Micellar Liquid Chromatography Research Articles

Evaluating the effectiveness of different household washing techniques for removal of insecticides from spinach and chickpea leaves by micellar liquid chromatography

In the past few decades, the employment of green analytical approaches in chromatographic method development has attracted the analytical separation community. The greenness of the developed method depends upon the toxicity of solvents and the amount of generated post-analysis waste generated. In this concern, micellar liquid chromatography (MLC) is a simple and rapid technique that generates very low toxic waste compared to traditional chromatographic pesticide detection methods. Here, MLC method has been validated and applied for the determination of monocrotofos (MCF), imidacloprid (ICP), dimethoate (DM) and profenofos (PFF) in spinach and chickpea leaves. The optimized mobile phase was 0.065 M SDS-2 % 1-propanol, 0.01 M NaH2PO4 buffered to pH 7. A C18 column was used for separation with a flow rate of 1 mL/min. The developed method has been validated following the guidelines of SANTE/11,312/2021 and ICH guidelines for; limit of quantification (0.05–0.20 mg/kg), linearity (r2> 0.997–0.999), precision (<6.3 %), accuracy (96.3 %–99.8 %) and robustness (<6) in real samples. ICP and MCF, apart from DM and PFF, were detected in the present work. After detecting insecticides in spinach and chickpea leaves both were washed with different household chemicals i.e. normal, lukewarm, common salt, lemon juice water and commercial ozonizer. Based on five washing techniques with insecticide concentration time intervals reduction rates were calculated for each washing treatment. The results show that lemon juice, common salt water, and ozonizer can be used as washing techniques for the reduction of superficial and systematic residues of ICP and MCF. Common salt and lemon juice water were better for washing over vinegar and potassium permanganate (KMnO4) as they enhance the colour of the green leafy vegetables and are available in every Indian kitchen. They can be easily used by lower socioeconomic classes who cannot afford KMnO4 and vinegar.

Eco-friendly micellar ultra-performance liquid chromatography (UPLC) method for simultaneous determination of non-communicable diseases drugs in bulk powder, spiked human plasma and in individual dosage forms

Eco-friendly micellar ultra-performance liquid chromatography (UPLC) method for simultaneous determination of non-communicable diseases drugs in bulk powder, spiked human plasma and in individual dosage forms

A comprehensive exploration of diverse green analytical techniques and their influence in different analytical fields

AbstractThe primary goals of green analytical chemistry (GAC) involve the development of novel analytical technologies or the modification of existing methods to incorporate procedures with reduced reliance on hazardous chemicals. Several approaches can be employed to achieve these objectives, including the utilization of environmentally friendly solvents and reagents, shortening chromatographic separation times, and the miniaturization of analytical devices. The review summarizes the evolution of GAC with its specific principles and with special emphasis on recent applications in modern era. A detailed exploration of the choice of green solvents and their practical utilization in green liquid chromatography (LC) techniques for environmentally friendly analysis of pharmaceuticals has been discussed. An ordinary description with a tabular illustration of various modern green analytical techniques like micellar LC, superheated water chromatography, high‐speed LC, enhanced fluidity LC, two‐dimensional LC, and aspects of gas chromatography has been thoroughly provided for concepts with their mechanism and limitations. GAC provides potential applications for food analysis, environmental analysis of pharmaceutically active compounds in the aquatic environment with special emphasis on strategies for greening LC and impurity profiling, and so on. Numerous benefits, limitations, and challenges that are linked to the field of GAC are highlighted in this review. We pay special attention to new green approaches, such as automation, miniaturization, direct analysis techniques, and some initiatives like the National Environmental Methods Index Label for Analytical Eco‐Scale assessment. In spite of many promising advancements, the understanding is still limited and challenging.

