Analysis of phytochemical profile of Terminalia arjuna bark extract with antioxidative and antimicrobial properties

Objective

To investigate phytochemical screening, antimicrobial activity and qualitative thin layer chromatographic separation of flavonoid components, antioxidant activity and total flavonoid compound of Terminalia arjuna.

Methods

For phytochemical screening, some common and available standard tests were done. Antimicrobial bioassay was done through agar well diffusion method. Detection of antioxidant activity and flavonoid compounds were done through thin layer chromatography. Total antioxidant activity was measured by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) in colorimetric method. Aluminum chloride colorimetric method was used for total flavonoid determination.

Results

Phytochemical screening showed the active compounds presence in high concentration, such as phytosterol, lactones, flavonoids, phenolic compounds and tannins and glycosides. The antimicrobial activity of extract showed that greater inhibition zone against Gram negative bacteria than Gram positive bacteria. This methanolic extract showed a promising antioxidant activity, as absorption of DPPH redicles decreased in DPPH free radical scavenging assay. Flavonoids components having antioxidant property present in the methanol extract at a level of 199.00 mg quercetin equivalent/g of dried methanol extract in colorimetric method.

Conclusions

The Terminalia arjuna bark extract revealed the presence of bio-active constituents which are known to exhibit medicinal as well as physiological activities.     

Antioxidant potential of Rumex vesicarius L.: in vitro approach

ObjectiveTo assess in-vitro antioxidant activity of different fraction and perform high performance thin layer chromatography fingerprint analysis of most active fraction of Rumex vesicarius L. (R. vesicarius).MethodsIn the present study, acetone, ethyl acetate, n-butanol, and methanol extracts of R. vesicarius were evaluated for radical scavenging activity by studying the inhibition of the level of lipid peroxidation induced by Fe(++)/ascorbate, DNA sugar damage, scavenging of hydrogen peroxide, diphenylphosphine DPPH radical scavenging activity, total phenolic content, total flavonoids content and total proanthocyanidin. High performance thin layer chromatography finger print profiling of R. vesicarius L. was also done.ResultsLipid peroxidation induced by the iron/ascorbate system, hydrogen peroxide, diphenylphosphine and DNA sugar damage were inhibited by the addition of different extract of R. vesicarius. Among them, methanolic extract showed maximum efficacy. The methanolic extract showed the highest total phenolic, total flavonoids and total proanthocyanidin contents.ConclusionsThe results suggest that the extracts can be a vital source of phytochemical antioxidants.     

GC-MS analysis,determination of total phenolics,flavonoid content and free radical scavenging activities of various crude extracts of Moringa peregrina (Forssk.) Fiori leaves

ObjectiveTo perform phytochemical screening, estimate total phenolics, flavonoids and to evaluate antioxidant potential of Moringa peregrina (M. peregrina) leaves.MethodsThe dried powdered leaves of M. peregrina (150 g) were extracted exhaustively by Soxhlet with ethanol and then fractionated into hexane, chloroform, ethy alacetate and methanol. All the prepared extracts were also analyzed by gas chromatography-mass spectrometry to identify and characterize the chemical compounds present in the crude extracts. Folin- Ciocalteu reagent and aluminium chloride colorimetric methods were used to estimate total phenolic and flavonoid content of extracts. Hydrogen peroxide and 1,1 diphenyl -2-picrylhydrazyl were used to determine in vitro antioxidant activity.ResultsPhytochemical analysis of ethanol extract showed presence of major classes of phytochemicals. Gas chromatography-mass spectrometry results revealed presence of 19 phytoconstituents in hexane extract, 6 in ethyl acetate and 7 compounds in methanolic extract. Methanol extract was found to contain the highest phenolic content and flavonoids. In vitro antioxidant activities of all crude extracts were significant and comparable with the standard ascorbic acid.ConclusionsResults of this study show that the leaves of M. peregrina are the rich source of phenolic compounds that can play an important role in preventing the progression of many diseases.     

