Chemical analysis and in vitro antimicrobial effects and mechanism of action of Trachyspermum copticum essential oil against Escherichia coli

Objective: To find a natural plant essential oil (EO) with excellent antimicrobial effect on food-borne bacteria and to explore the mechanism of its antimicrobial function against Escherichia colt (E. cal). Methods: The antimicrobial activity of seven EOs against Gramnegative E. coli ATCC 8739 and Gram-positive Staphylococcus aureus ATCC 6538 was investigated using agar disk diffusion method, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of each EO was determined using the broth dilution method. The chemical composition of the Trachyspermum copticum (T. copticum) EO was analyzed using gas chromatography mass spectrometry (GC/MS). In order to explore the mechanism of the antimicrobial action, 1 MIC and 2 MIC of T. copticum. EO was added to a suspension of E. coli, the growth curve and the scanning electron microscopy (SEM) analysis of E. coli, and the release of cell constituents and protein and potassium ions from the bacterial cell were measured. Results: The T. optician EO had the best antimicrobial activity against the test bacteria, and 10 compounds accounting for 94.57% of the total oil were identified, with the major components being thymol (46.22%), p-cymene (19.03%), and y-terpinene (22.41%). The addition of I MIC T. copticum. EO significantly inhibited the growth of E. coli and increased the release of cell constituents and protein and potassium ions from the bacterial cells. Scanning electron micrographs showed that T. copticum EO caused most of the E. co/i cell membranes to collapse and rupture, leading to cell death. Conclusions: These results indicate that T. copticum EO is a good natural antimicrobial agent for food-borne pathogens.

Isolation and identification of Salmonella from curry samples and its sensitivity to commercial antibiotics and aqueous extracts of Camelia sinensis(L.) and Trachyspermum ammi(L.)

Objective:To isolate Salmonella from curry samples and to evaluate the drug sensitivity of the food-borne Salmonella and its susceptibility to specific plant extracts.Methods:Salmonella was isolated from the curry samples by standard microbiological methods and was confirmed by biochemical tests.The antibiotic susceptibility test was conducted by disc diffusion method using commercially available antibiotics such as ampicillin,tetracycline,chloramphenicol,kanamycin,and penicillin.In addition,the susceptibility of the food-borne Salmonella was also evaluated against the aqueous extracts of Camelia sinensis(L.) Theaceae(tea leaves) and the Trachyspermum ammi(L.) Apiaceae(ajwain or omum seeds).Results:Out of fifty curry samples,only seven samples were identified to have Salmonella contamination.The Salmonella isolates showed a significant drug resistance pattern except for kanamycin.The plant extracts showed a considerable antibacterial activity against the isolates,indicating the presence of antimicrobial principle which can be exploited after complete pharmacological investigations.Conclusions:The present study demonstrates the occurrence of Salmonella in the curry samples,and shows significant drug resistance against most of the commercially available antibiotics,except kanamycin.Antimicrobial effect of the plant extracts against the food-bone Salmonella suggests that dietary including medicinal herbs would be one strategy to manage food borne pathogens.

Microwave-Assisted Rapid Extracellular Biosynthesis of Silver Nanoparticles Using Carom Seed (Trachyspermum copticum) Extract and in Vitro Studies

Microwave-assisted rapid extracellular biosynthesis of silver nanoparticles was carried out by using carom seed (Trachyspermum copticum) extract as the reducing agent. The reaction mixture containing AgNO3 and carom seed extract when exposed to microwave irradiation resulted in reducing silver ions to bio-functionalized silver nanoparticles of size 6- 50 nm. The AgNP were characterized by UV-vis spectroscopy (UV-vis), X-ray diffraction (XRD), energy dispersive X-ray analysis (EDAX), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM) and atomic force microscopy (AFM). Themogravimetric analysis (TGA) and fourier transform infrared spectroscopy (FTIR) are used to understand the possible mechanism of biosynthesis. In this study, we have also investigated the antimicrobial and antioxidant activities of bio-functionalized AgNP. The antibacterial activity is investigated by measuring the zone of inhibition and antioxidant study is done using 1,1-diphenyl-2-picryl hydrazyl (DPPH) method.