Koncept ekološki prihvatljivih hromatografskih metoda - studija slučaja na primeru razdvajanja dronedaron hidrohlorida i njegovih degradacionih proizvoda

Recently, concern about the environmental impact of drug analysis methods has increased significantly. Reversed-phase high-performance liquid chromatography (RP-HPLC), the predominant technique in drug analysis, relies heavily on organic solvents such as acetonitrile, which is known for its chromatographic efficiency, but also for its toxicity and flammability. To address these concerns, it is essential to minimize the use of toxic organic solvents. The aim of this study is to explore greener RP-HPLC modifications and evaluate their applicability in the pharmaceutical industry. Methods were developed for the separation of dronedarone hydrochloride and its degradation products based on experimental design, including micellar liquid chromatography (MLC), b-cyclodextrin (CD) modified RP-HPLC and ultra-high performance liquid chromatography (UHPLC). The eco-friendliness of these methods was assessed using the analytical eco-scale score, green analytical procedure index (GAPI) and analytical greenness (AGREE). AGREE appears to be the most suitable, as it revealed the greatest differences between the compared methods, as well as insights into critical aspects of the methods. UHPLC and b-CD modified RP-HPLC have been shown to be superior to MLC, and both methods can be a good choice, depending on whether the ease of implementation or energy efficiency is considered to be a more important criterion.

Use of green analysis methods to determine the ionization constant values of isoxazolyl penicillins

In this study, an environmentally friendly analysis of penicillinase-resistant antibiotics oxacillin, dicloxacillin, and cloxacillin was carried out by liquid chromatography methods using ethanol solvent that is not harmful to human health and the environment. Ionization constant (wwpKa) values of isoxazolyl penicillins in ethanol–water binary mixtures containing 20–35% (v/v) ethanol at 37 °C were obtained by green reverse phase liquid chromatography and micellar liquid chromatography methods known as green chromatography. The developed methods are based on a model that relates the chromatographic behavior of the identified analytes to pH. The linear relationship between the microscopic solvatochromic parameter ETN and the retention values of the compounds was determined depending on the change of organic solvent in the mobile phase. Using the obtained chromatographic data, conditions that ensure the simultaneous separation of the compounds for both methods were determined. The relationship between the calculated sspKa values and the macroscopic properties of ethanol was investigated. The agreement between the pKa (wwpKa) values in the water medium determined by this approach was determined by t-test at a 95% confidence level for both methods. The wwpKa values of the compounds at 25 °C were also estimated using Abraham solute parameters without any experimental work.

Green method for determination of four anti-viral drugs using micellar liquid chromatography: Application to dosage form analysis

A micellar liquid chromatographic (MLC) method was developed to separate and analyze four anti-viral drugs. The method utilized a Thermo Hypersil ODS C18 HPLC column with an isocratic elution mode employing a mobile phase consisting of sodium dodecyl sulfate (0.15 M, pH 3), isopropanol and triethylamine (79.7:20:0.3, v/v/v). The Design Expert 8® software was used to optimize four factors simultaneously and the developed quadratic D-optimal model could achieve maximum resolution between peaks while minimizing the run time. The method was found linear over the concentration ranges of 10–100 μg/mL for sofosbuvir, daclatasvir, and 20–200 μg/mL for ledipasvir and velpatasvir, and successfully applied to the analysis of sofosbuvir and velpatasvir in tablets. The MLC method was compared with a previously reported high-performance liquid chromatography (HPLC) method using the analytical GREEnness metric approach (AGREE), and was found to exhibit greener performance due to the use of safer reagents. The results indicate the high potential of micellar liquid chromatography in the separation of quaternary mixtures with the aid of response surface methodologies.