新疆香鳞毛蕨药材质量研究

目的：建立新疆产香鳞毛蕨的质量控制方法,并评价其质量,为该植物资源的开发利用提供科学依据。方法采用性状鉴别、显微鉴别、水分、灰分、浸出物、薄层鉴别和总间苯三酚含量测定等研究方法。结果新疆香鳞毛蕨的根茎横切面有分体中柱3-7个,周韧型。粉末黄棕色,孢子囊环带,薄壁细胞呈多角形或类圆形,含淀粉粒;管胞孔纹、梯纹。3批香鳞毛蕨水分、灰分、酸不溶灰分和浸出物含量分别为9．46％-10．22％、8．46％-9．52％、2．89％-3．56％、24．53％-32．33％,总间苯三酚含量为5．16％-8．10％,RSD＝7．14％。结论建立的方法适用于香鳞毛蕨药材的质量控制。     

A comparative study of the antioxidant,antimicrobial, cytotoxic and thrombolytic potential of the fruits and leaves of Spondias dulcis

Objective

To investigate the antioxidant, antimicrobial, cytotoxic and thrombolytic property of the fruits and leaves of Spondias dulcis (S. dulcis).

Methods

Methanolic extracts of fruits and leaves of S. dulcis were partitioned with chloroform and dichloromethane. The antioxidant potential of the crude extract and partitioned fractions were evaluated in terms of total phenolic content, total flavonoid content, DPPH radical scavenging potential, reducing potential and total antioxidant capacity by specific standard procedures. The antimicrobial activity was evaluated using disc diffusion method. The cytotoxicity was evaluated by using brine shrimp lethality bioassay and compared with vincristine sulfate. The thrombolytic activity was compared with streptokinase.

Results

The methanolic fruit extract exhibited the highest phenolic content, flavonoid content and antioxidant capacity, among the other extracts, with the highest DPPH radical scavenging activity at a concentration of 10 µg/mL (IC₅₀: 1.91 µg/mL) and maximum reducing power at a concentration of 100 µg/mL (EC₅₀: 3.58 µg/mL). Though all extract showed moderate antimicrobial activity against the bacterial strains, weak or no activity against fungus. The range of LC₅₀ value of all extracts was 1.335-14.057 µg/mL which was far lower than the cut off index for cytotoxicity. All extracts exhibited statistically significant (P<0.001) thrombolytic activity.

Conclusions

Our study suggested that S. dulcis exhibits antimicrobial activities against a wide variety of strains while it possesses significant antioxidant, cytotoxic and thrombolytic activity.     

广东紫荆皮的质量标准研究及其易混品辨析

目的 研究广东紫荆皮的质量标准.方法 采用2015年版<中国药典>中方法对广东紫荆皮药材中的水分、灰分及浸出物进行了测定,对广东紫荆皮药材薄层鉴别的条件及指标成分的质量分数测定条件进行了优化.结果 测得广东紫荆皮药材中水分不得超过15.5％,总灰分不得超过9.24％,70％(体积分数)乙醇热浸出物不得少于27.5％;以羽扇豆醇、没食子酸为对照品的薄层鉴别斑点清晰、分离度好.羽扇豆醇的质量分数测定在0.2160～10.80μg内与峰面积呈良好的线性关系,r=0.9999,平均回收率为96.9％(RSD=2.1％,n=6),样品平均质量分数为12.40 mg/g;而其混淆品未检出此成分.结论 本方法简便、专属性强、重复性好,能区分广东紫荆皮及其混淆品,为评价广东紫荆皮药材的质量标准提供科学依据.     

地肤子与其伪品灰菜子的鉴别

目的 :研究地肤子与灰菜子的鉴别 ,为准确用药提供依据。方法 :用放大镜观察二者的原药材外形 ;用显微镜观察二者的粉末显微特征 ,并用薄层色谱法对二者进行对比试验。结果 :地肤子与灰菜子在放大镜下观察 ,有比较明显的区别。薄层色谱的差别很大。结论 :灰菜子不能作地肤子使用     

Phytochemical and in vitro biological investigations of methanolic extracts of Enhydra fluctuans Lour.

ObjectiveTo study the phytochemical and biological properties (antioxidant, anthelmintic and thrombolytic) of methanolic extracts of Enhydra fluctuans Lour., a plant belonging to the Asteraceae family.MethodsThe phytochemical evaluation was carried out by qualitative analysis. In vitro antioxidant activity of extract was studied using free radical scavenging assay, ability of reduction, total phenol and total flavonoid contents determination assays. The anthelmintic activity was determined using paralysis and death time of Pheretima posthuma (earthworm) and thrombolytic activity by clot disruption assay.ResultsThe phytochemical evaluation showed significant presence of flavonoids, triterpenes, carbohydrate, reducing sugars, saponins, phenols, diterpenes, protein and tannin. The antioxidant activity was found significant [IC₅₀=(135.20±0.56) μg/mL] as compared to ascorbic acid [(130.00±0.76) µg/mL]. The reducing power was increased with concentration. Total phenol and total flavonoid contents were (153.08±0.38) mg/mL and (172.04±0.56) mg/mL respectively. The paralysis and death time of earthworms for different concentrations of extract were determined and compared with albendazole. The results showed that 10 mg/mL of the crude extract had similar effect with albendazole. Additionally, the crude extract showed a concentration depended relationship with its anthelmintic property. The clot lysis activity of crude extract was compared to the standard streptokinase's clot lysis (40.13%) activity and found significant (31%).ConclusionsThe study proves that the crude methanolic extract of Enhydra fluctuans Lour. has significant antioxidant, anthelmintic and thrombolytic activity containing wide range of phytochemicals.     

