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Anna Petruczynik
Department of Inorganic Chemistry, Medical University of Lublin, Chodzki 4a, 20-093 Lublin, Poland

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Journal article
Published: 20 March 2021 in Molecules
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Melanoma is an enormous global health burden, and should be effectively addressed with better therapeutic strategies. Therefore, new therapeutic agents are needed for the management of this disease. The aim of this study was the investigation of cytotoxic activity of some isoquinoline alkaloid standards and extracts obtained from Sanguinaria canadensis—collected before, during, and after flowering—against three different human melanoma cells (A375, G361, SK-MEL-3). The cytotoxicity of these extracts was not previously tested on these melanoma cell lines. Determination of alkaloid contents was performed by HPLC-DAD using Polar RP column and mobile phase containing acetonitrile, water, and 1-butyl-3-methylimidazolium tetrafluoroborate. The cytotoxicity of alkaloid standards was investigated by determination of cell viability and calculation of IC50 values. Significant differences were observed in the alkaloids content and cytotoxic activity of the extracts, depending on the season of collection of the plant material. In the Sanguinaria canadensis extracts high contents of sanguinarine (from 4.8543 to 9.5899 mg/g of dry plant material) and chelerythrine (from 42.7224 to 6.8722 mg/g of dry plant material) were found. For both of these alkaloids, very high cytotoxic activity against the tested cell lines were observed. The IC50 values were in the range of 0.11–0.54 µg/mL for sanguinarine and 0.14 to 0.46 µg/mL for chelerythrine. IC50 values obtained for Sanguinaria canadensis extracts against all tested cell lines were also very low (from 0.88 to 10.96 µg/mL). Cytotoxic activity of alkaloid standards and Sanguinaria canadensis extracts were compared with the cytotoxicity of anticancer drugs—etoposide, cisplatin, and hydroxyurea. In all cases except the one obtained for cisplatin against A375, which was similar to that obtained for Sanguinaria canadensis after flowering against the same cell line, IC50 values obtained for anticancer drugs were higher than the IC50 values obtained for sanguinarine, chelerythrine, and Sanguinaria canadensis extracts. Our results showed that Sanguinaria canadensis extracts and isoquinoline alkaloids, especially sanguinarine and chelerythrine, could be recommended for further in vivo experiments in order to confirm the possibility of their application in the treatment of human melanomas.

ACS Style

Tomasz Tuzimski; Anna Petruczynik; Tomasz Plech; Barbara Kaproń; Anna Makuch-Kocka; Małgorzata Szultka-Młyńska; Justyna Misiurek; Bogusław Buszewski. Determination of Cytotoxic Activity of Sanguinaria canadensis Extracts against Human Melanoma Cells and Comparison of Their Cytotoxicity with Cytotoxicity of Some Anticancer Drugs. Molecules 2021, 26, 1738 .

AMA Style

Tomasz Tuzimski, Anna Petruczynik, Tomasz Plech, Barbara Kaproń, Anna Makuch-Kocka, Małgorzata Szultka-Młyńska, Justyna Misiurek, Bogusław Buszewski. Determination of Cytotoxic Activity of Sanguinaria canadensis Extracts against Human Melanoma Cells and Comparison of Their Cytotoxicity with Cytotoxicity of Some Anticancer Drugs. Molecules. 2021; 26 (6):1738.

Chicago/Turabian Style

Tomasz Tuzimski; Anna Petruczynik; Tomasz Plech; Barbara Kaproń; Anna Makuch-Kocka; Małgorzata Szultka-Młyńska; Justyna Misiurek; Bogusław Buszewski. 2021. "Determination of Cytotoxic Activity of Sanguinaria canadensis Extracts against Human Melanoma Cells and Comparison of Their Cytotoxicity with Cytotoxicity of Some Anticancer Drugs." Molecules 26, no. 6: 1738.

Journal article
Published: 04 February 2021 in Molecules
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Melanoma is a serious form of skin cancer that begins in cells known as melanocytes. While it is less common than the other forms of skin cancer, melanoma is more dangerous because of its ability to spread to other organs more rapidly if it is not treated at an early stage. The number of people diagnosed with melanoma has increased over the last few decades. The most widely used treatments include surgery, chemotherapy, and radiation therapy. The search for new drugs to treat various cancers is one of the most important challenges of modern scientific research. Some isoquinoline alkaloids found in different plant species have strong cytotoxic effects on various cancer cells. We tested the effect of isoquinoline alkaloids and extracts obtained from various parts of Mahonia aquifolium collected in various vegetation seasons on human melanoma cancer cells and our data indicated that investigated extract induced significant reduction in cell viability of Human malignant melanoma cells (A375), human Caucasian malignant melanoma cell line (G361), and human malignant melanoma cell line (SKMEL3 cancer cell lines in a dose- and time-dependent manner. Differences in cytotoxic activity were observed for extracts obtained from various parts of Mahonia aquifolium. Significant differences were also obtained in the alkaloids content and cytotoxic activity of the extracts depending on the season of collection of plant material. Our investigations exhibit that these plant extracts can be recommended for further in vivo experiments in order to confirm the possibility of their use in the treatment of human melanomas.

ACS Style

Tomasz Tuzimski; Anna Petruczynik; Barbara Kaproń; Anna Makuch-Kocka; Małgorzata Szultka-Młyńska; Justyna Misiurek; Grażyna Szymczak; Bogusław Buszewski. Determination of Cytotoxic Activity of Selected Isoquinoline Alkaloids and Plant Extracts Obtained from Various Parts of Mahonia aquifolium Collected in Various Vegetation Seasons. Molecules 2021, 26, 816 .

AMA Style

Tomasz Tuzimski, Anna Petruczynik, Barbara Kaproń, Anna Makuch-Kocka, Małgorzata Szultka-Młyńska, Justyna Misiurek, Grażyna Szymczak, Bogusław Buszewski. Determination of Cytotoxic Activity of Selected Isoquinoline Alkaloids and Plant Extracts Obtained from Various Parts of Mahonia aquifolium Collected in Various Vegetation Seasons. Molecules. 2021; 26 (4):816.

