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Investigation of motilin's surface adsorption on glass and polypropylene surfaces using a simple and specific HPLC method

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JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 2025年

作者： Karaca, Sakine Atila ugur, duygu yeniceli Anadolu Univ Fac Pharm Dept Analyt Chem Eskisehir Turkiye

The nonspecific binding of peptides to material surfaces is a well-known phenomenon that makes measurement of these substances challenging. Peptide adsorption can be reduced with suitable materials or additives, but the complex nature of biomolecules makes it challenging to find an universal sample preparation method. This study focused on investigating the surface adsorption behavior of motilin to enhance the sensitivity and precision of analytical methods. A simple and specific HPLC method was developed for the quantification of motilin. Using this chromatographic method, it was discovered that motilin binds to both polypropylene and glass surfaces at high levels, with recovery rates of 32 and 24%, respectively. To mitigate this, the effect of protein (bovine serum albumin), salt (sodium chloride), and surfactant (Tween 80) on surface adsorption was investigated. Although all additives reduced adsorption, only Tween 80 achieved nearly 100% recovery with consistent results (RSD <= 2%). Consequently, the sample preparation procedure developed using Tween 80 was integrated into the HPLC method, and the method was validated according to the International Conference on Harmonization guideline [ICH Q2(R1)]. This method is expected to have potential applications in analytical chemistry, drug development, biotechnology, and the pharmaceutical industry for providing accurate and dependable peptide measurements.

关键词： Motilin surface adsorption binding HPLC peptide analysis

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Chemometrically assisted optimization and validation of a new HPLC method for the determination of paliperidone in pharmaceuticals

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JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 2018年第3期41卷 129-134页

作者： Karaca, Sakine Atila ugur, duygu yeniceli Anadolu Univ Dept Analyt Chem Fac Pharm TR-26470 Eskisehir Turkey

Paliperidone is an antipsychotic drug, which is used for the acute and maintenance treatment of schizophrenia. In this study, a new method was developed for the determination of Paliperidone in its extended-release tablets. Face-centered central composite design was applied for optimization of the method. Factors were decided as acetonitrile content, pH of the mobile phase and buffer concentration through preliminary studies. Optimal flow rate (1mL/min), column temperature (35 degrees C) and internal standard (Bupropion) were also determined during preliminary studies. Retention factors and tailing factors of Paliperidone and Bupropion were selected as responses. Derringer's desirability function was applied for simultaneously optimization of these four responses. Optimal conditions were predicted as phosphate buffer (pH:3, 23mM): acetonitrile (76:24, v:v). Developed method was validated in terms of linearity, detection and quantification limits, accuracy, precision, specificity and robustness. Method was found linear in the concentration range of 0.125-100 mu g/mL. Mean equation of the calibration curve was y=0.0807 x - 0.0102 (R-2=0.9999). Accuracy and precision of the method was evaluated with recovery values (98-102%) and relative standard deviation values (<2%), respectively. All other parameters were found acceptable. The method was successfully applied for the determination of Paliperidone in its extended-release tablets. [GRAPHICS] .

关键词： Experimental design high-performance liquid chromatography (HPLC) paliperidone pharmaceuticals validation

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Determination of Iloperidone in Pharmaceuticals by Validated High-Performance Liquid Chromatography

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ANALYTICAL LETTERS 2018年第1-2期51卷 24-32页

作者： Karaca, Sakine Atila ugur, duygu yeniceli Anadolu Univ Dept Analyt Chem Fac Pharm TR-26470 Eskisehir Turkey

Iloperidone is an antipsychotic drug used for treatment of acute schizophrenia in adults. In this study, a simple and specific method was developed for determination of iloperidone in tablets. The separation was performed on a C18 column using pH 3 20 mM phosphate buffer and acetonitrile as the mobile phase. The optimum conditions were 1 ml/min flow rate, 35 degrees C column temperature, and 2 mu L injection volume. Carbamazepine was used as an internal standard. Iloperidone was monitored at 274 nm with a diode array detector. The method was validated according to the literature guidelines and was linear from 0.5 to 100 mu g/ml of iloperidone. The detection and quantification limits were 0.0909 and 0.3030 mu g/ml, respectively. The method was acceptable in terms of accuracy, precision, specificity, robustness, and stability and employed for the analysis of tablets.

关键词： High-performance liquid chromatography (HPLC) iloperidone pharmaceuticals tablets

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Development of a simple HPLC method for the quantitation of vortioxetine in pharmaceuticals using DoE approach

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CHEMICAL PAPERS 2020年第5期74卷 1541-1549页

作者： Karaca, Sakine Atila Rashidova, Nurana ugur, Alper ugur, duygu yeniceli Anadolu Univ Fac Pharm Dept Analyt Chem TR-26470 Eskisehir Turkey Eskisehir Tech Univ Fac Engn Dept Mat Sci & Engn TR-26555 Eskisehir Turkey