Micellar enhanced chromatographic separation of selected hazardous chemical present in hair dye and their detection in formulations and swab, including assessment of damage caused to cuticle of hair shaft

Hydroquinone (HQ), resorcinol (RS), m-aminophenol (m-AMP) and p-phenylenediamine (p-PPD) are aromatic compounds which are generally used in hair dyes to provide different colours to hair. In European Union the concentrations of HQ, RS, m-AMP and p-PPD is regulated in hair dyes and other cosmetic products by EU commission regulation EU/2019/831. This legislation is generally exercised because all these compounds are toxic and may cause severe allergies when used regularly. However in India no such regulations exist to monitor these toxic compounds in hair dyes therefore in this study a simple, rapid, economical and ecofriendly micellar liquid chromatographic (MLC) technique has been developed which can monitor all the selected toxic compounds simultaneously. HQ and RS are positional isomers and are difficult to be separated by HPLC whereas with the developed MLC method it was well separated and detected. The developed MLC technique has been applied to detect and quantify selected analytes in oxidative and non-oxidative hair dyes and swab samples from the scalp. The simultaneous separation of selected analytes was performed in mobile phase 0.09 M SDS, 0.01 M NaH2PO4–2% v/v 1-butanol at pH 7 running through C18 column under isocratic mode at 1 mL/min. flow rate. All the analytes were eluted within 6 min. The present method has been validated following the EURCHEM Guideline, 2014 in terms of calibration range (0.08–15 µg/mL), limit of detection (0.01–0.09 µg/mL), limit of quantification (0.08–0.35 µg/mL), accuracy (<5.6%), precision (91–105%) and robustness (<5.8%). The selected compounds in hair dye formulation were found in the range of 0.06–12.2 µg/mL (when diluted 25 times). Hair dyes persistence study was conducted up to 10 days from the day of application on the scalp, suggesting that the dyes were not completely washed off and were retained on the scalp for more than one week. SEM analysis of dyed hair revealed that hair are severely damaged due to use of dyes. The advantage of the developed method is that it could easily be adopted by quality control and cosmetic laboratories for quality control and check for the simultaneous separation of positional isomers together with two other aromatic compounds.

Chromatographic behavior of polar compounds in the transition between reversed-phase and submicellar/micellar liquid chromatography

Micellar liquid chromatography (MLC) is a reversed-phase liquid chromatography (RPLC) mode employing a surfactant, typically sodium dodecyl sulfate (SDS), which is biodegradable, in an aqueous or aqueous-organic mobile phase above its critical micellar concentration (CMC). Under these conditions, substantial alterations in retention, selectivity and peak profile occur in comparison to RPLC using conventional hydro-organic mobile phases. These changes arise from the presence of micelles in the mobile phase and the adsorption of surfactant on the stationary phase. In this work, the chromatographic performance of polar compounds of diverse nature is investigated using mobile phases containing SDS below and above its CMC in the presence of acetonitrile. Three nucleosides, four diuretics, and four sulfonamides, all of biomedical and pharmaceutical significance and possessing distinct acid-base characteristics, were chosen as probe compounds. Modifications in retention and peak shape were investigated using experimental conditions that enable the transition from submicellar to micellar conditions. A comparison was also made with conventional RPLC. Analysing the polar compounds with RPLC under low submicellar or micellar conditions significantly decreases the consumption of organic solvent in comparison to hydrophilic liquid chromatography (HILIC).

A method to determine two antibiotics prescribed to treat nosocomial infections in plasma and urine by micellar liquid chromatography

Combined prescription of the antimicrobial drugs linezolid and meropenem is a common strategy to treat multidrug-resistant nosocomial infections. We propose an innovative method to determine these two drugs in plasma and urine, based on micellar liquid chromatography. Both biological fluids were diluted in mobile phase, filtered and directly injected, without any extraction step. Using a C18 column and a mobile phase of 0.1 M sodium dodecyl sulfate – 10 % methanol, phosphate buffered at pH 3, running under isocratic mode, both antibiotics were eluted without overlapping in<15 min. Detection was by absorbance: 255 nm for linezolid and 310 nm for meropenem. The influence of sodium dodecyl sulfate and methanol concentration on retention factor was established for both drugs using an interpretative approach assisted by chemometrics. The procedure was successfully validated following the guidelines of 2018 Bioanalytical Method Validation Guidance for Industry in terms of: linearity (determination coefficients over 0.99990), calibration range (1 – 50 mg/L), instrumental and method sensitivity, trueness (bias of −10.8 to + 2.4%), precision (relative standard deviation of < 10.2%), dilution integrity, carry-over effect, robustness and stability. It should be emphasized that the method uses low volumes of toxic and volatile solvents and can be achieved in a short period. The procedure was found useful for routine analysis, as it was cost-affordable, more eco-friendly and safer than hydroorganic HPLC, easy-to-handle and highly sample-throughput. Finally, it was applied to incurred samples of patients taking this medication.

