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Öğe Analysis of non-carcinogenic health risk assessment of elemental impurities in vitamin C supplements(Mashhad University of Medical Sciences, 2023) Canbolat, FadimeObjective(s): Elemental impurity exposure that may occur in the use of supplements has the potential to pose a risk to human health. Vitamin C supplements are among the most commonly used supplements on a daily basis and in the long-term due to the pharmacological properties of vitamin C. In this study, we aimed to evaluate the non-carcinogenic health risk of elemental impurities that may cause contamination in orally administered vitamin C supplements. Materials and Methods: Ten elemental impurities (Cd, Pb, As, Hg, Co, V, Ni, Cr, Sb, and Sn) in 12 supplements were analyzed using ICP-MS. The estimated daily intake (EDI), hazard quotient (HQ), and hazard index (HI) values of elemental impurities were calculated for non-carcinogenic risk assessment. Cancer risk (CR) was additionally calculated for elemental impurities with carcinogenic properties detected in the samples. Results: Low levels of Cr and Hg were detected in some samples. While the HQ values of sample 1, sample 2, sample 8, and sample 9 for Hg were calculated as 0.054, 0.096, 0.064, and 0.086, respectively, the HQ values of sample 5, sample 10, and sample 11 for Cr were calculated as 0.011, 0.017, and 0.014, respectively. Since only Hg or only Cr was detected in samples with elemental impurity, the HI values in the samples are the same as the HQ values. Since the HQ and HI values calculated from the samples are not≥1, there is no elemental impurity at a level that will hazard human health through supplement use. Other carcinogenic elements were not detected in the samples except Cr. In sample 5, sample 10, and sample 11, the CR values for Cr were 1.767.10-5, 2.571.10-5, and 2.089.10-5, respectively. In probability simulation, while HQ and CR values of Cr did not exceed the allowable value, the HQ level for Hg in the 95% slice was higher than the allowable value. Conclusion: There is no risk to human health and there is no critical difference between the supplements considering the elemental pollutant content among the vitamin C supplements of different trademarks. However, in order to keep the Hg level, which has a potential risk capacity, at low limits, it is recommended that the necessary risk-reducing measures be taken by the authorities and further studies be carried out.Öğe Characterization, biological activity, and anticancer effect of green-synthesized gold nanoparticles using Nasturtium officinale L.(BMC, 2024) Yayintas, Ozlem Tonguc; Demir, Neslihan; Canbolat, Fadime; Ayna, Tuelay Kilicaslan; Pehlivan, MelekBackground Nanostructured materials used have unique properties and many uses in nanotechnology. The most striking of these is using herbal compounds for the green synthesis of nanoparticles. Among the nanoparticle types used for green synthesis, gold nanoparticles (AuNPs) are used for cancer therapy due to their stable structure and non-cytotoxic. Lung cancer is the most common and most dangerous cancer worldwide in terms of survival and prognosis. In this study, Nasturtium officinale (L.) extract (NO), which contains biomolecules with antioxidant and anticancer effects, was used to biosynthesize AuNPs, and after their characterization, the effect of the green-synthesized AuNPs against lung cancer was evaluated in vitro. Methods Ultraviolet-visible (UV-Vis) spectrophotometry, scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDS), multiple analysis platform (MAP), and Fourier transform infrared (FT-IR) spectroscopy analyses were performed to characterize the AuNPs prepared from the N. officinale plant extract. Moreover, the antioxidant activity, total phenolic and flavonoid contents and DNA interactions were examined. Additionally, A549 lung cancer cells were treated with 2-48 mu g/mL Nasturtium officinale gold nanoparticles (NOAuNPs) for 24 and 48 h to determine the effects on cell viability. The toxicity of the synthesized NOAuNPs to lung cancer cells was determined by the 3-(4,5-dimethylthiazol-2-il)-2,5-diphenyltetrazolium bromide (MTT) assay, and the anticancer effect of the NOAuNPs was evaluated by apoptosis and cell cycle analyses using flow cytometry. Results The average size of the NPs was 56.4 nm. The intensities of the Au peaks from EDS analysis indicated that the AuNPs were synthesized successfully. Moreover, the in vitro antioxidant activities of the NO and NOAuNPs were evaluated; these materials gave values of 31.78 +/- 1.71% and 31.62 +/- 