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In vitro anti-Trypanosoma cruzi activity of methanolic extract of Bidens pilosa and identification of active compounds by gas chromatography-mass spectrometry analysis

  • Gabriel Enrique Cazares-Jaramillo;Zinnia Judith Molina-Garza;Itza Eloisa Luna-Cruz;Luisa Yolanda Solis-Soto;Jose Luis Rosales-Encina;Lucio Galaviz-Silva
    • Parasites, Hosts and Diseases
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    • v.61 no.4
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    • pp.405-417
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    • 2023
  • Chagas disease, caused by Trypanosoma cruzi parasite, is a significant but neglected tropical public health issue in Latin America due to the diversity of its genotypes and pathogenic profiles. This complexity is compounded by the adverse effects of current treatments, underscoring the need for new therapeutic options that employ medicinal plant extracts without negative side effects. Our research aimed to evaluate the trypanocidal activity of Bidens pilosa fractions against epimastigote and trypomastigote stages of T. cruzi, specifically targeting the Brener and Nuevo León strains-the latter isolated from Triatoma gerstaeckeri in General Terán, Nuevo León, México. We processed the plant's aerial parts (stems, leaves, and flowers) to obtain a methanolic extract (Bp-mOH) and fractions with varying solvent polarities. These preparations inhibited more than 90% of growth at concentrations as low as 800 ㎍/ml for both parasite stages. The median lethal concentration (LC50) values for the Bp-mOH extract and its fractions were below 500 ㎍/ml. Tests for cytotoxicity using Artemia salina and Vero cells and hemolytic activity assays for the extract and its fractions yielded negative results. The methanol fraction (BPFC3MOH1) exhibited superior inhibitory activity. Its functional groups, identified as phenols, enols, alkaloids, carbohydrates, and proteins, include compounds such as 2-hydroxy-3-methylbenzaldehyde (50.9%), pentadecyl prop-2-enoate (22.1%), and linalool (15.4%). Eight compounds were identified, with a match confirmed by the National Institute of Standards and Technology (NIST-MS) software through mass spectrometry analysis.

Structural evaluation of degradation products of Loteprednol using LC-MS/MS: Development of an HPLC method for analyzing process-related impurities of Loteprednol

  • Rajesh Varma Bhupatiraju;Bikshal Babu Kasimala;Lavanya Nagamalla;Fathima Sayed
    • Analytical Science and Technology
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    • v.37 no.2
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    • pp.98-113
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    • 2024
  • The current investigation entails the characterization of five degradation products (DPs) formed under different stress conditions of loteprednol using liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, this study developed a stable high-performance liquid chromatography (HPLC) method for evaluating loteprednol along with impurities. The method conditions were meticulously fine-tuned which involved the exploration of the appropriate solvent, pH, flow of the mobile phase, columns, and wavelength. The method conditions were carefully chosen to successfully resolve the impurities of loteprednol and were employed in subsequent validation procedures. The stability profile of loteprednol was exposed to stress degradation experiments conducted under five conditions, and DPs were structurally characterized by employing LC-MS/MS. The chromatographic resolution of loteprednol and its impurities along with DPs was effectively achieved using a Phenomenex Luna 250 mm C18 column using 0.1 % phosphoric acid, methanol, and acetonitrile in 45:25:30 (v/v) pumped isocratically at 0.8 mL/min with 243 nm wavelength. The method produces an accurate fit calibration curve in 50-300 ㎍/mL for loteprednol and LOQ (0.05 ㎍/mL) - 0.30 ㎍/mL for its impurities with acceptable precision, accuracy, and recovery. The stress-induced degradation study revealed the degradation of loteprednol under basic, acidic, and photolytic conditions, resulting in the formation of seven distinct DPs. The efficacy of this method was validated through LC-MS/MS, which allowed for the verification of the chemical structures of the newly generated DPs of loteprednol. This method was appropriate for assessing the impurities of loteprednol and can also be appropriate for structural and quantitative assessment of its degradation products.

