hplc
در نشریات گروه علوم پایه-
Gemcitabine is a potent cytotoxic agent classified as a pyrimidine nucleoside antimetabolite, widely used to treat various metastatic and prostatic cancers. Approved by the FDA in 1996, it is indicated for the treatment of non-small cell lung cancer, pancreatic cancer, and breast cancer. Advanced stages of pancreatic adenocarcinoma (II, III, or metastatic stage IV), where regional resection is no longer viable, gemcitabine often serves as the first-line treatment. Functioning as a prodrug, gemcitabine requires phosphorylation by deoxycytidine kinase to become its active form within cells, enabling its therapeutic effects. Extensive literature sourced from databases such as Google Scholar, Taylor & Francis, Science Direct, and PubMed has documented various analytical methods for estimating gemcitabine, either as a standalone drug or in combination with other agents. These methods span techniques such as UV spectrophotometry, Spectrofluorimetry, Fourier-transform infrared spectroscopy, High-Performance Liquid Chromatography (HPLC), High-Performance Thin-Layer Chromatography (HPTLC), and advanced hyphenated techniques. In addition, this review evaluates the greenness profiles of reported analytical methods (2003-2023) based on the National Environmental Method Index (NEMI), aligning with the Sustainable Development Goals (SDGs) 2030. The findings emphasize the importance of developing more robust, efficient, and eco-friendly methods, particularly through sorbent-based microextraction techniques, which could significantly enhance biomedical research. This review provides a valuable roadmap for researchers, encouraging the adoption of greener analytical approaches using eco-friendly solvents for the gemcitabine determination in biological matrices and drug products.
Keywords: Gemcitabine, UV, HPLC, UPLC, Biological Fluids -
Tridax procumbens is a medicinal plant extensively utilized in traditional medicine because of its antibacterial, anti-inflammatory, and wound-healing properties. Extraction and phytochemical analysis of T. procumbens seeks to determine its bioactive constituents, including flavonoids, alkaloids, and terpenoids. Apigenin is a significant flavonoid that has been isolated to assess its therapeutic capabilities. Since this plant possesses potential anti-inflammatory properties, we aimed to isolate and develop apigenin as one of its key bioactive constituents. Ultrasonic-assisted Soxhlet extraction of T. procumbens leaves and bark with a hydroalcoholic solvent (70% ethanol, 30% water) facilitated optimum recovery of phytoconstituents, followed by physicochemical, microbiological, and phytochemical evaluations to ascertain its composition, purity, and bioactive potential. Comprehensive investigations, including organoleptic assessment, heavy metal identification, and quantitative phytochemical profiling, have validated the purity and appropriateness of the extract for pharmaceutical use. Quantitative examination of T. procumbens extract identified several bioactive phytochemicals, including carbohydrates (11.38%), alkaloids (11.89%), and flavonoids (8.49%), underscoring potential therapeutic and nutritional advantages. An optimized HPLC approach utilizing a 70:30 methanol-water mobile phase and C18 column significantly enhanced the chromatographic separation and detection of Apigenin. This work offers significant insights into the composition and possible therapeutic qualities of T. procumbens extract. Future studies are needed to evaluate the synergistic activities of apigenin with other bioactive compounds in T. procumbens aimed at maximizing its therapeutic values. Moreover, in vivo studies and clinical trials are needed to test the extract's effectiveness and safety for its use in medicine. Further investigation on the molecular mechanisms explaining the anti-inflammatory activities of these plants could provide opportunities for creating new therapeutic methods.Keywords: Tridax Procumbens, Phytochemical Analysis, HPLC, Phytoconstituents, Ultrasonic Extraction
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Sustainable Extraction and Purification of Phytochemicals: A Review of Green Solvents and Techniques
The importance of phytochemicals in medicine, nutraceuticals, and cosmetics has recently drawn considerable attention toward their sustainable extraction and purification. This review presents the progress in green solvents and methodologies underlined by the drawbacks associated with conventional methods. Phytochemicals are bioactive substances derived from plants that are important for medicinal and commercial purposes. However, most classical methods of extraction and purification involve hazardous solvents and energy-intensive operations, raising environmental and safety concerns. Green solvents, including ionic liquids, deep eutectic solvents, and bio-based solvents, offer environmentally friendly alternatives with high efficiency and low toxicity. These include Supercritical fluid extraction (SFE), Ultrasound-Assisted Extraction (UAE), Microwave-Assisted Extraction (MAE), Pressurized liquid extraction (PLE), and Enzyme-Assisted Extraction (EAE), all of which are advanced green extraction methodologies. These methods afford higher extraction yields while simultaneously reducing the energy consumption and waste generation. Sophisticated purification strategies, including chromatographic techniques, membrane-based purification, and adsorption-desorption procedures, were evaluated for their compatibility with eco-friendly extraction processes. A comparison of the various techniques shows their efficacy, cost, and environmental friendliness. The discussion covers future perspectives, which pinpoint that the development of green solvent formulations, integration of extraction and purification systems, and overcoming of regulatory and scalability challenges are highly needed. This study will be able presents a comprehensive review of sustainable methodologies for the extraction and purification of phytochemicals, which would further help in devising greener, safer, and more efficient approaches in natural product research.
