广西姜黄挥发油两种提取方法的比较研究

以广西姜黄那坡县种植的姜黄为原料,以水蒸汽蒸馏法和以石油醚为溶剂的索氏提取法,分别提取挥发油,并用GC-MS分析法对这些以不同加工途径获取的姜黄挥发油进行成分分析,比较了不同的加工办法对挥发油的加工得率、主要成分和含量的影响。研究表明:广西那坡的姜黄挥发油主要成分为:α-姜黄烯、芳姜黄酮、(-)-姜烯、β-倍半水芹烯、β-姜黄酮、α-姜黄酮、4-(1,5-二甲基-4-己烯基)-2-环己烯酮、β-没药烯。     

索氏法提取姜黄挥发油及其成分的研究

本文以广西那坡县种植的姜黄为原料,分别以沸点为30-60℃,60-90℃的石油醚以及石油醚与乙醚的混合物作为溶剂,采用索氏提取法提取挥发油,并采用GC-MS分析法研究了这几种姜黄挥发油的主要成分,比较了不同的加工办法对挥发油的加工得率、主要成分、含量的影响。研究表明：用索氏提取法提取的广西姜黄挥发油其得率在7％以上,主要成分为：α-姜黄烯、（-）-姜烯、β-倍半水芹烯、芳姜黄酮、β-姜黄酮、α-姜黄酮、β-没药烯。     

姜黄根茎中姜黄素类成分含量的产地差异及其与环境因子的CCA分析

采用HPLC法对产自四川崇州和犍为、广东四会、广西玉林和博白及金秀、云南马关的姜黄(Curcuma longaL.)根茎中姜黄素类成分含量进行测定,并利用典范对应分析方法(CCA)研究了不同产地姜黄根茎中姜黄素类成分含量与地理-气候因子及根际土壤养分因子间的相关性。结果表明:不同产地根际土壤中有机质、全N、全P和全K含量分别为14.03～32.79、0.39～0.92、0.56～1.55和2.29～9.23 g.kg-1,根际土壤养分含量差异较明显;姜黄多生长在中性偏酸、水肥性能良好的土壤中。姜黄根茎中姜黄素、去甲氧基姜黄素和双去甲氧基姜黄素含量及姜黄素类成分总含量的平均值分别为1.53%、0.42%、0.67%和2.61%;不同产地姜黄根茎中姜黄素类成分含量有显著差异,且同一产地采自不同采样点及不同采样时间的样品姜黄素类成分的含量也有一定差异。姜黄素类成分总含量以广西博白产姜黄根茎最高(4.29%)、广东四会产姜黄根茎最低(1.73%)。CCA分析结果表明:在经度、纬度、海拔、年均气温、极端最高温、极端最低温、年降水量、日照时数和无霜期等地理-气候因子中,年均气温和极端最低温与姜黄素类成分含量极显著正相关;而在pH值及有机质、全N、全P、全K含量等根际土壤养分因子中,有机质含量与姜黄素类成分含量极显著正相关。分析结果显示:影响姜黄根茎中姜黄素类成分含量的主要环境因子是年均气温、极端最低温和根际土壤的有机质含量。     

姜黄属花卉种质资源表型性状分析

依据姜黄属(Curcuma)花卉育种目标，对16份姜黄属植物的11个数量性状及7个质量性状进行因子和聚类分析评价。结果表明，姜黄属植物表型性状复杂多变，供试材料数量性状变异系数介于26.70%～95.44%，叶片主脉绿色，上苞片藕粉或玫红色，晕斑绿色，唇瓣紫色和旗瓣白色为姜黄属典型表型性状。主成分分析表明，9个重要观赏性状归于3个主成分中。根据表型性状各主成分进行聚类分析，供试16份种质被分为三大类群，第Ⅰ类群11个品种均为姜荷花(Curcuma alismatifolia)，其中‘清迈粉’的上苞片面积最大，观赏性最突出，但株型偏小。第Ⅱ类群4个品种，花型花色极具特色，株型更大，抗性强，以所罗门姜黄(C. soloensis)表现最优。第Ⅲ类群的女皇郁金(C. petiolata)生长势最为强健。在三个类群之间开展杂交育种有望获得兼具观赏性和生长势的后代。     