0.46%, respectively, in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay at 200 g/mL and values of 25.89 +/- 1.90% and 33.81 +/- 0.62%, respectively, in the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay. The NO and NOAuNPs gave values of 0.389 +/- 0.027 and 0.308 +/- 0.005, respectively, in the ferrous ion reducing antioxidant capacity (FRAP) assay and values of 0.078 +/- 0.009 and 0.172 +/- 0.027, respectively, in the copper ion reducing antioxidant capacity (CUPRAC) assay. When the DNA cleavage activities of NO and the NOAuNPs were evaluated via hydrolysis, both samples cleaved DNA starting at a concentration of 25 g/mL in the cell culture analysis, while the nanoformulation of the NO components gave greater therapeutic and anticancer effects. We determined that the Au nanoparticles were not toxic to A549 cells. Moreover, after treatment with the half-maximal inhibitory concentration (IC50), determined by the MTT assay with A549 cells, we found that at 24 and 48 h, while the necrosis rates were high in cells treated with NO, the rates of apoptosis were greater in cells treated with NOAuNPs. Notably, for anticancer treatment, activating apoptotic pathways that do not cause inflammation is preferred. We believe that these results will pave the way for the use of NOAuNPs in in vitro studies of other types of cancer. Conclusion In this study, AuNPs were successfully synthesized from N. officinale extract. The biosynthesized AuNPs exhibited toxicity to and apoptotic effects on A549 lung cancer cells. Based on these findings, we suggest that green-synthesized AuNPs are promising new therapeutic agents for lung cancer treatment. However, since this was an in vitro study, further research should be performed in in vivo lung cancer models to support our findings and to explain the mechanism of action at the molecular level.Öğe Chitosan Nanoparticles Loaded with Quercetin and Valproic Acid: A Novel Approach for Enhancing Antioxidant Activity against Oxidative Stress in the SH-SY5Y Human Neuroblastoma Cell Line(Mdpi, 2024) Canbolat, Fadime; Demir, Neslihan; Yayintas, Ozlem Tonguc; Pehlivan, Melek; Eldem, Asli; Ayna, Tulay Kilicaslan; Senel, MehmetBackground: Multiple drug-delivery systems obtained by loading nanoparticles (NPs) with different drugs that have different physicochemical properties present a promising strategy to achieve synergistic effects between drugs or overcome undesired effects. This study aims to develop a new NP by loading quercetin (Que) and valproic acid (VPA) into chitosan. In this context, our study investigated the antioxidant activities of chitosan NPs loaded with single and dual drugs containing Que against oxidative stress. Method: The synthesis of chitosan NPs loaded with a single (Que or VPA) and dual drug (Que and VPA), the characterization of the NPs, the conducting of in vitro antioxidant activity studies, and the analysis of the cytotoxicity and antioxidant activity of the NPs in human neuroblastoma SH-SY5Y cell lines were performed. Result: The NP applications that protected cell viability to the greatest extent against H2O2-induced cell damage were, in order, 96 mu g/mL of Que-loaded chitosan NP (77.30%, 48 h), 2 mu g/mL of VPA-loaded chitosan NP (70.06%, 24 h), 96 mu g/mL of blank chitosan NP (68.31%, 48 h), and 2 mu g/mL of Que- and VPA-loaded chitosan NP (66.03%, 24 h). Conclusion: Our study establishes a successful paradigm for developing drug-loaded NPs with a uniform and homogeneous distribution of drugs into NPs. Chitosan NPs loaded with both single and dual drugs possessing antioxidant activity were successfully developed. The capability of chitosan NPs developed at the nanometer scale to sustain cell viability in SH-SY5Y cell lines implies the potential of intranasal administration of chitosan NPs for future studies, offering protective effects in central nervous system diseases.Öğe Comparison of Normal saline, Activated Charcoal and Intravenous Lipid Emulsion in a Rat Model of Colchicine Overdose: Experimental Study(Assoc Pharmaceutical Teachers India, 2024) Celikmen, Mustafa Ferudun; Sarikaya, Sezgin; Ozucelik, Dogac Niyazi; Canbolat, Fadime; Sumer, Engin; Keles, Elif cigdem; Celikmen, Deniz Sema MaktavAim: The aim of this study is to investigate the effects of Normal Saline (NS), Activated Charcoal (AC) and Intravenous Lipid Emulsion (ILE) in colchicine poisonings that resulted in death. Study Design: The research is an experimental study carried out in a medical school animal laboratory. Materials and Methods: 24 female Sprague-Dawley rats were divided into 4 equal groups. After giving Colchicine (1 mg/kg, PO) to all