Keywords: Extraction, Phytochemical, Purification Methods, Phenols, HPLC -
Increasing use of amphetamine as a highly abused psychotropic agent has irreversible side effects and damages to both the consumer and the community. In this study, different solid-phase extraction methods in urine samples as well as various quantitative determination amphetamine methods were compared systematically. Web of Science, Scopus, and PubMed databases were searched up to April 12, 2022, for relevant articles. The random-effects model was used to estimate the pooled measurements and 95% confidence intervals (95% CI). All analyses were performed with STATA version 15.0. Bias publication was performed for available studies reported in the literature. After applying limit of detection (LOD) criteria, solid phase extraction, and amphetamine, 36 articles remained. Among the collected studies, 16 were included in the meta-analysis. This meta-analysis revealed that the minimum detection (i.e., the limit of detection or LOD) value for the solid-phase extraction method coupled to high-performance liquid chromatography/UV (HPLC/UV) quantification technique is equal to 7.88 ng/mL (95% CI: 3.35–12.40), while for the gas chromatography/mass (GC/MS) method, the LOD value is equal to 2.75 ng/mL (95% CI: 0.77–4.73). Despite the use of different absorbents and their varied efficiency, the methods based on GC/MS can detect the analyte with higher sensitivity than that of HPLC/UV.
Keywords: Amphetamine, HPLC, GC, LOD, Meta-Analysis -
A genetic mutation in the beta-globin gene causes the common illness known as Sickle Cell Disease (SCD). "Sickle Cell Disease (SCD) is caused by a genetic mutation in the beta-globin gene, leading to the synthesis of hemoglobin S." The mutation can manifest in either homozygous or heterozygous form, potentially in combination with another mutation in the hemoglobin gene. Various approaches were created to accurately identify Sickle Cell Disease with high sensitivity and specificity. The present study aimed to determine the frequency of various genotypes of Sickle Cell Disease (SCD) in Kurdistan, Iraq, using hematological testing, hemoglobin isolation, and genetic testing. One hundred patients were examined individually. The Duhok-Jin center in Kurdistan, Iraq, provided the data in 2019 and 2020. The analysis was conducted in 2020. The procedures used for screening include a complete blood count, while confirmatory tests involve hemoglobin separation and genetic tests. The cohort of 100 participants in the study had a mean age of 15.8. 61% of the patients were diagnosed with sickle cell anemia, whereas the remaining 39% were identified with S/β-thalassemia. "In individuals with sickle cell anemia, the concentrations of HbS, HbF, and HbA2 were measured as 65.0 ± 15.04, 19.52 ± 17.3, and 2.65 ± 0.8, respectively. In individuals with S/β-thalassemia, the concentrations were 63.03 ± 13.44, 19.95 ± 12.0, and 4.63 ± 0.9, respectively." This investigation demonstrates that combining hematologic tests and HPLC with a confirmatory test, such as DNA analysis, can lead to an accurate diagnosis of Sickle Cell Disease.Keywords: Sickle Cell Anemia, HPLC, PCR-RFLP
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مقدمه
الیگوزواسپرمی یکی از دلایل ناباروری در مردان است که براساس معیارهای سارمان جهانی بهداشت، با تعداد کم اسپرم در مقایسه با حالت نرموزواسپرمی همراه است. از طرفی اسفنگولیپیدها تنظیم کننده های مهم بسیاری از فرایندهای سلولی مانند تمایز و آپوپتوز سلول هستند. از مهمترین متابولیت های اسفنگولیپیدی میتوان به سرامیدها و اسفنگوزین اشاره کرد که از بین رفتن حالت پایدار در میزان سرامید میتواند منجر به آپوپتوز شود. هدف از این مطالعه بررسی تغییرات انواع مختلف سرامید و اسفنگوزین در نمونه های الیگوزواسپرمی از مردان نابارور است.
مواد و روش هادر این مطالعه نمونه سیمن از زوج های مراجعه کننده به مرکز درمان ناباروری الزهرا، رشت جمع آوری شدند (10=n). نمونه ها براساس پارامترهای سازمان جهانی بهداشت آنالیز شدند و گروه های الیگوزواسپرمی (اسپرم کمتر از 15 میلیون در هر میلی لیتر سیمن) و نرموزواسپرمی وارد مطالعه شدند. مقدار انواع سرامیدها (Cer14, Cer16, Cer18 و Cer 20) و اسفنگوزین با استفاده از روش کروماتگرافی مایع با کارایی بالا اندازه گیری شد.
نتایجیافته ها نشان داد که تفاوت معنی داری در مقدار اسفنگوزین در گروه الیگوزواسپرمی در مقایسه با نرموزواسپرمی وجود دارد (05/0>p). همچنین مقدار انواع سرامیدها در گروه الیگوزواسپرمی در مقایسه با گروه کنترل به طور معنی داری افزایش داشت (0001/0>p).
بحث و نتیجه گیریدر مجموع نتایج نشان میدهد که متابولیت های اسفنگولیپیدی نقش بسیار مهمی در کمیت اسپرماتوژنز ایفا می کند و تغییرات در آن میتواند منجر به کاهش باروری در مردان شود. از این این متابولیت ها میتواند معیار خوبی برای ارزیابی های باروری و همچنین اهداف دارویی قرار گیرند.
کلید واژگان: اسپرم، ناباروری مردان، سرامید، اسفنگوزین، HPLC.BackgroundOligozoospermia is one of the causes of infertility in men, characterized by a low sperm count compared to normozoospermia according to World Health Organization criteria. Sphingolipids are crucial regulators of many cellular processes, such as cell differentiation and apoptosis. The most important sphingolipid metabolites include ceramides and sphingosine, where an imbalance in ceramide levels can lead to apoptosis. This study aims to investigate the changes in various types of ceramides and sphingosine in oligozoospermia samples from infertile men.
