Optimization of microwave-assisted extraction of bioactive alkaloids from lotus plumule using response surface methodology

In this work,a fast and efficient microwave-assisted extraction(MAE) method was developed to extract main bioactive alkaloids from lotus plumue.To optimize MAE conditions,three main factors were selected using univariate approach experiments,and then central composite design(CCD).The optimal extraction conditions were as follows:methanol concentration of 65%,microwave power of 200 W,and extraction time of 260 s.A high performance liquid chromatography–diode array detector(HPLC–DAD) method was established to quantitatively analyze these phytochemicals in different lotus plumule samples and in different part of lotus.Chromatographic separation was carried out on an Agilent Zorbax Extend-C_(18) column(4.6 mm×150 mm,3.5 μm).Gradient elution was applied with the mobile phase constituted with 0.1% triethylamine in water(A)and acetonitrile(B):40%-70% B at 0-8 min,70%-100% B at 8–9 min,100% B for 2 min,and then equilibrated with 40% B for 2 min.

Genotoxicity and Safety Pharmacology Studies of Indole Alkaloids Extract from Leaves of Alstonia scholaris (L.) R. Br.

Indole alkaloids extract(IAAS)was prepared from leaves of Alstonia scholaris(L.)R.Br.,an evergreen tropical plant widely distributed throughout the world.This plant has been used historically by the Dai ethnic people of China to treat respiratory diseases.This study evaluated the genotoxicity and safety pharmacology of IAAS to support clinical use.The bacterial reverse mutation(Ames)test,in vitro mammalian chromosomal aberration test,and in vivo mammalian erythrocyte micronucleus(MN)test were performed to evaluate genotoxicity.Mice were administered IAAS(240,480,or 960 mg/kg bw)once orally to observe adverse central nervous system effects.Furthermore,beagle dogs were administered IAAS(10,30,60 mg/kg bw)once via the duodenum to evaluate its effects on the cardiovascular and respiratory systems.IAAS with or without S9-induced metabolic activation showed no genotoxicity in the Ames test up to 500μg/plate,in the mammalian chromosomal aberration test up to 710μg/mL,or in the MN test up to 800 mg/kg bw.No abnormal neurobehavioral effects were observed in mice following treatment with up to 960 mg/kg bw of IAAS.Moreover,blood pressure,heart rate,electrocardiogram parameters,and depth and rate of breathing in anesthetized beagle dogs did not differ among the IAAS doses or from the vehicle group.These data indicated that IAAS did not induce mutagenicity,clastogenicity,or genotoxicity,and no pharmaco-toxicological effects were observed in the respiratory,cardiovascular,or central nervous systems.Our results increased understanding of safety considerations associated with IAAS,and may indicate that IAAS is a possible drug candidate.

Matrix Solid-phase Dispersion Extraction of Alkaloids from the Roots of Aconitum kusnezoffii Reichb

Matrix solid-phase dispersion（MSPD） was developed for the extraction of four alkaloids, including aconitine, mesaconitine, hypaconitine and deoxyaconitine, from the roots ofAconitum kusnezoffii Reichb. The determination of the analyte was carried out by high performance liquid chromatography with UV detection. The alkaline alumina was used as sorbent. The mixture of acetonitrile and water was used as elution solvent. Several extraction parameters, such as type of sorbent, the ratio of sample to solid support material, type of the elution solvent and the volume of the elution solvent were tested. Mean recoveries ranged from 93.16% to 102.73%, with relative standard deviations from 0.27% to 4.17%. With the extraction efficiency and time expenditure taken into account, MSPD extraction should be a comparatively good method.