Green micellar stability-indicating high-performance liquid chromatography method for determination of rupatadine fumarate in the presence of its main impurity desloratadine: Oxidative degradation kinetics study.

A green micellar stability-indicating high-performance liquid chromatography method was developed for rupatadine fumarate determination in existence with its main impurity desloratadine. Separation was attained using Hypersil ODS column (150×4.6mm, 5μm), the micellar mobile phase consisted of 0.13M sodium dodecyl sulfate, 0.1M disodium hydrogen phosphate adjusted by phosphoric acid to pH 2.8 and 10% n-butanol. The column was maintained at 45◦ C and detection was carried out at 267nm. A linear response was achieved over the range of 2-160μg/ml for rupatadine and 0.4-8μg/ml for desloratadine. The method was applied for rupatadine determination in alergoliber tablets and alergoliber syrup without the interference of methyl paraben and propyl paraben present as main excipients. Rupatadine fumarate revealed pronounced susceptibility to oxidation; further study of oxidative degradation kinetics was carried out. Rupatadine was found to follow pseudo-first-order kinetics when exposed to 10% H2 O2 at 60 and 80°C and the activation energy was found to be 15.69Kcal/mol. At a lower temperature (40°C), degradation kinetics regression was best fitted as a polynomial quadratic relationship, thus rupatadine oxidation at a lower temperature tends to adopt a second-order kinetics rate. Oxidative degradation product structure was revealed using infrared and found to be rupatadine N-oxide at all temperature values.

Micellar liquid chromatography as a sustainable tool to quantify three statins in oral solid dosage forms

A method based on micellar liquid chromatography has been developed to determine rosuvastatin, lovastatin and simvastatin in oral solid dosage forms. Samples were solved in mobile phase up to the target concentration, filtered and directly injected. The three statins were resolved in 30 min, using an aqueous solution of 0.10 M sodium dodecyl sulfate – 7.0% 1-butanol, buffered at pH 3 with 0.01 M phosphate salt as mobile phase, running under isocratic mode at 1 mL/min through a C18 column. Detection was at 240 nm. The effect of sodium dodecyl sulfate on elution strength was more important than that of the organic solvent. The procedure was successfully validated by the guidelines of the International Council for Harmonization in terms of: specificity, linearity (r2 > 0.990), calibration range (1.5 - 15 mg/L for rosuvastatin, 0.5–10 mg/L for lovastatin and simvastatin), limit of detection (0.4, 0.2 and 0.15 mg/L for rosuvastatin, lovastatin and simvastatin, respectively), trueness (98.8–101.7%), precision (<2.7%), carry-over effect, robustness, and stability. Values were inside the acceptance criteria of the Methods, Method Verification and Validation, Food and Drug Administration-Office of Regulatory Affairs, thus ensuring the reliability of the results. The main feature was the low proportion of organic solvent used, thus making the procedure sustainable and green. Besides, it was easy-to-conduct and with high sample-throughput, and then useful for routine analysis in pharmaceutical quality control. Finally, it was applied to commercial pharmaceutical preparations.

