Accurate Ultra-performance Liquid Chromatography Research Articles

Development of a Stability Indicating UPLC Method for the Determination of Tirbanibulin in Bulk and Its Pharmaceutical Dosage Form.

The primary goal of this study was to create and validate a simple, precise, sensitive, and accurate ultra-performance liquid chromatography (UPLC) method for estimating tirbanibulin in pure and dosage form. A UPLC technique was developed using a Waters Acquity UPLC Phenyl (100 x 2.1 mm, 1.7 µm) column. The developed technique was validated in accordance with the International Conference on Harmonization (ICH) guidelines. Tirbanibulin was separated chromatographically with high resolution using the mobile phase acetonitrile: buffer (30:70 v/v) at 0.5 mL/min, 5 µL injection volume, and 220 nm wavelength. The validated technique was found to be linear in the 1-15 µg/mL range. The detection and quantification limits for tirbanibulin were 0.03 and 0.1 µg/mL, respectively. The percentage relative standard deviation was less than 2%, demonstrating the precision of the developed technique. Furthermore, the recovery rate was nearly 100%, confirming the accuracy of the method. Minor modifications to the chromatographic conditions demonstrated the robustness of the method. The developed analytical method was precise, simple, reproducible, and sensitive. Consequently, it can be used to determine tirbanibulin.

Detection of lenalidomide metabolites in urine to discover drug-resistant compounds

Lenalidomide is the first-line drug for the clinical treatment of multiple myeloma. However, its efficacy differs significantly among patients. Clinically, after lenalidomide treatment, few patients’ conditions worsened, whereas others remained stable or improved. To clarify the reasons for this difference in efficacy, 20 patients with multiple myeloma who received maintenance treatment with lenalidomide were retrospectively included in this study. Lenalidomide metabolic compounds were detected in patient urine using mass spectrometry. A rapid and accurate ultra-performance liquid chromatography–time-of-flight tandem mass spectrometry (UPLC-TOF-MS/MS) method was used to characterize metabolites in the urine of different patients. Eleven metabolites, including four new compounds, were identified and characterized in all the samples. Among these, two metabolites were found to have obvious discrepancies in different groups of patients. One metabolite named Denitrified-2 glutarimide, a new potential compound, was only detected in the urine of ineffective and stable patients, whereas the other metabolite named 5-Hydroxy-lenalidomide was found only in the urine of effective patients.

METHOD DEVELOPMENT AND VALIDATION OF FLURBIPROFEN SODIUM BY USING UPLC

A simple, precise, and accurate ultra-performance liquid chromatography was developed for flurbiprofen sodium on Acquity BEHC18, (50 x 2.1 mm, 1.7 µm) using acetonitrile: water: glacial acetic acid as the mobile phase. The flow rate was 0.2 mL min-1 and effluent was monitored at 254 nm. The retention time for flurbiprofen sodium was found to be 2.3. Developed method was validated for precision, accuracy, linearity range, robustness and ruggedness as per ICH guideline

Simultaneous determination of cis- and trans-palmitoleic acid in rat serum by UPLC–MS/MS

Palmitoleic acid, a monounsaturated fatty acid which could affect glucose and lipid metabolism and reduce insulin resistance has two isomers, i.e. cis-palmitoleic acid (cPOA) and trans-palmitoleic acid (tPOA). However, the pharmacokinetic, metabolic transformation and structure–activity relationship of the two isomers have not been reported. A precise and accurate ultra performance liquid chromatography–tandem mass spectroscopy (UPLC–MS/MS) method was developed to determine cPOA and tPOA simultaneously. Both the cPOA and tPOA were administered i.g. (intragastric gavage) to rats at 75 mg/kg. Serum samples were collected and analyzed for the two isomers by UPLC–MS/MS on a reverse-phase BDS C18 column equilibrated and eluted with water (A) and acetonitrile (B) at a flow rate of 0.3 mL/min. The calibration curves for cPOA and tPOA were linear over the range 0.1–12 μg/mL. Analytes were monitored by selected-reaction monitoring in negative electrospray ionization mode. The Tmax of cPOA was 0.94 ± 0.44 h and the Cmax 8.17 ± 1.97 μg/L, and the Tmax and Cmax of tPOA were 1.50 ± 0.98 h and 14.77 ± 11.91 μg/L, respectively. AUC0–24 h of cPOA and tPOA were 59.45 ± 29.83 and 113.88 ± 72.25 mg/L·h. The method was applied in pharmacokinetic study of cPOA and tPOA in rat serum successfully. Besides, the concentrations of cPOA and tPOA in rat serums were observed fluctuating with a consistent trend, which may be due to reciprocal bio-convert in the body.