Liver cytochrome P450 system as affected by endophyte-infected tall fescue seed extracts and ergot alkaloids

Endophyte infected tall fescue (E+) is the base diet for nearly all beef cattle in the southern USA. It has been linked to a variety of toxicological conditions due to the presence of large numbers of ergot alkaloids. This study was designed to investigate the effects of E+ seed extract and selected ergot alkaloids on the detoxification pathway by cytochrome P450 (CYP3A4) enzyme system. Tests were performed using the P450- Glo CYP3A4 enzyme activity kit (Promega, WI), according to the manufacturer’s manual. Luminescence was measured using a single tube TD20/20 luminometer. Endophyte infected tall fescue seed was extracted with 50/50 methanol/25 mM ammonium carbonate, cleaned and concentrated on Strata-X reversed phase column (Phenomenex). The extracts were evaluated on an HPLC, and then tested using a serial dilution method. Commercially available ergonovine (EN), ergocorine (ER), bromocryptine (BC) and ergocryptine (EC) were tested individually using 0 to 44 nM concentrations. Seed extract of E+ produced a significant (P P < 0.05) dose dependent manner with EC being most potent, followed by ER, BC, and then EN (70%, 40%, 30% and 10% at 44 nM concentration). The similarity of the inhibition curves of seed extract to that of the commercially available ergot alkaloids suggests a related mode of action and that the use of such ergot alkaloids and CYP3A4 assay is a good model to study the toxicity of tall fescue. Furthermore, it provides the foundation to identify the individual toxic components of purified endophyte infected tall fescue extract.

Comparison of Extraction Methods for Total Alkaloids from Abrus Cantoniensis Hance

[Objectives] To compare different extraction methods for total alkaloids from Abrus Cantoniensis Hance taking chloroform as solvent. [Methods] The ethanol extraction method,ultrasonic extraction method,acid water extraction method,and Soxhlet extraction method were used to extract total alkaloids,and the total alkaloids of Abrus Cantoniensis Hance were measured at 238 nm wavelength. [Results] The extraction rate of ethanol extraction method was 14. 76 mg/100 g; the extraction rate of ultrasonic extraction method was 56. 85 mg/100 g; the extraction rate of acid water extraction method was 56. 63 mg/100 g; the extraction rate of Soxhlet extraction method was 58. 19 mg/100 g.[Conclusions]Results indicate that Soxhlet extraction method can obtain the highest extraction rate.

基于UPLC-Triple-TOF-MS技术初步分析雅安产道地中药材川牛膝中生物碱成分

采用乙醇提取、成盐、二氯甲烷萃取等方法初步富集川牛膝总生物碱,确定最优提取分离工艺;通过超高液相色谱串联三重四级杆飞行时间质谱法,采用Waters XSelect CSH-C_(18)色谱柱(2.1 mm×150 mm,1.7μm),梯度洗脱,流速为0.3 mL/min,测试波长254 nm,柱温为50℃,进样量为3μL,采用电喷雾离子源正离子模式,扫描范围为m/z 100~1500,并采用全扫描模式对样品数据进行收集,根据高分辨质谱结合二级质谱所得信息对川牛膝中生物碱类化学成分进行快速鉴定,通过对比分子离子峰和碎片离子的质荷比、Scifinder和Reaxy数据库及相关文献,推测出两种生物碱成分。研究建立的生物碱类化学成分提取与分析方法,为提高雅安产道地中药材川牛膝的质量控制水平及后续阐明药效物质基础提供了实验依据。

天茄子乙酸乙酯部位化学成分研究

采用脂多糖诱导小鼠单核巨噬细胞RAW 264.7炎症模型导向研究天茄子(Ipomoea turbinata seeds)的抗炎活性成分。采用硅胶、Sephadex LH-20、MCI、ODS柱色谱及半制备型高效液相等方法对乙酸乙酯活性部位进行分离纯化,结合现代波谱技术与理化性质确定化合物结构。从中共分离鉴定了15个化合物,分别为华佗豆甲碱(1)、华佗豆丙碱-4′-O-β-D-(6-O-反式香豆酰基)葡萄糖苷(2)、1,8,15,22-四氮杂环二十八烷-2,9,16,23-四酮(3)、1,8,15,22,29-五氮杂环三十五烷-2,9,16,23,30-五酮(4)、绿原酸甲酯(5)、绿原酸(6)、3,5-二咖啡酰基奎宁酸(7)、3,4-二咖啡酰基奎宁酸(8)、咖啡酸(9)、3,5-二羟基桂皮酸(10)、4-甲氧基肉桂酸(11)、6-氧-(E)-对羟基桂皮酰基-1-β-乙基-葡萄糖甙(12)、E-3-(3,4-二羟基苯亚甲基)-5-(3,4-二羟基苯基)二羟基呋喃-2-酮(13)、丁香脂素-4-O-β-D-吡喃葡萄糖苷(14)和松脂素-4-O-β-D-葡萄糖苷(15)。化合物3~8和10~15为首次从该植物中分离得到,其中8个化合物(3~5、10~13和15)为首次从旋花科中分离得到。活性筛选结果表明,咖啡酰奎宁酸类(5~8)和苯丙酸类成分(9~12)显示有较好的抑制NO释放活性,在50μmol/L浓度下抑制率为29.07%~40.30%。