Chicago/Turabian Style

Tomasz Tuzimski; Anna Petruczynik; Barbara Kaproń; Anna Makuch-Kocka; Małgorzata Szultka-Młyńska; Justyna Misiurek; Grażyna Szymczak; Bogusław Buszewski. 2021. "Determination of Cytotoxic Activity of Selected Isoquinoline Alkaloids and Plant Extracts Obtained from Various Parts of Mahonia aquifolium Collected in Various Vegetation Seasons." Molecules 26, no. 4: 816.

Journal article
Published: 05 January 2021 in Molecules
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Isoquinoline alkaloids may have a wide range of pharmacological activities. Some of them have acetylcholinesterase activity inhibition. Nowadays, neurodegenerative disorders such as Alzheimer’s disease have become a serious public health problem. Searching for new effective compounds with inhibited acetylcholinesterase activity is one of the most significant challenges of modern scientific research. The aim of this study was the in vitro investigation of acetylcholinesterase activity inhibition of extracts obtained from Sanguinaria canadensis collected before, during and after flowering. The acetylcholinesterase activity inhibition of these extracts has not been previously tested. The aim was also to quantify selected alkaloids in the investigated extracts by high performance liquid chromatography (HPLC). The analyses of alkaloid content were performed using HPLC in reversed phase (RP) mode using Polar RP column and mobile phase containing acetonitrile, water and ionic liquid (IL). The acetylcholinesterase activity inhibition of the tested plant extracts and respective alkaloid standards were examined using high performance liquid chromatography with diode-array detector (HPLC-DAD) for the quantification of 5-thio-2-nitro-benzoic acid, which is the product of the reaction between the thiocholine (product of the hydrolysis of acetylthiocholine reaction) with Ellman reagent. The application of the HPLC method allowed for elimination of absorption of interfering components, for example, alkaloids such as sanguinarine and berberine. It is revealed that the HPLC method can be successfully used for the evaluation of the acetylcholinesterase inhibitory activity in samples such as plant extracts, especially those containing colored components adsorbing at wavelength in the range 405–412 nm. The acetylcholinesterase inhibition activity synergy of pairs of alkaloid standards and mixture of all investigated alkaloids was also determined. Most investigated alkaloids and all Sanguinaria canadensis extracts exhibited very high acetylcholinesterase activity inhibition. IC50 values obtained for alkaloid standards were from 0.36 for berberine to 23.13 µg/mL for protopine and from 61.24 to 89.14 µg/mL for Sanguinaria canadensis extracts. Our investigations demonstrated that these plant extracts can be recommended for further in vivo experiments to confirm their acetylcholinesterase activity inhibition.

ACS Style

Tomasz Tuzimski; Anna Petruczynik. Application of HPLC-DAD for In Vitro Investigation of Acetylcholinesterase Inhibition Activity of Selected Isoquinoline Alkaloids from Sanguinaria canadensis Extracts. Molecules 2021, 26, 230 .

AMA Style

Tomasz Tuzimski, Anna Petruczynik. Application of HPLC-DAD for In Vitro Investigation of Acetylcholinesterase Inhibition Activity of Selected Isoquinoline Alkaloids from Sanguinaria canadensis Extracts. Molecules. 2021; 26 (1):230.

Chicago/Turabian Style

Tomasz Tuzimski; Anna Petruczynik. 2021. "Application of HPLC-DAD for In Vitro Investigation of Acetylcholinesterase Inhibition Activity of Selected Isoquinoline Alkaloids from Sanguinaria canadensis Extracts." Molecules 26, no. 1: 230.

Review
Published: 03 September 2020 in Molecules
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Therapeutic drug monitoring (TDM) is a tool used to integrate pharmacokinetic and pharmacodynamics knowledge to optimize and personalize various drug therapies. The optimization of drug dosing may improve treatment outcomes, reduce toxicity, and reduce the risk of developing drug resistance. To adequately implement TDM, accurate and precise analytical procedures are required. In clinical practice, blood is the most commonly used matrix for TDM; however, less invasive samples, such as dried blood spots or non-invasive saliva samples, are increasingly being used. The choice of sample preparation method, type of column packing, mobile phase composition, and detection method is important to ensure accurate drug measurement and to avoid interference from matrix effects and drug metabolites. Most of the reported procedures used liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) techniques due to its high selectivity and sensitivity. High-performance chromatography with ultraviolet detection (HPLC-UV) methods are also used when a simpler and more cost-effective methodology is desired for clinical monitoring. The application of high-performance chromatography with fluorescence detection (HPLC-FLD) with and without derivatization processes and high-performance chromatography with electrochemical detection (HPLC-ED) techniques for the analysis of various drugs in biological samples for TDM have been described less often. Before chromatographic analysis, samples were pretreated by various procedures—most often by protein precipitation, liquid–liquid extraction, and solid-phase extraction, rarely by microextraction by packed sorbent, dispersive liquid–liquid microextraction. The aim of this article is to review the recent literature (2010–2020) regarding the use of liquid chromatography with various detection techniques for TDM.

ACS Style

Tomasz Tuzimski; Anna Petruczynik. Review of Chromatographic Methods Coupled with Modern Detection Techniques Applied in the Therapeutic Drugs Monitoring (TDM). Molecules 2020, 25, 4026 .

AMA Style

Tomasz Tuzimski, Anna Petruczynik. Review of Chromatographic Methods Coupled with Modern Detection Techniques Applied in the Therapeutic Drugs Monitoring (TDM). Molecules. 2020; 25 (17):4026.

Chicago/Turabian Style

Tomasz Tuzimski; Anna Petruczynik. 2020. "Review of Chromatographic Methods Coupled with Modern Detection Techniques Applied in the Therapeutic Drugs Monitoring (TDM)." Molecules 25, no. 17: 4026.