Application of micellar liquid chromatography to model ecotoxicity of pesticides. Comparison with immobilized artificial membrane chromatography and n-octanol-water partitioning

The potential of Micellar Liquid Chromatography (MLC) to model ecotoxicological endpoints for a series of pesticides was investigated. To exploit the flexibility in MLC conditions, different surfactants were employed and retention mechanism was tracked and compared to Immobilized Artificial Membrane (IAM) chromatographic retention and n-octanol- water partitioning, logP. Neutral polyoxyethylene (23) lauryl ether (Brij-35), anionic sodium dodecyl sulfate (SDS) and cationic cetyltrimethylammonium bromide (CTAB) were used in presence of PBS at pH=7.40 and acetonitrile as organic modifier when necessary. Similarities/ dissimilarities between MLC retention and IAM or logP were investigated by Principal Component Analysis (PCA) and Liner Solvation Energy Relationships (LSER). LSER revealed that hydrogen bonding acidity is the most important factor for differentiation between MLC and IAM or logP. The impact of hydrogen bonding is exemplified in the relationships of MLC retention factors with IAM or logP, which necessitate the inclusion of a relevant descriptor. PCA further revealed that MLC retention factors are clustered together with IAM indices and logP within a broader ellipse formed by ecotoxicological endpoints, involving LC50/ EC50 values of six aquatic organisms namely Rainbow Trout, Fathead Minnow, Bluegill Sunfish, Sheepshead Minnow, Eastern Oyster and Water Flea as well as LD50 values of Honey Bee, thus justifying their use to construct relevant models. Satisfactory specific models for individual organisms, as well as general fish models, were obtained, in most cases, upon combination of MLC retention factors with Molecular Weight (MW) or/ and hydrogen bond parameters. All models were evaluated and compared to previously reported IAM and logP based models using an external validation data set. Predictions with Brij-35 and SDS based models were comparable, although slightly inferior than those obtained with IAM, while they were in all cases better than those obtained with logP. CTAB led to a satisfactory prediction model for Honey Bee, but it was found less suitable for aquatic organisms.

An easy and green assay to determine albendazole and ivermectin in veterinary preparations by micellar liquid chromatography.

A procedure to determine albendazole and ivermectin in veterinary formulations, like tablet, bolus, oral suspensions, and injections by micellar liquid chromatography, has been developed. Sample preparation was a batch solid-to-liquid extraction in mobile phase, consisting of a stirring step (15min), followed by ultrasonication (15min) and filtration of the obtained supernatant, to reach a target concentration of 2mg/L for both analytes. Using a mobile phase of 0.15M sodium dodecyl sulfate-6% 1-pentanol buffered at pH 3 with a 0.01M phosphate salt, running at 1mL/min through a C18 column, both drugs were resolved in less than 10min. Absorbance detection wavelength was 292nm. Procedure was validated by the guidelines of the International Council on Harmonization in terms of specificity, calibration range (0.025-5mg/L), trueness (97.8%-102.6%), precision (<2.2%), and system suitability. The method was found easy-to-handle, low cost, safe, green, and with high sample-throughput, thus useful for routine analysis. Therefore, it represents a valuable alternative for quality control of veterinary formulations. It was applied to samples of veterinary formulations purchased from local chemists and veterinarians, and label claims were inside the acceptance criteria (95%-105%).

Analysis of canthin-6-one alkaloids derived from Eurycoma spp. by micellar liquid chromatography and conventional high-performance liquid chromatography: a comparative evaluation.

Extracts of Eurycoma longifolia Jack (EL) and Eurycoma harmandiana Pierre (EH) contain numerous bioactive compounds and varying matrices that are challenging to separate using chromatographic techniques. Herein, micellar liquid chromatography (MLC) was used to analyze canthin-6-one alkaloids contained in these extracts, and the achieved performance was compared with that of a conventional high-performance liquid chromatography (HPLC) method. The optimal mobile phase of MLC corresponded to 15 : 85 (v/v) acetonitrile : water (pH 3) containing 110 mM sodium dodecyl sulfate and 10 mM NaH2PO4. The retention times of canthin-6-one-9-O-β-d-glucopyranoside, 9-hydroxycanthin-6-one, canthin-6-one, and 9-methoxycanthin-6-one were 4.78/15.42, 17.64/24.11, 32.84/38.27, and 39.04/39.86 min, respectively, in the cases of isocratic MLC and conventional HPLC. In both cases, the analyte resolution exceeded 1.5. The MLC elution behavior of the examined analytes was largely determined by their hydrophobicity and ionization. The sensitivity, precision, accuracy, and per-run acetonitrile consumption of the MLC method were comparable to those of the conventional HPLC method. However, the latter method exhibited higher performance for application to EL and EH samples, particularly those with low analyte concentrations and varying sample matrices. Overall, the analysis of canthin-6-one alkaloids using MLC was limited to trace analytes due to interference by the matrix.