Antifungal activities of coumarins isolated from Angelica gigas and Angelica dahurica against Plant pathogenic fungi (당귀와 백지로부터 분리한 Coumarin계 물질들의 식물병원균에 대한 항균활성)

  • Ryu, Shi-Yong;Kim, Young-Sup;Kim, Heung-Tae;Kim, Seong-Ki;Choi, Gyung-Ja;Kim, Jeoung-Seob;Lee, Seon-Woo;Heor, Jung-Hee;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Pesticide Science
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    • v.5 no.3
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    • pp.26-35
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    • 2001
  • In order to search potent antifungal substances from domestic plants, 40 plants cultivated in Korea were collected. After extracting with methanol (MeOH) and concentrating to dryness, the MeOH extracts were screened for in vivo antifungal activity against six plant diseases at a concentration of $2000{\mu}g/mL$. Fourteen extracts showed disease-controlling activity more than 90% against at least one of the 6 plant diseases tested; eight, seven, and three extracts controlled more than 90% the development of rice blast, tomato late blight, and wheat leaf rust, respectively. However, none of the extracts exhibited in vivo antifungal activity more than 90% against rice sheath blight, tomato gray mold, and barley powdery mildew. From the MeOH extracts of Angelica gigas and A. dahurica showing potent controlling activity against rice blast, 1 and 2 antifungal substances, respectively, were isolated by solvent partitioning and column chromatography. The three compounds were identified to be coumarins, namely, decursin, imperatorin, and isoimperatorin, by mass spectrometry and NMR spectroscopy. They were examined for in vitro and in vivo antifungal activities together with umbelliferone (7-bydroxycournarin) and scopoletin (6-methoxy-7-hydroxycoumarin) containing a free hydroxyl group at position 7 to investigate the structure-activity relationship. In vitro, most of 50% growth inhibitory concentrations ($IC_{50}$) were over $200{\mu}g/mL$, indicating that they have relatively weak antifungal activity. The antifungal activity of decursin and scopoletin, containing cyclic alkoxy groups instead of free hydroxyl group at position 7, was stronger than umbelliferone and scopoletin. Especially, decursin and imperatorin showed potent antifungal activities against Pythium ultimum and Magnaporthe grisea, respectively, with $IC_{50}$ values less than $25{\mu}g/mL$. In vivo, decursin and imperatorin showed potent antifungal activity against rice blast, whereas other coumarins hardly controlled the development of 6 plant diseases tested. These results suggest that the antifungal activity of 7-hydroxycoumarin derivative is substantially increased when the hydroxyl group at position 7 is protected by a stable cyclic alkoxy grouping.

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Antioxidant Activity of the Oven-dried Paprika Powders with Various Colors and Phycochemical Properties and Antioxidant Activity of Pork Patty Containing Various Paprika Powder (파프리카의 색이 열풍 건조한 파프리카 분말을 첨가한 돈육 분쇄육의 이화학적 특성과 항산화 활성 평가)

  • Shim, Yong Woo;Chin, Koo Bok
    • Food Science of Animal Resources
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    • v.33 no.5
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    • pp.626-632
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    • 2013
  • This study was performed to determine the antioxidant activity of the oven-dried paprika powder as affected by the color differences of paprika and to evaluate physicochemical characteristics and antioxidant activity of pork patties with various levels of paprika powders. The total phenolic contents of the paprika were not affected by color and solvent (p>0.05). The methanol extracted paprika powder showed higher DPPH radical scavenging activity than water extracted counterpart, and no differences were observed at concentration of 0.5% as compared to the reference (ascorbic acid) (p>0.05). In all treatments, the iron chelating ability increased with increasing concentrations. At a concentration of 1.0%, methanol extracts of orange paprika (MOP) and water extracts of red paprika (WRP) were not different from the reference, (ethylendiaminetetraacetic acid, EDTA). The paprika color and extraction solvent didn't affect reducing power of paprika powder at each concentration (p>0.05). Pork patties with red paprika powder were higher redness values than those with orange ones, regardless of addition level. The addition of red paprika increased the yellowness, and patties with 1.0% orange paprika powder showed the highest value. TBARS values were decreased with increasing paprika powder, especially, patties with 1.0% paprika powder were lower TBARS than those with 0.5% paprika powder, resulting in similar to those with ascorbic acid (p>0.05). Although the microbial counts increased with storage time, paprika powders did not inhibit microorganisms during storage. In conclusion, paprika powders could be used as a natural antioxidant in meat products, regardless of paprika color.