Journal article
Published: 29 August 2020 in Toxins
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Quinolizidine alkaloids exhibit various forms of biological activity. A lot of them were found in the Leguminosae family, including Laburnum and Genista. The aim of the study was the optimization of a chromatographic system for the analysis of cytisine and N-methylcytisine in various plant extracts as well as an investigation of the cytotoxic activities of selected alkaloids and plant extracts obtained from Laburnum anagyroides, Laburnum anagyroides L. quercifolium, Laburnum alpinum, Laburnum watereri, Genista germanica, and Genista tinctoria against various cancer cell lines. The determination of investigated compounds was performed by High Performance Liquid Chromatography with Diode Array Detection (HPLC-DAD), while High Performance Liquid Chromatography coupled with Quadrupole Time-of-Flight–Mass Spectrometry (HPLC-QTOF-MS) was applied for the qualitative analysis of plant extracts. The retention, separation selectivity, peaks shape, and systems efficiency obtained for cytisine and N-methylcytisine in different chromatographic systems were compared. The application of columns with alkylbonded and phenyl stationary phases led to a very weak retention of cytisine and N-methylcytisine, even when the mobile phases containing only 5% of organic modifiers were used. The strongest retention was observed when hydrophilic interaction chromatography (HILIC) or especially when ion exchange chromatography (IEC) were applied. The most optimal system in terms of alkaloid retention, peak shape, and system efficiency containing an strong cation exchange (SCX) stationary phase and a mobile phase consisted of 25% acetonitrile and formic buffer at pH 4.0 was applied for investigating alkaloids analysis in plant extracts. Cytotoxic properties of the investigated plant extracts as well as cytisine and N-methylcytisine were examined using human tongue squamous carcinoma cells (SCC-25), human pharyngeal squamous carcinoma cells (FaDu), human triple-negative breast adenocarcinoma cell line (MDA-MB-231), and human breast adenocarcinoma cell line (MCF-7). The highest cytotoxic activity against FaDu, MCF-7, and MDA-MB cancer cell lines was observed after applying the Genista germanica leaves extract. In contrast, the highest cytotoxic activity against SCC-25 cell line was obtained after treating with the seed extract of Laburnum watereri. The investigated plant extracts exhibit significant cytotoxicity against the tested human cancer cell lines and seem to be promising for further research on its anticancer activity.

ACS Style

Anna Petruczynik; Karol Wróblewski; Justyna Misiurek; Tomasz Plech; Karolina Szalast; Krzysztof Wojtanowski; Tomasz Mroczek; Grażyna Szymczak; Monika Waksmundzka-Hajnos; Piotr Tutka. Determination of Cytisine and N-Methylcytisine from Selected Plant Extracts by High-Performance Liquid Chromatography and Comparison of Their Cytotoxic Activity. Toxins 2020, 12, 557 .

AMA Style

Anna Petruczynik, Karol Wróblewski, Justyna Misiurek, Tomasz Plech, Karolina Szalast, Krzysztof Wojtanowski, Tomasz Mroczek, Grażyna Szymczak, Monika Waksmundzka-Hajnos, Piotr Tutka. Determination of Cytisine and N-Methylcytisine from Selected Plant Extracts by High-Performance Liquid Chromatography and Comparison of Their Cytotoxic Activity. Toxins. 2020; 12 (9):557.

Chicago/Turabian Style

Anna Petruczynik; Karol Wróblewski; Justyna Misiurek; Tomasz Plech; Karolina Szalast; Krzysztof Wojtanowski; Tomasz Mroczek; Grażyna Szymczak; Monika Waksmundzka-Hajnos; Piotr Tutka. 2020. "Determination of Cytisine and N-Methylcytisine from Selected Plant Extracts by High-Performance Liquid Chromatography and Comparison of Their Cytotoxic Activity." Toxins 12, no. 9: 557.

Journal article
Published: 27 May 2020 in Molecules
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Background: Determination of psychotropic drugs in clinical study is significant, and the establishment of methodologies for these drugs in biological matrices is essential for patients’ safety. The search for new methods for their detection is one of the most important challenges of modern scientific research. The methods for analyzing of psychotropic drugs and their metabolites in different biological samples should be based on combining a very efficient separation technique including high-performance liquid chromatography (HPLC), with a sensitive detection method and effectively sample preparation methods. Objective: Retention, peaks symmetry and system efficiency of vortioxetine on Hydro RP, Polar RP, HILIC A (with silica stationary phase), HILIC-B (with aminopropyl stationary phase), and ACE HILIC-N (with polyhydroxy stationary phase and SCX columns were investigated. Various mobile phases containing methanol or acetonitrile as organic modifiers and different additives were also applied to obtained optimal retention, peaks shape, and systems efficiency. The best chromatographic procedure was used for simultaneous analysis of vortioxetine and its metabolites in human serum, urine and saliva samples. Methods: Analysis of vortioxetine was performed in various chromatographic systems: Reversed phase (RP) systems on alkylbonded or phenyl stationary phases, hydrophilic interaction liquid chromatography (HILIC), and ion-exchange chromatography (IEC). Based on the dependence of log k vs the concentration of the organic modifier, log kw values for vortioxetine in various chromatographic systems were determined and compared with calculated log P values. Solid phase extraction (SPE) method was applied for sample pre-treatment before HPLC analysis. HPLC-QTOF-MS method was applied for confirmation of presence of vortioxetine and some its metabolites in biological samples collected from psychiatric patient. Conclusions: Differences were observed in retention parameters with a change of the applied chromatographic system. The various properties of stationary phases resulted in differences in vortioxetine retention, systems’ efficiency, and peaks’ shape. Lipophilicity parameters were also determined using different HPLC conditions. The most optimal systems were chosen for the analysis of vortioxetine in biological samples. Both serum and urine or saliva samples collected from patients treated with vortioxetine can be used for the drug determination. For the first time, vortioxetine was detected in patient’s saliva. Obtained results indicate on possibility of application of saliva samples, which collection are non-invasive and painless, for determination and therapeutic drug monitoring in patients.

ACS Style

Anna Petruczynik; Karol Wróblewski; Krzysztof Wojtanowski; Tomasz Mroczek; Dariusz Juchnowicz; Hanna Karakuła-Juchnowicz; Tomasz Tuzimski. Comparison of Various Chromatographic Systems for Identification of Vortioxetine in Bulk Drug Substance, Human Serum, Saliva, and Urine Samples by HPLC-DAD and LC-QTOF-MS. Molecules 2020, 25, 2483 .

AMA Style

Anna Petruczynik, Karol Wróblewski, Krzysztof Wojtanowski, Tomasz Mroczek, Dariusz Juchnowicz, Hanna Karakuła-Juchnowicz, Tomasz Tuzimski. Comparison of Various Chromatographic Systems for Identification of Vortioxetine in Bulk Drug Substance, Human Serum, Saliva, and Urine Samples by HPLC-DAD and LC-QTOF-MS. Molecules. 2020; 25 (11):2483.