不同产地生姜主要活性成分的比较分析北大核心CSCD

为全面评价生姜质量,通过GC-MS和HPLC法分别对不同产地生姜样品中挥发油和姜辣素进行比较分析。结果从9种生姜挥发油中分别鉴定了24~28种化学成分,占挥发油总量的85.31%~93.67%,主要为β-水芹烯(9.96%~20.15%)、莰烯(3.54%~14.53%)、柠檬醛(6.52%~12.49%)、β-柠檬醛(3.96%~7.44%)、姜烯(13.81%~19.74%)、姜黄烯(tr^5.52%)、β-倍半水芹烯(5.08%~6.22%)和β-红没药烯(2.63%~3.34%)等单萜和倍半萜类活性成分。9种生姜样品中6-姜辣素含量在0.067%~0.391%之间,均达到了现行中国药典不低于0.05%的规定。可见不同产地生姜主要活性成分含量差异较大。故以生姜的主要特征性有效成分姜烯、姜黄烯与6-姜酚共同作为其质量评价指标,将更加科学合理。     

不同产地生姜主要活性成分的比较分析

为全面评价生姜质量,通过GC-MS和HPLC法分别对不同产地生姜样品中挥发油和姜辣素进行比较分析。结果从9种生姜挥发油中分别鉴定了24~28种化学成分,占挥发油总量的85.31%~93.67%,主要为β-水芹烯(9.96%~20.15%)、莰烯(3.54%~14.53%)、柠檬醛(6.52%~12.49%)、β-柠檬醛(3.96%~7.44%)、姜烯(13.81%~19.74%)、姜黄烯(tr~5.52%)、β-倍半水芹烯(5.08%~6.22%)和β-红没药烯(2.63%~3.34%)等单萜和倍半萜类活性成分。9种生姜样品中6-姜辣素含量在0.067%~0.391%之间,均达到了现行中国药典不低于0.05%的规定。可见不同产地生姜主要活性成分含量差异较大。故以生姜的主要特征性有效成分姜烯、姜黄烯与6-姜酚共同作为其质量评价指标,将更加科学合理。     

姜黄地上部分化学成分的研究

为了解姜黄(Curcuma longa L.)地上部分的化学成分,采用硅胶、葡聚糖凝胶柱色谱和高效液相色谱从姜黄地上部分分离得到14个化合物。通过波谱分析,分别鉴定为槲皮素3-O-α-L-鼠李糖苷(1)、山柰酚3-O-α-L-鼠李糖(1→2)-α-L-鼠李糖苷(2)、橙皮素7-O-α-L-鼠李糖(1→6)-β-D-葡萄糖苷(3)、1,7-二(4-羟基苯基)庚烷-4E,6E-二烯-3-酮(4)、1,7-二(4-羟基苯基)庚烷-1E,4E,6E-三烯-3-酮(5)、3-羟基-4-甲氧基肉桂酸(6)、对羟基苯甲醛(7)、香草醛(8)、4-羟基-3-甲氧基苯甲酸(9)、异香草酸(10)、4-(1-羟基-1-甲基乙基)苯甲酸(11)、R-6-羟基-6-甲基-3-(2-羟基异丙基)-2-烯环己酮(12)、6,9-二羟基-4,7-巨豆二烯-3-酮(13)和β-胡萝卜苷(14)。化合物1、2、3、12和13首次从该植物中分离得到。经HPLC比较分析,姜黄地上部分缺乏姜黄药材的主要功能成分姜黄素。     

固相萃取-HPLC法测定生姜不同品种和器官中姜黄素含量

用固定萃取-HPLC法研究了生姜不同品种、同一品种不同产地及不同器官中姜黄素含量.生姜干粉用4倍量75%乙醇提取2次,提取液过C18固相萃取柱,80%的乙醇洗脱,HPLC测定姜黄素含量.13个不同品种或产地的生姜中,山东、潮州和湖北产的山东大肉姜姜黄素含量分别为0.76、1.11和0.75 mg/100 g干重,广西白肉姜、云南黄姜、潮州南姜、安徽菜姜、四川姜、四川小黄姜、梅县水姜、清远火姜、广州疏轮大肉姜和有机栽培的广州大肉姜的姜黄素含量分别为1.5450、1.08、0.84、0.92、1.16、1.00、2.63、2.86、2.20和5.01 mg/100 g干重.有机栽培的广州大肉姜的姜肉、姜皮和地上部茎叶中姜黄素含量分别为4.49、1.2和0.41 mg/100 g干重.但生姜中姜黄素含量远低于贵州产姜黄的含量(2857 mg/100 g干重).结果表明,生姜中姜黄素含量主要取决于品种,栽培地理位置对其有较少的影响.有机栽培可大大提高姜黄素含量.     