黄连生物碱超声辅助低共熔溶剂提取工艺优化

目的优化黄连生物碱超声辅助低共熔溶剂提取工艺。方法以葡萄糖-乳酸(1∶5)为低共熔溶剂。在单因素试验基础上,以提取时间、含水量、液料比为影响因素,生物碱得率为评价指标,Box-Behnken响应面法优化提取工艺。结果最佳条件为提取时间35 min,含水量26%,液料比33∶1,生物碱得率为7.22%。结论该方法稳定可靠,切实可行,可为黄连开发利用提供参考。

博落回生物碱的提取工艺及其在动物养殖中的研究进展

博落回生物碱是中兽药博落回的功能性成分,其提取工艺多种多样,利用传统的酸提碱沉法即可得到粗提物,工艺简单易于工业化生产,而液膜法、内部沸腾法、超声波提取技术、超临界流体萃取技术、亚临界流体萃取技术等新方法、新技术结合萃取法、结晶法、色谱法等分离纯化技术的应用,对提高中药提取率、活性成分提取率及获取高纯度有效成分等具有显著作用和优势。以血根碱和白屈菜红碱为主要成分开发的新中兽药和药物饲料添加剂产品在改善消化道健康、增强机体免疫力、促进生长、提高生产性能及替代抗生素改善动物健康等方面效果显著。文章综述了博落回生物碱的提取、分离纯化、生物活性及其在动物养殖中的应用进展,以期为博落回生物碱的深层次开发及进一步应用提供参考。

两面针粗生物碱的富集

目的探索两面针对体外人红系白血病细胞(HEL细胞)增殖活性有抑制作用的粗生物碱富集方法,为两面针药材抗肿瘤活性研究提供参考。方法以不同浓度(60%、70%、80%、90%、100%)乙醇对两面针药材的干燥根进行提取,以最高提取率确定最佳提取条件;通过静态、动态吸附实验考察不同类型(HPD 100、XDA-5、D101、HPD 722、HPD 400)大孔树脂对两面针药材的吸附量确定上样量,富集两面针粗生物碱;采用噻唑蓝(MTT)法研究各组粗生物碱对HEL细胞增殖活性的影响。结果两面针药材经5种浓度乙醇提取,其中60%乙醇提取浸膏的得率最高[(11.937±0.002)%];静态、动态吸附实验表明,5种大孔树脂对两面针药材吸附量为1:500(两面针浸膏:大孔树脂);不同型号大孔树脂富集得到的粗生物碱对HEL细胞的增殖活性影响不同,两面针粗生物碱样品浓度为80μg·mL-1时,抑制率分别为(59.62±3.46)%(XDA-5)、(55.81±5.16)%(HPD 100)、(59.33±1.33)%(D101)、(79.63±1.02)%(HPD 722)、(83.97±0.38)%(HPD 400),HPD 400型大孔树脂富集的两面针粗生物碱抑制HEL细胞增殖作用最显著(P<0.01)。结论大孔树脂富集两面针粗生物碱工艺简单、可行,HPD 400型大孔树脂富集的两面针粗生物碱对HEL细胞抑制作用显著。

黄花棘豆总生物碱提取工艺及抗斜纹夜蛾活性研究

试验旨在优化黄花棘豆中总生物碱的提取工艺并进行抗斜纹夜蛾活性研究。在单因素试验基础之上,设计Box-Behnken响应面试验优化总生物碱的提取工艺,采用浸虫浸叶法探究总生物碱抗斜纹夜蛾活性。结果显示,最佳提取工艺条件为:乙醇浓度64.00%、提取温度81.00℃、提取2次、提取时间1.50 h、液料比11.00 mL·g^(-1),总生物碱提取率为1.79 mg·g^(-1)。此工艺条件下获取的总生物碱对3日龄斜纹夜蛾幼虫的LC_(50)值为42.10 mg·mL^(-1)。