Antioxidant activities of Indigofera cassioides Rottl. Ex. DC. using various in vitro assay models

Objective

To evaluate the antioxidant potential of methanolic leaf extract of Indigofera cassioides (MEIC) using various in vitro antioxidant assay systems.

Methods

Antioxidant and free radical scavenging activity of MEIC was assayed by using different in vitro models like ABTS, DPPH, nitric oxide, superoxide, hydrogen peroxide and hydroxyl radical. Reductive ability of the extract was tested by the complex formation with potassium ferricyanide. Further total phenol and flavonoid contents of the crude extract were also determined. Rutin and ascorbic acid were used as standards.

Results

MEIC exhibited potent and concentration dependent free radical scavenging activity in all the tested models. Reductive ability was also found to increase with increase in MEIC concentration. Total phenol and flavonoid content determination showed that the extract is rich in phenols and flavonoids.

Conclusions

All the results of the in vitro antioxidant assays reveal potent antioxidant and free radical scavenging activity of the leaves of Indigofera cassioides, equivalent to that of standard ascorbic acid and rutin. This potent antioxidant activity may be attributed to its high phenolic and flavonoid contents.     

龙葵总提取物对多发性骨髓瘤U266细胞株的作用

目的:研究龙葵总提取物对U266细胞的细胞毒作用及其机制.方法:U266细胞株与龙葵总提取物共同培养,用CCK-8试剂检测细胞毒作用;用流式细胞仪测定细胞周期及凋亡.结果:龙葵总提取物对U266细胞有细胞毒作用,半数抑制(质量)浓度约为117 mg/L;龙葵总提取物影响U266细胞周期,G0/G1期细胞减少,S期、G2/M期细胞增加,凋亡细胞增加.用Annexin V/PI染色及TFAR19染色流式细胞仪检测,均显示细胞凋亡与龙葵总提取物有剂量依赖关系.结论:龙葵总提取物对U266细胞有体外细胞毒作用,其作用机制部分是诱导细胞凋亡.     

藜芦(Veratrum nigrum L.)生品与炮制品中藜芦新碱(veratrosine)含量和毒性差异研究

目的:探讨中药藜芦(Veratrum nigrum L.)的炮制机理研究。方法:利用HPLC-UV方法测定生物碱藜芦新碱(veratrosine)在生品和炮制品中的含量差异以及利用急性毒性实验,考察生品和炮制品毒性差异。结果:藜芦新碱(veratrosine)质量浓度在0 mg/L~2000 mg/L内与峰面积呈良好的线性关系,r=0.9998(n=7);平均回收率为98.6%,RSD为3.17%(n=5)。藜芦水提物灌胃给小鼠的LD50为4.2 g/kg,95%可信限为3.4 g/kg~5.4 g/kg;米泔制藜芦水提物灌胃给小鼠的LD50为5.3 g/kg,95%可信限为4.8 g/kg~6.1 g/kg。醋制藜芦水提物灌胃给小鼠的LD50为3.2 g/kg,95%可信限为2.5 g/kg~3.9 g/kg。结论:藜芦醋制后会使生物碱藜芦新碱(veratrosine)溶出率增高,毒性增强;藜芦米泔制后会使生物碱藜芦新碱(veratrosine)溶出率降低,毒性减弱。     

龙葵药材薄层鉴别研究

目的研究中药龙葵的薄层鉴别方法。方法依据现行《中国药典》2010年版一部附录ⅥB试验方法考察了不同提取方法、薄层板、展开剂、显色剂下龙葵的薄层鉴别方法。结果用硅胶G薄层板．二氯甲烷一甲醇一氨水（8：1．5：0．4）为展开剂,以10％硫酸乙醇试液显色,再置紫外灯（365nm）下检视斑点,分离效果最佳。结论本实验研究可作为龙葵药材质量标准的修订内容。     

鼠妇虫质量标准研究

目的 建立鼠妇虫的质量标准,为控制该药材质量提供检验依据.方法 对鼠妇虫进行性状、薄层鉴别,杂质、水分、灰分和浸出物的测定.结果 制定了鼠妇虫的性状和薄层鉴别法,确定了鼠妇虫中杂质、水分、灰分和浸出物的限量.结论 首次系统的对鼠妇虫的质量标准进行了研究,为完善鼠妇虫质量控制方法,建立该药材国家标准奠定了实验基础.     