姜黄标准汤剂UPLC指纹图谱的建立和4种成分定量分析研究CSCD

采用超高效液相色谱法(ultra-high performance liquid chromatography, UPLC)建立姜黄标准汤剂指纹图谱并同时测定4种有效成分的含量,为姜黄标准汤剂质量标准的制定提供依据。建立姜黄标准汤剂指纹图谱,采用质谱分析和对照品比对,对共有指纹峰进行指认,借助化学计量学方法对16批样品指纹图谱进一步处理和分析。同时,对姜黄标准汤剂中姜黄素、去甲氧基姜黄素、双去甲氧基姜黄素和阿魏酸的含量进行测定。结果显示,姜黄标准汤剂具有10个共有峰,指认出其中8个成分;聚类分析(hierarchical cluster analysis, HCA)和主成分分析(principal component analysis, PCA)将16批样品归为两类,正交偏最小二乘法-判别式分析(orthogonal partial least squares-discriminant analysis, OPLS-DA)共找到6个差异性标志物。含量测定结果显示,云南大理的姜黄原料制备的标准汤剂姜黄素、去甲氧基姜黄素、双去甲氧基姜黄素及阿魏酸的含量要高于四川宜宾和乐山的样品。该方法准确可靠,能够为姜黄标准汤剂的质量评价提供参考。     

发光二级管不同光质对姜黄光合特性和姜黄素含量的影响

以发光二级管(LED)为光源研究不同光质对姜黄生理特性的影响。结果表明:红光、蓝光和红蓝组合光处理均提高了姜黄素含量;红光有利于提高叶绿素a和总叶绿素含量、可溶性糖含量,蓝光有利于提高类胡萝卜素含量、SOD和POD活性、蛋白质含量,绿光有利于提高类胡萝卜素含量和蛋白质含量,黄光有利于提高蛋白质含量。红光/蓝光=7:3时姜黄的总叶绿素含量、类胡萝卜素含量、SOD活性、Fv/Fo和姜黄素含量分别比对照显著提高14.23%、23.60%、18.18%、22.60%和66.65%;说明红光/蓝光=7∶3时最适合于姜黄的生长和姜黄素的生成。     

Evaluation of antioxidant activities and antimutagenicity of turmeric oil: a byproduct from curcumin production

Curcumin removed turmeric oleoresin (CRTO) was extracted with hexane concentrated to get turmeric oil, and that was fractionated using silica gel column chromatography to obtain three fractions. These fractions were analyzed by GC and GC-MS. Turmeric oil contained aromatic turmerone (31.32%), turmerone (15.08%) and curlone (9.7%), whereas fractions III has aromatic turmerone (44.5%), curlone (19.22%) and turmerone (10.88%) as major compounds Also, oxygenated compounds (5,6,8-10) were enriched in fraction III. Turmeric oil and its fractions were tested for antioxidant activity using the beta-carotene-linoleate model system and the phosphomolybdenum method. The fraction III showed maximum antioxidant capacity. These fractions were also used to determine their protective effect against the mutagenicity of sodium azide by means of the Ames test. All the fractions and turmeric oil exhibited a markedly antimutagenicity but fraction III was the most effective. The antioxidant effects of turmeric oil and its fractions may provide an explanation for their antimutagenic action.     

Chemical composition of turmeric oil--a byproduct from turmeric oleoresin industry and its inhibitory activity against different fungi

Curcumin, the yellow coloring pigment of turmeric is produced industrially from turmeric oleoresin. The mother liquor after isolation of curcumin from oleoresin known as curcumin removed turmeric oleoresin (CRTO) was extracted three times with n-hexane at room temperature for 30 min to obtain turmeric oil. The turmeric oil was subjected to fractional distillation under vacuum to get two fractions. These fractions were tested for antifugal activity against Aspergillus flavus, A. parasiticus, Fusarium moniliforme and Penicillium digitatum by spore germination method. Fraction II was found to be more active. The chemical constituents of turmeric oil, fraction I and fraction II were determined by GC and identified by GC-MS. Aromatic turmerone, turmerone and curlone were major compounds present in fraction II along with other oxygenated compounds.     

In vitro induction, screening and detection of high essential oil yielding somaclones in turmeric (Curcuma longa L.)

