0000000000053124
AUTHOR
Laura Escuder-gilabert
Biopartitioning micellar separation methods: modelling drug absorption
The search for new pharmacologically active compounds in drug discovery programmes often neglects biopharmaceutical properties as drug absorption. As a result, poor biopharmaceutical characteristics constitute a major reason for the low success rate for candidates in clinical development. Since the cost of drug development is many times larger than the cost of drug discovery, predictive methodologies aiding the selection of bioavailable drug candidates are of profound significance. This paper has been focussed on recent developments and applications of chromatographic systems, particularly those systems based on amphiphilic structures, in the frame of alternative approaches for estimating t…
Enantioresolution in electrokinetic chromatography-complete filling technique using sulfated gamma-cyclodextrin. Software-free topological anticipation
Few papers have tried to predict the resolution ability of chiral selectors in capillary electrophoresis for the separation of the enantiomers of chiral compounds. In a previous work, we have used molecular information available on-line to establish enantioresolution levels of basic compounds using highly sulfated β-CD (HS-β-CD) as chiral selector in electrokinetic chromatography-complete filling technique (EKC-CFT). The present study is a continuation of this previous work, introducing some novelties. In this work, the ability of sulfated γ-cyclodextrin (S-γ-CD) as chiral selector in EKC-CFT is modelled for the first time. Thirty-three structurally unrelated cationic and neutral compounds …
Reliability of the retention factor estimations in liquid chromatography.
The retention factor is one of the most universally used parameters in chromatography. However, large differences in the experimental retention factor values are observed when the same compound is injected in a given stationary/mobile phase system under intermediate precision conditions. Conventional protocols for estimating retention factors have problems that mainly arise from difficulties in the hold-up time measurements and the omission of the existence of extra-column times by practicing chromatographers. In the present paper, three different approaches for estimating retention factors are tested: (i) classical retention factor estimations based on the gross hold-up time, (ii) based on…
Modeling the chiral resolution ability of highly sulfated β-cyclodextrin for basic compounds in electrokinetic chromatography
Abstract Despite the fact that extensive research in the field of enantioseparations by capillary electrophoresis has been carried out, it is difficult to predict whether a concrete chiral selector would be useful for the separation of a racemic compound. Hence, several experimental effort is necessary to test the abilities of individual chiral selectors, usually by trial and error procedures. Thus, the enantioseparation of a new racemate becomes a time- and money-consuming task. In this work, the ability of highly sulfated β-cyclodextrin (HS-β-CD) as chiral selector in electrokinetic chromatography (EKC) is modeled for the first time, using exclusively directly-available structural data of…
Biopartitioning micellar chromatography: an in vitro technique for predicting human drug absorption.
The main oral drug absorption barriers are fluid cell membranes and generally drugs are absorbed by a passive diffusion mechanism. Biopartitioning micellar chromatography (BMC) is a mode of micellar liquid chromatography that uses micellar mobile phases of Brij35 under adequate experimental conditions and can be useful to mimic the drug partitioning process in biological systems. In this paper the usefulness of BMC for predicting oral drug absorption in humans is demonstrated. A hyperbolic model has been obtained using the retention data of a heterogeneous set of 74 compounds, which shows predictive ability for drugs absorbed by passive diffusion. The model obtained in BMC is compared with …
Chromatographic retention–activity relationships for prediction of the toxicity pH-dependence of phenols
Abstract An investigation of the use of the chromatographic retention (log k ) as an in vitro approach for modeling the pH-dependence of the toxicity to Guppy of phenols is developed. A data set of 19 phenols with available experimental toxicity–pH data was used. The importance of the mechanism of toxic action (MOA) of phenols was studied. log k data at three pH values were used for the phenols classification and two groups or ‘MODEs’ were identified. For one ‘MODE’ a quantitative retention–activity relationship (QRAR) model was calculated. Finally, the model was used to assess the toxicity to Guppy of phenols at different pH values. The results of this investigation suggest that chromato…
Development of predictive retention-activity relationship models of non-steroidal anti-inflammatory drugs by micellar liquid chromatography: comparison with immobilized artificial membrane columns.