阿育魏实果醋发酵工艺优化及其抗氧化活性研究

试验以阿育魏实为原料,进行阿育魏实果醋的研制并对其抗氧化活性进行评价。通过单因素试验探究阿育魏实发酵液的加入量、醋酸菌的接种量、发酵时间和发酵温度对果醋酸度和品质的影响,并在单因素基础上以总酸的含量为考察指标,采用正交试验优化阿育魏果醋发酵条件,最终获得最优发酵条件:初始酒精度为6.0%vol、阿育魏实发酵液加入量为80m L、接种量为0.06g/m L、发酵时间为6d、发酵温度29℃时。在此条件下,果醋酸度为52.34g/L,感官评价分为94.00分。抗氧化实验结果表明,Fe^(3+)还原能力、ATBS^(+)·清除能力、DPPH清除能力、·OH清除率、酪氨酸酶二酚酶活力的抑制率分别为76.54%、82.74%、62.14%、60.14%、41.24%,说明阿育魏实-果醋具有一定的抗氧化能力。

星点设计-效应面法优化阿育魏实总黄酮提取工艺及其抗氧化活性研究

以新疆不同产地阿育魏实为研究对象、总黄酮得率为评价指标,采用星点设计-效应面法优化阿育魏实总黄酮提取工艺,通过测定DPPH自由基清除率和Fe^(3+)还原能力评价其抗氧化活性。结果表明,固定超声功率为250 W、温度为40℃,在乙醇体积分数为81%、料液比为1∶26(g∶mL)、提取时间为39 min的最优条件下,总黄酮得率实际值(2.494%)与预测值(2.444%)相接近;不同产地阿育魏实总黄酮得率有所差异,其中,新疆东部哈密地区阿育魏实总黄酮得率普遍较高;不同产地阿育魏实总黄酮均具有一定的抗氧化能力,对DPPH自由基的清除能力和Fe^(3+)的还原能力均随浓度增加呈上升趋势。

阿育魏实化学成分的研究Ⅱ

目的对阿育魏实[Trachyspermum ammi(L.)Sprague]的化学成分进行研究。方法阿育魏实乙醇提取物的正丁醇部分通过多次硅胶柱色谱和反复凝胶柱色谱分离纯化,根据光谱数据和理化性质确定各化合物的结构。结果分离到5个已知化合物,分别是槲皮素(Ⅰ),山柰酚(Ⅱ),山柰酚-3-O-芸香糖苷(Ⅲ),东莨菪内酯(Ⅳ),对香豆酸(Ⅴ)。结论以上化合物均为首次从该种植物中分离得到。

电感耦合等离子光谱法测定阿育魏实中的微量元素

目的建立电感耦合等离子体原子发射光谱法测定阿育魏实中的微量元素。方法用硝酸-高氯酸进行消解,电感耦合等离子体原子发射光谱法（ICP-AES）测定阿育魏实样品中B、Ba、Cu、Sr、Ti、Zn、Mn、P、Ni、Mg、Ca、Co、C、Al、Fe等多种微量元素的含量。结果其相对标准偏差为0.42%~2.8%,回收率为96.2%~101.6%,维药阿育魏实中含比较丰富的Ca、Mg、Fe、K元素,微量元素在其中可能起到一定的药理作用。结论 ICP-AES法具有测定简单,快速,灵敏度高,准确的特点,适用于阿育魏实中微量元素的测定。

阿育魏实挥发油成分的GC-MS研究

目的:搞清维吾尔族药材阿育魏实的挥发油中的主要化学成分。方法:采用水蒸气蒸馏法制备阿育魏实挥发油,用气相色谱-质谱联用技术对阿育魏实的挥发油成分进行研究。结果:阿育魏实挥发油的总收率为7.8%,从中共鉴定出8个化合物,其相对含量之和占挥发油总量的99.99%,其中主要成分为麝香草酚(Thymol,57.45%),1-Methyl-2-(1-methylethyl)benzen(e16.72%),2.6.6-Trimethyl-bicyclo[3.1.1]hept-2-en(e23.69%)。结论:该研究提供了一种阿育魏实挥发油成分的分析方法,并且发现阿育魏实可作为麝香草酚的重要资源植物。

阿育魏实化学成分的预试验研究

目的研究阿育魏实所含化学成分。方法采用化学成分系统预试验辅助专项预试验法。结果阿育魏实中生物碱、黄酮、糖类、氨基酸、皂苷、挥发油、酚性成分、有机酸、油脂、甾体类成分的系统预试实验结果为阳性,生物碱和黄酮的辅助专项预试实验结果为阳性。结论阿育魏实中含有生物碱、黄酮、糖类、氨基酸、皂苷、挥发油、酚性成分、有机酸、油脂、甾体类等成分。

维药阿育魏实总黄酮抗氧化活性研究

[目的]评价阿育魏实总黄酮的抗氧化能力。[方法]采用羟自由基、超氧阴离子自由基、亚硝基自由基、DPPH自由基、ABTS自由基体系对阿育魏实总黄酮抗氧化的能力进行研究。[结果]在质量浓度0.025～0.500 mg/ml的范围内,阿育魏实总黄酮对羟自由基、超氧阴离子自由基、亚硝基自由基、DPPH自由基、ABTS自由基的清除率分别为31.68%～99.01%、27.45%～90.52%、11.62%～86.40%、41.31%～92.21%和17.53%～62.67%,且存在明显的量效关系。IC50分别为0.038 49、0.053 51、0.271 60、0.026 17和0.033 87 mg/ml。[结论]阿育魏实总黄酮具有很好的体外抗氧化活性,是一种新型天然抗氧化物质来源。