Simultaneous Analysis of Topotecan and Capsaicin by Micellar Liquid Chromatography

Objectives: This study aimed to develop a micellar liquid chromatography-based simultaneous estimation method for the analysis of topotecan and capsaicin. Materials and Methods: The Liquid chromatographic system consisted of the Shimadzu Prominence HPLC model (Shimadzu Corp., Kyoto, Japan) containing an LC-20AT isocratic pump, an autosampler SIL-20AC, and a diode array detector SPD-M20 A (190–800 nm) was used. The drugs were analyzed at a wavelength of 230 nm using sodium dodecyl sulfate, buffer salt (0.01 M sodium dihydrogen phosphate), and n-butanol (7%) as mobile phase. Results: The developed Micellar liquid chromatography-Photodiode Array Detection (MLC-PDA) method successfully eluted the topotecan and capsaicin with retention times of 6.2 min and 16.3, respectively. The developed method has displayed the limits of detection i.e. 0.05 and 0.98 μg/mL and limits of quantification i.e 0.08 and 1.25 μg/mL for topotecan and capsaicin, respectively. Statistical analysis further demonstrated that the developed method is linear, exact, accurate, and specific for the analysis of topotecan and capsaicin. The method was verified as per the ICH guidelines. Conclusion: The developed MLC-PDA method was found to be precise, simple, and reproducible. The developed MLC-PDA method successfully estimated the topotecan and capsaicin from the combination of drugs. Micellar chromatography is an accurate, rapid, cost-effective, and less hazardous method as compared to HPLC.

Combined Micellar Liquid Chromatography Technique and QSARs Modeling in Predicting the Blood-Brain Barrier Permeation of Heterocyclic Drug-like Compounds.

The quantitative structure-activity relationship (QSAR) methodology was used to predict the blood-brain permeability (log BB) for 65 synthetic heterocyclic compounds tested as promising drug candidates. The compounds were characterized by different descriptors: lipophilicity, parachor, polarizability, molecular weight, number of hydrogen bond acceptors, number of rotatable bonds, and polar surface area. Lipophilic properties of the compounds were evaluated experimentally by micellar liquid chromatography (MLC). In the experiments, sodium dodecyl sulfate (SDS) as the effluent component and the ODS-2 column were used. Using multiple linear regression and leave-one-out cross-validation, we derived the statistically significant and highly predictive quantitative structure-activity relationship models. Thus, this study provides valuable information on the expected properties of the substances that can be used as a support tool in the design of new therapeutic agents.

Establishment of Green Mixed Micellar Liquid Chromatography with Deep Eutectic Solvents as Mobile Phase Modifiers and Evaluation of Greenness

Establishment of Green Mixed Micellar Liquid Chromatography with Deep Eutectic Solvents as Mobile Phase Modifiers and Evaluation of Greenness

Micellar and sub-micellar chromatography with a cocamidopropyl betaine surfactant