The predictive and interpretative capability of quantitative chromatographic retention-biological activity models is supported by the fact that under adequate experimental conditions the solute partitioning into chromatographic system can emulate the solute partitioning into lipid bilayers of biological membranes, which is the basis for drug and metabolite uptake, passive transport across membranes and bioaccumulation. The use of micellar solutions of Brij35 as mobile phases in reversed-phase liquid chromatography has proven to be valid to predict some biological activities of different kinds of drugs. In this study, quantitative retention-activity relationship (QRAR) models to describe som…
Experimental-Like Affinity Constants and Enantioselectivity Estimates from Flexible Docking
Experimental-like affinity constants and enantioselectivity estimates, not predicted so far computationally, were obtained using a novel flexible modeling/docking combined strategy. The S- and R-warfarin-human serum albumin (HSA, site I) complexes were used as an interaction model. The process for a verified estimation includes the following: (i) ionized open chain forming at physiological pH (a recent focus); (ii) conformational search (molecular mechanics and Monte Carlo methods); (iii) rigid protein-flexible ligand docking (GlideXP) generating low energy paired S- and R-poses; (iv) graphical comparison against the X-ray crystal structure (unsatisfactory verification step); (v) quantum po…
Chromatographic multivariate quality control of pharmaceuticals giving strongly overlapped peaks based on the chromatogram profile
In the present paper, the simultaneous quantification of two analytes showing strongly overlapped chromatographic peaks (alpha = 1.02), under the assumption that both available equipment and training of the laboratory staff are basic, is studied. A pharmaceutical preparation (Mutabase) containing two drugs of similar physicochemical properties (amitriptyline and perphenazine) is selected as case of study. The assays are carried out under realistic working conditions (i.e. routine testing laboratories). Uncertainty considerations are introduced in the study. A partial least squares model is directly applied to the chromatographic data (with no previous signal transformation) to perform quali…
Direct chromatographic study of the enantioselective biodegradation of ibuprofen and ketoprofen by an activated sludge
[EN] The quantification of the enantiomeric fraction (EF) during the biodegradation process is essential for environmental risk assessment. In this paper the enantioselective biodegradation of ibuprofen, IBU, and ketoprofen, KET, two of the drugs most consumed, was evaluated. Biodegradation experiments were performed in batch mode using a minimal salts medium inoculated with an activated sludge (collected from a Valencian Waste Water Treatment Plant) and supplemented with the racemate of each compound. The inoculum activity was verified using fluoxetine as reference compound. The experimental conditions used (analyte concentration and volume of inoculum) were chosen according to OECD guidel…
Uncertainty-Based Internal Quality Control. Harmonization Considerations
Three main quality aspects for analytical laboratories are internal method validation, internal quality control (IQC), and sample result uncertainty. Unfortunately, in the past they have been used in a nonharmonized way. The most universal IQC tool is the mean chart, but some criteria used to fix their control limits do not fit the real nature of analytical results. A new approach for fixing these limits is proposed (the u-approach). The key is the combined uncertainty, u, obtained from the method validation information, also used for estimating the sample result uncertainty. A comparative study on "in-control" simulated, bibliographic, and real laboratory data suggests that the u-approach …
Fast evaluation of enantioselective drug metabolism by electrophoretically mediated microanalysis: application to fluoxetine metabolism by CYP2D6.
In this work, a capillary electrophoretic methodology for the enantioselective in vitro evaluation of drugs metabolism is applied to the evaluation of fluoxetine (FLX) metabolism by cytochrome 2D6 (CYP2D6). This methodology comprises the in-capillary enzymatic reaction and the chiral separation of FLX and its major metabolite, norfluoxetine enantiomers employing highly sulfated β-CD and the partial filling technique. The methodology employed in this work is a fast way to obtain a first approach of the enantioselective in vitro metabolism of racemic drugs, with the additional advantage of an extremely low consumption of enzymes, CDs and all the reagents involved in the process. Michaelis-Men…
Analysis of pharmaceutical preparations containing local anesthetics by micellar liquid chromatography and spectrophotometric detection
An HPLC procedure for the determination of six local anesthetics, bupivacaine, lidocaine, mepivacaine, procaine, propanocaine and tetracaine, in pharmaceutical silane ODS-2 C18 analytical column and spectrophotometric detection at 230 nm were used. The chromatographic tographic behaviour of local anesthetics with different micellar eluents of sodium dodecyl sulphate (SDS) is described. Selection of the adequate composition of the micellar mobile phase (SDS and 1-propanol concentrations) for the analysis of pharmaceuticals was studied. Adequate retention was achieved with an eluent containing 0.15 M SDS +10% 1-propanol at pH 3. Application of the proposed method to the analysis of eight phar…