Materials and MethodsIn this study, semen samples were collected from couples visiting the Alzahra Infertility Treatment Center in Rasht (n=10). The samples were analyzed based on World Health Organization parameters, and oligozoospermia (sperm count less than 15 million per milliliter of semen) and normozoospermia groups were included in the study. The levels of various ceramides (Cer14, Cer16, Cer18, and Cer20) and sphingosine were measured using high-performance liquid chromatography.
ResultsThe findings showed a significant difference in the level of sphingosine in the oligozoospermia group compared to the normozoospermia group (p<0.05). Additionally, the levels of various ceramides were significantly increased in the oligozoospermia group compared to the control group (p<0.0001).
ConclusionOverall, the results indicate that sphingolipid metabolites play a crucial role in the quantity of spermatogenesis, and alterations in these metabolites can lead to reduced fertility in men. Therefore, these metabolites can serve as good criteria for fertility assessments and pharmaceutical targets.
Keywords: Sperm, Male Infertility, Ceramide, Sphingosine, HPLC -
Plants naturally produce various types of secondary metabolites known as polyphenols. The plant's production process of polyphenols can be altered due to environmental stressors such as climate change and the accumulation of heavy metals in the soil or water it relies on. Phragmites australis, also called the common reed, is a perennial grass species that thrives in wetland and riparian habitats. This study aims to quantify the impact of seasonal variations and fluctuations in heavy metal concentrations on the production process and yield of plant products in the selected plant. Additionally, it seeks to evaluate the plant's ability to remediate pollutants through phytoremediation. To achieve this objective, soil and plant samples were collected from three distinct locations in the vicinity of Samawah city. Among these, two areas were identified as being polluted with high levels of heavy metals, while the third area was found to be uncontaminated. The sampling was conducted over the course of a year, encompassing all four seasons, in order to assess the impact of seasonal variations. The quantity of heavy metals, specifically lead, chromium, mercury, and cadmium, present in plant samples and the sediment surrounding the plant was determined using atomic absorption spectrometry. Polyphenols were quantified using high-performance liquid chromatography (HPLC), while photosynthesis pigments were determined using spectrophotometry. The findings revealed notable correlations between specific polyphenols and distinct heavy metals, indicating possible interactions and adaptations of the plant in reaction to stress caused by metals. The results showed that Apigenine-7-O-glucoside exhibited the highest polyphenol concentration, which was more than 9000 mg/kg DW in both autumn and spring at the Samaweh site, followed by caffeic acid showed the highest amount (more than 3000 mg/kg DW) in all seasons at the Kader site. Furthermore, the findings indicated that the plant's phytoremediation capacity varies across various seasons and regions. There was no discernible pattern in the levels of polyphenols or the efficacy of heavy metal removal in response to climate change. The findings of this study indicate that the Phragmatis australis plant may possess phytoremediation capabilities.
Keywords: Common Reed, Environmental Contamination, HPLC, Phytoremediation, Al Muthanna -
Coffee contamination by Ochratoxin A (OTA) is becoming the main global emerging threat to its quality. The main objective of this study was to determine the levels of Ochratoxin A in coffee beans and brewed coffee samples. Twenty samples of coffee beans and brewed were each randomly collected from coffee stores and street coffee vendors, respectively. The levels of OTA in the coffee samples were determined using High-Performance Liquid Chromatography (HPLC) with a Fluorescence Detector (FD). The calibration curve for the analyte was linear in the concentration range of 1mg/L - 15mg/L (R2 = 0.997) and 1mg/kg - 15 mg/kg for green coffee beans (R2 = 0.998). The mean concentration of OTA in brewed and coffee beans composite samples was found to be 1.92 μg/L ± 0.07μg/L (n=10) and 1.52 μg/kg ± 0.28 μg/kg (n=10), respectively. The Limit of Detection (LOD) and Limit of Quantification (LOQ) of the method calculated from the calibration curves were 0.76 μg/kg, 0.69 μg/L and 2.31 μg/kg, 2.07 μg/kg for green coffee bean and brewed samples, respectively. The precision of the method expressed in percent Relative Standard Deviations (RSD) of the recoveries was less than 20 % and the mean recovery was 76.28 % – 106.57%. Even though the concentrations of OTA in studied samples didn’t exceed the maximum residue limit set by international organizations, a positive result signals coffee pollution by these toxic substances and careful monitoring is essential to improve the quality of coffee and halt their health and economic effects.Keywords: Mycotoxins, Ochratoxin A, Coffee, HPLC, Immunoaffinity Column
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مکمل های غذایی از محصولات مهم و پرکاربرد در حوزه سلامت هستند که خود همین مکمل ها، طبقه بندی ها و زیر مجموعه های خاص خودشان را دارند امروزه منابع گیاهی به عنوان جانشین مواد شیمیایی در داروها و مکمل های غذایی مورد توجه روزافزون قرارگرفته است، لذا تحقیق حاضر با هدف بررسی ترکیبات دارویی و آنالیز مکمل غذایی فرموله شده از غلات انجام شد.
بعد از تهیه نمونه های گیاهی و فرمولاسیون، عصاره گیری به روش ماسراسیون و شناسایی ترکیبات تشکیل دهنده آن توسط دستگاه کروماتوگرافی مایع با کارایی بالا HPLC انجام گرفت و سپس آنالیز و عناصر معدنی نیتروژن، فسفر، پتاسیم، کلسیم، منیزیم، سدیم، آهن، روی، منگنز، مس و کادمیوم طبق روش های استاندارد A.O.A.C به روش هضم تر با استفاده از اسید نیتریک، کلریدریک و اسید پرکلریدریک صورت گرفت و به وسیله دستگاه فوتومتر شعله ای و اسپکتروفتومتر جذب اتمی اندازه گیری و با یکدیگر مقایسه شد.