新疆产阿育魏实挥发油及不同极性部位抗氧化活性研究

采用2,2'连氨-双-(3-乙基苯并噻唑啉-6-磺酸)二胺盐(ABTS)和二苯代苦味酰基自由基(DPPH),还原能力法对阿育魏实抗氧化活性进行研究,并测定了各样品中总酚和总黄酮含量。结果表明:阿育魏实挥发油和甲醇提取物具有明显的抗氧化能力,清除DPPH.IC50为0.22mg/mL和0.16mg/mL,ABTS.IC50为3.82mg/mL和3.20mg/mL。在3mg/mL时,挥发油和不同溶剂提取物中甲醇提取物表现出较好的还原能力。挥发油和提取物抗氧化活性与酚类和黄酮类化合物具有很大相关性。

新疆阿育魏实中微量元素含量测定

采用微波消解技术处理样品,电感耦合等离子体原子发射光谱（ICP-AES）测定阿育魏实中Cu,Zn,Fe,K,Ca,Na,Mg,Mn,Sr,Cr和Ni 11种微量元素含量。各微量元素的检出限为0.0001~0.0210μg/mL;各元素标准曲线线性关系良好,相关系数在0.9835~0.9999范围内;加标回收率在92.23%~103.20%,测定结果准确可靠。结果表明：阿育魏实富含多种有益微量元素,将其开发成为药食两用功能调味品,具有广阔的前景。

阿育魏实挥发油成分的GC-MS分析

采用水蒸气蒸馏的方法提取了阿育魏实挥发油 ,并利用气相色谱 -质谱联用技术对其化学成分做了分析 ,共鉴定出麝香草酚、p_伞花烃、萜品烯、α_崖柏烯、β_月桂烯和香芹酚等53个化合物。研究阿育魏实挥发油的成分 ,对民族药品质提升及原料的质量保证具有十分重要的意义。

维吾尔药阿育魏实化学成分的分离与鉴定

目的研究新疆维吾尔药材阿育魏实中的化学成分。方法采用硅胶柱色谱及Sephadex LH-20分离化合物,运用现代波谱技术确定所得化合物结构。结果从阿育魏实乙醇提取物分离得到5个化合物,分别鉴定为4-羟基薄荷-1-烯-7-酸(4-hydroxymenth-1-en-7-oic acid,1)、E-3-(3,4-二羟基苯基)-丙烯酸3-(3,4-二羟基苯基)丙酯[E-3-(3,4-dihydroxyphenyl)propyl 3-(3,4-dihydroxyphe-nyl)acrylate,2]、6-羟基麝香草酚6-O-β-D-葡萄糖苷(6-hydroxythymol 6-O-β-D-glucopyranoside,3)、麝香草氢醌6-O-6'-乙酰基-β-D-葡萄糖苷(thymohydroquinone 6-O-6'-acetyl-β-D-glucopyranoside,4),对羟基桂皮酸(4-hydroxycinnamic acid,5)。结论化合物1、2为新天然产物,化合物4首次从伞形科植物分离得到。

维药阿育魏实总多糖的提取工艺研究

[目的]通过正交试验探讨提取维药阿育魏实总多糖的最佳提取工艺,为资源的合理开发利用提供科学依据.[方法]以水为溶剂回流提取,采用苯酚-浓硫酸法显色,采用紫外分光光度法（UV）测定阿育魏实中总多糖的含量,并进行方法学验证.[结果]维药阿育魏实总多糖的最佳提取工艺为提取温度90℃、提取时间3h、料液比1∶50、提取次数1次;标准品溶液和供试品溶液均在488 nm处有最大吸收,总多糖溶液浓度在0.002 5 ～0.012 5 mg/ml线性关系良好（r=0.999 3）;精密度试验、稳定性试验、重复性试验和加样回收率试验的RSD值分别为0.6％、0.5％、0.069％和0.9％,同时得到阿育魏实多糖平均提取率可达3.41 mg/g.[结论]该方法具有操作简单、重复性好、灵敏度高,适用于阿育魏实中总多糖含量测定.

