Rupatadine: A Review On Analytical Method Development And Validation For Quantification Of Bulk And Pharmaceutical Dosage Form By Liquid Chromatography

Abstract

Through its interaction with certain receptors, rupatadine, a non-sedative, selective, and long-acting H1 antihistamine, has antagonistic PAF effects. We have conducted a thorough literature review of numerous journals pertaining to analytical and pharmaceutical chemistry. Additionally, we have examined instrumental analytical methods developed and employed for the purpose of identifying drugs in bulk pharmaceuticals, formulations, and biological fluids, either individually or in combination with other drugs. The most recent analytical techniques, such as liquid chromatography, RP HPLC, HPTLC, and HPLC, are covered in this review.

Keywords

Introduction

       
           
       
Figure-2. Mechanism of action of anti-allergic agents.

Rupatadine exhibits potent antagonistic action on PAF and histamine H1 receptors. Studies conducted in vivo and in vitro on a variety of animal species, including dogs, rats, mice, guinea pigs, rabbits, and rabbits, showed this activity. Furthermore, rupatadine has a profile as an anti-allergic medication with several advantageous effects, including the suppression of cytokine release, the inhibition of neutrophil and eosinophil migration, and the inhibition of mast cell degranulation. Rupatadine binds to the histamine H1 receptor with a strong affinity. Using the guinea-pig ileum functional test, the antihistamine activity of rupatadine was compared with various first- and second-generation antihistamines [22]. It turned out to be more effective than diphenhydramine, hydroxyzine, cetirizine, loratadine, and terfenadine. In vitro, a few metabolites of rupatadine also have antihistamine properties. In vitro, desloratadine and rutadine exhibit comparable antihistamine potency (binding assay Kiapp = 22 nM for desloratadine and 26 nM for rutadine, respectively).[2]

Analytical Method Validation

Validation is a method-based approach to ensure a method's suitability as a quality control tool for analytical measurements. Any analytical measurement's goal is to produce accurate, dependable, and consistent results. The use of validated analytical techniques is crucial to accomplishing this objective. An analytical method consists of techniques, methods, procedures, and protocols. Analytical method validation includes the determination of accuracy, precision, specificity, detection limit, quantitation limit, linearity, range, and robustness. The results from method validation can be used to moderate the quality, reliability, and consistency of analytical results, which is an integral part of any good analytical practice. The majority of laws and standards of quality governing laboratories also entail the validation of analytical procedures.

Analytical Methods for Rupatadine

IP-2022 (Rupatadine fumarate tablets) - Liquid chromatography- Mobile phase: Buffer (pH 2.8):acetonitril:methanol (30:30:40), Colum: 25cm×4.6mm(5µm), Flow rate: 1.0 mL/min, Wavelength: 210nm.[3]

Reported Methods for Rupatadine

Kumar N et al

An accurate, precise, simple, and selective stability-indicating gradient reverse phase ultra performance liquid chromatographic method has been developed and validated for the determination of montelukast and rupatadine in pharmaceutical formulations in the presence of degradation products. This method is useful for measuring rupatadine and its metabolites. Using a mobile phase containing a gradient mixture of solvent A (0.02 M KH2PO4, pH 3.0) and B (90:10 v/v mixture of acetonitrile and water), the chromatographic separation was carried out on an Acquity BEH C8 column (100 mm x 2.1 mm I.D., 1.7 ?m) at a flow rate of 0.5 mL/min. At 240 nm in wavelength, the detection was done. The drug product was exposed to stress conditions like acid, base, oxidative, hydrolytic, thermal, and photolytic degradation in order to demonstrate stability, showing the method's capabilities. The breakdown products of montelukast and rupatadine were clearly separated. The new method's specificity, linearity, accuracy, precision, and robustness were all validated in accordance with ICH recommendations. The procedure's validation produced satisfactory results. This stability-indicating method can be applied to regular manufacturing sample analysis as well as evaluating the stability of montelukast and rupatadine tablets.[6]