با توجه به نتایج حاصل از آنالیز عصاره مشاهده شد که نمونه مورد مطالعه دارای 31/76 درصد ترکیبات فنلی می باشد که از این میزان 41/2 درصد آن را گالیک اسید تشکیل می دهد و بیشترین ماده متعلق به آلفا لینولنیک اسید با 80/26 درصد می باشد، همچنین دارای 51/43 میلی گرم بر گرم امگا 3، 44/58 میلی گرم بر گرم امگا 6 و 24/19 میلی گرم بر گرم امگا 9 و همچنین ویتامین های مختلفی از جمله 4/52 میلی گرم ویتامین ث و 6/63 میکرو گرم ویتامین کا بود.
بر اساس بررسی ترکیبات دارویی و آنالیز مکمل غذایی فرموله شده از غلات و نتایج حاصل از آنالیز عصاره ها مشاهده شد که مکمل غذایی مورد مطالعه دارای ترکیبات شاخص فنلی و نیز مواد معدنی ماکرو و میکرو و ویتامین های بسیار موثر می باشدکلید واژگان: آنالیز، ترکیبات دارویی، HPLC، پودر غذایی، غلات.A Quarterly Publication The Application of Chemistry in Environment, Volume:14 Issue: 56, 2024, PP 19 -27Food supplements are one of the most important and widely used products in the field of health, which have their own special supplements, classifications and subsets. Today, plant sources are receiving more and more attention as substitutes for chemicals in medicines and food supplements, so this research aims to The investigation of medicinal compounds and the analysis of food supplements formulated from cereals were carried out. After preparation of plant samples and formulation, extraction by maceration method and identification of its constituent compounds was done by high-performance liquid chromatography-HPLC device, and then analysis and mineral elements of nitrogen, phosphorus, potassium, calcium, magnesium, sodium, Iron, zinc, manganese, copper and cadmium were measured by flame photometer and atomic absorption spectrophotometer using a more digestible method using nitric acid, hydrochloric acid and perchloric acid according to AOC standard methods and compared with each other. According to the results of the extract analysis, it was observed that the studied sample has 31.76% of phenolic compounds, of which 41.2% is gallic acid, and the most substance belongs to alpha-linolenic acid with It is 26.80 percent, it also has 43.51 mg/g of omega 3, 58.44 mg/g of omega 6 and 19.24 mg/g of omega 9, as well as various vitamins from The content was 52.4 mg of vitamin C and 63.6 micrograms of vitamin K. Based on the review of medicinal compounds and the analysis of food supplements formulated from grains and the results of extract analysis, it was observed that the studied food supplement has phenolic index compounds as well as macro and micro minerals and very effective vitamins.
Keywords: Analysis, Medicinal Compounds, HPLC, Food Powder, Cereals -
A novel, high-performance, recyclable heterogeneous catalyst functionalized with melamine was advantageously prepared and rhodanine on magnetic silica was successfully carried out. After that, the Cu ions were coordinated with the existing ligands decorated on the magnetic silica surface. This catalyst was identified by different analyses and acted as a recoverable catalyst for xanthene synthesis. Its advantages include the solvent-free conditions of the reaction, the high efficiency of the products in a short period of time, as well as its simple collection from the reaction environment. Moreover, the prepared sorbent has been developed for the extraction of diazinon from different real samples through magnetic solid phase extraction (MSPE). Under optimal conditions, the linear range for measuring diazinon was 0.5-200 µg L-1. The coefficient of determination (R2) was 0.9985. The limit of detection (LOD) of the method was 0.08 μg L-1. Relative standard deviation values (RSD%) for the concentration of 10.0 μg L-1 and 100 μg L-1 were between 5.3% and 5.9%. Finally, the proposed method was used to determine diazinon in orange and agricultural water. The results showed that the percentage of relative recovery of the selected toxin in the studied real samples was between 92 and 105.Moreover, the prepared sorbent has been developed for the extraction of diazinon from different real samples through magnetic solid phase extraction (MSPE). The parameters affecting the extraction were optimized by response surface methodology (RSM). The optimal extraction conditions were 15 min for contact time, 7 mg for sorbent amount, and 0% (w/v) for NaCl concentration.Moreover, 20 µL of methanol was used to elute the extracted analyte. Under optimal conditions, the linear range for measuring diazinon was 0.5-200 µg L-1. The coefficient of determination (R2) was 0.9985. The limit of detection (LOD) of the method was 0.08 μg L-1. Relative standard deviation values (RSD%) for the concentration of 10.0 μg L-1 and 100 μg L-1 were between 5.3% and 5.9%. Finally, the proposed method was used to determine diazinon in orange and agricultural water. The results showed that the percentage of relative recovery of the selected toxin in the studied real samples was between 92 and 105.Keywords: Heterogeneous catalyst, Melamine, nanocatalyst, Magnetic nanoparticles, MSPE, Diazinon, HPLC