Stability Indicating Quality by Design-based Development and Validation of Bilastine and Montelukast by Ultra Performance Liquid Chromatography Method

Background: The current work, which was conducted under the quality by design (QbD) paradigm, aims to establish the optimization of ultra-performance liquid chromatography (UPLC) using the design of experiments and response surface theory, such as central composite design (CCD), in order to achieve a good separation and resolution. Methods: The mobile phase was composed of methanol (20:80% v/v) and 0.1% tri fluro amine (TFA) buffer, and chromatographic separation was performed on column phenomenex C18 (50 mm x 4.6 mm x 2.5 m) at a flow rate of 0.4 mL/ min. Montelukast (MON) and bilastine (BIL) were both detected at 260 nm. The suggested method was approved in accordance with International Council for Harmonization (ICH) recommendations. Results: The created technique successfully separates both BIL and MON with a chromatographic resolution of 6.4. The technique was linear for BIL and MON concentrations of 0.5–3.0 μg/mL and 0.25–1.5, respectively, and recovery rates ranged from 99–100%. The drug compounds had distinct degradation products that were identified in stress patterns. Conclusion: For the quantification of BIL and MON in the combination tablet, a precise, simple, reliable, and accurate UPLC method was designed. According to the method validation, it was possible to successfully separate two medicines from their degradants utilizing aQbD techniques.

Ultra-Performance Liquid Chromatographic and Densitometric Methods for Sensitive Determination of Xipamide and Triamterene in Pure and Pharmaceutical Dosage Forms.

Validated ultra-performance liquid chromatography (UPLC) and thin-layer chromatography (TLC) densitometric methods were prescribed for determination of antihypertensive components. To establish and validate rapid and accurate UPLC and TLC densitometric methods for determination of Xipamide and Triamterene in pure and dosage forms. The first method, UPLC, depended on using an Agilent Zorbax Eclipse Plus C8 (50 mm × 2.1 mm, 1.8 μm) column, a mobile phase composed of acetonitrile-water (70 + 30, v/v) adjusted by acetic acid to obtain pH 3, 0.2 mL/min flow rate, and UV detection at 231.4 nm. The second method was a TLC densitometric method. Separation was achieved by using toluene-methanol-ethyl chloride-acetic acid (7 + 2 + 1 + 0.2, v/v/v) as the mobile phase, pre coated silica gel plates as the stationary phase, and UV detection at 300.0 nm. The obtained results were validated and statistically compared with official and reported methods. The obtained results showed high accuracy and reproducible results with excellent mean recoveries for both drugs. The UPLC method showed shorter retention time for both Xipamide (0.88 min) and Triamterene (0.63 min), a lower detection limit of less than 0.055 µg/mL for both drugs with high selectivity, decreased injection volume (1 µL), and a lower flow rate than any HPLC method. Both proposed methods were sensitive, selective, and effectively applied to pure and dosage forms (Epitens®). Unprecedented sensitive, rapid, and reproducible UPLC and TLC methods were developed for selective determination of mixtures of Xipamide and Triamterene, with LOD os less than 0.076 µg/mL for both drugs.