Nogueira, Daniele Rubert, et al

The quantification of rupatadine in pharmaceutical dosage forms using reversed-phase liquid chromatography (RP-LC) has been validated. Using a Gemini C18 column (150 mm × 4.6 mm I.D.) kept at 30 °C, the LC technique was performed. Ammonium acetate buffer (pH 3.0; 0.01 M) containing 0.05% 1-heptanesulfonic acid–acetonitrile (71.5:28.5, v/v) was used as the mobile phase. The flow rate was set at 1.0 mL min^?1 and detected at 242 nm using a photodiode array (PDA). The chromatographic separation had a linear range of 0.5–400 ?g mL?1 (r 2 = 0.9999) and was obtained with a retention period of 5.15 min. Degradation studies demonstrated the method's specificity and stability-indicating ability while also demonstrating the absence of excipient influence. With a bias of less than 0.58%, the accuracy was 100.39%. The quantitation and detection limits were 0.5 ?g mL?1 and 0.01 ?g mL?1, respectively. Additionally, satisfactory findings for precision, sensitivity, and robustness were shown by method validation. The suggested approach was used to analyze drug dose forms and guarantee therapeutic efficacy.[7]

HK Trivedi et al

For the purpose of identifying rupatadine (RUPA) and its related compounds in pharmaceutical dosage forms, a photodiode array detector in conjunction with a simple, sensitive, and repeatable reversed-phase high-performance liquid chromatography (RP-HPLC) approach was developed. The Hypersil BDS (150 x 4.6 mm, 5 ?m) column was used for chromatographic separation, and the mobile phase consisted of a gradient combination of a solvent (methanol) and a buffer (acetate buffer, pH 6.0). The temperature of the column oven was kept at 50°C, the flow rate was 1.0 mL/min, and the eluted chemicals were seen at 264 nm for the associated substances and test. In 15.0 minutes, the new approach was able to separate RUPA from its four recognized and three unknown impurities. Rupatadine underwent deterioration under stress conditions including acidic, basic, oxidative, photolytic, thermal, and hydrolytic. Under oxidative stress conditions, rupatadine was shown to degrade extensively; under acid, base, hydrolytic, thermal, and photolytic stress conditions, it degraded somewhat. Every impurity was clearly separated from the primary peak and from each other, demonstrating the method's capacity to indicate stability. The International Conference on Harmonization (ICH) requirements were followed in the validation of the created approach. The RP-HPLC method that has been developed and verified is compatible with LC-MS and has the potential to identify eluted unknown impurities of RUPA.[8]

MM Amer et al

To determine rupatadine fumarate in the presence of its primary contaminant, desloratadine, a high-performance liquid chromatographic method was devised that indicates green micellar stability. A 150 × 4.6 mm, 5 ?m Hypersil ODS column was used to achieve separation. The micellar mobile phase included 10% n-butanol, 0.1 M disodium hydrogen phosphate that had been pH 2.8-adjusted with phosphoric acid, and 0.13 M sodium dodecyl sulfate. The column was kept at 45 °C, and 267 nm was used for detection. For rupatadine, a linear response was obtained over the range of 2-160 ?g/ml, and for desloratadine, 0.4-8 ?g/ml. The technique was used to determine the amount of rupatadine in alergoliber syrup and tablets without the interference of propyl and methyl paraben, which are the primary excipients.[9]

