrokinetic chromatography versus reversed phase liquid chromatography for the determination of coumarin in curry.

Coumarin is a phytotoxin found in the popular spice cinnamon, which is used to flavor many Asian curry dishes. In this work, we developed and compared the analytical performance of reversed-phase liquid chromatography (RP-LC) and sweeping-micellar electrokinetic chromatography (MEKC) methods for the determination of coumarin in complex curry (gravy) samples. Using a matrix matched sample (curry after solvent extraction with methanol and diluted with 100 mM phosphoric acid), the intra-day and inter-day repeatability of retention/migration time and (corrected) peak area for both methods were acceptable (%RSD (n=6) ≤ 5%). The linear range and limit of quantitation (LOQ) were an order of magnitude better in RP-LC (RP-LC linear range = 0.11-108 mg/kg, LOQ = 0.11 mg/kg) (Sweeping-MEKC linear range = 2.16-216 mg/kg, LOQ = 2.16 mg/kg). However, the limit of detection (S/N=3) and LOQ in sweeping-MEKC was 0.65 mg/kg and 2.16 mg/kg, which were sufficient to report the levels of coumarin ≥ the European limit of 2 mg/kg in foods. During the analysis of 25 curry samples, relatively similar results for sweeping-MEKC and RP-LC were obtained for 6 samples that contained coumarin >LOQ of sweeping-MEKC. Interferences in RP-LC lead to significant overestimation of coumarin levels in 3 samples. Coumarin levels above the EU limit was found in 6 curry samples using the more selective sweeping-MEKC. This work should also raise public awareness on the presence of potentially high levels of coumarin in some foods.

Bile Salts in Chiral Micellar Electrokinetic Chromatography: 2000-2020.

Bile salts are naturally occurring chiral surfactants that are able to solubilize hydrophobic compounds. Because of this ability, bile salts were exploited as chiral selectors added to the background solution (BGS) in the chiral micellar electrokinetic chromatography (MEKC) of various small molecules. In this review, we aimed to examine the developments in research on chiral MEKC using bile salts as chiral selectors over the past 20 years. The review begins with a discussion of the aggregation of bile salts in chiral recognition and separation, followed by the use of single bile salts and bile salts with other chiral selectors (i.e., cyclodextrins, proteins and single-stranded DNA aptamers). Advanced techniques such as partial-filling MEKC, stacking and single-drop microextraction were considered. Potential applications to real samples, including enantiomeric impurity analysis, were also discussed.

A stationary pseudophase semi-permanent coating for open-tubular capillary liquid chromatography and electrochromatography.

We describe the chromatographic and electrochromatographic separation of small neutral and charged analytes using a fused silica capillary with a stationary pseudophase semi-permanent coating of didodecyldimethyl ammonium bromide (DDAB) aggregates. The coating was prepared by flushing the capillary with a DDAB solution that was rinsed out with the mobile phase. Our studies (i.e., electroosmotic flow measurements by capillary electrophoresis, chromatographic retention of a neutral probe and atomic force microscopy) suggested the formation of DDAB patchy admicelle, complete admicelle, or larger aggregates at the solid surface - liquid interface inside the capillary, depending on the concentration of DDAB used in coating the capillary. The analytical figures of merit for open tubular liquid chromatography (OT-LC, pressure driven) and open tubular capillary electrochromatography (OT-CEC, voltage driven) using a capillary coated with 0.5 mM DDAB and mobile phase/background solution of 25 mM borate buffer at pH 9.5 with 10% MeOH were the following: LOD = 3.0-5.0 µg/mL (OT-LC) and 2.5-5.0 µg/mL (OT-CEC); linearity R2 > 0.99 (peak area (OT-LC) and corrected peak area (OT-CEC)), intraday and interday repeatability%RSD < 5% (n = 12) for retention/migration time, peak area (OT-LC) and corrected peak area (OT-CEC). The reversed-phase and anion-exchange property of the stationary pseudophase was studied by the addition of organic solvents and sodium chloride to the mobile phase, respectively. We also demonstrate the increase in the ks of the tested analytes by implementing successive multiple ionic layer (SMIL) coating strategies with DDAB in combination with a cationic and/or anionic polyelectrolyte. The use of a stationary pseudophase coating is potentially an easy alternative way to conduct open-tubular liquid chromatography and electrochromatography.

Analytical Separation of Carcinogenic and Genotoxic Alkenylbenzenes in Foods and Related Products (2010-2020).

Alkenylbenzenes are potentially toxic (genotoxic and carcinogenic) compounds present in plants such as basil, tarragon, anise star and lemongrass. These plants are found in various edible consumer products, e.g., popularly used to flavour food. Thus, there are concerns about the possible health consequences upon increased exposure to alkenylbenzenes especially due to food intake. It is therefore important to constantly monitor the amounts of alkenylbenzenes in our food chain. A major challenge in the determination of alkenylbenzenes in foods is the complexity of the sample matrices and the typically low amounts of alkenylbenzenes present. This review will therefore discuss the background and importance of analytical separation methods from papers reported from 2010 to 2020 for the determination of alkenylbenzenes in foods and related products. The separation techniques commonly used were gas and liquid chromatography (LC). The sample preparation techniques used in conjunction with the separation techniques were various variants of extraction (solvent extraction, liquid-liquid extraction, liquid-phase microextraction, solid phase extraction) and distillation (steam and hydro-). Detection was by flame ionisation and mass spectrometry (MS) in gas chromatography (GC) while in liquid chromatography was mainly by spectrophotometry.