Pharmacokinetic Study of Zhebeirine in Mouse Blood by Ultra- Performance Liquid Chromatography/tandem Mass Spectrometry

Introduction: In this study, a precise, rapid and accurate ultra-performance liquid chromatography- tandem mass spectrometry (UPLC-MS/MS) method for the quantification of zhebeirine in mouse blood was developed, and pharmacokinetics of zhebeirine was studied for the first time after intravenous and oral administration. Methods: The mobile phase consisted of acetonitrile-0.1% formic acid, with a flow rate at 0.4 mL/m during 4 min run time. MRM modes of m/z 414.5→81.0 for zhebeirine and m/z 430.2→412.2 for 3- dehydroverticine (internal standard) were utilized to perform quantitative analysis. Protein in mouse blood was directly precipitated with acetonitrile for sample preparation. Results: The linear range was 1-3000 ng/mL with r>0.995, and LLOQ was 1 ng/mL. The intra-and inter-day precision of zhebeirine in mouse blood was less than 13%. The accuracy ranged from 91.2% to 112.5%, while the matrix effects were between 84.8% and 106.4%. Conclusion: The UPLC-MS/MS was successfully applied to the pharmacokinetic study on zhebeirine after intravenous and oral administration, and the bioavailability was determined to be 22.8%.

Pharmacokinetic Comparisons of Eight Active Components from Raw Farfarae Flos and Honey-Processed Farfarae Flos after Oral Administration in Rats by UHPLC-MS/MS Approaches.

Farfarae flos (FF) is widely used for cough over thousands of years in China, but little is known about their pharmacokinetics properties. This study was aimed to establish a rapid and accurate ultraperformance liquid chromatography with triple-quadrupole tandem mass spectrometry method for compare pharmacokinetics studies of eight active compounds after oral administration between raw and honey-processed farfarae flos extracts. Optimum separation was performed on a Thermo Hypersil GOLD C18 column (100 mm × 2.1 mm, 1.9 µm particles size) with a gradient elution of acetonitrile as mobile phase A and 0.3% formic acid aqueous solution as mobile phase B. The flow rate was set as 0.3 mL/min and separated for 34.0 minutes. Electrospray ionization in the negative ion mode and selected reaction monitoring were used to identify and separate active components. The results met the acceptance criteria and showed that this method exhibited good linear, precision, accuracy, and stability. The extraction recoveries ranged from 81.54% to 104.48%, and the matrix effects ranged from 81.94% to 103.02%. These results show that the validated method could be successfully applied to evaluate the pharmacokinetic study in rats after oral administration of raw farfarae flos (R-FF) and honey-processed farfarae flos (H-FF).

Ultra-performance liquid chromatography–tandem mass spectrometric determination of ramipril in human plasma

Purpose: To develop a sensitive and accurate ultra-performance liquid chromatography–tandem mass spectrometric (UPLC-MS) method for quantification of ramipril in human plasma.Methods: Ramipril was extracted from biological fluid using equal volumes of n-hexane and propanol (1:1, v/v), and then chromatographed in a suitable C18 column with methanol: 0.1 % HCOOH (4: 1, v/v) as mobile phase. Atorvastatin was used as an internal standard for the chromatographic separation and quantification. The method was validated according to the United States Food and Drug Administration guidelines for standard indices.Results: Ramipril was determined in the concentration range 0.05 and 1000 ng/mL the validation procedure exhibited a correlation coefficient of 0.9979 + 0.002 (p = 0.05). The studied drug was quantified with lower ceiling of 0.05 ng/mL, and showed an accuracy of 105.00 %.Conclusion: A sensitive UPLC-MS analytical method has been successfully developed for the quantification of ramipril in human plasma. This method can be applied efficiently for the quantification of ramipril in bioavailability and pharmacokinetic studies.
 Keywords: Liquid chromatography–tandem mass, Ramipril, Stability, Biological fluids, Plasma