Leaf of turmeric contains an essential oil used extensively in perfumery, pharmaceuticals and aromatherapy. Five somaclones were induced in turmeric on MS media with varying amounts of plant growth regulators. All somaclones were subsequently transferred to the field. Essential oil was extracted from leaves of in vitro and ex vitro grown plants and subjected to quantitative and qualitative evaluation. A positive correlation was established between the leaf oil content and oil constituent of in vitro grown and field transferred somaclones. Somaclones (C2, C4, C5) containing 0.16–0.18 % oil in vitro retained normal oil content (0.48–0.5 %) in the field. Similarly in vitro grown somaclones C3 and C7 with 0.36 and 0.25 % oil content retained proportionately increased oil yields of 1 % and 0.76 under ex vitro condition. GC–MS analysis of the oil revealed similar spectrum of constituents both among in vitro and ex vitro grown plants with alpha-phellandrene as major one. Thus the novel method of in vitro screening could be applied for rapid identification of high essential oil yielding turmeric genotypes thereby reducing labour, cost and time required in conventional ex vitro screening of somaclones.     

黔产莪术油的品质分析

该研究应用比重瓶法、折光率测定法、旋光度测法及电感耦合等离子体发射光谱法,分别对莪术油的相对密度、折光率、比旋度、重金属含量进行了测定,同时采用HPLC法测定莪术油中成分含量以及建立指纹图谱进行相似度的比较分析。结果表明:黔产莪术油的相对密度为0.987 0,比旋度为+24.146 8°,折光率为1.509,指纹图谱与中药制剂指纹图谱对照,相似度为0.976,总重金属含量总和10 mg·kg~(-1),砷盐未检测出。检测的各项指标均符合2015版中国药典莪术油项下的相关规定,该研究方法可行、数据可靠。该研究结果确认了黔产莪术油有较高的安全性及良好的品质,可作为药材原料使用,为黔产莪术油的综合开发利用及质量控制提供了科学依据。     

莪术油对Hela细胞迁移及侵袭的初步实验研究

目的:探讨莪术油对宫颈癌细胞系Hela细胞迁移和侵袭的影响。方法:宫颈癌细胞系Hela细胞培养后分实验组及对照组,实验组以125μg/m L浓度的莪术油作用48 h,对照组细胞不进行任何处置。细胞划痕实验检测莪术油对Hela细胞迁移的影响;Transwell小室实验检测莪术油对Hela细胞侵袭的影响。结果:0 h处理后实验组、对照组划痕宽度无统计学差异(P0.05);48 h后实验组划痕宽度明显大于对照组的划痕宽度(4.33±0.58 m vs 2.17±0.29 m,P0.05)。实验组48 h后的穿透细胞数明显少于对照组(26.2±1.3个vs 62.2±2.3个,P0.05)。结论:莪术油可抑制宫颈癌细胞的迁移与侵袭。     

Influence of solid state fermentation by Trichoderma spp. on solubility,phenolic content,antioxidant, and antimicrobial activities of commercial turmeric

The influence of solid state fermentation (SSF) by Trichoderma spp. on the solubility, total phenolic content, antioxidant, and antibacterial activities of turmeric was determined and compared with unfermented turmeric. The solubility of turmeric was monitored by increase in its phenolic content. The total phenolic content of turmeric extracted by 80% methanol and water after SSF by six species of Trichoderma spp. increased significantly from 2.5 to 11.3–23.3 and from 0.5 to 13.5–20.4 GAE/g DW, respectively. The antioxidant activities of fermented turmeric were enhanced using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS), and ferric ion-reducing antioxidant power (FRAP) assays. The antibacterial activity of fermented turmeric against human-pathogenic bacteria Escherichia coli, Streptococcus agalactiae, Staphylococcus aureus, Entreococcus faecalis, Methicillin-Resistant S. aureus, Klebsiella pneumonia, and Pseudomonas aeruginosae showed a broad spectrum inhibitory effect. In conclusion, the results indicated the potentials of using fermented turmeric as natural antioxidant and antimicrobial material for food applications.     

Bioactivities of the leaf essential oil of Curcuma longa (var. ch-66) on three species of stored-product beetles (Coleoptera)

Essential oil extracted from the leaves of turmeric, Curcuma longa L., was investigated for contact and fumigant toxicity and its effect on progeny production in three stored-product beetles, Rhyzopertha dominica F. (lesser grain borer), Sitophilus oryzae L. (rice weevil), and Tribolium castaneum Herbst (red flour beetle). Oviposition-deterrent and ovicidal actions of C. longa leaf oil were also evaluated against T. castaneum. The oil was insecticidal in both contact and fumigant toxicity assays. The adults of R. dominica were highly susceptible to contact action of C. longa leaf oil, with LD50 value of 36.71 microg/mg weight of insect, whereas in the fumigant assay, adults of S. oryzae were highly susceptible with LC50 value of 11.36 mg/liter air. Further, in T. castaneum, the C. longa oil reduced oviposition and egg hatching by 72 and 80%, respectively at the concentration of 5.2 mg/cm2. At the concentration of 40.5 mg/g food, the oil totally suppressed progeny production of all the three test insects. Nutritional indices indicate >81% antifeedant action of the oil against R. dominica, S. oryzae and T castaneum at the highest concentration tested.     