无根藤中阿朴菲类生物碱的化学成分分析

为了寻找无根藤(Cassytha filiformis)中具有降糖活性的阿朴菲类生物碱,以无根藤为研究对象,通过酸溶碱沉法提取,乙酸乙酯萃取获取总生物碱,并通过多种色谱方法进行分离和纯化,再通过波谱学技术鉴定化合物的结构,最后采用葡萄糖氧化酶法评价所得阿朴菲类生物碱对HL-7702细胞葡萄糖消耗活性。鉴定结果发现13个阿朴菲类生物碱,分别鉴定为:无根藤米丁(1)、无根藤米里丁(2)、小唐松草宁碱(3)、新木姜子素(4)、N-metilseconeolitsine(5)、N-methyl-2,3,6-trimethoxymorphin-andien-7-one(6)、10-O-甲基汝兰碱(7)、无根藤定碱(8)、norneolitsine(9)、O-甲基无根藤碱(10)、无根藤碱(11)、filiformine(12)、降南天竹碱(13),其中,化合物5和13首次从该植物中分离得到。细胞葡萄糖消耗活性结果显示,化合物3和11显著促进HL-7702细胞葡萄糖消耗,具有研究开发成降糖先导化合物的潜在价值。

高效液相色谱法同时检测茶叶提取物中9种儿茶素类和3种生物碱类化合物

建立了以Kromasil 100-5-C_(18)为分析柱,甲醇与0.01%(体积分数)磷酸水为流动相梯度洗脱的高效液相色谱方法,同时定量检测茶叶提取物中的9种儿茶素类和3种生物碱类化合物。结果显示,在一定的质量浓度范围内,12种化合物的峰面积与质量浓度呈良好的线性关系,方法定量限为2.5~25 mg/L,回收率为94%~103%,相对标准偏差(RSD)≤2.7%。方法精密度好、准确度高,分析时间短,检测效率高,可以作为研究茶叶提取物的可靠方法,也可推广用于茶原料的产品质控。

响应面法优化甜石榴皮生物碱提取工艺及其抗氧化活性评价

以甜石榴皮为原料,采用超声波辅助法提取生物碱。以生物碱得率为指标,在单因素试验的基础上,通过响应面法优化甜石榴皮生物碱的提取工艺,并评价其抗氧化活性。结果表明:最佳提取工艺为乙醇体积分数32%、超声时间51 min、超声温度39℃、料液比1∶17(g/mL),在此条件下生物碱得率为2.35%。抗氧化试验表明,在一定质量浓度范围内,甜石榴皮生物碱对DPPH自由基和羟基自由基的清除率最高分别为82.32%、64.36%,IC50分别为0.368、0.590 mg/mL,具有较好的抗氧化活性。

超声微波协同提取药桑叶生物碱及其保肝活性研究

本研究以药桑叶为原料,采用单因素结合响应面法优化了药桑叶生物碱的超声微波协同提取工艺,并采用四氯化碳致小鼠急性肝损伤模型评价了药桑叶提取物(MNLE)的保肝活性。结果表明药桑叶生物碱的最佳提取工艺为:料液比1∶54 g/mL,乙醇浓度79%,微波功率300~0 W(0~7 min),超声功率493 W,提取时间29 min,提取温度70℃,在此条件下生物碱提取率可达11.325 mg/g。药效学试验结果显示,与模型组相比,MNLE能明显降低血清谷丙转氨酶(ALT)、谷草转氨酶(AST)水平以及肝组织丙二醛(MDA)和一氧化氮(NO)水平(P<0.05或P<0.01),升高肝组织超氧化物歧化酶(SOD)和谷胱甘肽(GSH)水平(P<0.05或P<0.01),肝组织病理学变化也得到了明显改善。结果表明MNLE具有较好的保肝作用,其作用机制与其抗氧化活性相关。