Fit-for-purpose chromatographic method for the determination of amikacin in human plasma for the dosage control of patients
In this paper, a simple, rapid and sensitive method based on liquid chromatography with fluorimetric detection (HPLC-FLD) for the determination of amikacin (AMK) in human plasma is developed. Determination is performed by pre-column derivatization of AMK with ortho-phtalaldehyde (OPA) in presence of N-acetyl-L-cysteine (NAC) at pH 9.5 for 5 min at 80 °C. In our knowledge, this is the first time that NAC has been used in AMK derivatization. Derivatization conditions (pH, AMK/OPA/NAC molar ratios, temperature and reaction time) are optimized to obtain a single and stable, at room temperature, derivative. Separation of the derivative is achieved on a reversed phase LC column (Kromasil C18, 5 μ…
Characterizing the interaction between enantiomers of eight psychoactive drugs and highly sulfated-β-cyclodextrin by counter-current capillary electrophoresis
The estimation of apparent binding constants and limit mobilities of the complexes of the enantiomers that characterize the interaction of enantiomers with chiral selectors, in this case highly sulfated β-cyclodextrin, was approached using a simple and economic electrophoretic modality, the complete filling technique (CFT) in counter-current mode. The enantiomers of eight psychoactive drugs, four antihistamines (dimethindene, promethazine, orphenadrine and terfenadine) and four antidepressants (bupropion, fluoxetine, nomifensine and viloxazine) were separated for the first time for this cyclodextrin (CD). Estimations of thermodynamic and electrophoretic enantioselectivies were also performe…
Chromatographic evaluation of the toxicity in fish of pesticides
Abstract Ecotoxicity assessment is essential before placing new chemical substances on the market. An investigation of the use of the chromatographic retention (log k) in biopartitioning micellar chromatography (BMC) as an in vitro approach to evaluate the toxicity in fish of pesticides (acute toxicity levels as pLC50) is proposed. A heterogeneous data set of 85 pesticides from six chemical families with available experimental fish toxicity data (ECOTOX database from U.S. Environmental Protection Agency (EPA)) was used. For pesticides exhibiting non-polar narcosis mechanism in fish (non-specific toxicity), more reliable models and precise pLC50 estimations are obtained from log k (quantitat…
Quantitative retention- and migration-toxicity relationships of phenoxy acid herbicides in micellar liquid chromatography and micellar electrokinetic chromatography
Abstract The need to obtain a tool for estimation of toxicity parameters for chemicals supports, the postulation of predictive models as an alternative to conventional classical assays. The use of micellar solutions of Brij35 as mobile phases in reversed phase liquid chromatography has proven to be valid in predicting some biological activities of different kinds of drugs. In this paper, the correlations between retention of phenoxy acids using different concentrations of Brij35 as micellar mobile phase in micellar liquid chromatography (MLC) and migration in micellar electrokinetic chromatography (MEKC) with several toxicity parameters are studied. Adequate correlations retention- and migr…
Chromatographic estimation of the soil-sorption coefficients of organic compounds
Abstract Soil-sorption coefficient ( K OC ) assessment is essential before placing a new chemical substance on the market. Since experimental K OC measurements are tedious, time-consuming and costly, alternative approaches to estimating K OC from other descriptors have been proposed. We review the use of chromatographic retention (log k ) values, obtained in different types of chromatographic systems, as descriptors to establish predictive log K OC -log k models. We highlighted critical aspects of such models and performed a comparative study using data in the literature. We compare two strategies to deal with the large variability of experimental log K OC data. We contrast the results of t…
Reversed phase liquid chromatography for the enantioseparation of local anaesthetics in polysaccharide-based stationary phases. Application to biodegradability studies.
[EN] A comprehensive study on the chiral separation of bupivacaine, mepivacaine, prilocaine and propanocaine with eight commercial polysaccharide-based chiral stationary phases (CSPs) in reversed phase conditions compatible with MS detection is performed. Methanol and acetonitrile are used as organic modifiers. Retention and resolution values obtained for each compound in the different CSPs and mobile phases are compared. The polysaccharide-based CSPs tested present different enantioselectivity towards the analytes. From the results, the experimental conditions for determining the enantiomers of bupivacaine, mepivacaine, prilocaine and propanocaine in saline aqueous samples using MS detecti…
Comparison between sodium dodecylsulphate and cetyltrimethylammonium bromide as mobile phases in the micellar liquid chromatography determination of non-steroidal anti-inflammatory drugs in pharmaceuticals.