Determination and pharmacokinetics and bioavailability of O-demethyl nuciferine in mice by UPLC–MS/MS

In this study, a precise, rapid, and accurate ultra-performance liquid chromatography–tandem mass spectrometer (UPLC–MS/MS) method for the quantitation of O-demethyl nuciferine in mouse blood was developed, and pharmacokinetics of O-demethyl nuciferine was studied for the first time after sublingual injection and gavage. The study was performed with an UPLC ethylene bridged hybrid (UPLC BEH) (2.1 mm × 50 mm, 1.7 μm) column at 30 °C, using diazepam as the internal standard (IS). The mobile phase consisted of acetonitrile–10 mmol/L ammonium acetate (containing 0.1% formic acid), with a flow rate of 0.4 mL/min for 4 min run time. Multiple reaction monitoring (MRM) modes of m/z 282.1→219.0 for O-demethyl nuciferine and m/z 296.2→265.1 for IS were utilized to conduct quantitative analysis. Protein in mouse blood was directly precipitated with acetonitrile for sample preparation. The linear range was 1–500 ng/mL with r > 0.995, and the lower limits of quantification (LLOQ) was 1 ng/mL. The intra- and inter-day precision of O-demethyl nuciferine in mouse blood were RSD < 14% and RSD < 15%, respectively.r The accuracy ranged from 89.0% to 110.7%, with a recovery higher than 88.9%, while the matrix effect was between 103.1% and 108.7%. We further applied this UPLC–MS/MS method to the pharmacokinetic study on O-demethyl nuciferine after sublingual injection and gavage and determined the bioavailability to be 6.4%.

Determination of ethanamizuril, a novel triazine coccidiostat, in rat plasma by ultra-performance liquid chromatography system-tandem mass spectrometry and its application in a toxicological study.

Ethanamizuril is a new triazine compound that has the potential to be a novel anticoccidial drug. Toxicological studies in experimental rats were performed to understand the safety profile of ethanamizuril for drug product development. In this study, a novel, selective and accurate ultra-performance liquid chromatography tandem mass spectrometry method has been developed for the determination of ethanamizuril concentrations in rat plasma. With 4-nitro-o-cresol as an internal standard, sample pretreatment involved a one-step extraction with acetonitrile of 100 μL plasma. The detection was carried out by electrospray ionization mass spectrometry in negative ion mode with selected ion recording. The standard curves were linear (r2 ≥ 0.999) over the concentration range of 0.1-100 μg/mL. The relative standard deviations of intra- and inter-day precisions were less than 8.4 and 8.87%, respectively. The mean extraction recovery of ethanamizuril from rat plasma was 97.68-102.57%. The method was fully validated and successfully applied to monitor plasma concentrations of ethanamizuril in a short-term toxicity study and two-generation reproduction toxicity study. The result of the study confirmed that the elimination of ethanamizuril in rats is slow.

Simultaneous Determination of Methocarbamol and Paracetamol in the Presence of Three Related Substances by Ultra Performance Liquid Chromatography

Introduction:A rapid, and accurate Ultra Performance Liquid Chromatography (UPLC) method was developed to simultaneously analyze Methocarbamol, Paracetamol and the related substancesMaterials and Methods:Waters ACQUITY UPLC® BEH Phenyl C18 column was used in conjunction with UV detection at 225nm. Gradient elution with 0.05M, pH 6 phosphate buffer and acetonitrile flow at 0.3mL /min rate were used to separate the substances. The retention times for 4-Aminopheno, Paracetamol, Guaifenesin, Methocarbamol, and 4-Chloroacetanilide were 1.319 minute, 2.224 minute, 4.467 minute, 4.769 minute and 5.433 minute respectively. The concentration was linear in the range of 2-100 µg/ml for Methocarbamol, and 1-100 µg/mL for Paracetamol. The percentage recoveries were between 99.28±1.23% to 100.57±0.99% for Methocarbamol, and between 99.08±1.23% to 101.23±1.39% for Paracetamol.Results and Discussion:The validated optimal protocol is robust and accurate for simultaneous analysis of Methocarbamol, Paracetamol and the related substances, applicable for bulk powder as well as pharmaceutical formulation.Conclusion:In this paper, a highly sensitive, accurate, and precise UPLC method with UV-Vis detection was developed and validated for quality control of MET and PAR in bulk as well as in pharmaceutical preparations.