A Shirkhedkar et al

For the purpose of analyzing rupatadine fumarate in both its bulk and tablet formulations, a high-performance thin-layer chromatographic technique that is straightforward, accurate, selective, and stability-indicating has been designed and validated. Toluene, methanol, and triethylamine 4:1:0.2 (v/v) were utilized as the mobile phase and aluminum foil TLC plates coated with silica gel 60F254 as the stationary phase. For rupatadine fumarate, a compact band (RF 0.61 ± 0.02) was discovered. At 264 nm, densitometric measurement was carried out in absorbance mode. The 400–1400 ng band?1 range showed a good linear association (r2 = 0.9992 ± 0.0001) between peak area and concentration, according to linear regression analysis. The slope and intercept had mean values ± SD of 2.5471 ± 0.005 and 1055.2 ± 4.20, respectively. The robustness, precision, and recovery of the approach were all verified. The quantitation and detection limits were 201.91 ng band?1 and 66.63 ng band?1, respectively. Rupatadine fumarate was broken down by oxidation, photochemical and thermal degradation, hydrolysis in both acidic and alkaline solutions, and all of these processes together. The technique allows for accurate, precise, and repeatable drug analysis, as demonstrated by statistical analysis. It can be applied to the quantitative analysis and identification of rupatadine fumarate in tablet formulations and in bulk drugs.[10]

RV Rele et al

Ion pair extractive spectrophotometric methods have received a lot of attention for the quantitative measurement of many pharmaceutical preparations because extractive spectrophotometric methods are popular since their sensitivity in drug assay is high. These suggested techniques are extractive spectrophotometric techniques that use chloroform as a solvent to determine the amount of rupatadine in tablet formulations. The resulting color ion-pair complexes are incredibly stable. By adjusting one parameter at a time while holding the other parameters constant, the operating conditions of these approaches were determined by watching the impact on the color species' absorbance. For these techniques, the different parameters that were needed to achieve maximal color development were optimized.

Accurate, simple and sensitive extractive spectrophotometric techniques has been developed for estimating rupatadine from pharmaceutical dose forms as rupatadine fumarate. The procedures relied on the drug's ability to create colored complexes in an acidic solution with reagents such as bromocresol green, Eriochrome black-T, and solo chrome dark blue. Under experimental conditions, the ion-associated complexes were generated and quantitatively extracted in chloroform. The measurements of absorbance were made at 416, 511, and 527 nm, in that order. The suggested techniques underwent statistical validation. Standard addition techniques were used to recover the procedures. For procedures I, II, and III, the linearity was determined to be 1–12 µg/ml, 2.5–50 µg/ml, and 100–600 µg/ml, respectively. High procedure precision is shown by low standard deviation and percentage RSD. Therefore, routine estimate of rupatadine as rupatadine fumarate in tablets can benefit from these methods.[11]

A Jani et al

The detection of rutadine and monteclast in pharmaceutical compositions has been made possible by the development and validation of a accurate, cost effective and simple stability indicating RP-HPLC method. Under a range of stress conditions, this approach can provide an assay of both drugs in the presence of their degraded products. Rupatadine degraded modestly under alkaline conditions, but montelukast was significantly vulnerable to photodegradation and acidic conditions. Techniques: HibarR 250-4, C-18 columns (250mm ×4.6mm,5um) were used for the chromatographic separation. A mobile phase of methanol: water (90:10v/v) with 0.1% triethylamine pH 3.41 adjusted with ortho phosphoric acid was used, with a flow rate of 1 ml/min. The discovered detection wavelength was 260 nm. Findings: Rupatadine and Montelukast were shown to have retention times of 4.31 and 11.59 minutes, respectively. For both medications, the technique was found to be linear over a range of 15–4 ?g/ml, with correlation coefficients (r2) of 0.999 and 0.996 for montelukast and rupatadine, respectively. Rupatadine and Montelukast yielded percentage recoveries of 99.49–100.25% and 99.52–100.53%, respectively, for both medications. It was discovered that the method's %RSD for accuracy and precision was less than 2%. In conclusion, the method's specificity, linearity, accuracy, precision, and robustness were all validated in accordance with the ICH criteria. The developed HPLC method is capable of resolving both drugs' degradant peaks. Thus, this approach is indicative of stability. Rupatadine and Montelukast's combination dosage form can be routinely analyzed using the proposed approach.[12]