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The dried roots and rhizomes of various Rheum spp (R. officinale Baill, R. palmatum L., R. tanguticum Maxim, R. undulatum L, R. emodi Wall. Ex Mesin. etc.) of Polygonaceae family are collectively known as Rhubarb. It is one of the popular herbs used in Chinese medicinal system. Rhubarb (Da-Huang) is not only used as purgative drug in Chinese pharmacopoeia since ancient time, but also well-documented in Korean and Japanese ethnomedical preparations for its various applications. The current research works on Rhubarb elucidated the various pharmacological activities including anticancer, antimicrobial, anti-infammatory, hepatoprotective, gastrointestinal regulating, cardiovascular protecting etc. Some hydroxy anthraquinonoids viz., aloe - emodin, emodin, physcion, rhein, chrysophanol and their glycosides are the mainly responsible for the versatile bio-activities of rhubarb. In fact, these constituents are referred as the ‘taxonomic markers’ for the respective plants. In this regard, multidisciplinary approach for rapid and simultaneous phytochemical analysis and biological screening of these plants should be adopted. Several extraction and analytical (chromatographic as well as electrochemical) techniques are reported in literature for the separation, identification and estimation of these plant secondary metabolites. Some of these methods may provide novel approach for the quality assessment of this widespread herbal drug. In this review article, some recent reports on various qualitative and quantitative methods for detection and estimation of rhubarb anthraquinonoids are summarized which may provide a novel pathway for the study of these active quality markers in this traditional Chinese medicine.Keywords: Rhubarb, Traditional Medicines, Bioactive Anthraquinonoids, Quality Control, HPTLC, HPLC, UPLC, HSCCC, Mec, CZE
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Linum album as one of the endemic perennial plants in Iran is a natural source of lignan compounds. In the present study, the seeds of L. album were collected from its natural habitats in four regions of Iran (Aliabad, Jowkar, Dasht-e Arzhan, Taleqan). The effects of ecotype and gibberellic acid (GA) pretreatment were evaluated on germination traits of L. album seeds in a factorial based on completely randomized design with four replications. Also, the capacity of studied ecotypes in regard to secoisolariciresinol (SECO), podophyllotoxin (PTOX) and 6-methoxy-podophyllotoxin (6-MPTOX) production was assayed under in vitro condition by high performance liquid chromatography (HPLC) based on completely randomized design with four replications. In all studied ecotypes of L. album, seed germination percentage, germination rate and seedling growth was significantly improved in response to GA pretreatment. The obtained sterilized seedlings were used as in vitro explants that were successfully propagated in MS medium without any growth regulators. The in vitro proliferation rate in ecotype of Jowkar in terms of shoot number and plantlet biomass was significantly higher than the other ecotypes. The highest SECO and PTOX contents were extracted in the Dasht-e Arzhan plantlets which were higher (up to four times) than the other ecotypes. Also, the plantlets of Dasht-e Arzhan, Taleqan and Jowkar ecotypes showed the highest content of 6-MPTOX. According to germination, morphological and phytochemicals traits, the ecotype of Dasht-e Arzhan can be considered as a good candidate for breeding programs of improving PTOX production in this plant.Keywords: biotechnology, HPLC, Iranian Flax, Lignan, Podophyllotoxin
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Hair loss and hirsutism have been major complaints due to increased concentrations of dihydrotestosterone. The plant Platycladus orientalis, with 5-alpha reductase inhibitor properties, has been used to treat these disorders. Its formulation with lipophilic carriers in SLN possesses high loading capacity and greater permeability to hair follicles. The present study aimed to determine the content of active ingredients in the extract of P. orientalis L. and to prepare and characterize the solid lipid nanoparticles (SLN) of Platycladus orientalis L. extract as a 5-alpha reductase inhibitor. The total methanolic extract was obtained following the maceration technique. This preparation was analyzed by HPLC using Quercetin and Cedrol as standard components. SLNs were prepared by high-shear homogenization and ultrasound. Four Glucire-GMS-Compritol-Precirol lipids and three poloxamer-tween80-Labrasol surfactants were further used in the formulations. Particle size, zeta potential, nanoparticle morphology, encapsulation percentage, crystal structure, physical stability, size, and zeta potential were studied 0, 3, and 6 months after preparation. Within 1-7 days after preparation, formulations containing GMS and compritol lipids became solid and jelly. Meanwhile, the formulations with Precirol as the lipid and Poloxamer as the surfactant with 0.3% extract exhibited desirable properties such as average particle size (192 nm), the encapsulation of the extract inside the nanoparticles was almost 71%, and good zeta potential. This formulation containing precirol as a lipid, poloxamer as a surfactant, and 0.3% plant extract exhibited greater 5-alpha reductase inhibitor activity, and it can be recommended to treat hair loss and hirsutism.Keywords: Cedrol, hair loss, hirsutism, HPLC, solid-lipid nanoparticles
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The current study describes the in-situ growth of ZnS/MoS2 on cellulose paper using the thin-film microextraction (TFME) method. On cellulose paper, ZnS/MoS2 was grown using a simple, easy, and inexpensive hydrothermal method. The digoxin was quantified after TFME using high-performance liquid chromatography-UV detection (HPLC-UV). The effect of effective parameters such as extraction time, extraction temperature, agitation speed, and sample ionic strength was investigated, and the best conditions were selected. Under ideal conditions, a calibration curve with good linearity (R2=0.987) and a low limit of detection (LOD) in the range of 0.01–100 ng mL-1 was obtained. The detection limits were 0.03 ng mL-1. For the digoxin, the relative standard deviations (RSDs) were 5.3%. The method was successfully applied to the study of urine and plasma. Relative recoveries were found to range between 96% and102 %. The proposed method provided a straightforward, efficient, and environmentally friendly way for determining digoxin in real-world samples.