Validated UPLC/MS/MS assay for quantitative bioanalysis of elbasvir in rat plasma and application to pharmacokinetic study.

Rapid, sensitive, selective and accurate ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the quantification of elbasvir (ELB) in rat plasma with deuterated elbasvir (ELB-D6) as internal standard (IS).Sample preparation was done by protein precipitation using acetonitrile containing 50 ng/mL IS. Chromatographic separation was achieved by an UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) column with a gradient mobile phase consisting of acetonitrile-water (containing 5.0mM ammonium acetate with 0.01% acetic acid, pH 4.5) as mobile phase at a flow rate of 0.3 mL/min for 3 min. ELB was monitored using positive electrospray triple quadrupole mass spectrometer (Waters Xevo TQ-S) via multiple reaction monitoring (MRM) mode. The monitored transitions were set at m/z 882.51→656.42 and m/z 888.49→662.43 for ELB and ELB-D6, respectively. The achieved lower limit of quantification was 1.0 ng/mL. The validated method had an excellent linearity in the range of 1.0-2000 ng/mL (r(2)>0.996). Recovery efficiency at three levels QC concentrations of 2.0 (low), 160 (medium) and 1600 (high) ng/mLwas in the range of 98.29-106.40% for ELB. Matrix effect was found to be minimal. The intra- and inter-day precisions were less than 7.01%. The intra- and inter-day accuracies were determined to be within ±6.23% for all accuracy measurements. The validated simple and rapid UPLC-MS/MS method was successfully used to the pharmacokinetics study of ELB in rats, providing its applicability in relevant preclinical studies.

Elucidating the structure of carbon nanoparticles by ultra-performance liquid chromatography coupled with electrospray ionisation quadrupole time-of-flight tandem mass spectrometry

A fast and accurate ultra-performance liquid chromatography coupled with electrospray ionisation quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-Q-TOF-MS/MS) method was developed for the separation and structural elucidation of fluorescent carbon nanoparticles (CNP). The CNP was synthesised from microwave-assisted pyrolysis of citric acid (CA) and 1,2-ethylenediamine (EDA). By using UPLC separation, the CNP product was well separated into ten fractions within 4.0 min. Based on high-accuracy MS and MS/MS analyses, the CNP species were revealed to display six kinds of chemical formulas, including (C10H20N4O5)n, (C8H12N2O5)n, (C16H22N4O9)n, (C6H8O7)n, (C14H18N2O11)n, and (C14H16N2O10)n. In particular, our study revealed for the first time that the CNP species exist as supramolecular clusters with their individual monomers units linked together through non-covalent bonding forces. These findings clearly indicated the usefulness of UPLC-ESI-Q-TOF-MS/MS in identifying the chemical composition of CNP product. It is anticipated that our proposed methodology can be applied to study the structure-property relationships of CNP, facilitating in the production of CNP with desirable spectral features.