animals, different treatments (NS, AC, ILE) were given to 3 groups. Group 4 was not treated. Colchicine blood samples (0.8 mL) were taken from the vena jugularis externa at 4, 8 and 24 hr and evaluated by liquid chromatography. Results: While the blood colchicine level decreased in all groups at 24 hr, the highest decrease was observed in the AC group. Compared to the 4 th hr, blood colchicine levels at 8 hr decreased by 15.49% in the NS group, 64.55% in the AC group and 34.56% in the ILE group. Blood colchicine levels at 24 hr decreased by 66.1% in the NS group, 73.12% in the AC group, 37.73% in the ILE group and 59.17% in the untreated group compared to the 8 th hr. Conclusion: AC administration is very effective in lowering blood colchicine levels. ILE can be used with NS and AC as an early treatment option for colchicine overdoses. However, further studies are needed for more effective methods in the treatment of colchicine overdose.Öğe Development of chitosan nanoparticle loaded with Tricholoma fracticum extract and evaluation of in vitro antioxidant activity(Wiley, 2024) Canbolat, Fadime; Acar, Ismail; Tezel, Ruhiye NilayThe objective of this study was to develop Tricholoma fracticum extract-loaded chitosan nanoparticles (TFNPs) by ionic gelation method and to evaluate their in vitro antioxidant activity. Phenolic and flavonoid contents in the T. fracticum extract were measured spectrophotometrically and chromatographically. Characterisation of NPs was evaluated by field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), ZETA analysis, Fourier-transform infrared spectroscopy (FT-IR), UV-visible spectroscopy (UV-Vis) and thermogravimetric analysis (TGA). In vitro antioxidant capacity was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2 '-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. The phenolic and flavonoid contents in the T. fracticum extract were measured as 7.1 +/- 0.3 mg Gallic Acid Equivalent/g extract and 5.5 +/- 0.6 mg Quercetin Equivalent/g extract, respectively. The particle size, polydispersity index (PDI) and zeta potential of Blank NP1 and TFNP were 265.5 +/- 15.8 nm, 0.4, 38.7 +/- 4.0 mV and 333.2 +/- 16.3 nm, 0.4, 37.0 +/- 4.1 mV, respectively. The highest antioxidant activity was observed in TFNP, followed by T. fracticum extract, chitosan and blank NP, respectively. The preserved or enhanced antioxidant activity observed in the encapsulated T. fracticum extract indicates the potential for loading similar mushroom extracts onto chitosan and thus preserving their bioactive properties.Öğe Elemental Impurity Analysis in Five Different Types of Coffee: Assessment of Carcinogenic and Non-carcinogenic Risks(2024) Canbolat, FadimeIn the study, the levels of Cd, Pb, As, and Hg elemental impurities potentially present in coffee bean samples from Indonesia, Kenya, Colombia, Guatemala, and Türkiye were determined using chromatographic analysis, and the analysis results were utilized to assess the risks on human health. The risk assessment of coffee was calculated for one or three servings per day for 365 days a year. Exposure to coffee consumption was calculated according to age groups of young adults and middle-aged adults (20-65 years). When Cd, Pb, As and Hg levels in coffee samples were analyzed, Cd in coffee samples was found to be in the range of approximately 3.70 - 5.89 µg/kg, Pb in the range of 25.68-41.11 µg/kg, As in the range of 1.45-6.64 µg/kg and Hg in the range of 1.06-5.06 µg/kg. Hazard Index (HI) values for all elements in the assessment of non-carcinogenic risks were found to be <1.0. When the cancer risk (CR) value was calculated for Cd, Pb, and As, it was found that the CR value did not exceed the United States Environmental Protection Agency (USEPA) criteria in all coffee samples in both scenarios. Considering the assessment of the health risks of elemental impurities in five different coffee bean samples, it was concluded that all samples' CR and HI values did not exceed the USEPA criteria.