Chicago/Turabian Style

Anna Petruczynik; Karol Wróblewski; Krzysztof Wojtanowski; Tomasz Mroczek; Dariusz Juchnowicz; Hanna Karakuła-Juchnowicz; Tomasz Tuzimski. 2020. "Comparison of Various Chromatographic Systems for Identification of Vortioxetine in Bulk Drug Substance, Human Serum, Saliva, and Urine Samples by HPLC-DAD and LC-QTOF-MS." Molecules 25, no. 11: 2483.

Journal article
Published: 31 December 2019 in Open Chemistry
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Retention, separation selectivity and system efficiency of selected basic psychotropic drugs (clozapine, aripiprazole, vortioxetine and zolpidem) and drug metabolites (desmethylclozapine, clozapine N-oxide and dehydroaripiprazole) on Hydro RP, Phenyl-Hexyl and Polar RP columns were studied. Mobile phases containing methanol or acetonitrile as organic modifiers, acetate buffer at pH 3.5 and addition of diethylamine (DEA) as a silanol blocker were applied. Significant differences in the retention, peak shapes and systems’ efficiency of the investigated compounds were obtained depending on the tested chemically bonded stationary phases with various ligands. Based on the obtained results the Phenyl-Hexyl column was selected for analysis of the drugs and their metabolites in human serum and saliva samples. Solid phase extraction (SPE) was applied for sample pre-treatment. The best SPE-HPLC-DAD procedure was used for simultaneous analysis of clozapine, aripiprazole and their metabolites in body fluids. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was applied for confirmation of the presence of the investigated compounds in biological samples. The lower limit of quantification (LLOQ) of clozapine obtained using the proposed method was 10 ng/mL. The validated method for determining the presence of clozapine and its main metabolite was successfully applied in therapeutic drug monitoring.

ACS Style

Karol Wróblewski; A. Petruczynik; T. Tuzimski; K. Prajsnar; D. Przygodzka; G. Buszewicz; H. Karakuła-Juchnowicz; J. Róg; J. Morylowska-Topolska; M. Waksmundzka-Hajnos. Optimization of chromatographic systems for analysis of selected psychotropic drugs and their metabolites in serum and saliva by HPLC in order to monitor therapeutic drugs. Open Chemistry 2019, 17, 1361 -1373.

AMA Style

Karol Wróblewski, A. Petruczynik, T. Tuzimski, K. Prajsnar, D. Przygodzka, G. Buszewicz, H. Karakuła-Juchnowicz, J. Róg, J. Morylowska-Topolska, M. Waksmundzka-Hajnos. Optimization of chromatographic systems for analysis of selected psychotropic drugs and their metabolites in serum and saliva by HPLC in order to monitor therapeutic drugs. Open Chemistry. 2019; 17 (1):1361-1373.

Chicago/Turabian Style

Karol Wróblewski; A. Petruczynik; T. Tuzimski; K. Prajsnar; D. Przygodzka; G. Buszewicz; H. Karakuła-Juchnowicz; J. Róg; J. Morylowska-Topolska; M. Waksmundzka-Hajnos. 2019. "Optimization of chromatographic systems for analysis of selected psychotropic drugs and their metabolites in serum and saliva by HPLC in order to monitor therapeutic drugs." Open Chemistry 17, no. 1: 1361-1373.

Journal article
Published: 01 December 2019 in Acta Chromatographica
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In reversed-phase liquid chromatography, in case of the absence of additives, cationic basic compounds give rise to asymmetrical and broad peaks as a result of interactions of analyte cations with residual free silanols on silica-based stationary phases. Ionic liquids, added to the mobile phase, have been suggested as alternatives to amines to block the activity of free silanols. The different parameters affecting the retention behavior, symmetry of peak, system efficiency, and separation selectivity of selected psychotropic drugs, especially the effect of concentration of ionic liquid, kind and concentration of organic modifiers of mobile phases, and kind of stationary phases were investigated. The most selective and efficient systems are used for separations of psychotropic drug standards' mixture and for determination of selected psychotropic drugs in human serum.

ACS Style

A. Petruczynik; Karol Wróblewski; M. Waksmundzka-Hajnos. Application of mobile phases containing ionic liquid for the analysis of selected psychotropic drugs by HPLC–DAD and HPLC–MS. Acta Chromatographica 2019, 31, 255 -261.

AMA Style

A. Petruczynik, Karol Wróblewski, M. Waksmundzka-Hajnos. Application of mobile phases containing ionic liquid for the analysis of selected psychotropic drugs by HPLC–DAD and HPLC–MS. Acta Chromatographica. 2019; 31 (4):255-261.

Chicago/Turabian Style

A. Petruczynik; Karol Wróblewski; M. Waksmundzka-Hajnos. 2019. "Application of mobile phases containing ionic liquid for the analysis of selected psychotropic drugs by HPLC–DAD and HPLC–MS." Acta Chromatographica 31, no. 4: 255-261.

Journal article
Published: 02 October 2019 in Toxins
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Alkaloids have protective functions for plants and can play an important role in living organisms. Alkaloids may have a wide range of pharmacological activities. Many of them have cytotoxic activity. Nowadays, cancer has become a serious public health problem. Searching for effective drugs with anticancer activity is one of the most significant challenges of modern scientific research. The aim of this study was the investigation of cytotoxic activity of extracts obtained from Corydalis lutea root and herb, Dicentra spectabilis root and herb, Fumaria officinalis, Macleaya cordata leaves and herb, Mahonia aquifolia leaves and cortex, Meconopsis cambrica root and herb on FaDu, SCC-25, MCF-7, and MDA-MB-231 cancer cell lines. The cytotoxic activity of these extracts has not been previously tested for these cell lines. The aim was also to quantify selected alkaloids in the investigated extracts by High Performance Liquid Chromatography (HPLC). The analyses of alkaloid content were performed using HPLC in reversed phase (RP) mode using Polar RP column and mobile phase containing acetonitrile, water and ionic liquid (IL). Cytotoxic effect of the tested plant extracts and respective alkaloid standards were examined using human pharyngeal squamous carcinoma cells (FaDu), human tongue squamous carcinoma cells (SCC-25), human breast adenocarcinoma cell line (MCF-7), human triple-negative breast adenocarcinoma cell line (MDA-MB-231). All investigated plant extracts possess cytotoxic activity against tested cancer cell lines: FaDu, SCC-25, MCF-7, and MDA-MB-231. The highest cytotoxic activity against FaDu, SCC-25, and MCF-7 cell lines was estimated for Macleaya cordata leaf extract, while the highest cytotoxic activity against MDA-MB-231 cell line was obtained for Macleaya cordata herb extract. Differences in cytotoxic activity were observed for extracts obtained from various parts of investigated plants. In almost all cases the cytotoxic activity of investigated plant extracts, especially at the highest concentration against tested cell lines was significantly higher than the activity of anticancer drug etoposide. Our investigations exhibit that these plant extracts can be recommended for further in vivo experiments to confirm their anticancer activity.