The retention behaviour of non-steroidal anti-inflammatory drugs (NSAIDs) using micellar mobile phases of sodium dodecylsulphate (SDS) is studied and compared with that observed with micellar mobile phases of cetyltrimethylammonium bromide (CTAB). A liquid chromatographic procedure for the determination of acemetacin, diclofenac, indomethacin, ketoprofen, naproxen and tolmetin in pharmaceutical preparations is described. The proposed system uses a Kromasil C18 analytical column and a solution of 0.15 M SDS at pH 3 with 10% 1-propanol as mobile phase. Under these conditions, the studied NSAIDs elute between 6 and 10 min at a 1 mL min(-1) flow rate. Limits of detection (LOD) are lower than 0.…
Biopartitioning micellar chromatography: An alternative high-throughput method for assessing the ecotoxicity of anilines and phenols
An investigation of the use of the chromatographic retention (log k) as an in vitro approach for modelling the toxicity to Fathead Minnows of anilines and phenols is developed. A data set of 65 compounds with available experimental toxicity data was used. Log k data at three pH values were used for the compounds classification and two groups or 'MODEs' were identified. For one 'MODE' a quantitative retention-activity relationship (QRAR) model was calculated. Finally, it was used to estimate the toxicity to Fathead minnows of anilines and phenols for which experimental data are not available. These estimations were compared to those obtained from another toxicity (to Tetrahymena pyriformis) …
Modelling the enantioresolution capability of cellulose tris(3,5-dichlorophenylcarbamate) stationary phase in reversed phase conditions for neutral and basic chiral compounds
[EN] To the best of our knowledge, the prediction of the enantioresolution ability of polysaccharides-based stationary phases in liquid chromatography for structurally unrelated compounds has not been previously reported. In this study, structural information of neutral and basic compounds is used to model their enantioresolution levels obtained from an immobilised cellulose tris(3,5-dichlorophenylcarbamate) stationary phase in reversed phase conditions. Thirty-four structurally unrelated chiral drugs and pesticides, from seven families, are studied. Categorical enantioresolution levels (RsC, 0 = no baseline enantioresolution and 1 = baseline enantioresolution) are established from the expe…
Potential of biopartitioning micellar chromatography as an in vitro technique for predicting drug penetration across the blood–brain barrier
The blood-brain barrier (BBB) is considered to be the main barrier to drug transport into the central nervous system (CNS). The BBB restricts the passive diffusion of many drugs from blood to brain. The ease with which any particular drug diffuses across the BBB is determined largely by the molecular features of drugs, and it is therefore possible to predict the BBB permeability of a drug from its molecular structure. Biopartitioning micellar chromatography (BMC), a mode of micellar liquid chromatography that uses micellar mobile phases of Brij35 in adequate experimental conditions, can be useful in mimicking the drug partitioning process into biological systems. Retention in BMC depends on…
Modelling bioconcentration of pesticides in fish using biopartitioning micellar chromatography.