Öğe Evaluation of Lipid Emulsion-Mediated Sequestration and Redistribution of the Highly Lipophilic Carbamazepine in the Plasma of Rats(2024) Ekşioğlu, Merve; Gürsoy, Deniz Algedik; Sümer, Engin; Canbolat, Fadime; Sarıkaya, SezginAim: The idea that intravenous lipid emulsion (ILE) may serve as a \"reservoir\" for lipophilic drugs has emerged in research as an intravascular \"lipid sink\" effect. Carbamazepine (CBZ) is a widely used anticonvulsant. This compound has a neutral and highly lipophilic structure and can easily cross body membranes. In this study, our hypothesis focused on the potential efficacy of ILE in modulating blood carbamazepine concentrations. Material and Methods: 22 adult Sprague-Dawley rats were divided into four groups. All groups received CBZ at a dose of 20 mg/kg orogastrically. The first group was the control group. In the second group (activated charcoal group), activated charcoal (AC) was administered orogastrically at a dose of 1 g/kg five minutes after orogastric administration of carbamazepine. The third group (lipid group) received ILE at a dose of 3 ml/kg/min at the fifth minute. The fourth group was the saline group, in which 16 ml/kg of 0.9% NaCl was infused at the fifth minute. Blood samples of 0.5 ml were collected at 0, 4, 8, and 24 hours. Plasma was separated by centrifugation (4000 rpm, 10 minutes) and stored at -80oC for determination of CBZ concentrations. An Agilent 6410B HP-1200 LC series (USA) liquid chromatography system was used for analysis. Quantitative analysis was performed in the multiple reaction mode with electrospray positive ionization (ES+). Results: At the 8th hour of orogastric CBZ administration, CBZ concentration was significantly lower in the activated charcoal group than in the lipid and saline groups (p: 0.021; p: 0.023; p<0.05, respectively). There was no significant difference in CBZ concentrations between the other groups at 8 hours (p>0.05). In the lipid group, the increase in CBZ plasma concentrations was statistically significant at 4 and 8 hours compared to 0 hours (p: 0.005; p: 0.005, respectively). Conclusion: In the lipid group, plasma CBZ concentrations increased at 4 and 8 hours in plasma samples from which lipids were separated by differential centrifugation. In the lipid group, no effects favoring drug-lipid sequestration on the plasma distribution of CBZ were observed.Öğe Evaluation of the Antidepressant Effect of Propolis in Chronic Unpredictable Mild Stress-Induced Depression Model in Rats(Galenos Publ House, 2024) Taskiran, Ali; Canbolat, Fadime; Yucelli, Sena Nur; Cevreli, BurcuAim: In this study, the antidepressant effect of propolis was investigated in a model of chronic unpredictable depression in rats. Materials and Methods: Wistar-Albino male rats were used in the study and were divided into four groups as propolis, stress, stress + propolis, and control groups. Eight animals were assigned to each group. The experimental protocol was applied to the stress groups for 60 days, and the animals were exposed to different stressors. Propolis extract (100 mg/kg) was administered orally to propolis and stress + propolis groups throughout the experimental protocol. As a result of depression modeling, the Forced Swimming Test, Sucrose Preference Test, and Elevated Plus Maze Test were applied for behavioral evaluation. Twenty-four hour urine samples were collected for quantitative analysis of serotonin 5-hydroxytryptamine (5-HT) and its metabolite 5-hydroxy indole acetic acid (5-HIAA) in urine by liquid chromatography-tandem mass spectrometry method. The animals were sacrificed as a result of the experiment process. Results: It was seen that there was a statistical difference for behavioral tests between the groups (p<0.05). The administration of propolis to rats under stress has been shown to alter sugar consumption in rats (p<0.05). For Forced Swimming Test, there was a statistical difference between the stress group and the other groups. For 5-HT and 5-HIAA levels, there was no significant difference between the groups (p>0.05). Conclusion: The findings have shown that propolis extract may help to prevent depression, thanks to its antidepressant-like effects.Öğe Evaluation of the Antidepressant Effect of Propolis in Chronic Unpredictable Mild Stress-induced Depression Model in Rats(Galenos Publ House, 2023) Taskiran, Ali; Canbolat, Fadime; Yucelli, Sena Nur; Cevreli, BurcuAim: In this study, the antidepressant effect of propolis was investigated in a model of chronic unpredictable depression in rats. Materials and Methods: Wistar-Albino male rats were used in the study and were divided into four groups as propolis, stress, stress+propolis, and control groups. Eight animals were assigned to each group. The experimental protocol was applied to the stress groups for 60 days, and the animals were exposed to different stressors. Propolis extract (100 mg/kg) was administered orally to propolis and stress+propolis groups throughout the experimental protocol. As a result of depression modeling, the Forced Swimming Test, Sucrose Preference Test, and Elevated Plus Maze Test were applied for behavioral evaluation. 