VK Redasani et al

The simultaneous measurement of rupatadine fumarate (RPT) and montelukast sodium (MNT) was achieved by the development and validation of a accurate, sensitive, simple and selective high performance liquid chromatography (HPLC) technique. With a mobile phase of methanol: acetonitrile: buffer (40: 30: 30), (pH 3 with H3PO4) at a flow rate of 1.0 mL/min and column oven temperature of 40°C, isocratic chromatographic separation was obtained at 270 nm using a Hypersil BDS C8 (250 mm × 4.6 mm, 5 ?m) column. The method's selectivity and reproducibility for the simultaneous estimate of RPT and MNT are demonstrated by statistical analysis. The approach is useful for stability indicator since it can successfully separate medicines from their breakdown products. According to ICH criteria, the devised approach was validated in terms of precision, accuracy, specificity and linearity.[13]

AS Sutar et al

Design of Experiment aided stability suggesting that response surface approach was used in the design, development, and optimization of RP-HPLC for the simultaneous determination of Rupatadine fumarate and Montelukast sodium. Acetonitrile: Phosphate buffer (75:25) v/v, pH adjusted to 4.0, flow rate of 1 ml/min, and UV detection at 246 nm on RP-C18 column were used to achieve separation. The stress degradation investigations were carried out in accordance with established protocols. The procedure was certified in compliance with legal mandates. The validation results were found to be within the given range. Rupatadine was eluted at 13.25 minutes and montelloukast at 3.99 minutes, respectively. The new approach is suitable as evidenced by the well-resolved character of the stress degradation products from the drug peak. The creation of an experiment methodology can aid in the quick and cost-effective optimization of the mobile phase, saving time for the method's development. The new method is sensitive and accurate, and it can be used as a stability indicating method to identify degradation products in routine drug analyses.[14]

R Khatun et al

For the purpose of more accurately and precisely analyzing rupatadine in tablet and bulk dosage form, a quick, sensitive, reversed phase high performance liquid chromatographic technique has been designed and validated. Using a 150 × 4.6 mm, 5µm analytical column with an orthophosphoric acid-adjusted pH of 3.0, 0.02 M phosphate buffer, HPLC-grade methanol, and acetonitrile at a 45:30:25% v/v ratio, the chromatographic separation was accomplished. A photodiode array detector was used to measure the wavelength of the detector at 242 nm, the column oven temperature at 25ºC, and the flow rate of 0.5 ml/min. Rupatadine's retention time was 7.94 minutes throughout the 13-minute total run time. With a r2 value of 1.0, the standard curves were linear over the concentration range of 20–70 µl. For rupatadine, the theoretical plate was 7468 and the tailing factor was 1.21. For the characteristics of linearity, accuracy, stability of the solution, precision of the system, specificity, and robustness, the suggested method was validated in accordance with ICH guidelines. It is suitable for routine quality control analysis of rupatadine in tablet dosage form. All validation parameters of the analysis technique produced satisfactory results with an appropriate correlation coefficient and a decreased percentage RSD, indicating that the created approach can be utilized for stability, quality control and other research. The proposed RP-HPLC has been validated in accordance with the recommendation of ICH guidelines. The method is precise, simple, cost effective, less time consuming, accurate, and convenient to use.[15]