HPLC测定莪术油葡萄糖注射液中四种成分的含量

采用HPLC法测定了莪术油葡萄糖注射液中四种成分(莪术二酮、莪术醇、牻牛儿酮、呋喃二烯)含量。本法以十八烷基硅烷键合硅胶为填充剂,流动相为甲醇-水(85∶15 V/V),流速为0.8mL/min,柱温:30℃,检测波长为215nm。在该色谱条件下,分别建立莪术二酮、莪术醇、牻牛儿酮、呋喃二烯的线性回归方程。结果表明,该法的精密度、重复性、稳定性良好(RSD<2.0%),平均回收率均大于96%(RSD<3.0%,n=5),通过测定得到莪术油葡萄糖注射液中四种主分的含量分别为67,26,20,45μg/mL。HPLC法准确、可靠、重复性良好,可用于莪术油葡萄糖注射液的质量控制。     

Aromatic‐turmerone attenuates invasion and expression of MMP‐9 and COX‐2 through inhibition of NF‐κB activation in TPA‐induced breast cancer cells

Recent evidence suggests that breast cancer is one of the most common forms of malignancy in females, and metastasis from the primary cancer site is the main cause of death. Aromatic (ar)‐turmerone is present in Curcuma longa and is a common remedy and food. In the present study, we investigated the inhibitory effects of ar‐turmerone on expression and enzymatic activity levels of 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA)‐induced matrix metalloproteinase (MMP)‐9 and cyclooxygenaase‐2 (COX‐2) in breast cancer cells. Our data indicated that ar‐turmerone treatment significantly inhibited enzymatic activity and expression of MMP‐9 and COX‐2 at non‐cytotoxic concentrations. However, the expression of tissue inhibitor of metalloproteinase (TIMP)‐1, TIMP‐2, MMP‐2, and COX‐1 did not change upon ar‐turmerone treatment. We found that ar‐turmerone inhibited the activation of NF‐κB, whereas it did not affect AP‐1 activation. Moreover, The ChIP assay revealed that in vivo binding activities of NF‐κB to the MMP‐9 and COX‐2 promoter were significantly inhibited by ar‐turmerone. Our data showed that ar‐turmerone reduced the phosphorylation of PI3K/Akt and ERK1/2 signaling, whereas it did not affect phosphorylation of JNK or p38 MAPK. Thus, transfection of breast cancer cells with PI3K/Akt and ERK1/2 siRNAs significantly decreased TPA‐induced MMP‐9 and COX‐2 expression. These results suggest that ar‐turmerone suppressed the TPA‐induced up‐regulation of MMP‐9 and COX‐2 expression by blocking NF‐κB, PI3K/Akt, and ERK1/2 signaling in human breast cancer cells. Furthermore, ar‐turmerone significantly inhibited TPA‐induced invasion, migration, and colony formation in human breast cancer cells. J. Cell. Biochem. 113: 3653–3662, 2012. © 2012 Wiley Periodicals, Inc.     

Efficacy of bioactive compounds from Curcuma longa L. against mosquito larvae

To identify larvicidal compounds from the ethanolic extracts of Curcuma longa root, the active compounds were isolated using activity‐guided fractionation with column chromatography and identified based on nuclear magnetic resonance (NMR) and mass spectrometry (MS) data. The dipping method was used to determine the larvicidal activities of each compound against 4th‐instar larvae of Culex pipiens pallens. Two compounds were isolated and identified, ar‐turmerone and 8‐hydroxyl‐ar‐turmerone. The two compounds exhibited larvicidal activities against the 4th‐instar larvae of C. pipiens pallens after 24 hr of treatment with LC₅₀ values of 138.86 and 257.68 ppm, respectively. The larvicidal activities of ar‐turmerone and 8‐hydroxyl‐ar‐turmerone against C. pipiens pallens are reported herein for the first time. The elucidation of the structure of these phytochemicals and their insecticidal activities are important for assessing the potential of this plant as a botanical insecticide.