Effect of Yacon (Smallanthus sonchifolius H. Robinson) Extracts on Herbicidal, Fungicidal, and Insecticidal Activities (야콘(Smallanthus sonchifolius H. Robinson) 추출물의 제초, 살균 및 살충활성 효과)

  • Yun, Young-Beom;Kim, Jin-Hwa;Jang, Se-Ji;Kim, Do-Ik;Kwon, Oh-Do;Kuk, Yong-In
    • Korean Journal of Weed Science
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    • v.32 no.2
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    • pp.98-106
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    • 2012
  • This research has been carried out to find out the highest effect on insecticidal, fungicidal and herbicidal activities in leaves, stem and tuber extracts of yacon (Smallanthus sonchifolius) and extraction methods such as water, boiling water and methanol. Characteristics of potential herbicidal components among extraction methods were investigated by solvent fractions such as hexane, chloroform, ethyl acetate, butanol, and water. Generally, methanol extract was the best on inhibition of germination rate, plant height, and root length in cucumber and barley. On the other hand, the inhibition effect on growth in cucumber and barley was the best in tuber among plant parts of yacon. Inhibition of germination rate, plant height, and root length in cucumber and barley in solvent fractions was the best in water fraction, but there were no differences in other fractions. Digitaria sanguinalis L. and Solanum nigrum L. by 5 and 10% extractions of yacon tuber were controlled by more than 70~80% and 95~100%, respectively. However, there was no inhibition effect on foliar treatment in cucumber and barley as affected by 5 and 10% extractions of yacon tuber. Mortality of green peach aphid (Myzus persicae Sulzer) was 50% at 3 days after treatment of 5% extracts of yacon leaves. Mortality of brown plant hopper (Nilaparvata lugens Stal) was only 24% in 5% extracts of stems and leaves with midrib, but was 57% in 5% extracts of leaves without midrib. There was no fungicidal effect on anthracnose (Colletotrichum truncatum), wilt disease (Fusarium oxysporum), verticillium wilt (Verticillium dahliae), bacterial blight (Xanthomonus oryzae) in 5% extracts of yacon leaves.

Method Development for the Profiling Analysis of Endogenous Metabolites by Accurate-Mass Quadrupole Time-of-Flight(Q-TOF) LC/MS (LC/TOFMS를 이용한 생체시료의 내인성 대사체 분석법 개발)

  • Lee, In-Sun;Kim, Jin-Ho;Cho, Soo-Yeul;Shim, Sun-Bo;Park, Hye-Jin;Lee, Jin-Hee;Lee, Ji-Hyun;Hwang, In-Sun;Kim, Sung-Il;Lee, Jung-Hee;Cho, Su-Yeon;Choi, Don-Woong;Cho, Yang-Ha
    • Journal of Food Hygiene and Safety
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    • v.25 no.4
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    • pp.388-394
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    • 2010
  • Metabolomics aims at the comprehensive, qualitative and quantitative analysis of wide arrays of endogenous metabolites in biological samples. It has shown particular promise in the area of toxicology and drug development, functional genomics, system biology and clinical diagnosis. In this study, analytical technique of MS instrument with high resolution mass measurement, such as time-of-flight (TOF) was validated for the purpose of investigation of amino acids, sugars and fatty acids. Rat urine and serum samples were extracted by selected each solvent (50% acetonitrile, 100% acetonitrile, acetone, methanol, water, ether) extraction method. We determined the optimized liquid chromatography/time-of-flight mass spectrometry (LC/TOFMS) system and selected appropriated columns, mobile phases, fragment energy and collision energy, which could search 17 metabolites. The spectral data collected from LC/TOFMS were tested by ANOVA. Obtained with the use of LC/TOFMS technique, our results indicated that (1) MS and MS/MS parameters were optimized and most abundant product ion of each metabolite were selected to be monitorized; (2) with design of experiment analysis, methanol yielded the optimal extraction efficiency. Therefore, the results of this study are expected to be useful in the endogenous metabolite fields according to validated SOP for endogenous amino acids, sugars and fatty acids.