紫堇属植物苄基异喹啉类成分研究进展

全球紫堇属Corydalis约400余种,广泛分布于北温带和非洲南部地区。在我国,该属多种物种作为民间用药具有上千年历史。紫堇属植物富含异喹啉生物碱,对该类成分通常以回流、超声加热、湿法超微粉碎法和酶解辅助技术等方法提取,并使用溶剂分离法、硅胶柱色谱法、反向填料柱色谱法、HPLC法、离子交换树脂法、高效逆流色谱法和活性引导技术等方法进行分离纯化法。紫堇属生物碱有效部位或单体成分都具有较强药理活性,现代药理学表明,该属所含生物碱类成分对心脑血管、抗肿瘤、镇痛、消炎、保肝和抗血小板凝集等均具有显著的药理作用。本文对已报道的紫堇属植物生物碱类成分的结构类型、特征、药理作用以及提取分离方法研究进行综述,为植物药用化学成分富集和安全用药提供借鉴。

猪毛菜总生物碱提取工艺优化研究

以猪毛菜总生物碱提取率为指标,通过单因素试验和响应面试验优化猪毛菜总生物碱提取工艺。以DPPH和羟基自由基清除率确认猪毛菜总生物碱抗氧化能力。结果表明,猪毛菜总生物碱最优提取工艺为液料比36∶1(mL∶g)、超声提取时间60 min、复合酶含量2%、酶解时间1.4 h、乙醇体积分数57%,在最优条件下,猪毛菜总生物碱提取率可达0.6982%。抗氧化活性实验表明,随着猪毛菜生物碱浓度增加,抗氧活性随之增加,当猪毛菜生物碱质量浓度为0.5 mg/mL时,DPPH自由基和羟基自由基清除率分别达到84.34%和75.69%。本研究优化了猪毛菜总生物碱的提取工艺,初步分析猪毛菜生物碱抗氧化活性,为猪毛菜有效药用成分的分析研究和功能性食品的开发提供科学依据。

响应面法优化低共熔溶剂提取吴茱萸生物碱

通过响应面法优化超声法辅助低共熔溶剂提取吴茱萸中生物碱的提取工艺。制备多种低共熔溶剂,通过供氢体、受氢体以及醇类碳链长度和羟基数量的筛选,采用最佳低共熔溶剂体系对吴茱萸生物碱进行提取,探讨低共熔溶剂的摩尔比、含水量(体积分数)以及提取时间、提取温度和料液比对生物碱提取率的影响。结果表明:以乙酰丙酸和1,4-丁二醇为受氢体和供氢体制备低共熔溶剂,在摩尔比2:1、含水量30%、提取时间50 min、提取温度30℃、料液比1:40 g/mL超声提取条件下,吴茱萸生物碱提取率达2.52%,是传统有机溶剂乙醇提取的3.2倍,低共熔溶剂在提取生物碱成分方面具有一定发展潜力。

红花中总生物碱回流提取工艺优化

目的 筛选红花中总生物碱最优回流提取工艺。方法 以盐酸五羟色胺为对照品,红花中总生物碱含量为指标,采用单因素实验,考察了温度、时间、料液比和甲醇体积分数等对红花中总生物碱含量的影响。通过正交试验,对红花中总生物碱的提取工艺进行了优化,并对最优提取工艺进行了实验验证。结果 最佳工艺条件为温度65℃,液料比(v/w)50∶1,甲醇体积分数40%,提取时间60 min。在优化的工艺条件下,总生物碱平均含量为35.99 mg/g,RSD为0.26%(n=3)。结论 本研究所得最优提取工艺简单、易行,工艺参数可靠,提取效果良好,为红花中总生物碱进一步开发利用提供了实验依据。

超声-酸性乙醇协同提取荷叶生物碱的工艺优化

在单因素实验的基础上,通过正交实验考察了pH值、料液比、超声时间、乙醇体积分数对荷叶生物碱提取率的影响。确定超声-酸性乙醇协同提取荷叶生物碱的最佳工艺为:pH值2、料液比1∶18(g∶mL)、超声时间40 min、乙醇体积分数70%,在此条件下,荷叶生物碱提取率可达3.22%。该工艺具有操作简便、提取率高、耗时短等优点,可应用于荷叶生物碱的提取。