ACS Style

Anna Petruczynik; Tomasz Plech; Tomasz Tuzimski; Justyna Misiurek; Barbara Kaproń; Dorota Misiurek; Małgorzata Szultka-Młyńska; Bogusław Buszewski; Monika Waksmundzka-Hajnos. Determination of Selected Isoquinoline Alkaloids from Mahonia aquifolia; Meconopsis cambrica; Corydalis lutea; Dicentra spectabilis; Fumaria officinalis; Macleaya cordata Extracts by HPLC-DAD and Comparison of Their Cytotoxic Activity. Toxins 2019, 11, 575 .

AMA Style

Anna Petruczynik, Tomasz Plech, Tomasz Tuzimski, Justyna Misiurek, Barbara Kaproń, Dorota Misiurek, Małgorzata Szultka-Młyńska, Bogusław Buszewski, Monika Waksmundzka-Hajnos. Determination of Selected Isoquinoline Alkaloids from Mahonia aquifolia; Meconopsis cambrica; Corydalis lutea; Dicentra spectabilis; Fumaria officinalis; Macleaya cordata Extracts by HPLC-DAD and Comparison of Their Cytotoxic Activity. Toxins. 2019; 11 (10):575.

Chicago/Turabian Style

Anna Petruczynik; Tomasz Plech; Tomasz Tuzimski; Justyna Misiurek; Barbara Kaproń; Dorota Misiurek; Małgorzata Szultka-Młyńska; Bogusław Buszewski; Monika Waksmundzka-Hajnos. 2019. "Determination of Selected Isoquinoline Alkaloids from Mahonia aquifolia; Meconopsis cambrica; Corydalis lutea; Dicentra spectabilis; Fumaria officinalis; Macleaya cordata Extracts by HPLC-DAD and Comparison of Their Cytotoxic Activity." Toxins 11, no. 10: 575.

Journal article
Published: 20 September 2019 in Molecules
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Background: Plants are an important origin of natural substances that the raw material for various pharmaceutical and therapeutic applications due to the presence of phytochemicals, such as alkaloids. Alkaloids, which are found in different plant species, possess numerous biological activities. Some alkaloids have strong cytotoxic effects on various cancer cells. The search for new drugs to treat various cancers is one of the most important challenges of modern scientific research. Objective: This study aimed to investigate of cytotoxic activity of extracts that were obtained from Chelidonium Majus; Berberis sp.; Thalictrum foetidum containing various alkaloids on selected cancer cell lines. The aim was also the quantification of selected alkaloids in the investigated extracts by HPLC. Methods: The analysis of alkaloids contents were performed while using HPLC in reversed phase (RP) mode using Polar RP column and mobile phase containing acetonitrile, water, and ionic liquid. The cytotoxic effect of the tested plant extracts and respective alkaloids’ standards were examined while using human pharyngeal squamous carcinoma cells (FaDu), human tongue squamous carcinoma cells (SCC-25), human breast adenocarcinoma cell line (MCF-7), and human triple-negative breast adenocarcinoma cell line (MDA-MB-231). Conclusion: All of the investigated plant extracts possess cytotoxic activity against cancer cell lines: FaDu, SCC-25, MCF-7, and MDA-MB-231. The highest cytotoxic activity against FaDu and MDA-MB-231 cells was observed for Chelidonium majus root extract, while the highest cytotoxic activity against SCC-25 and MCF-7 cells was estimated for the Thalictrum foetidum root extract. There obtained significant differences in the cytotoxic activity of extracts that were obtained from the roots and herbs of Chelidonium majus and Thalictrum foetidum. Based on these results, investigated plant extracts can be recommended for further investigations of anticancer activity.

ACS Style

Anna Petruczynik; Tomasz Tuzimski; Tomasz Plech; Justyna Misiurek; Karolina Szalast; Grażyna Szymczak. Comparison of Anticancer Activity and HPLC-DAD Determination of Selected Isoquinoline Alkaloids from Thalictrum foetidum, Berberis sp. and Chelidonium majus Extracts. Molecules 2019, 24, 3417 .

AMA Style

Anna Petruczynik, Tomasz Tuzimski, Tomasz Plech, Justyna Misiurek, Karolina Szalast, Grażyna Szymczak. Comparison of Anticancer Activity and HPLC-DAD Determination of Selected Isoquinoline Alkaloids from Thalictrum foetidum, Berberis sp. and Chelidonium majus Extracts. Molecules. 2019; 24 (19):3417.

Chicago/Turabian Style

Anna Petruczynik; Tomasz Tuzimski; Tomasz Plech; Justyna Misiurek; Karolina Szalast; Grażyna Szymczak. 2019. "Comparison of Anticancer Activity and HPLC-DAD Determination of Selected Isoquinoline Alkaloids from Thalictrum foetidum, Berberis sp. and Chelidonium majus Extracts." Molecules 24, no. 19: 3417.