Development and Validation of UPLC Method for the Determination of Duloxetine Hydrochloride and Its Impurities in Active Pharmaceutical Ingredient

A suitable, rapid, sensitive and accurate ultra-performance liquid chromatography (UPLC) method was developed for the quantitative determination of Duloxetine hydrochloride and its impurities in active pharmaceutical ingredient. Chromatographic separation was achieved on shim-pack XR-ODS II (3.0 × 100 mm, 2.2 μm), and the gradient eluted within a period of time, that is, 15 minutes. The eluted compounds were monitored at 230 nm. The flow rate was 0.9 ml/min and the column oven temperature was maintained at 40oC. The resolution of Duloxetine hydrochloride and 12 impurities (potential impurity, process related impurity and degradation products) were greater than 1.3. The correlation coefficient (r2>0.99) values indicated clear correlations between the investigated compound concentrations and their peak areas within the quantitation limit to 200% level. The performance of the method was validated according to the present ICH guidelines for specificity, quantitation limit, detection limit, linearity, accuracy, precision, ruggedness and robustness. The recoveries obtained (93.28-102.41%) ensured the accuracy of the developed methods.

Chemical Prospection of Important Ayurvedic Plant Tinospora cordifolia by UPLC-DAD-ESI-QTOF-MS/MS and NMR

A rapid, sensitive, and accurate ultra-performance liquid chromatography coupled with mass spectrometric method (UPLC-MS) was developed and validated for simultaneous determination of four bioactive compounds, syringin (3), cordifolioside A (4), magnoflorine (6) and tinocordiside (10) in the stem of Tinospora cordifolia. The analysis was performed using an Acquity C18 column and gradient elution of 0.05% formic acid in water and acetonitrile at a detection wavelength of 267 nm in 5 min. A high correlation coefficient (r2 > 0.998) indicated good correlation between investigated compounds concentration and their peak area within the test ranges. The LODs for compounds 3, 4, 6 and 10 were 1.95, 0.97, 3.90 and 0.97 ng/mL, respectively, and LOQs were 6.64, 3.20, 12.87 and 3.20 ng/mL, respectively. The overall intra- and inter-day variations of the four compounds were less than 1%. The variation of these four bioactive compounds in T. cordifolia hosted on fifteen different trees was also determined. The compounds (3, 4, 6 and 10) were found in high amount in the T. cordifolia hosted on Azadirachta indica and Mangifera indica as compared with other plants. Twelve compounds were identified on the basis of their mass and UV-vis spectra. The NMR fingerprinting of the extract revealed the presence of alkaloids, fatty acid methyl esters, polysaccharides and marker components of T. cordifolia.

Photocatalytic degradation of four non-steroidal anti-inflammatory drugs in water under visible light by P25-TiO2/tetraethyl orthosilicate film and determination via ultra performance liquid chromatography electrospray tandem mass spectrometry

Photocatalytic degradation of four non-steroidal anti-inflammatory drugs (NSAIDs) — salicylic acid, ibuprofen, naproxen and diclofenac in water has been investigated using a novel P25-TiO2/tetraethyl orthosilicate (TEOS) film on glazed ceramic surface under a visible light incubator. Influences of main parameters, such as adsorption, stir, initial pH, amount of P25-TiO2/TEOS and irradiation time has been studied. Results show that the degradations of four NSAIDs obtained maximum and equilibrium effect within 10h of irradiation at pH 6.0 condition. The maximum degradation effects of salicylic acid, ibuprofen, naproxen and diclofenac under optimum conditions are 76%, 85%, 94% and 65%, respectively. The photocatalytic degradation reactions follow first order kinetics. Naproxen is selected as the representative to analyze the photocatalytic products, and a probable photocatalytic degradation pathway of naproxen is put forward. To support the degradation kinetics study a rapid, sensitive and accurate ultra performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) method has been developed.