A novel HPLC method was proposed for the quantitation of vortioxetine, which is a new antidepressant drug, in tablets. Separation was performed on a Zorbax Eclipse Plus C18 (3.5 mu m particles, 4.6 x 50 mm) column at 35 degrees C with 1.0 mL/min flow rate and chlorpromazine was selected as internal standard. Box-Behnken design, a design of experiment (DoE) method, was applied to explore the influences of mobile phase pH, acetonitrile content and buffer concentration on chromatographic separation of vortioxetine and chlorpromazine. The optimized conditions were acetate buffer (25 mM, pH:4):acetonitrile (63.5:36.5, v/v). Retention times of chlorpromazine and vortioxetine were 3.8 and 4.6 min, respectively, and a good chromatographic separation was obtained under optimal conditions. Validation studies of the developed method were conducted according to the International Conference on Harmonization (ICH) guideline. The method was linear in the concentration range of 0.5-50 mu g/mL. The detection and quantitation limits were 0.080 and 0.264 mu g/mL, respectively. Recovery values within 98-102% and relative standard deviation values lower than 2% indicate the precision and accuracy of the method. The method was applied to vortioxetine tablets with no interference. [GRAPHICS] .

关键词： Vortioxetine HPLC-DAD Determination DoE Tablet analysis

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Development of a sensitive HPLC method with fluorescence detection for determination of motilin in human plasma

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BIOANALYSIS 2023年第4期15卷 219-229页

作者： Karaca, Sakine Atila Temel, Tuncer ugur, duygu yeniceli Anadolu Univ Fac Pharm Dept Analyt Chem TR-26470 Eskisehir Turkiye Eskisehir Osmangazi Univ Fac Med Dept Internal Med TR-26040 Eskisehir Turkiye

Background: Motilin is a peptide-structured gastrointestinal system hormone. In this study, a sensitive HPLC-fluorescence detection method was developed and validated for the quantification of motilin in human plasma. Materials & methods: Optimization processes were carried out with the experimental design methodology. Analyses were performed on a C8 column (4.6 x 150 mm, 3.5 mu m particles) using water and acetonitrile containing trifluoroacetic acid as the mobile phase. Results & conclusion: The method was linear from 2 to 200 ng/ml of motilin. The assay variability was less than 5%. The limit of quantification was found to be 1.84 ng/ml. The applicability of the developed method was successfully demonstrated by quantifying the levels of motilin in human plasma samples.

关键词： chromatography experimental design motilin peptide quantification

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A sensitive capillary LC-UV method for the simultaneous analysis of olanzapine, chlorpromazine and their FMO-mediated N-oxidation products in brain microdialysates

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TALANTA 2017年 162卷 268-277页

作者： Hendrickx, Stijn ugur, duygu yeniceli Yilmaz, Isil Tan Sener, Erol Van Schepdael, Ann Adams, Erwin Broeckhoven, Ken Cabooter, Deirdre Katholieke Univ Leuven Pharmaceut Anal Dept Pharmaceut & Pharmacol Sci Herestr 49 B-3000 Leuven Belgium Anadolu Univ Dept Analyt Chem Fac Pharm TR-26470 Eskisehir Turkey Vrije Univ Brussel Dept Chem Engn Pleinlaan 2 B-1050 Brussels Belgium

A specific and sensitive capillary liquid chromatography-ultraviolet detection (cap-LC-UV) method in combination with a micro-extraction by packed sorbent (MEPS) sample clean-up procedure has been developed and validated for the simultaneous analysis of chlorpromazine, olanzapine and their flavin-containing monooxygenase (FMO) mediated N-oxides in rat brain microdialysates. Chromatographic separation was obtained on an Acclaim Pepmap RP C18 column with an ID of 300 mu m. An injection volume of 20 L was used to inject the largely aqueous samples and was shown to have no influence on the obtained peak shape of the compounds of interest. Optimal conditions for MEPS extraction were obtained on a mixed-mode M1 (80% C8, 20% SCX) cartridge after diluting microdialysate samples with phosphate buffer pH 2.5 (1:3 v/v). The method was validated and lower limits of quantification (LLOQ) were determined at 0.5 nM for all compounds. Linearity was demonstrated between the LLOQ and 1 mu M for all compounds (R-2 > 0.995). MEPS recoveries were between 92% and 98%, with intra- and interday variabilities below 15%. The applicability of the developed method was successfully demonstrated by analysing rat brain microdialysates. The capillary LC-UV method in combination with MEPS sample treatment provides a simple, sensitive method to quantify all compounds of interest in 45 min and can be applied for routine therapeutic monitoring and pharmacokinetic studies of olanzapine, chlorpromazine and their respective N-oxides.