M Farooqui et al

A precise, cost effective and simple stability-indicating RP-HPLC method has been developed and validated for use in determining the amount of rutadine fumarate present in pharmaceutical tablets. Under various stress circumstances, the devised approach provided discrete detection and determination of rupatadine in the presence of degradant products. For chromatographic separation, a Spheris orb CN, 250 x 4.6mm, 5µ (Water, Ireland) was employed. Prior to use, the mobile phase was filtered and degassed. It was made up of a 40:60 ratio of acetonitrile to phosphate buffer pH 4.4, flowing at a rate of 1.5 milliliters per minute. A PDA detector was used to carry out the detection at 242 nm. The medication's susceptibility to UV, heat, and acid-base hydrolysis was demonstrated by subjecting the drug solution to stress degradation. The active ingredient in the drug was clearly isolated from the degradants. The technique was also validated in compliance with ICH standards. Therefore, it was discovered that the developed approach was both stability indicating and specific. Based on the findings regarding column and mobile phase variations, the Spherisorb CN (Cyano) column and the mobile phase specified in the chromatography conditions were deemed the most appropriate columns for validation and optimization, with the ultimate goal of a better overall result, improved sensitivity, quicker analysis, and resolution. The results of the stress research show that the medication is prone to degradation by heat, UV light, hydrogen peroxide, and acid-base hydrolysis. More polarity than the analyte itself is indicated by a lower RT of the degraded components, whereas less polarity is indicated by a higher RT of the degraded components. The active medicinal ingredient is isolated from the degradants. Therefore, it was discovered that the developed approach was both specific and stability indicating.[16]

A Almahri  et al

This work used two proven methods to determine rupatadine through the first application of spectrofluorimetric and resonance Rayleigh scattering techniques. The suggested techniques relied on the easy development of an association complex in an acidic media between erythrosin B reagent and rupatadine. The quenching impact of rupatadine on the fluorescence intensity of erythrosin B at 556 nm (excitation = 530 nm) was the basis for the spectrofluorimetric determination. On the other hand, the resonance Rayleigh scattering (RRS) technique depended on an increase in erythrosin B's resonance Rayleigh scattering spectrum at 344 nm following the addition of rupatadine. The developed methods yielded linear results spanning ranges 0.15?2.0 ?g/ml and 0.1?1.5 ?g/ml, respectively, with spectrofluorimetric method and RRS method detection limits of 0.030 ?g/ml and 0.018 ?g/ml. The International Council for Harmonization's criteria were followed in the validation of both procedures, and all reaction conditions for the formation of rupatadine-erythrosin B were optimized experimentally. The rupatadine concentration in its pharmaceutical tablet dosage form was estimated using the described procedures with satisfactory recoveries. Additionally, in compliance with US Pharmacopeia criteria, a content uniformity test for the commercial rupatadine tablets was successfully conducted using the recommended spectroscopic methods.

Through electrostatic attraction and hydrophobic forces, rupatadine and erythrosine B can react to produce an ion-pair complex in a slightly acidic media. This process will determine a new spectral resonance. Spectrophotometric analysis and Rayleigh scattering Techniques for estimating rupatadine were created and verified using high sensitivity, simplicity, speed, and selectivity. Additionally, the sample The preparation procedures for both approaches were straightforward and unnecessary. laborious procedures. Furthermore, following dilution, the generated complex was assessed using Distilled water was used for both techniques. Ultimately, the suggested techniques were effectively utilized to examine rupatadine in the dose form of pharmaceutical tablets and to evaluate the consistency of RPT's content in its dosage form as pharmaceutical tablets.[17]

Raja T et al

A precise, simple, accurate and simple reversed-phase high performance liquid chromatography (RP-HPLC) method has been developed and validated for the simultaneous estimation of Montelukast Sodium and Rupatadine Fumarate in their combination dosage form. The proposed method relies on employing a Symmetry C-8 analytical column (150 x 4.6 mm; 5 ?) to separate the two medicines in reversed-phase mode. Acetonitrile: phosphate buffer pH 4.7 adjusted with o-phosphoric acid (60:40, v/v) made up the ideal mobile phase. The UV detection wavelength was set at 254 nm. The mobile phase flow rate was 1.2 mL min-1. For Rupatadine Fumarate and Montelukast Sodium, the retention periods were 3.22 and 10.67 minutes, respectively. The approach was verified in compliance with ICH regulations. It was discovered to be repeatable and accurate. For both Rupatadine Fumarate and Montelukast Sodium, linearity was achieved in the concentration range of 100-300 ?g mL-1, with correlation coefficients of 0.999 and 0.999, respectively. For both drugs, the mean percent recovery of triplicate samples at each level was found to be between 98.7% and 99.5%, with an RSD of less than 2.0%. The suggested approach can be effectively used for pharmaceutical dosage forms and bulk production quality control.[18]