Ecotoxicity assessment is essential before placing new chemical substances on the market. An investigation of the use of the chromatographic retention (log k) in biopartitioning micellar chromatography (BMC) as an in vitro approach to evaluate the bioconcentration factor (BCF) of pesticides in fish is proposed. A heterogeneous set of 85 pesticides from six chemical families was used. For pesticides exhibiting bioconcentration in fish (experimental log BCF > 2), a quantitative retention-activity relationships (QRAR) model is able to perform precise log BCF estimations of new pesticides. Considering the present data, the results based on log k seem to be more reliable than those from availabl…
Trimeprazine is enantioselectively degraded by an activated sludge in ready biodegradability test conditions
[EN] A great number of available pharmaceuticals are chiral compounds. Although they are usually manufactured as racemic mixtures, they can be enantioselectively biodegraded as a result of microbial processes. In this paper, a biodegradability assay in similar conditions to those recommended in OECD tests of enantiomers of trimeprazine (a phenothiazine employed as a racemate) is carried out. Experiments were performed in batch mode using a minimal salts medium inoculated with an activated sludge (collected from a Valencian Waste Water Treatment Plant, WWTP) and supplemented with the racemate. The concentration of the enantiomers of trimeprazine were monitored by means of a chiral HPLC metho…
Monod-based ‘single-data’ strategy for biodegradation screening tests
Environmental contextObtaining biodegradation data over time can be difficult, especially when dealing with environmental compartments of increasing complexity. We evaluated the possibility of obtaining a full biodegradation depletion curve from a single biodegradation-time experimental measurement, and found that environmental information related to potential chemical persistence can be derived. The applicability of this ‘single-data’ strategy is illustrated using simulated and experimental data for several compounds. AbstractInformation obtained from biodegradability tests, e.g. half-life (t50) or kinetics parameters, is relevant in environmental risk assessment of new chemicals. In thes…
On-line monitoring of food fermentation processes using electronic noses and electronic tongues: A review
Fermentation processes are often sensitive to even slight changes of conditions that may result in unacceptable end-product quality. Thus, close follow-up of this type of processes is critical for detecting unfavorable deviations as early as possible in order to save downtime, materials and resources. Nevertheless the use of traditional analytical techniques is often hindered by the need for expensive instrumentation and experienced operators and complex sample preparation. In this sense, one of the most promising ways of developing rapid and relatively inexpensive methods for quality control in fermentation processes is the use of chemical multisensor systems. In this work we present an ov…
Emerging approaches to estimate retention factors in high performance liquid chromatography.
The retention factor is one of the most universally used parameters in chromatography. The errors associated with the conventional ways to determine the retention factor of compounds in liquid chromatography are studied and compared with those corresponding to new approaches. The later avoid the use of extra-column time and hold-up time values, which have proven to be tedious and ambiguous. Simulations and real data, used to examine the accuracy of four different approaches (two classic and two new), suggest that the new approaches could be considered more satisfactory than the classic ones.
Review: Highlights in recent applications of electronic tongues in food analysis
This paper examines the main features of modern electronic tongues (e-tongues) and their most important applications in food analysis in this new century. The components of an e-tongue (automatic sampler, array of chemical sensors, and data processing system) are described. Applications commented include process monitoring, freshness evaluation and shelf-life investigation, authenticity assessment, foodstuff recognition, quantitative analysis, and other quality control studies. Finally, some interesting remarks concerning the strengths and weaknesses of e-tongues in food analysis are also mentioned.
Simultaneous Determination of Pyridoxine and Riboflavin in Energy Drinks by High-Performance Liquid Chromatography with Fluorescence Detection
Energy drinks, as familiar consumer products, have been widely used in laboratory courses to help promote student interest, as well as to connect lecture concepts with laboratory work. Energy drinks contain B vitamins: pyridoxine (vitamin B6) and riboflavin (vitamin B2) of which amounts are high enough to be of concern. In this work, a fast and inexpensive high-performance liquid chromatography (HPLC) coupled with fluorescence detection method for determining pyridoxine and riboflavin simultaneously in energy drinks is developed. It takes advantage of the native fluorescence of B vitamins and provides high selectivity and sensitivity with low background noise. The method is suitable for und…
Quality control of pharmaceuticals containing non-steroidal anti-inflammatory drugs by micellar liquid chromatography
A liquid chromatographic procedure is described for the determination of acemetacin, diclofenac, indomethacin, ketoprofen, nabumetone, naproxen, piketoprofen, and tolmentin in pharmaceutical preparations. The compounds were separated on a Kromasil C18 analytical column, with a guard column of similar characteristics; the mobile phase was 0.06 M cetyltrimethylammonium bromide, at pH 7, containing 10% 1-butanol. At a flow rate of 1 mL min−1 the elution time of the most retained compound was 23 min. Limits of detection were between 0.01 μ g mL−1 for diclofenac and 0.2 μ g mL−1 for naproxen. The proposed method enables the determination of non-steroidal anti-inflammatory drugs in pharmaceutical…
Microseparation techniques for the study of the enantioselectivity of drug-plasma protein binding.