Comparative study
Published: 16 July 2019 in Molecules
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Background: Identification and quantitative determination of cytisine, especially in biological samples and pharmaceutical formulations, is still a difficult analytical task. Cytisine is an alkaloid with a small and very polar molecule. For this reason, it is very weakly retained on reversed phase (RP) stationary phases, such as commonly used alkyl-bonded phases. The very weak retention of cytisine causes it to be eluted together with the components of biological matrices. Objective: Comparison and evaluation of various chromatographic systems for analysis of cytisine in different matrices—serum, saliva and pharmaceutical formulation—by high performance liquid chromatography (HPLC) with diode array (DAD), fluorescence (FLD) and mass spectrometry (MS) detection. Methods: The analyses were performed using HPLC in reversed phase (RP), hydrophilic interaction liquid chromatography (HILIC) and ion exchange chromatography (IEC) modes. Different sample pre-treatment methods were tested: Protein precipitation (with acetone, methanol (MeOH) or acetonitrile (ACN), and solid phase extraction (SPE) using cartridges with octadecyl (C18), hydrophilic-lipophilic balanced copolymer (HLB) or strong cation exchange sorbents (Strata X-C). Conclusion: Significant differences were observed in retention parameters with a change of the used chromatographic system. The various properties of stationary phases resulted in differences in analyte retention, peaks’ shape and systems’ efficiency. The weakest retention was observed using RP systems; however, the use of the Polar RP phase can be an alternative for application in green chromatography. In the strongest retention was observed using a strong cation exchange (SCX) phase. The most optimal systems were chosen for the analysis of cytisine in the pharmaceutical preparation, serum and saliva after sample pre-treatment with the new SPE procedure. Due to the sensitivity, the use of HPLC-DAD or HPLC-FLD is the most optimal for drug analysis in pharmaceutical preparations, whereas HPLC-MS is suitable for analysis of cytisine in biological samples.

ACS Style

Karol Wróblewski; Anna Petruczynik; Tomasz Tuzimski; Dominika Przygodzka; Grzegorz Buszewicz; Patrycjusz Kołodziejczyk; Piotr Tutka. Comparison of Various Chromatographic Systems for Analysis of Cytisine in Human Serum, Saliva and Pharmaceutical Formulation by HPLC with Diode Array, Fluorescence or Mass Spectrometry Detection. Molecules 2019, 24, 2580 .

AMA Style

Karol Wróblewski, Anna Petruczynik, Tomasz Tuzimski, Dominika Przygodzka, Grzegorz Buszewicz, Patrycjusz Kołodziejczyk, Piotr Tutka. Comparison of Various Chromatographic Systems for Analysis of Cytisine in Human Serum, Saliva and Pharmaceutical Formulation by HPLC with Diode Array, Fluorescence or Mass Spectrometry Detection. Molecules. 2019; 24 (14):2580.

Chicago/Turabian Style

Karol Wróblewski; Anna Petruczynik; Tomasz Tuzimski; Dominika Przygodzka; Grzegorz Buszewicz; Patrycjusz Kołodziejczyk; Piotr Tutka. 2019. "Comparison of Various Chromatographic Systems for Analysis of Cytisine in Human Serum, Saliva and Pharmaceutical Formulation by HPLC with Diode Array, Fluorescence or Mass Spectrometry Detection." Molecules 24, no. 14: 2580.

Journal article
Published: 01 January 2019 in Journal of AOAC INTERNATIONAL
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Ionic liquids (ILs) are a class of unique substances composed purely by cation and anions. Because of the unique properties of ILs (e.g., electric conductivity, low volatility, thermostability, and tenability), their use has attracted considerable interest, including an increasing number of publications on their use in analytical separation techniques. ILs applied as extraction solvents in liquid–liquid extraction methods and promote interactions between the analyte and solvent because of their unique chemical functional groups. In solid-phase extraction, ILs are usually used for the modification of stationary phases. ILs in LC are applied as mobile phase additives to obtain a better peak shape and high system efficiency for the analysis of ionic, especially basic, compounds. This review highlights some of the applications of ILs to extraction and analysis by LC of xenobiotics in food, environmental, and biological samples.

ACS Style

Tomasz Tuzimski; Anna Petruczynik. Ionic Liquids Applied to Extraction of Xenobiotics from Food, Environmental, and Biological Samples and for Analysis by Liquid Chromatography. Journal of AOAC INTERNATIONAL 2019, 102, 3 -22.

AMA Style

Tomasz Tuzimski, Anna Petruczynik. Ionic Liquids Applied to Extraction of Xenobiotics from Food, Environmental, and Biological Samples and for Analysis by Liquid Chromatography. Journal of AOAC INTERNATIONAL. 2019; 102 (1):3-22.

Chicago/Turabian Style

Tomasz Tuzimski; Anna Petruczynik. 2019. "Ionic Liquids Applied to Extraction of Xenobiotics from Food, Environmental, and Biological Samples and for Analysis by Liquid Chromatography." Journal of AOAC INTERNATIONAL 102, no. 1: 3-22.

Articles
Published: 21 July 2018 in Journal of Liquid Chromatography & Related Technologies
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A HPLC procedure on strong cation exchange column (SCX) has been developed for the analysis of selected alkaloids. The retention, separation selectivity, symmetry of peaks and system efficiency were examined in various eluent systems containing different kind or concentration of buffers, addition of various organic modifiers. The retention factors as a function of a concentration of buffers and the percentages of acetonitrile in the eluents were investigated. The influence of column temperature on chromatographic parameters was also examined. The best chromatographic systems were selected for the separation of extracts’ components. HPLC procedure on SCX column with mobile phases containing acetonitrile and buffer at pH 2.5 has been developed for the analysis of berberine and magnoflorine in selected plant materials from genus Thalictrum, Magnolia and Calatheae. The application of ion-exchange chromatographic system lets to obtain the good system efficiency and most symmetrical peaks of investigated alkaloids. Berberine and magnoflorine were quantified on SCX column with mobile phase containing acetonitrile, phosphate buffers and addition of chlorides. The content of alkaloids in extracts obtained from different plant species and different parts of plants was investigated. The influence of the vegetation period on alkaloids’ level was examined.

ACS Style

Anna Petruczynik; Justyna Misiurek; Justyna Dyjach; Mariola Kołsut; Dorota Misiurek; Monika Waksmundzka-Hajnos. Optimization of ion-exchange systems for isoquinoline alkaloids analysis in plant materials. Journal of Liquid Chromatography & Related Technologies 2018, 41, 761 -769.

AMA Style

Anna Petruczynik, Justyna Misiurek, Justyna Dyjach, Mariola Kołsut, Dorota Misiurek, Monika Waksmundzka-Hajnos. Optimization of ion-exchange systems for isoquinoline alkaloids analysis in plant materials. Journal of Liquid Chromatography & Related Technologies. 2018; 41 (12):761-769.

Chicago/Turabian Style

Anna Petruczynik; Justyna Misiurek; Justyna Dyjach; Mariola Kołsut; Dorota Misiurek; Monika Waksmundzka-Hajnos. 2018. "Optimization of ion-exchange systems for isoquinoline alkaloids analysis in plant materials." Journal of Liquid Chromatography & Related Technologies 41, no. 12: 761-769.