Keywords: Thin film microextraction, Cellulose paper, ZnS, MoS2, HPLC -
در این مطالعه، یک جاذب جدید بر اساس نسل دوم پلی (آمیدوآمین) مغناطیسی پوشش داده شده با سیلیس Fe3O4@SiO2@PAMAM سنتز شد. . سپس، از میکروسکوپی الکترون روبشی با گسیل اثر میدانی (FE-SE)، طیف بینی پراش پرتوی ایکس (XRD)، طیف بینی مادون قرمز - تبدیل فوریه (FT- IR) ،آنالیز حرارتی وزن سنجی (TGA)، میکروسکوپ الکترونی عبوری (TEM) برای بررسی ریخت شناسی و ساختاری جاذب تهیه شده، استفاده گردید. برای بررسی کارآیی روش، میکونازول، کلوتریمازول و تیکونازول به عنوان سه گونه ی آزمایشی هدف انتخاب شدند و به منظور بررسی کارآیی روش پیشنهادی در آنالیز نمونه های حقیقی، نمونه های بیولوژیکی، مانند: ادرار و پلاسما مورد ارزیابی قرارگرفتند. تحت شرایط بهینه، گستره ی خطی غلظتی برای میکونازول دربازه ی µg L-1 1-200،برای کلوتریمازول در بازه ی µg L-1 1-500 و برای تیکونازول در بازه ی µg L-1 1-200 با ضریب تعیین (2r) دربازه ی 9871/0 تا 9977/0 در اندازه گیری آن ها توسط HPLC-UV محاسبه شدند. LOD روش برای میکونازول، کلوتریمازول و تیکونازول، به ترتیب µg L-116/0، µg L-118/0 و µg L-1 14/0 و LOQ روش نیز برای برای میکونازول، کلوتریمازول و تیکونازول ، به ترتیب µg L-1 53/0، µg L-1 60/0 وµg L-1 46/0 محاسبه گردیدند. RSD% روش در یک روز برای گونه های مورد بررسی در محدوده ی 9/5-6/4 برآورد شدند.
کلید واژگان: استخراج فاز جامد مغناطیسی، دندریمر پلی آمیدو آمین، داروهای ضد قارچ، کروماتوگرافی مایع با کارایی بالا، طراحی آزمایشIn this study, an attempt was made to synthesize a new sorbent based on the second generation of silica coated magnetic poly(amidoamine) (Fe3O4@SiO2@PAMAM) to improve the performance of magnetic solid phase extraction (MSPE) of some antifungal drugs including miconazole, clotrimazole and ticonazole in various real samples such as urine and human plasma. The extracted analytes were measured by high performance liquid chromatography equipped with ultraviolet detection (HPLC-UV). Field emission-scanning electron microscopy (FE-SEM), X-ray diffraction analysis (XRD), Thermogravimetric analysis (TGA), Transmission electron microscopy (TEM) and Fourier transform-infrared spectroscopy (FT-IR) were used to study the morphology and structure of the prepared sorbent. The various factors such as: extraction time, sorbent amount, solvent desorption volume, desorption time, ionic strength and pH were studied and optimized. The method is validated according to ICH guidelines with respect to precision, accuracy, linearity, specificity, robustness, and limits of detection and quantification. Under the optimized condition, the linearity of the method was in the range of 1–500 µg L-1 (miconazole= 1-200 µg L-1, clotrimazole = 1-500 µg L-1 and ticonazole = 1-200 µg L-1). The obtained correlation coefficients (r^2) were between 0.9871-0.9977. The limits of detection (LODs) were also calculated to be 0.14-0.18 µg L-1 (miconazole= 0.16 µg L-1, clotrimazole = 0.18 µg L-1 and ticonazole=0.14 µg L-1). The limits of quantification (LOQs) were also in the range of 0.46-0.60 µg L-1 for the selected analytes. The relative standard deviations (RSDs%), were obtained in the range of 4.6 to 5.9%. Moreover, the calculated enrichment factors were between 85 and 93. The proposed method was also employed for the analysis of various real samples such as urine and plasma samples. The obtained recoveries indicated that the method was useful and applicable in complicated real samples
Keywords: Magnetic solid phase extraction, PAMAM, Antifungal, HPLC, Response surface methodology -
نشریه کیفیت و ماندگاری تولیدات کشاورزی و مواد غذایی، سال سوم شماره 2 (پیاپی 10، پاییز 1402)، صص 58 -64طب سنتی و مکمل امروزه در درمان بسیاری از اختلالات و بیماریها جایگاه ویژه ای پیدا کرده است. در ایران نیز بعلت اقلیم خاص و تنوع آب هوا گیاهان دارویی ارزشمندی رشد می کنند. گیاه زرشک (Berberis Vulgaris) یکی از مهمترین و شاخص ترین گونه های گیاهی است و دارای خواص درمانی بسیاری می باشد. بربرین بعنوان مهمترین آلکالویید زرشک بعنوان یک ماده طبیعی دارای خواص فارماکولوژیک در اندامهای گیاه بخصوص ریشه می توان یافت. ریشه های گیاه از شهر تهران جمع آوری شده و پس از خشک کردن در دمای محیط و دور از نور آفتاب، آسیاب شدند. عصاره متانولی جهت استخراج بربرین تهیه شده، پس از فیلتر کردن و جهت شناسایی بربرین از دستگاه HPLC با آشکارساز Diode Array استفاده گردید. کروماتوگرام حاصله نشان دهنده پیکهای ترکیبات عصاره ریشه زرشک و بخصوص بربرین در مقایسه با ماده استاندارد مرجع کاملا بصورت واضح جدا و شناسایی گردید. میزان بربرین ریشه زرشک با توجه به منحنی کالیبراسیون 44/1 درصد تعیین گردید. بر اساس نتایج بدست آمده و مروری بر خواص درمانی و دارویی زرشک بومی ایران، تعیین مقدار ماده موثره در زمینه کاربرد آن در پیشگیری و درمان اختلالات و بیماریهای مختلف بصورت عصاره استاندارد شده و یا بربرین استخراج شده حایز اهمیت می باشد.کلید واژگان: زرشک، بربرین، آلکالوییدها، کروماتوگرافی مایع، بربریس وولگاریسJournal of Quality and Durability of Agricultural and Food Products, Volume:3 Issue: 2, 2023, PP 58 -64Today, traditional and complementary medicine has a special place in the treating of many disorders and diseases. Valuable medicinal plants grow in Iran due to the special climate diversity. The barberry plant (Berberis vulgaris) is one of the most important and prominent plant species and has many therapeutic properties. Berberine, as the most important barberry alkaloid in the root, has a suitable concentration for extraction and utilization and introducing it as a natural substance with pharmacological properties is the aim of this research. The roots of the barberry plant were collected from Tehran city and ground after drying at ambient temperature and away from sunlight. Methanol was used to extract the berberine present in the root of this plant. After filtering, the methanolic extract was used to identify berberine using an HPLC device with a DAD detector. The resulting chromatogram shows the proper separation of barberry root extract and berberine peak peaks compared to the berberine standard, which was clearly separated and identified. The amount of berberine in barberry root was determined according to the calibration curve of 1.44%. Based on the obtained results and a review of the therapeutic and medicinal properties of native Iranian barberry, it is important to determine the amount of the effective substance in the context of its use in the prevention and treatment of various disorders and diseases.Keywords: Barberry, Berberine, Alkaloids, HPLC, B. vulgaris
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هدف
هدف از این تحقیق بررسی اثر عوامل تاثیرگذار بر القای ریشه مویین و بررسی میزان تولید آلکالوئیدهای ایندولی در ریشه های مویین تولید شده در اثر تلقیح گیاه پروانش با Agrobacterium rhizogenes بود.