Stereoselectivity in protein binding can have a significant effect on the pharmacokinetic and pharmacodynamic properties of chiral drugs. The investigation of enantioselectivity of drugs in their binding with human plasma proteins and the identification of the molecular mechanisms involved in the stereodiscrimination by the proteins represent a great challenge for clinical pharmacology. In this review, the separation techniques used for enantioselective protein binding experiments are described and compared. An overview of studies on enantiomer–protein interactions, enantiomer–enantiomer interactions as well as chiral drug–drug interactions, including allosteric effects, is presented. The c…
Evaluation of the enantioselective binding of imazalil to human serum albumin by capillary electrophoresis
In this work, a methodology for the evaluation of enantioselective binding of imazalil (IMA) enantiomers to human serum albumin (HSA) that does not require the separation of free and bound to HSA fractions is developed. This methodology comprises the incubation of IMA–HSA designed mixtures for 30 min directly in the capillary electrophoresis system and the subsequent direct injection and chiral separation of IMA employing highly sulfated β-cyclodextrin as chiral selector and the complete filling technique. Two mathematical approaches were used to estimate apparent affinity constants (K1), protein binding and enantioselectivity (ES) for both enantiomers of IMA. Moderate enantioselective bind…
Comparative modelling study on enantioresolution of structurally unrelated compounds with amylose-based chiral stationary phases in reversed phase liquid chromatography-mass spectrometry conditions
[EN] Polysaccharide-based chiral stationary phases (CSPs) are the most used chiral selectors in HPLC. These CSPs can be used in normal, polar organic and aqueous-organic mobile phases. However, normal and polar organic mobile phases are not adequate for chiral separation of polar compounds, for the analysis of aqueous samples and for MS detection. In these situations, reversed phase conditions, without the usual non-volatile additives incompatible with MS detection, are preferable. Moreover, in most of the reported chiral chromatographic methods, retention is too large for routine work. In this paper, the chiral separation of 53 structurally unrelated compounds is studied using three commer…
Enantioseparation of nuarimol by affinity electrokinetic chromatography-partial filling technique using human serum albumin as chiral selector
The present paper deals with the enantiomeric separation of nuarimol enantiomers by affinity EKC-partial filling technique using HSA as chiral selector. Firstly, a study of nuarimol interactions with HSA by CE-frontal analysis was performed. The binding parameters obtained for the first site of interaction were n(1) = 0.84; K(1) = 9.7 +/- 0.3x10(3 )M(-1) and the protein binding percentage of nuarimol at physiological concentration of HSA was 75.2 +/- 0.2%. Due to the moderate affinity of nuarimol towards HSA the possibility of using this protein as chiral selector for the separation of nuarimol using the partial filling technique was evaluated. A multivariate optimization approach of the mo…
Quantitative Retention−Structure and Retention−Activity Relationship Studies of Local Anesthetics by Micellar Liquid Chromatography
The retention of compounds in micellar liquid chromatography (MLC) is governed by hydrophobic and electrostatic forces. For ionic compounds, both interactions should be considered. The present report offers a novel retention model that includes the hydrophobicity of compounds and the molar fraction of the charged form of compounds and compares it with other previously reported models. High correlations between the logarithm of capacity factors and these structural parameters were obtained for local anesthetics with different degrees of ionization using a nonionic surfactant solution as mobile phase. Modeling the retention of compounds as a function of physicochemical parameters and experime…
Anticipating the impact of pitfalls in kinetic biodegradation parameter estimation from substrate depletion curves of organic pollutants
[EN] Accurate and reliable estimation of kinetic parameters of pollutant biodegradation processes is essential for environmental and health risk assessment. Common biodegradation models proposed in the literature, such as the nonlinear Monod equation and its simplified versions (e.g. Michaelis-Menten-like and first-order equations), are problematic in terms of accuracy of kinetic parameters due to the parameter correlation. However, a comparison between these models in terms of accuracy and reliability, related to data imprecision, has not been performed in the literature. This task is necessary, mainly because the model selection cannot be straightforward, as shown in this work. To facilit…
A 21st century technique for food control: electronic noses.