维吾尔药阿育魏实中一个单萜糖苷的谱学分析

目的:确证维吾尔药阿育魏实中一个单萜糖苷类化合物的结构。方法:采用硅胶柱色谱、开放式ODS柱色谱及高效液相色谱分离化合物,运用UV、IR、NMR、CD及MS技术确定该化合物结构。结果:对该化合物结构的谱学特征进行全归属。结论:该化合物鉴定为6-羟基麝香草酚3-O-β-D-吡喃葡萄糖苷,首次从该植物分离得到。

Synergistic Antioxidant Action of Oregano, Ajowan and Borage Extracts

Radical scavenging activity (RSA), antioxidant activity (AA), reducing power, total polyphenol (TP) and flavonoid contents (TFC) of oregano (Oreganum vulgare spp), ajowan (Trachyspermum ammi) and Indian borage (Plectranthus amboinicus) extracts were evaluated. Oregano exhibited maximum radical scavenging activity (88.2%, 82.3%) for aqueous and ethanolic extracts at 50 ppm concentration respectively, followed by ajowan (86.9%, 68.4%) and Indian borage (30.5%, 30.4%). Extracts of oregano and ajowan showed better antioxidant activity and reducing power than that of Indian borage. Aqueous extracts of oregano had highest TP (Gallic acid equivalents) and TFC (Catechin equivalents) of 27.7% and 50.6 % respectively compared to ajowan (6.7%, 24.4%) and Indian borage (4.2%, 5.5%). Synergistic studies showed that the addition of oregano extract appreciably enhanced the RSA of ajowan and Indian borage extracts even at 50 ppm concentration.

阿育魏实挥发性成分提取工艺参数筛选及其体外活性研究

非酒精性脂肪性肝病(Non-alcoholic fatty liver disease,NAFLD)是指肝细胞内以脂肪变性为主要特征的临床病理综合征,而资料显示阿育魏实具有一定的降脂活性,可以改善NAFLD。因此本研究采用Box-Benhnken响应面法筛选阿育魏实挥发性成分(Trachyspermum ammi(L.)Sprague volatile constituent,TSVC)提取参数并初步考察其体外活性研究。在单因素基础上,采用四因素三水平响应面法筛选阿育魏实挥发性成分的提取工艺参数,分别建立NAFLD和胰岛素抵抗细胞模型并进行干预,微量法检测细胞葡萄糖消耗量、TG、TC、AST、ALT,进行油红染色考察其改善非酒精性脂肪肝和胰岛素抵抗活性。当TSVC最佳提取条件为料液比9mL/g、浸泡时间12h、超声时间15min和提取时间6.5h,其提取率为2.12%;体外实验结果表明:与模型组相比,阿育魏实挥发性成分中、高剂量组降低细胞内的TG、TC、AST、ALT含量;增加胰岛素抵抗模型细胞的葡萄糖消耗量;油红染色结果中看出可以减少脂质沉积。总之,阿育魏实挥发性成分在一定浓度范围内可改善非酒精性脂肪肝和胰岛素抵抗,为其后期研究提供理论依据和实验基础。

维药阿育魏果中总黄酮提取工艺及含量测定

[目的]探讨阿育魏果中总黄酮的提取方法及含量测定方法。[方法]采用正交试验,以提取的总黄酮的含量为评价指标,紫外可见分光光度法测定总黄酮含量。[结果]超声法提取阿育魏果中总黄酮的含量为10.378%,相对标准偏差为2.38%(n=3)。最优工艺如下:时间为15 min,功率为150 W,料液比为1∶20,乙醇浓度为70%。上述4个因素对阿育魏果中总黄酮提取率的影响从大到小依次为乙醇浓度、料液比、超声功率和超声时间。[结论]超声提取阿育魏果中总黄酮快速且高效;紫外外分光光度法准确度较高,偏差较小。

计算机辅助策略研究阿育魏实有效成分与诱导癌细胞凋亡机制的关系

目的本研究的目的是通过计算机辅助的方法研究阿育魏实(Trachyspermum ammi (L.) Sprague)有效成分诱导癌细胞凋亡的机制。方法通过文献挖掘的方法确定阿育魏实候选小分子化合物,综合靶点预测、分子对接、网络药理学和分子动力学(MD)模拟等方法筛选阿育魏实化学成分中诱导癌细胞凋亡的活性成分并对其作用机制进行探讨。结果数据挖掘搜集到42个候选有效成分,随后运用靶点预测方法获得这些化合物的作用靶点,构建关键蛋白质的蛋白质-蛋白质相互作用网络(PPI)和mi RNA-m RNA作用网络。然后,通过分子对接和分子动力学模拟进行验证,最终筛选出化合物1、10、11、12和ERBB2, IGF1R, ESR1, NQO1, MMP9之间能较好的相互作用。结论计算机辅助筛选分析表明,阿育魏实中化合物1,10,11,12可以靶向癌细胞中的蛋白ERBB2,IGF1R,ESR1,NQO1,MMP9,其可能揭示和探索阿育魏实用于治疗癌症复杂的分子机制。