有梗石龙尾甲醇提取物抗志贺菌病作用的体内和体外研究

目的：探讨有梗石龙尾甲醇提取物抑制志贺菌病的作用。方法：首先将有梗石龙尾甲醇提取物做初步化学筛选。使用2种革兰阴性菌、2种革兰阳性菌,2种抗生素耐药的志贺杆菌和1种非抗生素耐药的志贺杆菌经纸片扩散法进行体外抗菌筛选。蓖麻油诱发 Wistar大鼠腹泻后,以洛哌丁胺作为标准对照药进行有梗石龙尾甲醇提取物止泻实验。通过人工胃模型测定有梗石龙尾甲醇提取物的体外抗酸作用,并且对其中和胃酸的效率、容量、体积和氢离子消耗进行评估。结果：有梗石龙尾甲醇提取物中含有酚类、黄酮类、生物碱等化学成分,以及脂肪和油脂。有梗石龙尾甲醇提取物被证明对包括抗生素耐药的志贺杆菌在内的4个菌株具有较强的抑制作用,并且其抗腹泻作用具有剂量依赖性。当服用最低剂量100mg/kg时,其抗腹泻效果比标准对照药洛哌丁胺更好（P〈0.01）。体内实验结果显示有梗石龙尾甲醇提取物具有轻微的抗酸作用。结论：有梗石龙尾甲醇提取物具有轻微的抗酸活性以及强效的抗菌、止泻作用,这对于开发抗痢疾、抗腹泻作用的草药具有一定的意义。     

桑托利纳蓍降低弗氏佐剂致炎症大鼠的血清白细胞介素6水平及痛觉敏感程度

目的：免疫系统涉及炎症的很多病因学及病理学机制。药用植物中的很多成分被认为具有非特异性的免疫调节作用。基于此并且根据白细胞介素6在炎症和疼痛产生中的作用,本研究探讨桑托利纳蓍（Achilleasantolina）的提取物对弗氏佐剂致炎症大鼠的作用。方法：提取桑托利纳蓍地上部分的甲醇提取物和脱脂提取物用于本次研究。通过辐射热测量和器官充盈度测量器分别衡量弗氏佐剂致炎症大鼠不同实验天数后的痛觉敏感程度和足肿胀程度,并使用大鼠白细胞介素酶联免疫吸附试剂盒测量大鼠血清白细胞介素6的水平。结果：桑托利纳蓍的甲醇提取物减少炎症大鼠足肿胀、降低痛觉敏感程度和血清白细胞介素6水平呈量效关系。剂量为200mg／kg的桑托利纳蓍甲醇提取物在预处理和短期治疗期间具有明显的抗痛觉过敏和抗炎作用,400和200mg／kg剂量的治疗作用没有明显差别。桑托利纳蓍的脱脂提取物对于弗氏佐剂所导致的大鼠炎症反应在实验的各个阶段均未显示出明显的治疗作用（P〉0．05）。短期治疗期间,200mg／kg的桑托利纳蓍甲醇提取物在减少炎症大鼠足肿胀、降低痛觉敏感程度和血清白细胞介素6水平方面均优于阳性对照药消炎痛（P〈0．01,P〈0．01,P〈0．05）。结论：桑托利纳蓍的甲醇提取物预处理用药和短期用药均有明显的抗炎和免疫调节作用。     