This work examines the main features of modern electronic noses (e-noses) and their most important applications in food control in this new century. The three components of an electronic nose (sample handling system, detection system, and data processing system) are described. Special attention is devoted to the promising mass spectrometry based e-noses, due to their advantages over the more classical gas sensors. Applications described include process monitoring, shelf-life investigation, freshness evaluation, authenticity assessment, as well as other general aspects of the utilization of electronic noses in food control. Finally, some interesting remarks concerning the strengths and weakn…
In-line capillary electrophoretic evaluation of the enantioselective metabolism of verapamil by cytochrome P3A4
Abstract In this paper a methodology for the in-line evaluation of enantioselective metabolism by capillary electrophoresis has been developed and applied to the study of verapamil metabolism by cytochrome P3A4. The developed methodology comprises an in-capillary reaction step carried out by electrophoretically mediated microanalysis and a separation step in which highly sulfated β-cyclodextrin with partial filling technique has been employed as chiral selector for verapamil and norverapamil enantiomers resolution, joining the advantages of both methodologies in a unique assay. Kinetic parameters of the enzymatic reaction (Km and Vmax) have been evaluated for both verapamil enantiomers by n…
Permeability and toxicological profile estimation of organochlorine compounds by biopartitioning micellar chromatography
This paper points out the usefulness of biopartitioning micellar chromatography (BMC) as a high-throughput primary screening tool providing key information about the oral absorption, skin permeability (Kp), brain–blood distribution coefficient (BB) and ecotoxicological parameters such as median lethal concentration (LC50) and bioconcentration factors of 15 organochloride compounds. The retention data of compounds in BMC conditions were interpolated in previously developed quantitative–retention activity relationships by our research group. Results show that the compounds studied readily cross the intestinal barrier (oral absorption >ercnt;) and the blood–brain barrier (log BB >p;0.4). In ad…
Electronic noses and tongues to assess food authenticity and adulteration
Abstract Background There is a growing concern for the problem of food authenticity assessment (and hence the detection of food adulteration), since it cheats the consumer and can pose serious risk to health in some instances. Unfortunately, food safety/integrity incidents occur with worrying regularity, and therefore there is clearly a need for the development of new analytical techniques. Scope and approach In this review, after briefly commenting the principles behind the design of electronic noses and electronic tongues, the most relevant contributions of these sensor systems in food adulteration control and authenticity assessment over the past ten years are discussed. It is also remar…
Improved accuracy of environmentally relevant parameter estimates derived from biodegradation assays.
Biodegradation assays involve both biodegradation and analytical processes which can be affected by systematic errors, among others. These errors can affect all the environmentally relevant parameters related to biodegradability, enantioselectivity (in the case of chiral compounds), kinetic parameters and persistence of chemicals. However, such impacts have never been well-characterized. In this work, calculations and models used for a long time are studied by simulating systematic errors at the 5% level, which affect independently the analytical calibration step and the biodegradation process. The impact of these errors is also compared with those obtained from an alternative approach: rec…
Biopartitioning micellar chromatography to predict ecotoxicity
Abstract The knowledge of chemical toxicity is necessary for risk assessment and ranking of chemicals according their hazard potential. The use of biopartitioning micellar chromatography has proven to be valid in predicting several biological activities of different kinds of compounds. In this paper, the relationships between retention of 66 organic pollutants and several ecotoxicity parameters are studied. Adequate correlations retention-ecotoxicity (LC 50 in fish and daphnia, EC 50 in green algae and daphnia, values of chronic toxicity in fish and green algae and bioconcentration factor in fish) of organic pollutants are obtained. These models are compared with the corresponding obtained …
Evaluation of enantioselective binding of propanocaine to human serum albumin by ultrafiltration and electrokinetic chromatography under intermediate precision conditions.
Abstract Stereoselectivity in protein binding can have a significant effect on the pharmacokinetic and pharmacodynamic properties of chiral drugs. In this paper, the enantioselective binding of propanocaine (PRO) enantiomers to human serum albumin (HSA), the most relevant plasmatic protein in view of stereoselectivity, has been evaluated by incubation and ultrafiltration of racemic PRO–HSA mixtures and chiral analysis of the bound and unbound fractions by electrokinetic chromatography using HSA as chiral selector. Experimental conditions for the separation of PRO enantiomers using HSA as chiral selector and electrokinetic chromatography have been optimised. Affinity constants and protein bi…
Chromatographic quantitation of the hydrophobicity of ionic compounds by the use of micellar mobile phases
Abstract Many biologically active compounds of interest in structure–activity relationships are ionic at physiological pH. However, ionic organic compounds are only weakly or not retained in conventional RPLC which impedes the chromatographic estimation of their hydrophobicity and the development of quantitative retention–activity relationship studies. The use of micellar mobile phases allows the retention of ionic compounds. Hydrophobic and electrostatic forces govern the retention of ionic compounds in micellar liquid chromatography. In this paper three different retention models log k–log P for ionic compounds are tested (P=partition coefficient). The retention model (log k=a log P+bα+c)…
Determination of procaine and tetracaine in plasma samples by micellar liquid chromatography and direct injection of sample
A liquid chromatographic procedure is proposed for the determination of procaine and tetracaine in plasma samples with direct injection. The method uses a Spherisorb octadecylsilane ODS-2 C18 analytical column and a micellar mobile phase containing 0.15 M sodium dodecyl sulphate, 0.5% triethylamine at pH 2.5 and 10% propanol. The UV detection was carried out at 300 nm. Plasma sample preparation required only adequate dilution with the mobile phase before injection into the chromatographic system. The proposed method allows the determination of procaine and tetracaine in plasma at therapeutic levels.