24-hour urine samples were collected for quantification analysis of serotonin (5-HT) and its metabolite 5-hydroxy indole acetic acid (5-HIAA) in urine by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. The animals were sacrificed as a result of the experiment process. The results obtained from the study were analyzed using the Statistical Package for the Social Sciences. Results: It was seen that there was a statistical difference for behavioral tests between the groups (p<0.05). The administration of propolis to rats under stress has been shown to alter sugar consumption in rats (p<0.05). For Forced Swimming Test, there was a statistical difference between the stress group and the other groups. For 5-HT and 5-HIAA levels, there was no significant difference between the groups (p>0.05). Conclusion: The findings have shown that propolis extract may help to prevent depression, thanks to its antidepressant-like effects.Öğe Evaluation of the Antigenotoxic Effect of Quercetin Against Antiepileptic Drug Genotoxicity by Comet Analysis(2023) Canbolat, Fadime; Kenanoglu, Nihan Akıncı; Yuksel, Tugba Nurcan; Berber, Ahmet AliValproic acid (VPA) is among the most commonly used antiepileptic drugs in childhood and adult epilepsy. Although VPA is well tolerated, it can cause life-threatening side effects. VPA has toxic and genotoxic effects. Antioxidants can reverse drugs' toxic and genotoxic effects. Therefore, our study aimed to evaluate the antigenotoxic protective effect of quercetin (QUE) against VPA genotoxicity by in vitro comet assay analysis method. Comet assay analysis was performed in five different groups. Group I; negative control (Sterile H2O), Group II; positive control (H2O2), Group III; VPA was applied in four different dose ranges, Group IV; QUE was applied in four different dose ranges, Group V; For the simultaneous combined administration of VPA and QUE, three different doses of VPA + four different doses of QUE were administered. Low-dose administration of QUE was more effective in ameliorating the damage caused by low-dose VPA (62.5 ?g/ml) administration. It is seen that the genotoxic damage caused by the application of 125 ?g/ml VPA can be eliminated by QUE at all doses. It was determined that different doses of QUE exhibited a significant antigenotoxic effect against damage caused by 125 µg/mL VPA (P<0.05). In our study, the curative effect of QUE on DNA damage was determined by in vitro comet analysis. Our analysis results showed that QUE ameliorates VPA-induced genetic damage.Öğe Evaluation of the bioactivities of turmeric spices of different origins(2023) Canbolat, Fadime; Ateş, Gülçin ÖzcanOur study aimed to compare the in vitro bioactivities of turmeric spice samples obtained from three different sources (India, Pakistan, and Indonesia). Our study involved the determination of total phenolic and flavonoid content, in vitro antioxidant activities, tyrosinase enzyme activity, and antimicrobial activity (Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Candida albicans) analyses in three different spice samples. Each spice sample was dissolved in 250 mL of ethanol and stirred on a magnetic stirrer for 36 hours. After filtering out the solid parts, the residues were dissolved again in 250 mL of ethanol separately. This process was repeated three times. After the filtration steps, all filtrates were combined. The remaining solvents in the filtrate were evaporated using an evaporator. The residue of the extracts was placed in Eppendorf tubes and stored in a freezer until use. It has been determined that turmeric of Indian origin, with its high phenolic and flavonoid content, exhibits more potent antioxidant and antityrosinase effects than those from Indonesia and Pakistan. However, antimicrobial activity could not be detected within the studied concentration range of 10 µg/mL to 400 mg/mL.Öğe Identification of the Candidate mGlu2 Allosteric Modulator THRX-195518 through In Silico Method and Evaluation of Its Neuroprotective Potential against Glutamate-Induced Neurotoxicity in SH-SY5Y Cell Line(Multidisciplinary Digital Publishing Institute (MDPI), 2024) Canbolat, Fadime; Kantarci-Carsibasi, Nigar; Işık, Sevim; Shamshir, Suhair Rami Mohammed; Girgin, MüntehaGlutamate (Glu) toxicity has been an important research topic in toxicology and neuroscience studies. In vitro and in vivo studies have shown that Group II metabotropic Glu2 (mGlu2) activators have cell viability effects. This study aims to determine a candidate ligand with high mGlu2 allosteric region activity among cytotoxicity-safe molecules using the in silico positioning method and to evaluate its cell viability effect in vitro. We investigated the candidate molecule’s cell viability effect on the SH-SY5Y