关键词： Olanzapine Chlorpromazine FMO Capillary LC Microdialysis MEPS

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Development of a sensitive and quantitative HPLC-FLD method for the determination of obestatin in human plasma

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JOURNAL OF PEPTIDE SCIENCE 2023年第11期29卷 e3511-e3511页

作者： Karaca, Sakine Atila Temel, Tuncer ugur, duygu yeniceli Anadolu Univ Fac Pharm Dept Analyt Chem Eskisehir Turkiye Eskisehir Osmangazi Univ Fac Med Dept Internal Med Eskisehir Turkiye Anadolu Univ Fac Pharm Dept Analyt Chem TR-26470 Eskisehir Turkiye

Obestatin is a gastrointestinal system peptide. The quantification of this peptide is conventionally performed using immunological techniques. In this study, a selective and sensitive HPLC method coupled with fluorescence detection for the quantitation of obestatin in human plasma was developed and validated. The separation was obtained on a C-18 (4.6 x 100 mm, 3.5-mu m particles) column using a mobile phase composed of acetonitrile and water, both including 0.1% trifluoroacetic acid. The developed method was found to be linear in the concentration range of 20 to 1000 ng/mL, with a coefficient of determination of 0.9982. The precision results were less than 10%, and the accuracy results were between 92% and 107%. The detection and quantification limit values were obtained as 2.8 and 9.4 ng/mL, respectively. Analyte solutions were found stable for 24 h at room temperature, three freeze-thaw cycles, and 2 weeks at -20 degrees C. The developed method was successfully used for the quantification of obestatin in human plasma samples. In conclusion, the developed method is sensitive and specific for measuring the plasma concentrations of obestatin.

关键词： chromatography experimental design obestatin peptide quantification

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Determination of Ochratoxin-A in the brain microdialysates and plasma of awake, freely moving rats using ultra high performance liquid chromatography fluorescence detection method

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JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 2019年 1125卷 121700-000页

作者： Kaya, Elif Mine Oncu Korkmaz, Orhan Tansel ugur, duygu yeniceli Sener, Erol Tuncel, A. Nese Tuncel, Muzaffer Eskisehir Tech Univ Sci Fac Dept Chem TR-26470 Eskisehir Turkey Eskisehir Osmangazi Univ Med Fac Dept Physiol TR-26480 Eskisehir Turkey Anadolu Univ Fac Pharm Dept Analyt Chem TR-26470 Eskisehir Turkey Lefke European Univ Fac Pharm Dept Analyt Chem Lefka Cyprus

In this study, a simple, efficient and rapid Ultra High Performance Liquid Chromatography method with fluorescence detection (UHPLC-FLD) has been developed and validated for the determination of Ochratoxin-A (OTA) in rat brain microdialysates and plasma samples. Six adult male wistar rats were used in the study and a single dose (5 mg/kg b.w.) of OTA was given by intraperitoneal (i.p.) injection. Rat blood and microdialysate samples were collected simultaneously after i.p. injection in awake freely moving rats, over a twelve-hour period. An UHPLC analysis was performed on a Zorbax Eclipse Plus C8 (150 mm x 3.0 mm ID x 1.8 mu m particles) column with a mobile phase of acetonitrile:water:phosphoric acid (50:50:0.1, v/v) using a flow rate of 0.6 mL/min. The fluorescence detector was set at 330 nm excitation and 460 nm emission wavelengths. Diflunisal (DIF) was used as an internal standard (IS). OTA and IS were separated within 5 min under these conditions. The method was validated in terms of linearity, precision, accuracy, limit of detection, limit of quantification, and stability. Calibration curves obtained with spiked biological matrices show good linearity with high correlation coefficients. The intra- and inter-day assay variability was < 5% for the OTA. The limit of detection and the limit of quantification values were found to be 0.490 ng/mL and 1.48 ng/mL for plasma;0.0900 ng/mL and 0.270 ng/mL for microdialysate samples, respectively. This method was successfully applied for the monitoring of OTA levels in the rat brain and plasma samples.

关键词： Ochratoxin-A UHPLC Rat plasma Rat brain Validation Microdialysis

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A novel capillary electrophoresis method for the quantification of asenapine in pharmaceuticals using Box-Behnken design

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CHEMICAL PAPERS 2020年第12期74卷 4443-4451页

作者： Aliyeva, Sona Atila Karaca, Sakine ugur, Alper Dal Pocan, Arin Gul yeniceli ugur, duygu Anadolu Univ Fac Pharm Dept Analyt Chem TR-26470 Eskisehir Turkey Eskisehir Tech Univ Fac Engn Dept Mat Sci & Engn TR-26555 Eskisehir Turkey

A new capillary electrophoresis (CE) method was presented for the determination of asenapine, an atypical antipsychotic drug, in pharmaceuticals. Box-Behnken design, an experimental design method, was used to investigate the effects of run buffer pH, run buffer concentration and applied potential on the separation of asenapine and granisetron (IS). The optimum conditions were phosphate buffer (15 mM, pH: 3.1) with 25.7 kV voltage and 20 celcius capillary temperature. The method was validated according to ICH guideline. A good linearity was obtained in the concentration range of 0.27-6.4 mu g/mL with LOD and LOQ values of 0.07 and 0.24 mu g/mL, respectively. The precision and accuracy of the method were satisfying with intra and interday recovery values of 97.8-100.8% and RSD less than 2%. The proposed CE method was applied to asenapine tablets successfully, for the first time.

关键词： Asenapine Capillary electrophoresis Determination Experimental design Tablet analysis

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