مواد و روش هااز گیاهچه استریل پروانش جهت تهیه ریزنمونه استفاده شد. اثر نوع محیط کشت، ریزنمونه، سویه باکتری و ژنوتیپ گیاه بر میزان القای ریشه مویین بررسی شد. میزان آلکالوئیدهای ایندولی با استفاده از دستگاه HPLC اندازه گیری شد. ماهیت تراریختگی ریشه های مویین با استفاده از روش PCR و پرایمرهای اختصاصی تایید گردید.
نتایجسویه A4 بهعنوان بهترین سویه A. rhizogenes جهت القای ریشه مویین و محیط کشت MS بهعنوان بهترین محیط کشت برای توسعه و استقرار کشت ریشه مویین در ژنوتیپ سفید پروانش تعیین شدند. نتایج حاصل از تکثیر اختصاصی ژن های rolB و virD نشان دهنده ماهیت تراریختی ریشه های مویین بود. بهطور کلی مقدار وین دولین و کاتارانتین ریشه های مویین بیشتر از ریشه معمولی و اندام هوایی گیاه پروانش بود. از طرفی، مقدار این آلکالوئیدها در اندام هوایی گیاه از ریشه معمولی بالاتر بود.
نتیجه گیریبهنظر می رسد با ورود ژن های rol از باکتری به ژنوم گیاه و ایجاد ریشه مویین و در نتیجه تغییر در میزان تولید هورمون های درونزا و پاسخ های سلول گیاهی به ورود ژن های باکتری، میزان و پروفایل تولید متابولیت های ثاتویه گیاه تغییر پیدا می کند و در نتیجه می توان با گرینش کلون هایی با افزایش تولید آلکالوئیدهای مهم دارویی و بهینه سازی شرایط تولید انبوه متابولیت ها در بیوراکتورها، به سمت تولید تجاری این ترکیبات از طریق کشت سلول و بافت گیاهی حرکت کرد.
کلید واژگان: آگروباکتریوم رایزوژنز پروانش، ریشه مویین، متابولیت های ثانویهAimHairy roots induced by inoculation with Agrobacterium rhizogenes are suitable for the production and accumulation of plant secondary metabolites. Genetically modified hairy roots produce valuable secondary metabolites to a greater amount and with more genetic stability than cell suspension cultures and normal roots. Catharanthus roseus is one of the most important medicinal plants that produces valuable secondary metabolites such as indole alkaloids. The purpose of this research was to investigate the effect of factors influencing the induction of hairy roots and to investigate the amount of production of secondary metabolites (vindoline and catharanthine) in the hairy roots produced by inoculation of C. roseus plant with A. rhizogenes.
Material and MethodsIn order to produce sterile C. roseus seedlings and to prepare explants, seeds cultured on MS culture medium. The effect of the type of culture medium (MS, 1/2 MS and B5), the type of explant (leaf and stem), the type of bacterial strain (A4, 1132 and 2656) and the type of plant genotype (white and red) on the amount of hairy root induction in a completely randomized design was investigated. Extraction of samples was done using methanol solvent, then the amount of indole alkaloids was calculated using HPLC method and standard curve. The nature of transgenic hairy roots was confirmed after DNA extraction from different samples using PCR method with specific primers (rolB and virD).
ResultsAccording to the results, the most efficient strain in transferring T-DNA to the explants and the appearance of hairy roots was related to A4 strain (53.8%). 1/2 MS culture medium with 77.69% of root induction was the best medium combination for the appearance of hairy roots. Also, white genotype (62% rooting) and leaf explant (77% rooting) were identified as the best genotype and explant for the production of hairy roots. After the specific amplification of the rolB gene and the lack of amplification of the virD gene in the PCR reaction in the samples, the molecular confirmation of the transgenic nature of the hairy root clones was done. Based on the results of HPLC experiment, the average amount of vindoline and catharanthine in hairy roots (0.24 and 0.615 mg/g dry weight) is higher than aerial parts (0.106 and 0.488 mg/g dry weight) and normal root (0.021 and 0.013 mg/g dry weight). The amount of these alkaloids in the aerial parts was higher than in the normal roots of the plant.
