PMID- 32872484 OWN - NLM STAT- MEDLINE DCOM- 20210617 LR - 20210617 IS - 2072-6651 (Electronic) IS - 2072-6651 (Linking) VI - 12 IP - 9 DP - 2020 Aug 29 TI - Determination of Cytisine and N-Methylcytisine from Selected Plant Extracts by High-Performance Liquid Chromatography and Comparison of Their Cytotoxic Activity. LID - 10.3390/toxins12090557 [doi] LID - 557 AB - 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. FAU - Petruczynik, Anna AU - Petruczynik A AUID- ORCID: 0000-0003-3294-3217 AD - Department of Inorganic Chemistry, Medical University of Lublin, Chodzki 4a, 20-093 Lublin, Poland. FAU - Wroblewski, Karol AU - Wroblewski K AUID- ORCID: 0000-0002-8918-7056 AD - Department of Experimental and Clinical Pharmacology, University of Rzeszow, Kopisto 2a, 35-959 Rzeszow, Poland. AD - Laboratory for Innovative Research in Pharmacology, University of Rzeszow, Kopisto 2a, 35-959 Rzeszow, Poland. FAU - Misiurek, Justyna AU - Misiurek J AUID- ORCID: 0000-0002-0791-4222 AD - Department of Inorganic Chemistry, Medical University of Lublin, Chodzki 4a, 20-093 Lublin, Poland. FAU - Plech, Tomasz AU - Plech T AUID- ORCID: 0000-0002-8162-8435 AD - Department of Pharmacology, Medical University of Lublin, Chodzki 4a, 20-093 Lublin, Poland. FAU - Szalast, Karolina AU - Szalast K AUID- ORCID: 0000-0003-4657-490X AD - Department of Pharmacology, Medical University of Lublin, Chodzki 4a, 20-093 Lublin, Poland. FAU - Wojtanowski, Krzysztof AU - Wojtanowski K AD - Department of Pharmacognosy with Medicinal Plant Unit, Medical University of Lublin, Chodzki 1, 20-093 Lublin, Poland. FAU - Mroczek, Tomasz AU - Mroczek T AD - Department of Pharmacognosy with Medicinal Plant Unit, Medical University of Lublin, Chodzki 1, 20-093 Lublin, Poland. FAU - Szymczak, Grazyna AU - Szymczak G AD - Botanical Garden of Maria Curie-Sklodowska University in Lublin, Slawinkowska 3, 20-810 Lublin, Poland. FAU - Waksmundzka-Hajnos, Monika AU - Waksmundzka-Hajnos M AD - Department of Inorganic Chemistry, Medical University of Lublin, Chodzki 4a, 20-093 Lublin, Poland. FAU - Tutka, Piotr AU - Tutka P AD - Department of Experimental and Clinical Pharmacology, University of Rzeszow, Kopisto 2a, 35-959 Rzeszow, Poland. AD - Laboratory for Innovative Research in Pharmacology, University of Rzeszow, Kopisto 2a, 35-959 Rzeszow, Poland. AD - National Drug and Alcohol Research Centre, University of New South Wales, Sydney, NSW 2031, Australia. LA - eng PT - Comparative Study PT - Journal Article DEP - 20200829 PL - Switzerland TA - Toxins (Basel) JT - Toxins JID - 101530765 RN - 0 (Alkaloids) RN - 0 (Antineoplastic Agents, Phytogenic) RN - 0 (Azocines) RN - 0 (Plant Extracts) RN - 0 (Quinolizines) RN - 53S5U404NU (cytisine) RN - TT0MW69NCI (N-methylcytisine) SB - IM MH - Alkaloids/*isolation & purification/pharmacology MH - Antineoplastic Agents, Phytogenic/*isolation & purification/pharmacology MH - Azocines/isolation & purification/pharmacology MH - Cell Survival/drug effects MH - *Chromatography, High Pressure Liquid MH - Dose-Response Relationship, Drug MH - Humans MH - MCF-7 Cells MH - Neoplasms/drug therapy/pathology MH - Plant Extracts/*isolation & purification/pharmacology MH - Quinolizines/isolation & purification/pharmacology MH - Spectrometry, Mass, Electrospray Ionization MH - Tandem Mass Spectrometry PMC - PMC7551552 OTO - NOTNLM OT - HPLC OT - N-methylcytisine OT - cytisine OT - cytotoxicity OT - plant extracts COIS- The author declares no conflict of interest. EDAT- 2020/09/03 06:00 MHDA- 2021/06/22 06:00 PMCR- 2020/09/01 CRDT- 2020/09/03 06:00 PHST- 2020/07/24 00:00 [received] PHST- 2020/08/24 00:00 [revised] PHST- 2020/08/27 00:00 [accepted] PHST- 2020/09/03 06:00 [entrez] PHST- 2020/09/03 06:00 [pubmed] PHST- 2021/06/22 06:00 [medline] PHST- 2020/09/01 00:00 [pmc-release] AID - toxins12090557 [pii] AID - toxins-12-00557 [pii] AID - 10.3390/toxins12090557 [doi] PST - epublish SO - Toxins (Basel). 2020 Aug 29;12(9):557. doi: 10.3390/toxins12090557.