Ionic liquids in electrokinetic chromatography.

There is an interest in the application of ionic liquids as additives into the separation media to improve achiral and chiral separations in electrokinetic chromatography (EKC). This review will critically discuss the developments on the use of ionic liquids in the different modes of EKC during the last five years (2015-mid 2020). A healthy number of 48 research articles searched through Scopus were categorised into two: ionic liquids as sole pseudophase (micelles, microemulsions, ligand exchange pseudophase or molecular pseudophase) and ionic liquids with pseudophase (achiral or chiral). More than half of the papers dealt with chiral separations that were mostly facilitated by another additive or pseudophase. The role of ionic liquids for improvement of separations were analysed, and we provided some recommendations for further investigations. Finally, the use of ionic liquids in different on-line sample concentration or stacking methods (i.e., field enhancement and sweeping) was briefly discussed.

High Performance Liquid Chromatography versus Stacking-Micellar Electrokinetic Chromatography for the Determination of Potentially Toxic Alkenylbenzenes in Food Flavouring Ingredients.

Alkenylbenzenes, including eugenol, methyleugenol, myristicin, safrole, and estragole, are potentially toxic phytochemicals, which are commonly found in foods. Occurrence data in foods depends on the quality of the analytical methodologies available. Here, we developed and compared modern reversed-phase high performance liquid chromatography (HPLC) and stacking-micellar electrokinetic chromatography (MEKC) methods for the determination of the above alkenylbenzenes in food flavouring ingredients. The analytical performance of HPLC was found better than the stacking-MEKC method. Compared to other HPLC methods found in the literature, our method was faster (total run time with conditioning of 15 min) and able to separate more alkenylbenzenes. In addition, the analytical methodology combining an optimized methanol extraction and proposed HPLC was then applied to actual food flavouring ingredients. This methodology should be applicable to actual food samples, and thus will be vital to future studies in the determination of alkenylbenzenes in food.

A simple apparatus for electrokinetic removal of sodium dodecyl sulfate from protein digests.

Sodium dodecyl sulfate (SDS) in proteomics samples needs to be removed and estimated prior to mass spectrometry (MS)-based analysis and to avoid MS ion-source contamination. Here, we describe an organic solvent free method to remove SDS using a simple apparatus that mainly consists of an agarose gel inside a 1 mL plastic micropipette tip and a voltage power supply with electrodes. A small volume of sample (e.g., 50 µL) is loaded on top of the gel and then voltage (cathode at the sample side) is applied with an acidic solution at the other end of the micropipette tip. Within 25 min, SDS was removed (e.g., ≥99% SDS in 3.5 mM SDS) and the peptides were retained in the sample solution. The strategy was compared to the commercially available and expensive Pierce spin column for the removal of SDS and recovery of peptides from a digested bovine serum albumin sample.

Metabolic Stability of New Mito-Protective Short-Chain Naphthoquinones.

Short-chain quinones (SCQs) have been identified as potential drug candidates against mitochondrial dysfunction, which is largely dependent on their reversible redox characteristics of the active quinone core. We recently synthesized a SCQ library of > 148 naphthoquinone derivatives and identified 16 compounds with enhanced cytoprotection compared to the clinically used benzoquinone idebenone. One of the major drawbacks of idebenone is its high metabolic conversion in the liver, which significantly restricts is therapeutic activity. Therefore, this study assessed the metabolic stability of the 16 identified naphthoquinone derivatives 1-16 using hepatocarcinoma cells in combination with an optimized reverse-phase liquid chromatography (RP-LC) method. Most of the derivatives showed significantly better stability than idebenone over 6 h (p < 0.001). By extending the side-chain of SCQs, increased stability for some compounds was observed. Metabolic conversion from the derivative 3 to 5 and reduced idebenone metabolism in the presence of 5 were also observed. These results highlight the therapeutic potential of naphthoquinone-based SCQs and provide essential insights for future drug design, prodrug therapy, and polytherapy, respectively.

Open-tubular admicellar electrochromatography of charged analytes.