A UPLC-MS/MS Method for Qualification of Quercetin-3-O-β- D-glucopyranoside-(4®1)-α-L-rhamnoside in Rat Plasma and Application to Pharmacokinetic Studies

A sensitive and accurate ultra-performance liquid chromatography coupled with triple quadrupole mass (UPLC-MS/MS) method was developed for the determination of quercetin-3-O-β-D-glucopyranoside-(4→1)-α-L-rhamnoside (QGR) in rat plasma using rutin as internal standard. Chromatographic separation was achieved on a Acquity BEH C18 column (100 mm × 2.1 mm, 1.7 μm) with a gradient elution of acetonitrile and 0.10% formic acid (v/v) at a flow rate of 0.4 mL/min. QGR and rutin were detected using electrospray negative ionization mass spectrometry in the multiple reaction monitoring (MRM) mode. The method demonstrated good linearity and did not show any endogenous interference with the QGR and rutin peaks. This method was successfully applied to a pharmacokinetic study of QGR in rats after intravenous (20 mg/kg) and oral (40 mg/kg) administration, and the results showed that the compound was poorly absorbed, with an absolute bioavailability of approximately 3.41%.

Determination of free and total warfarin concentrations in plasma using UPLC MS/MS and its application to a patient samples

Warfarin is routinely monitored by assessing its pharmacologic effects on the international normalized ratio. However, having a patient with INR not responding to increasing warfarin dose mandates a direct measurement of warfarin concentrations (total and free) for better patient clinical management of warfarin therapy. Therefore, a new fully validated specific, precise and accurate ultra-performance liquid chromatography tandem mass spectrometry was developed for the determination of free and total warfarin in human plasma. Free warfarin was measured in plasma filtrate, prepared by ultrafiltration, and sample pretreatment involved protein precipitation with acetonitrile. Linear response (r(2) ≥0.99) was observed over the studied range of free and total warfarin, with the lower limit of detection of 0.25 ng/mL. The intra- and inter-day precision (relative standard deviation) values were <10% and the accuracy (relative error) was ≤6.6 for free and total warfarin. There was no significant difference (p>0.05) between inter- and intra-day studies for the free and total warfarin, which confirmed the reproducibility of the assay method. The mean extraction efficiency was 88.6-107.2% of free and total warfarin. The assay was sensitive to follow warfarin pharmacokinetics (free and total) in a patient with resistance to warfarin up to 24 h after a daily dose of warfarin.

Pharmacokinetics of ketorolac loaded to polyethylcyanoacrylate nanoparticles using UPLC MS/MS for its determination in rats

Polyethylcyanoacrylate (PECA) nanoparticles (NPs) have been employed as biodegradable polymeric carriers for oral (PO) delivery of ketorolac. The nanoparticles were prepared by polymerization technique at room temperature in a continuous aqueous phase at pH 2.5. This polymerization technique was able to hold 76–96% of ketorolac and the drug loading was a monomer concentration dependent. The feasibility of PECA NPs as PO controlled drug delivery systems of ketorolac was investigated in two groups of rats which were given orally either ketorolac tromethamine solution (1.5 mg/kg) or the selected ketorolac NPs aqueous dispersion (1.6 mg/kg). Ketorolac plasma concentrations were measured by a new fully validated specific, precise and accurate ultra-performance liquid chromatography tandem mass spectrometry (UPLC MS/MS) assay. The detection was performed on Waters TQ detector via negative electrospray ionization in a multiple reaction monitoring mode. Linear response ( r 2 ≥ 0.995) was observed over the range of 10–10,000 ng/ml of ketorolac, with the lower limit of quantification of 5 ng/ml with 1 μl injection volume. The intra- and inter-day precision (relative standard deviation, R.S.D.) values were <10% and the accuracy (relative error) was ≤8 for ketorolac concentrations. The drug solution is rapidly absorbed, distributed, and eliminated and shows a monophasic elimination phase. The assay was sensitive to follow pharmacokinetics of ketorolac in rats up to 24 h after a PO dose of its aqueous solution or NPs suspension. After NPs administration the mean Cmax, 5.0 ± 1.3 mg/l, was attained at 1 h. The drug was successfully maintained around this elevated plasma drug concentration up to 6 h (>2 t 1/2), in rats. The AUC was significantly higher after the NPs suspension than the solution of ketorolac. This present study provides evidence that the sorption of ketorolac to PECA NPs could control the drug release/elimination in rats.