Fast-multivariate optimization of chiral separations in capillary electrophoresis: Anticipative strategies
Abstract The design of experiments (DOE) is a good option for rationally limiting the number of experiments required to achieve the enantioresolution (Rs) of a chiral compound in capillary electrophoresis. In some cases, the modeled Rs after DOE analysis can be unsatisfactory, maybe because the range of the explored factors (DOE domain) was not the adequate. In these cases, anticipative strategies can be an alternative to the repetition of the process (e.g. a new DOE), to save time and money. In this work, multiple linear regression (MLR)-steepest ascent and a new anticipative strategy based on a multiple response-partial least squares model (called PLS2-prediction) are examined as post-DOE…
On the measurement of consistent long-term retention factor values in micellar liquid chromatography
Abstract In the field of the quantitative structure–retention and retention–activity relationships (QRAR and QSRR) is crucial to obtain consistent retention factors (k). For this purpose, two unbiased approaches to estimate k are used: (i) the IUPAC approach (based on the extra-column time correction) and (ii) the ‘2-references’ approach (based on the k estimation respect to two prefixed reference k values). Three reference chemicals were selected attending to their retention time, chemical stability and non-ionic character. Consistent retention factor values for these references were estimated for C18 chromatographic columns and Brij35 solutions as mobile phases after statistical analysis.…
Cyclodextrins in capillary electrophoresis: Recent developments and new trends
Despite the fact that extensive research in the field of separations by capillary electrophoresis (CE) has been carried out and many reviews have been published in the last years, a specific review on the use and future potential of cyclodextrins (CDs) in CE is not available. This review focuses the attention in the CD-CE topic over the January 2013-February 2014 period (not covered by previous more general CE-reviews). Recent contributions (reviews and research articles) including practical uses (e.g. solute-CD binding constant estimation and further potentials; 19% of publications), developments and applications (mainly chiral and achiral analysis; 38 and 24% of publications, respectively…
Quantitative structure-retention relationships for ionic and non-ionic compounds in biopartitioning micellar chromatography
Quantitative structure–retention relationships, QSRRs, represent a powerful tool in chromatography. The objectives of QSRR studies are to predict the chromatographic retention behaviour of solutes based on their structural properties, to elucidate retention mechanisms, to optimize the separation of complex mixtures or to prepare experimental designs. In this paper, using the retention factors of 151 structurally unrelated solutes that cover a wide range of hydrophobicity, molecular size, hydrogen bonding properties and ionization degrees obtained in biopartitioning micellar chromatography (BMC) at different Brij35 micellar concentrations, several multivariate QSRR models are tested. It is d…
Toward a Quality Guide to Facilitate the Transference of Analytical Methods from Research to Testing Laboratories: A Case Study
Abstract At present, there is no single viewpoint that defines QA strategies in analytical chemistry. On the other hand, there are no unique protocols defining a set of analytical tasks and decision criteria to be performed during the method development phase (e.g., by a single research laboratory) in order to facilitate the transference to the testing laboratories intending to adapt, validate, and routinely use this method. This study proposes general criteria, a priori valid for any developed method, recommended as a provisional quality guide containing the minimum internal tasks necessary to publish new analytical method results. As an application, the selection of some basic internal qu…
Modified Gaussian models applied to the description and deconvolution of peaks in chiral liquid chromatography.
Abstract The description of the profiles of chromatographic peaks has been studied extensively, with a large number of proposed mathematical functions. Among them, the accuracy achieved with modified Gaussian models that describe the deviation of an ideal Gaussian peak as a change in the peak variance or standard deviation over time, has been highlighted. These models are, in fact, a family of functions of different complexity with great flexibility to adjust chromatographic peaks over a wide range of asymmetries and shapes. However, an uncontrolled behaviour of the signal may occur outside the region being fitted, forcing the use of different strategies to overcome this problem. In this wo…