Fundamental studies on the separation of cationic and anionic analytes in open-tubular admicellar electrochromatography (OT-AMEC) using cetyltrimethylammonium bromide (CTAB) and fused silica capillaries are presented. OT-AMEC was compared with open-tubular admicellar liquid chromatography (OT-AMLC) by running the two methods using the same mobile phases. The mobile phases were buffered at pH ≥ 6 and contained a low concentration (above the critical surface aggregation concentration and below the critical micelle concentration) of CTAB. The stationary pseudophase of CTAB admicelles were formed at the solid surface and liquid interface inside the capillary by simply conditioning the capillary with the mobile phase. Separations were performed in a 30 cm (21.5 cm to UV detector) long and 50 µm inner diameter capillary, using low pressure (50 mbar) in OT-AMLC and high voltage (15 kV at negative polarity) in OT-AMEC. The appropriate equations for the experimental estimation of retention factor (k) values of analytes were discussed. For anionic analytes, k in OT-AMEC were carefully determined by considering the observed interaction between CTAB monomers and tested analytes. The calculated k for each analyte was found similar in OT-AMLC and OT-AMEC, although the mechanism of retention was not entirely different due to the contribution of electrophoresis in OT-AMEC. Studies on the addition of a typical (i.e., acetonitrile) and atypical modifier (i.e., nonyl-ß-glucoside) into the mobile phase, and sample focusing with >10x improvement in peak height under isocratic conditions were also conducted.

Recent advancements in open-tubular liquid chromatography and capillary electrochromatography during 2014-2018.

This review critically discusses the developments on open-tubular liquid chromatography (OT-LC) and open-tubular capillary electrochromatography (OT-CEC) during 2014-2018. An appropriate Scopus search revealed 5 reviews, 4 theoretical papers on open-tubular format chromatography, 29 OT-LC articles, 68 OT-CEC articles and 4 OT-LC/OT-CEC articles, indicating a sustained interest in these areas. The open-tubular format typically uses a capillary column with inner walls that are coated with an ample layer or coating of solid stationary phase material. The ratio between the capillary internal diameter and coating thickness (CID/CT) is ideally ≤ 100 for appropriate chromatographic retention. We, therefore, approximated the CID/CT ratios and found that 22 OT-LC papers have CID/CT ratios ≤100. The other 7 OT-LC papers have CID/CT ratio >100 but have clearly demonstrated chromatographic retention. These 29 papers utilised reversed phase or ion exchange mechanisms using known or innovative solid stationary phase materials (e.g. metal organic frameworks), stationary pseudophases from ionic surfactants or porous supports. On the other hand, we found that 68 OT-CEC papers, 7 OT-LC papers and 4 OT-LC & OT-CEC papers have CID/CT ratios >100. Notably, 44 papers (42 OT-CEC and 2 OT-LC & OT-CEC) did not report the retention factor and/or effective electrophoretic mobility of analytes. Considering all covered papers, the most popular activity was on the development of new chromatographic materials as coatings. However, we encourage OT-CEC researchers to not only characterise changes in the electroosmotic flow but also verify the interaction of the analytes with the coating. In addition, the articles reported were largely driven by stationary phase or support development and not by practical applications.

Pressurized Hot Water Extraction and Capillary Electrophoresis for Green and Fast Analysis of Useful Metabolites in Plants.

The search for useful compounds from plants is an important research area. Traditional screening that involves isolation and identification/quantitation is tedious, time consuming, and generates a significant amount of chemical waste. Here, we present a simple, fast, and green strategy to assess ≥0.1% wt/wt quantities of useful compounds in plants/spices using pressurized hot water extraction using a household espresso machine followed by chemical analysis using capillary electrophoresis. Three demonstrations with polygodial, cinnamaldehyde, coumarin, and shikimic acid as target metabolites are shown. Direct analysis of extracts was by the developed micellar electrokinetic chromatography and capillary zone electrophoresis methods. The approach, which can be implemented in less developed countries, can process many samples within a day, much faster than traditional techniques that would normally take at least a day. Finally, 0.8-1.1% wt/wt levels of shikimic acid were found in Tasmanian-pepperberry and Tasmanian-fuschia leaves via the approach.

Admicelles in open-tube capillaries for chromatography and electrochromatography.

Surfactant bilayers or admicelles at the solid surface-liquid interface inside 50-200 µm inner diameter (i.d.) open-tube fused-silica capillaries were developed as 'soft' stationary pseudophases for the liquid chromatographic (LC) separations of neutral and charged analytes. Admicelles were formed in-situ from buffered aqueous mobile phases with cetytrimethylammonium bromide at concentrations between the critical surface aggregation concentration and critical micelle concentration, which were determined by electroosmotic flow measurements using capillary electrophoresis. There were no micelles in the mobile phase solution. Also, there was no solid phase that is classically required in LC. Pressure and voltage driven modes or open-tubular admicellar liquid chromatography (OT-AMLC) and electrochromatography, respectively were proposed based on the separation of neutral analytes. The parameters (i.e., pH, concentration of surfactant, salt, and methanol in the mobile phase and capillary i.d.) that affected the surprising chromatographic effect of admicelles at the interface were investigated. The analytical performance of OT-AMLC for small molecules were found acceptable. Applications to environmental water and biological (HepG cell line metabolism media) samples analysis with appropriate sample preparation procedures were also conducted. The use of pseudophases at the solid surface-liquid interface could be a viable solution to problems associated with the use of solid stationary or support materials in nano- and micro-liquid chromatography and electrochromatography.