Journal article
Published: 01 November 2017 in Journal of AOAC INTERNATIONAL
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An HPLC procedure on a polar reversed-phase column with mobile phases containing ionic liquid (IL) was developed for the analysis of selected alkaloids from different chemical groups. We aimed to obtain optimal conditions for the separation of alkaloids because widely used silica-based stationary phases exhibit a silanol effect, rendering analysis of basic analytes extremely difficult. Retention, separation selectivity, peak symmetry, and system efficiency were examined in various eluent systems containing different concentrations of IL and acetonitrile. The obtained results revealed substantial influence from the concentrations of IL, the organic modifier, and temperature on the retention behavior of the investigated alkaloids. The most selective and efficient chromatographic systems were applied for the analysis of several alkaloids in a plant extract.

ACS Style

Anna Petruczynik; Justyna Misiurek; Tomasz Tuzimski; Monika Waksmundzka-Hajnos. Application of Mobile Phases Containing Ionic Liquid for HPLC Analysis of Selected Isoquinoline Alkaloids. Journal of AOAC INTERNATIONAL 2017, 100, 1652 -1659.

AMA Style

Anna Petruczynik, Justyna Misiurek, Tomasz Tuzimski, Monika Waksmundzka-Hajnos. Application of Mobile Phases Containing Ionic Liquid for HPLC Analysis of Selected Isoquinoline Alkaloids. Journal of AOAC INTERNATIONAL. 2017; 100 (6):1652-1659.

Chicago/Turabian Style

Anna Petruczynik; Justyna Misiurek; Tomasz Tuzimski; Monika Waksmundzka-Hajnos. 2017. "Application of Mobile Phases Containing Ionic Liquid for HPLC Analysis of Selected Isoquinoline Alkaloids." Journal of AOAC INTERNATIONAL 100, no. 6: 1652-1659.

Journal article
Published: 01 September 2017 in Acta Chromatographica
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ACS Style

Karol Wróblewski; A. Petruczynik; B. Buszewski; Małgorzata Szultka-Młyńska; Hanna Karakula-Juchnowicz; M. Waksmundzka-Hajnos. Determination of vortioxetine in human serum and saliva samples by HPLC–DAD and HPLC–MS. Acta Chromatographica 2017, 29, 325 -344.

AMA Style

Karol Wróblewski, A. Petruczynik, B. Buszewski, Małgorzata Szultka-Młyńska, Hanna Karakula-Juchnowicz, M. Waksmundzka-Hajnos. Determination of vortioxetine in human serum and saliva samples by HPLC–DAD and HPLC–MS. Acta Chromatographica. 2017; 29 (3):325-344.

Chicago/Turabian Style

Karol Wróblewski; A. Petruczynik; B. Buszewski; Małgorzata Szultka-Młyńska; Hanna Karakula-Juchnowicz; M. Waksmundzka-Hajnos. 2017. "Determination of vortioxetine in human serum and saliva samples by HPLC–DAD and HPLC–MS." Acta Chromatographica 29, no. 3: 325-344.

Journal article
Published: 01 August 2017 in JPC – Journal of Planar Chromatography – Modern TLC
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Tomasz Tuzimski; Anna Petruczynik. Application of mobile phases containing ionic liquid for the separation of a mixture of ten selected isoquinoline alkaloids by 2D-TLC and identification of analytes in Rhizoma Coptidis (Huang Lian) Extract by TLC and HPLC—DAD. JPC – Journal of Planar Chromatography – Modern TLC 2017, 30, 245 -250.

AMA Style

Tomasz Tuzimski, Anna Petruczynik. Application of mobile phases containing ionic liquid for the separation of a mixture of ten selected isoquinoline alkaloids by 2D-TLC and identification of analytes in Rhizoma Coptidis (Huang Lian) Extract by TLC and HPLC—DAD. JPC – Journal of Planar Chromatography – Modern TLC. 2017; 30 (4):245-250.

Chicago/Turabian Style

Tomasz Tuzimski; Anna Petruczynik. 2017. "Application of mobile phases containing ionic liquid for the separation of a mixture of ten selected isoquinoline alkaloids by 2D-TLC and identification of analytes in Rhizoma Coptidis (Huang Lian) Extract by TLC and HPLC—DAD." JPC – Journal of Planar Chromatography – Modern TLC 30, no. 4: 245-250.

Journal article
Published: 01 June 2017 in Acta Chromatographica
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Three independent reversed phase high-performance liquid chromatography (HPLC) procedures with diode array detection (DAD) for the analysis of carbamazepine (CBZ), topiramate (TPM), and valproic acid (VPA) have been developed in order to determine drug penetration of the blood–brain barrier. Determination of CBZ was performed on C18 column with mobile phases containing methanol (55%, v/v), acetate buffer at pH 3.5 (20%, v/v), double distilled water (25%, v/v), and 0.025 M L−1 diethylamine (DEA). The mobile phase containing acetonitrile and water (8:2, v/v) or acetonitrile and phosphate-citrate buffer at pH 2.6 (1:1), respectively, for analysis of VPA and TPM was applied. Quantification of carbamazepine was performed at 285 nm without extraction procedure before the analysis. Determination of topiramate and valproic acid was performed using precolumn derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). FMOC-Cl is a suitable agent, which reacts with both primary and secondary amines and also with acidic groups. Topiramate was determined at 263 nm and valproic acid at 300 nm. The proposed procedures are simple, not time-consuming, and suitable for the determination of investigated compounds in mouse brain homogenates.

ACS Style

Karol Wróblewski; A. Petruczynik; I. Radzik; S. J. Czuczwar; M. Waksmundzka-Hajnos. Determination of selected antiepileptic drugs in mouse brain homogenates by HPLC—DAD. Acta Chromatographica 2017, 29, 219 -234.

AMA Style

Karol Wróblewski, A. Petruczynik, I. Radzik, S. J. Czuczwar, M. Waksmundzka-Hajnos. Determination of selected antiepileptic drugs in mouse brain homogenates by HPLC—DAD. Acta Chromatographica. 2017; 29 (2):219-234.