Antiadipogenic Effect of Vitis amurensis Root Methanol Extract and Its Solvent Fractions in 3T3-L1 Preadipocytes (머루근 추출물 및 분획물의 항비만 활성)

  • Park, Jung Ae;Jin, Kyong-Suk;Oh, You Na;Hyun, Sook Kyung;Choi, Yung Hyun;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.23 no.1
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    • pp.69-78
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    • 2013
  • Vitis amurensis Rupreche, a sort of grape, grows naturally in Asian countries. It is known for important biological effects such as antioxidation, anti-inflammation, neuroprotection, and angiogenesis inhibition. Although its root is used as a traditional folk medicine in Korea, the root's biological activities are poorly studied. In the present study, the effects of V. amurensis root methanol extract (VARM) and its solvent fractions on adipocyte differentiation and adipogenesis in 3T3-L1 preadipocytes were investigated. The VARM significantly suppressed adipocyte differentiation, lipid accumulation, and the triglyceride (TG) content of 3T3-L1 preadipocytes in a dose-dependent manner, without cytotoxicity. To identify active molecules, the VARM was fractionated with a series of organic solvents including dichloromethane ($CH_2Cl_2$), ethyl acetate (EtOAc), and n-butanol (n-BuOH). All the fractions also showed inhibition of lipid accumulation in the 3T3-L1 preadipocytes. The $CH_2Cl_2$ fraction showed the most powerful anti-obesity effect through the modulation of cytidine-cytidine-adenosine-adenosinethymidine (CCAAT)/enhancer binding protein ${\alpha}$ ($C/EBP{\alpha}$), $C/EBP{\beta}$, peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) gene and protein expression. Oleanolic acid was one of the main active compounds involved in the anti-obesity activity of the V. amurensis root. These results provide important new insight into the potential potent anti-adipogenic effect of the V. amurensis root and illustrate that one of the main compounds involved in this effect is oleanolic acid.

Oxidation Reaction of Hydrazobenzene by Activated Catalysts of Oxygen Adducted Tetradentate Schiff Base Cobalt(Ⅱ) Complexes in Methanol Solvent. (Ⅱ) (메탄올 용매에서 산소 첨가된 네자리 Schiff Base Cobalt(II) 착물들의 활성촉매에 의한 Hydrazobenzene의 산화반응 (제 2 보))

  • Ki-Kyung Chjo;Yong-Kook Choi;Sang-Bock Kim;Jong-Ki Park;Dong-Hwa Park
    • Journal of the Korean Chemical Society
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    • v.36 no.6
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    • pp.894-905
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    • 1992
  • Dioxygen binding and homogeneous catalytic oxidation of hydrazobenzene were investigated by employing tetradentate Schiff base Cobalt(II) complexes such as Co(II)(SED)$(Py)_2$, Co(II)(SOPD)$(Py)_2$ and Co(II)(SND)$(Py)_2$ in saturated oxygen methanol solvent. The major product of hydrazobenzene ($H_2$AB) oxidation by catalysts of superoxo type [Co(III)(SED)(Py)$O_2$] and [Co(III)(SOPD)(Py)$O_2$] complexes are trans-azobenzene (t-AB) and rate constants k for oxidation reaction was 7.692 ${\times}$ $10^{-2}$ M/sec for [Co(III)(SED)(Py)$O_2$] and 5.076 ${\times}$ $10^{-2}$ M/sec for [Co(III)(SOPD)(Py)$O_2$]. But cis-azobenzene (c-AB) are obtained as a major product with ${\mu}$-peroxo type [Co(III)(SED)(Py)]$_2O_2$ catalyst, and rate constant k is 1.266 ${\times}$ $10^{-2}$ M/sec. The rate constants of oxidation reaction has been studied spectrophotometrically and the rate law established. A mechanism involving a intermediate activated complexes of catalyst, hydrazobenzene and oxygen has been proposed. $H_2$AB + Co(II)(Schiff base)$(Py)_2$ + $O_2$ ${\rightleftharpoons}_{MeOH}^K$ Co(III)(Schiff base)(Py)$O_2$${\cdot}$$H_2$AB + Py $\longrightarrow^k$ Co(II)(Schiff base)$(Py)_2$ + t-AB + $H_2O_2$(Scchiff base : SED and SOPD). $H_2$AB + 2Co(II)(SND)$(Py)_2$ + $O_2$ ${\rightleftharpoons}_{MeOH}^K$ [Co(III)(SND)(Py)]$_2O_2$${\cdot}$H_2$AB + 2Py ${\longrightarrow}^k$ (Co(II)(SND)$(Py)_2$ + c-AB + $H_2O_2$.