NS Dighe et al

For the purpose of simultaneously estimating Rupatadine fumarate and Montelucast sodium from pharmaceutical dosage forms, a sensitive, simple, quick and selective isocratic reversed phase High Performance Liquid Chromatography method has been developed. The mobile phase consists of a mixture of methanol, acetonitrile, and buffer 40:30:30 (pH adjusted to 3.2 using ortho phosphoric acid) at a flow rate of 1.0 mL/min. A stationary phase was a 250 mm x 4.6 mm Hypersil BDS C8 column with 5µ particle sizes. Rupatadine fumarate and Montelucast sodium had retention times of 2.79 and 3.97 minutes, respectively. At 270 nm, the eluent was found. The suggested technique for the simultaneous determination of montelucid sodium and rutadine fumarate is quick, accurate, selective, and exact.

The experiment's modalities were successfully validated in accordance with the analytical techniques outlined in routine analysis and the recommendations established by the ICH. Recovery studies and preliminary analysis of the standard sample were used to validate the suggested approach. The suggested RP-HPLC method for the simultaneous determination of Rupatadine fumarate and Montelucast sodium is precise, linear, accurate and robust, according to the results obtained. Analysis of the combined dose tablet formulation has shown the usefulness of the suggested approaches. As a result, these constituents' quantitative determination in the formulation of combination dose tablets can be accomplished using the suggested method.[19]

DR Nogueria et al

Using nimesulide as the internal standard, a stability-indicating MEKC was developed and validated for the analysis of rupatadine in tablet dosage forms. A fused-silica capillary (50 ?m id; effective length, 40 cm) was employed to conduct the MEKC technique. The BGE was composed of a pH 10 solution of 25 mM anionic detergent SDS and 15 mM borate buffer. A 25 kV applied voltage was used to maintain the capillary temperature at 35°C. The injection was carried out in the hydrodynamic mode for five seconds at 50 mbar, and a photodiode array detector tuned to 205 nm was used for detection.In the range of 0.5–150 ?g/mL, the technique was linear (r2 = 0.9996). Degradation experiments, including those using MS, demonstrated the method's specificity and stability-indicating ability and demonstrated that there was no excipient interference and increase in cytotoxicity. 99.98?curacy was achieved with a bias of less than 1.06%. 0.1 and 0.5 ?g/mL were the LOD and LOQ, respectively. When the suggested approach was successfully used to analyze rupatadine quantitatively in pharmaceutical formulations, the results showed no statistically significant difference (p >0.05) when compared to an established RP-LC method.[20]

Y Tian et al

Using estazolam as the internal standard (IS), a selective, sensitive, rapid, & simple liquid chromatographic–tandem mass spectrometric (LC–MS/MS) technique was created and verified for the measurement of rupatadine in human plasma. After being extracted , the analytes were separated on a reverse phase C18 column using a mobile phase of methanol–ammonium acetate (pH 2.2; 5 mM) (50:50, v/v). The analytes were then analyzed by a triple-quadrupole mass spectrometer in the positive ion and multiple reaction monitoring (MRM) mode, with rupatadine being extracted at m/z 416 ? 309 and the IS being extracted at m/z 295 ? 267. The assay demonstrated a linear dynamic range for rupatadine in human plasma of 0.1–100 ng/ml. With a relative standard deviation of less than 20%, the lower limit of quantification (LLOQ) was set at 0.1 ng/ml.For concentrations over the range of the standard curve, acceptable precision and accuracy were attained. Rupatadine's pharmacokinetics have been effectively studied in healthy volunteers using the proven LC-MS/MS technology. In conclusion, a technique for measuring rupatadine in human plasma employing multiple reaction monitoring in the positive ionization mode of LC-MS/MS is presented. When measuring rupatadine at concentrations between 0.1 and 100 ng/ml in human plasma samples derived from pharmacokinetic, bioavailability, or bioequivalency studies, this technique has demonstrated sufficient sensitivity and acceptable precision.[21]