Chicago/Turabian Style

Karol Wróblewski; A. Petruczynik; I. Radzik; S. J. Czuczwar; M. Waksmundzka-Hajnos. 2017. "Determination of selected antiepileptic drugs in mouse brain homogenates by HPLC—DAD." Acta Chromatographica 29, no. 2: 219-234.

Journal article
Published: 03 April 2017 in Journal of Liquid Chromatography & Related Technologies
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Selected alkaloids were chromatographed on cyanopropyl-silica thin layers using nonaqueous and aqueous eluents containing various free silanol blocker agents. Because of the strong retention of these basic compounds, nonaqueous eluents containing medium polar diluents, strongly polar modifiers, or aqueous eluents containing organic modifier, water, and silanol blockers (ammonia, diethylamine, and ionic liquid) were required for analysis. Mobile phases containing addition of acids were also tested for separation of investigated alkaloids. The most selective and efficient systems were used in two-dimensional separations of isoquinoline alkaloids’ mixture and plant extracts.

ACS Style

A. Petruczynik; J. Misiurek; S. Serafin; M. Waksmundzka-Hajnos. The effect of chromatographic conditions on the separation of selected alkaloids on CN-silica layers. Journal of Liquid Chromatography & Related Technologies 2017, 40, 1 -7.

AMA Style

A. Petruczynik, J. Misiurek, S. Serafin, M. Waksmundzka-Hajnos. The effect of chromatographic conditions on the separation of selected alkaloids on CN-silica layers. Journal of Liquid Chromatography & Related Technologies. 2017; 40 (5-6):1-7.

Chicago/Turabian Style

A. Petruczynik; J. Misiurek; S. Serafin; M. Waksmundzka-Hajnos. 2017. "The effect of chromatographic conditions on the separation of selected alkaloids on CN-silica layers." Journal of Liquid Chromatography & Related Technologies 40, no. 5-6: 1-7.

Journal article
Published: 01 April 2017 in JPC – Journal of Planar Chromatography – Modern TLC
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In this study, authors propose a simple and cost-effective two-dimensional thin-layer chromatography (2D-TLC) method for the simultaneous determination of eleven standards alkaloids: allocryptopine (A), berberine (Be), boldine (Bo), chelidonine (Ch), glaucine (G), papaverine (Pa), emetine (E), columbamine (Col), magnoflorine (M), palmatine (Pal), and coptisine (Cop). Separation of the alkaloid mixture was achieved by 2D-TLC using an aqueous mobile phase (RP) in the first dimension (80% methanol + water + 0.05 mL−1 diethylamine) and a normal phase (NP) in the second dimension (18% methanol, 18% acetone in 63% diisopropyl ether containing 1% ammonia, v/v) on bilayer Multi-K CS5 plates. The composition of the mobile phases was optimized in terms of retention, separation selectivity, spots symmetry, and system efficiency. The procedure was evaluated in terms of natural samples analysis. Magnoflorine and berberine were identified in Thalictrum foetidum root extract. Additionally, the alkaloids in the extract sample were confirmed by high-performance liquid chromatography—diode-array detection method.

ACS Style

Tomasz Tuzimski; Anna Petruczynik; Justyna Misiurek. Separation of a mixture of eleven alkaloids by 2D-TLC on Multi-K CS5 plates and identification of analytes in Thalictrum foetidum root extract by TLC and HPLC—DAD. JPC – Journal of Planar Chromatography – Modern TLC 2017, 30, 142 -147.

AMA Style

Tomasz Tuzimski, Anna Petruczynik, Justyna Misiurek. Separation of a mixture of eleven alkaloids by 2D-TLC on Multi-K CS5 plates and identification of analytes in Thalictrum foetidum root extract by TLC and HPLC—DAD. JPC – Journal of Planar Chromatography – Modern TLC. 2017; 30 (2):142-147.

Chicago/Turabian Style

Tomasz Tuzimski; Anna Petruczynik; Justyna Misiurek. 2017. "Separation of a mixture of eleven alkaloids by 2D-TLC on Multi-K CS5 plates and identification of analytes in Thalictrum foetidum root extract by TLC and HPLC—DAD." JPC – Journal of Planar Chromatography – Modern TLC 30, no. 2: 142-147.

Articles
Published: 20 January 2017 in Journal of Liquid Chromatography & Related Technologies
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A simple and sensitive high-performance liquid chromatographic method with diode array detection is described for the quantification of some psychotropic drugs: fluoxetine, sertraline, alprazolam, perphenazine, zolpidem, and hydroxyzine in fortified human serum samples. The test compounds were extracted from human serum by solid-phase extraction using C18 extraction column and injected into C18 or Polar-RP analytical columns for separation. A mobile phase was composed of methanol, water, acetate buffer at pH 3.5, and diethylamine. The method was validated for the concentration range 0.4–5 µg mL−1 for zolpidem and 0.5–6 µg mL−1 for other drugs. Mean recoveries were from 87.79% to 107.94% with adequate precision (% RSD ≤2.1%). The full separation of all investigated drugs, good peaks’ symmetry, and simultaneously high system efficiency were obtained on Polar-RP column, which was used for the first time to analyze these drugs. System efficiency obtained on the column was significantly higher compared to that obtained on commonly used C18 column. The method seems to be suitable for the analysis of investigated drugs in human plasma for psychiatric patients in multiple drug overdoses as well as for control of pharmacotherapy, particularly in combination therapy.

ACS Style

Karol Wróblewski; Anna Petruczynik; Monika Waksmundzka-Hajnos. Separation and determination of selected psychotropic drugs in human serum by SPE/HPLC/DAD on C18 and Polar-RP columns. Journal of Liquid Chromatography & Related Technologies 2017, 40, 75 -82.

AMA Style

Karol Wróblewski, Anna Petruczynik, Monika Waksmundzka-Hajnos. Separation and determination of selected psychotropic drugs in human serum by SPE/HPLC/DAD on C18 and Polar-RP columns. Journal of Liquid Chromatography & Related Technologies. 2017; 40 (2):75-82.

Chicago/Turabian Style

Karol Wróblewski; Anna Petruczynik; Monika Waksmundzka-Hajnos. 2017. "Separation and determination of selected psychotropic drugs in human serum by SPE/HPLC/DAD on C18 and Polar-RP columns." Journal of Liquid Chromatography & Related Technologies 40, no. 2: 75-82.