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On the Utilization of Inactive BHC isomers -Synthesis of 3-(2,4,5-trichlorophenyl)-1-methyl urea as a herbicide- (BHC 이성질체(異性質體)의 활용(活用)에 관(關)한 연구(硏究) -제초제(除草劑)로서 3-(2,4,5-trichlorophenyl)-1- methyl urea의 합성(合成)-)

  • Lee, Kyu-Seung;Park, Chang-Kyu
    • Applied Biological Chemistry
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    • v.22 no.2
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    • pp.109-122
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    • 1979
  • Present study was carried out to reduce residual toxicity of BHC insecticides inherent in the organochlorine pesticides. For This end, r-isomer, the most potent insecticidal component among the BHC stereoisomers, was isolated and thus fortified by means of solvent precipitation. In parallel, 3-(2,4,5-trichlorophenyl)-1-methyl urea was prepared in good yield from technical BHC via 1,2,4-trichlorobenzene, 1,2,4,-trichloronitrobenzene, and 2,4,5-trichloroaniline. In addition, certain merit of the compound which make it possible to use as a herbicide is discussed. The results are summarized as follows; 1. Recrystallizing technical BHC from methanol-water binary solvent system, r-isomer was enriched to 49.7% at 95% recovery of r-isomer. 2. By partitioning technical BHC in 85% of methanolic solution into chloroform, r-isomer was fortified to 89.6% at 90.5% recovery of r-isomer. 3. Yield of 1,2,4-trichlorobenzene from technical BHC was greatly dependent upon concentration of alkalies and to less degree on the alkalies. 4. Surfactants, in particular cationic a quartenary ammonium salt, increased yield of 1,2,4-trichlorobenzene from technical BHC by alkaline hydrolysis. 5. Conversion of 1,2,4-trichlorobenzene to 2,4,5-trichloronitrobenzene was effected almost quantitatively utilizing $HNO_3-H_2SO_4$ nitrating agent at low temperature. 6. Yield of 91.4% was observed for the synthesis of 2,4,5-trichloroaniline by reducing 2,4,5-trichloronitrobenzene in the presence of iron turning and hydrochloric acid. 7. Overall yield based on BHC of 3-(2,4,5-trichlorophenyl)-1- methyl urea was 60.8%. 8. Inhibition effects, both germination and growth, 3-(2,4,5-trichlorophenyl)-1-methyl urea on several crops were found comparable to or more potent than those of $linuron{\circledR}\;and\;diuron{\circledR}$. In addition, it was also noted that susceptibility to the prepared compound depended upon the crops as well as specific part (shoots, roots) of the plant exposed to the chemicals.

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Antioxidative Activity of Aralia elata Shoot and Leaf Extracts (두릅 순 및 잎 추출물의 항산화 효과)

  • Cha, Jae-Young;Ahn, Hee-Young;Eom, Kyung-Eun;Park, Bo-Kyung;Jun, Bang-Sil;Cho, Young-Su
    • Journal of Life Science
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    • v.19 no.5
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    • pp.652-658
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    • 2009
  • The comparative activities of aqueous, ethanol, and methanol extracts from Aralia elata shoot (AES) and leaf (AEL) were tested by in vitro experimental models of linoleic acid peroxidation by thiocyanate and thiobarbituric acid (TBA) methods and scavenging activities of free radicals by DPPH (${\alpha}$,${\alpha}$'-diphenyl-${\beta}$-picrylhydrazyl). In addition, bio-active materials (phenolic compounds and minerals) were also measured. The extract yield of each solvent extracted from AES and AEL was 3.08% and 3.13% in aqueous, 0.58% and 0.66% in ethanol, and 0.81% and 1.73% in methanol, respectively. The highest extract yield was found in the aqueous extract from AEL. Major mineral contents (mg%) of AES and AEL were 575.7 and 759.3 in Ca, 353.5 and 330.0 in K, and 31.3 and 31.0 in Mg, respectively. The highest free radical scavenging activity was found in the aqueous extract by 28.69% at 0.1% additional level from AES and in the methanol extract by 92.36% at 0.1% additional level from AEL. Free radical scavenging activity was stronger in AEL than in AES. In antioxidative activities determined by thiocyanate and TBA methods against lipid peroxidation using linoleic acid, ethanol extracts from AEL showed the highest antioxidative activity at all treatment concentrations. These results may provide the basic data to understand the biological activities of bio-active materials derived from AES and AEL.