We have coated a typical C18 column with the pH cationic charge controllable zwitterionic synthetic surfactant, cocamidopropyl betaine (CAPB), in order to generate a mixed mode reversed phase weak ion exchange column. As determined by the Thomas model, the column has an adsorbed surfactant capacity of 0.557 mmoles. The addition of 8.8 × 10–4 M CAPB to the totally aqueous mobile phase ensured stability of the surfactant on the column and permitted separation of the four component sulfonamide mixture with micellar liquid chromatography (MLC) in under 11 min. Comparatively, with a dilute H2SO4 mobile phase with no CAPB, the separation time for the sulfonamide mixture reached an excessive run time of an hour on the bare C18 chains. With CAPB in the mobile phase (no organic solvent present), a seven component sulfonamide mixture could be separated in less than 45 min. A five component short chain carboxylic acid mixture, separated in 20 min, was used to examine the ion exchange character of the column in pH environments of 2.3 and 4.6. Three phase MLC equilibrium analysis was also done in these pH environments with the sulfa drug and carboxylic acid mixtures to determine partition coefficients. Finally, a quite high molecular weight (70,000) anionic polystyrene sulfonate polymer was characterized by MLC with only CAPB and variable pH mobile phases; the optimal pH was determined to be 5.6. A totally aqueous mobile phase without CAPB was not suitable for profiling this polymer.

Quantitative Retention (Structure)-Activity Relationships in Predicting the Pharmaceutical and Toxic Properties of Potential Pesticides.

The micellar liquid chromatography technique and quantitative retention (structure)–activity relationships method were used to predict properties of carbamic and phenoxyacetic acids derivatives, newly synthesized in our laboratory and considered as potential pesticides. Important properties of the test substances characterizing their potential significance as pesticides as well as threats to humans were considered: the volume of distribution, the unbonded fractions, the blood–brain distribution, the rate of skin and cell permeation, the dermal absorption, the binding to human serum albumin, partitioning between water and plants’ cuticles, and the lethal dose. Pharmacokinetic and toxicity parameters were predicted as functions of the solutes’ lipophilicities and the number of hydrogen bond donors, the number of hydrogen bond acceptors, and the number of rotatable bonds. The equations that were derived were evaluated statistically and cross-validated. Important features of the molecular structure influencing the properties of the tested substances were indicated. The QSAR models that were developed had high predictive ability and high reliability in modeling the properties of the molecules that were tested. The investigations highlighted the applicability of combined chromatographic technique and QS(R)ARs in modeling the important properties of potential pesticides and reducing unethical animal testing.

Micelle-Incorporated Liquid Chromatography in the Light of Green Chemistry: An Application for the Quality Control Analysis of Anti-Platelet Fixed-Dose Combinations.

Oral anti-platelet agents are the cornerstone of the treatment of multiple cardiovascular diseases and in the long-term prevention of their recurrence. In the present work, we report a method based on micellar liquid chromatography coupled with ultraviolet detection (MLC/UV), for the simultaneous quantification of combined anti-platelet therapy namely, clopidogrel bisulfate (CPS), aspirin (ASP), together with salicylic acid (SA), in their pharmaceutical dosage form. The incorporation of 0.1M polyoxyethylene 23 lauryl ether (Brij-35) as a surfactant into the mobile phase improved solute-mobile phase interaction allowing for minimal organic solvent utilization, enhanced resolution, and rapid analysis (7 min). Furthermore, we performed a comprehensive evaluation of the environmental impact caused by our procedures versus previously reported analytical procedures applied in the determination of CPS and ASP. The evaluation was made using the Eco-scale tool. The results of the developed method indicated the superiority of our procedures in terms of greenness without compromising the quality of performance characteristics. The method was linear in the range of 1-100 µg/mL with limits of detection of 0.28, 0.32, and 0.29 µg/mL for CPS, ASP, and SA, respectively. The developed method can also be utilized to test the purity and the stability of ASP in pharmaceutical formulations through monitoring SA as its main degradation product. The MLC/UV method was successfully applied to the quantitative analysis of CPS, ASP together with SA-as a main degradation product of ASP-in their pharmaceutical dosage form. The developed method was successfully applied for the determination of clopidogrel bisulfate (CPS), aspirin (ASP), together with salicylic acid (SA), in their pharmaceutical dosage form.