龙葵提取物对荷lewis肺癌小鼠肿瘤组织细胞周期调控相关蛋白及凋亡的影响

[目的]观察龙葵氯仿及正丁醇提取物对荷lewis肺癌小鼠肿瘤组织细胞周期调控相关蛋白、细胞凋亡及相关凋亡蛋白表达的影响。[方法]建立lewis肺癌模型,64只C57BL/6J小鼠随机分成模型组、环磷酰胺组、龙葵氯仿和正丁醇提取物高、中、低剂量组,给药15d后处死,剥离肿瘤组织,称重,计算肿瘤生长抑制率;TUNEL染色法检测细胞凋亡;免疫组织化学法检测PCNA、p53、p21和Bax、Bcl-2。[结果]龙葵氯仿及正丁醇提取物对荷瘤小鼠肿瘤增殖有一定抑制作用,其中氯仿提取物中剂量组和正丁醇提取物高剂量组的抑瘤率分别为38.93%和32.14%（P〈0.01或P〈0.05）。龙葵氯仿及正丁醇提取物可降低荷瘤小鼠肿瘤组织PCNA和P53阳性细胞表达率,分别为29.7%-58%和32%-56%,并提高P21阳性细胞表达率（20%-32%）（P〈0.01或P〈0.05）。荷瘤小鼠肿瘤组织细胞的凋亡率均不同程度升高（15%-38%）（P〈0.01或P〈0.05）。Bax阳性细胞表达率均有不同程度升高（11%-41%）,Bcl-2阳性细胞表达率则均有不同程度降低（23%-37%）（P〈0.01或P〈0.05）,并可不同程度上调荷瘤小鼠肿瘤组织Bax/Bcl-2的比值。[结论]龙葵正丁醇及氯仿提取物有一定抑制lewis肺癌的生长作用,其作用机制可能与对荷瘤小鼠肿瘤组织细胞周期调控相关蛋白（PCNA、p53、p21）表达的影响、上调肿瘤组织Bax蛋白的表达、下调Bcl-2蛋白的表达,上调Bax/Bcl-2的比值;促进肿瘤组织的细胞凋亡等相关。     

戈壁天冬中的木脂素和酚酸类化学成分

目的 研究天门冬属植物戈壁天冬中的化学成分。方法 使用硅胶柱色谱、凝胶柱色谱、制备液相色谱及薄层色谱等多种色谱分离技术对戈壁天冬全草95%乙醇提取物进行分离纯化,利用理化常数和多种波谱技术对化合物进行了结构鉴定。结果 从戈壁天冬中分离鉴定了9个化合物,其中6个是木脂素类化合物,分别为(+)-松脂素（1）,(-)-丁香脂素（2）,(+)-丁香脂素-4'-O-β-D-葡萄糖苷（3）,(-)-表丁香脂素（4）,(+)-nyasol（5）,(+)-4'-O-methylnyasol（6）;3个酚酸类化合物,分别为阿魏酸（7）,异阿魏酸（8）,咖啡酸（9）。结论 所有化合物均为首次从本植物中分离得到,其中化合物1~4和8为首次从天门冬属植物中分离得到。      

气相色谱法检测9种药材中有机氯农药的残留量

目的测定9种药材中有机氯农药的残留量。方法以正已烷／丙酮（8／2）混合溶剂,在Florisil固相萃取柱上净化提取,采用毛细管气相色谱柱分离样品和电子捕获检测器进行检测。结果11种农药能较好地分离,峰面积与其浓度均具有良好线性,两个水平的加样回收率分别为79．9％89．0％和86．3％-104．8％,相对标准偏差为1．8％7．1％,检测限为2g／kg;广藿香和三七中有机氯农药残留超过国家标准。结论所建GC-ECD法能满足药材中有机氯农药残留量的检测,药典应制订药材中农药残留量的标准。     

高效薄层层析法测定罗布麻烟中罗布麻浸膏的含量

利用高效薄层层析法,以罗布麻浸膏中的槲皮素为标准,测定罗布麻烟中罗布麻浸膏的含量。以甲苯:甲酸:乙酸乙酯:正己烷(2:0.2:1.5:0.2)作展开剂。可使得药烟中槲皮素与其它成分很好分离。分离后,用薄层扫描仪于波长410nm,参比波长750 nm扫描定量。本法需样品少,快速简便、灵敏准确。可适用于药烟生产厂家化验室检测使用。     

露兜簕的质量标准研究

目的：建立露兜簕的质量标准。方法：通过对露兜簕的性状、显微鉴别、薄层色谱鉴别的研究建立露兜簕的质量标准。结果：按照薄层色谱法用硅胶G薄层板为固定相,以环己烷-乙酸乙酯（6∶1）为展开剂,喷以1%三氯化铝乙醇溶液,在105℃加热5min,置紫外光灯（365nm）下检视。供试品色谱中,在与对照药材色谱图相应的位置上显相同颜色的荧光斑点。结论：本文建立的方法能够有效控制露兜簕的质量。