human neuroblastoma cell line by MTT analysis. In the study, LY 379268 (agonist) and JNJ-46281222 (positive allosteric modulator; PAM) were used as control reference molecules. Drug bank screening yielded THRX-195518 (docking score being −12.4 kcal/mol) as a potential novel drug candidate that has a high docking score and has not been mentioned in the literature so far. The orthosteric agonist LY 379268 exhibited a robust protective effect in our study. Additionally, our findings demonstrate that JNJ-46281222 and THRX-195518, identified as activating the mGlu2 allosteric region through in silico methods, preserve cell viability against Glu toxicity. Therefore, our study not only emphasizes the positive effects of this compound on cell viability against Glu toxicity but also sheds light on the potential of THRX-195518, acting as a mGlu2 PAM, based on in silico absorption, distribution, metabolism, excretion, and toxicity (ADMET) data, as a candidate drug molecule. These findings underscore the potential utility of THRX-195518 against both neurotoxicity and Central Nervous System (CNS) disorders, providing valuable insights.Öğe Protective Effects of Ramelteon on Acute Lung Injury in Endotoxin-Induced Sepsis in Rats(2023) Yüksel, Tuğba Nurcan; Köse, Duygu; Gürbüz, Muhammet Ali; Halıcı, Zekai; Canbolat, Fadime; Bozgeyik, EsraIntroduction: Sepsis is a life-threatening excessive systemic inflammatory reaction syndrome to infection that usually occurs in patients with bacteremia. The respiratory system is one of the structures most affected by acute organ damage. Melatonin plays an important role in re gulating various physiological functions of the body, including antioxidant and anti- inflammatory. Ramelteon (RAME) is the first melatonin receptor agonist confirmed for clinical use. The goal of this study is to determine the effects of RAME on endotoxin- induced septic lung injury in rats. Materials and Methods: Thirty-two female rats were separated randomly into four groups (n =8). Group healthy received intraperitoneal normal saline, group sepsis received intraperitoneally 10 mg/kg lipopolysaccharide (LPS), group sepsis+RAME2 received 10 mg/kg LPS plus 2mg/kg RAME, and group sepsis+RAME4 received 10 mg/kg LPS plus 4mg/kg RAME. RAME was administered by oral gavage 1 hour before LPS administration. The lung tissues were collected 12 hours after LPS administration and in vestigated molecularly (qRT- PCR analyses of Tumor Necrosis Factor-?, nuclear factor kappa-?, and interleukin 1-beta mRNA expression) and histopathologically (s taining with Harris Hematoxylin and Eosin Y). Results: TNF-?, NF- ??, and IL-1? levels significantly decreased dose-dependent in the septic rats following RAME administration. RAME admi nistration ameliorated histopathological injury in lung tissues due to sepsis. Conclusion: RAME ameliorated the inflammatory response in endotoxin-induced sepsis. These findings suggest that RAME can be a promising agent by contributing to alternative preventive treatment methods for sepsis with its anti- inflammatory effect.Öğe Validation of Colchicine Assay Method for Therapeutic Drug Monitoring in Human Plasma(2022) Canbolat, FadimeColchicine (COL) reduces the frequency of attacks in Familial Mediterranean Fever (FMF) patients and is effective in preventing and arresting renal amyloidosis in most patients. COL has a narrow therapeutic window. The blood concentration to achieve therapeutic effects can be determined by Therapeutic Drug Monitoring (TDM). However, the use of selective and sensitive analytical methods is necessary for achieving success with TDM. The purpose of this study is to develop and validate a new method for quantitative assay of COL in human plasma samples by liquid chromatograph- tandem mass spectrometry (LC-MS/MS). In our study, to 1ml plasma sample, 0.25 ml internal standard solution was added. The solution was extracted by liquid-liquid extraction (LLE). The method was validated according to the European Medicines Agency (EMEA). The total run time was 8 min in LC-MS/MS. The method has been validated over the 0.25 - 8.0 ng/mL calibration range for COL. It was seen that the method has been validated since the results of the analysis meet the EMEA criteria. In our study, COL plasma levels were found to be approximately 1.097±0.42 ng/ml in 40 FMF patients using an oral dose of 1.5-2 mg/day. A validated, rapid, simple, cost-effective, and sensitive LC-MS/MS method was developed and optimized for quantitation of COL in plasma. It has been thought that pharmacokinetic studies of COL in plasma can be performed easily using this validated method