J Wen et al

A sensitive method for measuring rupatadine and its metabolite desloratadine simultaneously in human plasma was developed using liquid chromatography/tandem mass spectrometry (LC–MS/MS). Following the addition of the internal standard (IS), diphenhydramine, plasma samples were extracted using a 1:1, v/v mixture of methyl tert-butyl ether and n-hexane. AUltimateTM AQ-C18 (4.6 mm × 100 mm, 5 ?m) column was used for the study, and its mobile phase included an 80/20 methanol/water combination with 0.0005% formic acid injected at 0.3 ml min?1. In multiple reaction monitoring mode, the analytes and the IS were identified in positive ionization mode, while their precursor ? product ion combinations were monitored at m/z 416 ? 309, 311 ? 259, and 256 ? 167, respectively.For rupatadine and desloratadine, the assay's linear ranges were 0.1–50 and 0.1–20 ng ml?1, respectively. Both desloratadine and rupatadine have great sensitivity and selectivity, with dependable quantitative lower limits of 0.1 ng ml?1. Less than 7.2% was the accuracy between and between runs. In quality control samples at three levels, the accuracy ranged for rupatadine and desloratadine, respectively, from ?9.2% to +6.4% and ?7.2% to +7.2%. The technique has been successfully used to investigate the pharmacokinetics of rupatadine and its primary metabolite in healthy Chinese volunteers who were given tablets containing 10, 20, and 40 mg of rupatadine orally.To simultaneously determine rupatadine and its active metabolite desloratadine in human plasma, a sensitive and selective LC–MS/MS technique has been developed and validated. The suggested technique demonstrated high sensitivity, achieving an LLOQ of 0.1 ng ml?1 for both desloratadine and rupatadine. One-step liquid-liquid extraction was used to pretreat plasma samples before they were subjected to an isocratic LC analysis. Additionally, each sample took 5.0 minutes to analyze in total.[22]

C Sun et al

For the purpose of simultaneously determining rupatadine (RT) and its two active metabolites, desloratadine (DT) and 3-hydroxydesloratadine (3-OH-DT), in human plasma, a simple LC–ESI–MS/MS method was created and verified. On a C18 column with gradient elution, the chromatographic separation was performed using methanol and 10 mM ammonium acetate with 0.1% (v/v) formic acid. For RT, DT, and 3-OH-DT, the corresponding lower limit of quantification (LLOQ) was 0.05, 0.035, and 0.035 ng/mL.Analytes' intra- and inter-day precision fell between 1.0 and 4.7% and 2.2 and 12.1%, respectively. Analytes' intra- and inter-day accuracy fell between -7.7% and 5.2% and -4.1% and 4.8%, respectively.The technique was effectively used in a pharmacokinetic investigation of rupatadine and its two metabolites, DT and 3-OH-DT, in healthy individuals after oral dosages of rupatadine fumarate tablets (10, 20, 40 mg) given singly and multiple (10 mg). For the simultaneous measurement of RT and its metabolites DT and 3-OH-DT in human plasma, a straightforward LC-MS/MS approach has been created and validated. With an LLOQ of 0.05 ng/mL for RT and 0.035 ng/mL for DT and 3-OH-DT, the method is very sensitive and appropriate for pharmacokinetic studies. Throughout the trial, there were no adverse events at any dose, and the medication was well taken with negligible side effects.[23]

ACKNOWLEDGEMENTS

We express our sincere thanks to the Department of Pharmaceutical Chemistry, College of Pharmacy, Madras Medical College (MMC), Chennai for providing necessary facilities for the research work.

CONFLICTS OF INTEREST

The author declares there is no conflict of interest.

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