ConclusionThe production of hairy roots in response to the inoculation of plant explants with A. rhizogenes is influenced by various factors such as the type of explant, the genotype of the plant, the strain of bacteria and the composition of culture medium. These factors must be optimized to establish effective hairy root cultur in a plant. It seems that with the introduction of rol genes from bacteria to the plant genome (hairy root) and as a result of the change in the amount of endogenous hormone production and the response of the plant cell to the entry of bacterial genes, the amount and profile of the production of the secondary metabolites of the plant will change. As a result, by selecting clones with increased production of medicinally important alkaloids and optimizing the conditions of large scale production of secondary metabolites in bioreactors, we can move towards the commercial production of these compounds through cell and plant tissue culture.
Keywords: Agrobacterium rhizogenes, Catharanthus roseus, HPLC, Secondary metabolites -
IntroductionEryngium plants represent the most diverse species within the Apiaceae family, with approximately 250 species. Numerous species of Eryngium, including Eryngium campestre and E. foetidum, have been used for centuries in traditional medicine. E. billardieri was selected for this study due to its purported applications in traditional Iranian medicine, as well as the absence of prior research on the chemical constituents within this plant.Materials and methodsThe aerial parts of the plant were extracted by n-hexane, dichloromethane, and methanol (MeOH) by a Soxhlet apparatus. The MeOH extract was exposed to C18 Sep-Pak fractionation cartridge by a step gradient of MeOH-H2O. Further purification was performed by preparative HPLC and the purified compounds were separated to be elucidated with H NMR and C NMR results. The essential oil was obtained by the Clevenger apparatus. The n-hexane extract was transformed into methyl ester through a process involving saponification and esterification and the obtained essential oil and fatty acids were analyzed using a flame ionization detector (FID) and a DB-1 capillary column.ResultsThe 40% methanol extract led to the identification of two compounds such as decursine and Chlorogenic acid. The essential oil composition analysis yielded the followingresultsapproximately 92% of the essential oil compounds were identified. The predominant compounds were isofenchol (36.85%), germacrene D (7.69%) and gurjunene (6.67%). The analysis of methyl esters revealed the presence of four fatty acid compounds: oleic acid with 27.2%, elaidic acid with 6.91%, palmitic acid with 4.62%, and stearic acid with 2.55% of the total area.ConclusionTwo substances (decursine and chlorogenic acid) were isolated from the methanol extract, and oleic acid was isolated and identified from the n-hexane extract. Concerning the phytochemical potential inherent in this genus and specifically in this plant, further extensive research in this domain is warranted.Keywords: HPLC, Essential oil, GC-MS, NMR
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Salvia mirzayanii Rech. & Esfand is an Iranian endemic plant belonging to the Lamiaceae family which has many pharmacological effects including antioxidant, anti-cholinesterase, antimicrobial, anticancer, anti-inflammatory and enhancing cognition and memory. It is considered as an endemic species of Iran, that is in danger of extinction. The bulks aromatic plants come from wild populations whose essential oils compositions as well as their biological properties are severely affected by the geographical location Therefore, the aim of the present study is to provide more information on the variation of essential oil composition of S. mirzayanii collected from four different geographical regions. The aerial parts of four populations of S. mirzayanii were collected from different natural habitats of Fars province in southwest of Iran. Chemical composition of S. mirzayanii were isolated by hydrodistillation using clevenger type apparatus and analyzed by GC-FID and GC/MS. The major compounds in four populations were α-terpinyl acetate, Eudesm-7(11)-en-4-ol, bicyclogermacrene, δ-cadinene and 1,8-Cineole in four populations. Polyphenolics content were identified by HPLC analysis. Predominant phenolic constituents in all extracts were chlorogenic acid, vaniline and rosmarinic acid, p-cumaric acid, trans-ferulic acid and quercetin. The DPPH radical inhibition was measured by using a micro-plate reader. The best antioxidant activity was related to ecotype3 with 1281.48 μg/mL and the highest amount of total phenol is related to ecotype1 and ecotype4.Keywords: Salvia mirzayanii, essential oil, Phenolic content, HPLC
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Harmaline and harmine, are the major ß-carbolines present in the seeds of the Peganum harmala L. These compounds are known as herbal active principals with potential use in pharmaceutical and medicine. To assess the growth inhibitory effect of phyto-alkaloids, harmaline and harmine, on cancer cell lines. The P. harmala L.’s alkaloids were extracted by acidic/basic extraction method and identified by two methods, Fourier Transform Infra-Red Spectroscopy (FTIR) and High-Performance Liquid Chromatography (HPLC). breast cancer cell lines, MDA_MB_231, were subjected to different concentration (1–100 μg. mL -1) of the P. harmala extract at different time courses (24h, 48h). Methylthiazol Tetrazolium (MTT) test, the half maximal inhibitory concentration (IC50) and the morphological changes through optical microscopy were evaluated cell lines, the P. harmala extract decreased cell viability in longer time exposure in a dose dependent manner. The more concentrated extract led to higher motility of MDA-MB-231 at 24h.It was observed that 30 μg. mL -1 is the minimum lethal dose that kills approximately 50% of cells at 24 hours in MDA-MB-231 cell line (IC50). The morphological observation ensured the apoptosis nature of P. harmala on cells as their membrane kept intact and no membrane permeabilization was observed. The results revealed that the P. harmala extracts decreased significantly growth rate and cell survival of cancer cell lines. higher growth inhibition of MDA-MB-231 cell line by the P. harmala extract was confirmed.Keywords: Cell viability, FT-IR, HPLC, Harmine, Harmaline
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