灵芝多糖的提取工艺研究

目的 确定灵芝多糖的最佳提取工艺.方法 以苯酚-硫酸法作为灵芝多糖的测定方法,以多糖提取量为考察指标,改进灵芝的提取工艺.对浸提过程中的温度、时间以及固液比3个因素进行单因素试验和正交试验.结果 灵芝多糖的最佳提取条件为浸提温度90℃,浸提时间2.0h,料液比1∶15;浸提温度对多糖提取率的影响最大,其次为浸提固液比和浸提时间.结论 该工艺可用于灵芝多糖的提取生产,为灵芝产品的开发提供了有益的参考.     

正交试验优选微乳超声提取灵芝中灵芝多糖工艺

目的 探讨优选微乳超声法提取灵芝中灵芝多糖工艺.方法 以超声功率、提取时间和药材与溶媒比作为考察因素,以灵芝多糖得率为考察指标,采用正交试验优选微乳超声提取灵芝中灵芝多糖工艺条件,并进行验证性试验.结果 最佳条件为超声功率500W、超声提取时间50min、药材与溶媒比1∶20时,是微乳超声提取灵芝中灵芝多糖工艺最佳条件.结论 应用微乳超声法提取灵芝中基础物质,可同时提取脂溶性和水溶性成分的特性,并具有省时环保和操作简便的优点,可为灵芝基础物质的研究提供科学依据.     

西藏灵芝多糖的提取工艺研究

目的优化西藏灵芝多糖的提取工艺。方法以蒽酮-硫酸法作为灵芝多糖含量的测定方法,以多糖提取得率为考察指标,对浸提过程中的温度、时间、次数以及料液比4个因素进行单因素试验和正交试验。结果灵芝多糖的最佳提取条件为提取温度100℃,浸提时间2 h,浸提次数3次,料液比为1∶30。结论该提取工艺条件稳定,重现性好,可为西藏灵芝多糖的大生产提供参考依据。     

BBD设计-效应面法优选灵芝多糖提取工艺

目的:优选灵芝多糖的提取工艺。方法:采用单因素试验考察料液比、浸提温度、浸提时间对灵芝多糖抗氧化活性的影响。以液料比、浸提温度、浸提时间为自变量,以灵芝多糖对1,1-二苯基-2-三硝基苯肼(DPPH)自由基的半数清除率的倒数(1/EC50)为因变量,通过对自变量各水平进行多元线性回归及二项式拟合,采用BBD设计-效应面法优选灵芝多糖提取工艺。结果:优选提取工艺为浸提温度63℃、浸提时间2 h、料液比1∶33(g/ml),在此条件下,灵芝多糖的1/EC50为0.732 2 ml/mg,与响应面拟合方程的预测值0.733 7ml/mg相当。结论:优选的提取工艺合理、可行,可用于灵芝多糖的提取。     

星点设计-效应面法优化鹿角灵芝微乳超声提取工艺

目的 探讨以微乳超声法同时提取鹿角灵芝中灵芝酸和灵芝多糖的最佳条件。方法 采用星点设计-效应面法,以物料溶剂比(X₁)、超声提取时间(X₂)和超声功率(X₃)作为考察因素,以灵芝酸A、灵芝酸C₂、灵芝多糖含量和提取物得率为考察指标,并对考察因素和考察指标总评归一值进行多元线性回归和二项式拟合,以效应面优选最佳的微乳超声提取工艺,并做验证试验。结果 微乳超声提取鹿角灵芝的最佳条件为物料溶剂比1∶18、超声提取时间59 min、超声功率513 W,此条件下灵芝酸A、灵芝酸C₂、灵芝多糖含量和提取物得率分别为0.647 8,0.108 5,11.20 mg·g^-1和34.28%。结论 微乳超声可同时提取鹿角灵芝中脂溶性和水溶性成分,并且节能、省时、操作简便,为工业化提取鹿角灵芝提供了新途径。     

用Folin-酚法测定灵芝产品中多糖肽含量

目的 建立灵芝和灵芝产品中多糖肽的含量测定方法.方法 采用紫外分光光度法,检测波长为750nm,用灵芝活性多糖肽GL-PP-T4为对照品,测定灵芝与灵芝产品中多糖肽的含量.结果 多糖肽GL-PP-T4在25μg·mL-1～250μg·mL-1的范围内呈现良好的线性,r为0.9994,平均回收率是99.45%,RSD为0.98%(n=5).且本法能排除制剂中淀粉添加剂的干扰.结论 本法稳定可靠、简便快速,可作为灵芝及其产品质量控制指标.     

灵芝孢子粉中多糖的制备工艺研究

目的：针对灵芝孢子粉多糖的制备工艺进行研究。方法：以多糖转移率为指标,比较布袋包煎和搅拌提取两种工艺;采用单因素法,考察搅拌提取中料液比和提取次数对多糖提取的影响;根据滤液的性状,比较静置取上清液、离心、布袋滤过和板框滤过等多种滤过方法,优化滤过工艺。结果：灵芝孢子粉多糖的最佳制备工艺条件为：按料液比1：30加水至提取罐中,煮至100℃,投料,保温搅拌提取2min,静置24min,取上清液,离心,滤液先通过（5μm滤袋）,再板框滤过,滤液浓缩至相对密度为1．10（60℃）,醇沉,取沉淀减压干燥,千膏得率为3．26％,多糖含量为25．1％,多糖转移率97％。结论：该工艺可行有效,具有实际生产意义。     

正交试验优选虫草多糖脂质体的制备工艺

目的 优选虫草多糖脂质体制备的最佳工艺.方法 采用逆向蒸发法制备发酵虫草菌粉多糖脂质体,以包封率为指标,以药脂比、卵胆比与水油比为因素,采用L9(34)正交试验设计对制备条件进行优选,鱼精蛋白测定发酵虫草菌粉多糖脂质体的包封率,透射电镜及马尔文激光粒度分析仪测定其形态和粒径.结果 逆向蒸发法制备发酵虫草菌粉多糖脂质体的最佳工艺为大豆卵磷脂与药物的比例为20:1,大豆卵磷脂与胆固醇的比例为2:1,乙醚与磷酸盐缓冲液的比例为3:1.结论 采用最佳工艺制备的发酵虫草菌粉多糖脂质体包封率较高,形态和粒径较均匀,重现性好.     

芝芪菌质提取物免疫活性研究

目的 探究芝芪菌质及其各部位免疫活性。 方法 芝芪菌质为灵芝菌丝-黄芪药渣的固体发酵复合体。通过小鼠碳粒廓清实验,鸡红细胞吞噬功能实验,小鼠血清溶血素、免疫球蛋白M(IgM)和免疫球蛋白G(IgG)的含量检测,以及小鼠脾淋巴细胞增殖实验,筛选了芝芪菌质水提液,芝芪菌质水浸液,芝芪菌质水提液经大孔树脂分离的部位(水洗脱液、40%乙醇洗脱液和95%乙醇洗脱液)的非特异性和特异性免疫活性。 结果 碳粒廓清实验中,K值最高为水浸液和水提液组的0.23,α值最高为水提液组的2.44。吞噬指数最高为水浸液和水提液组的0.36,鸡红细胞吞噬百分率最佳为水浸液组的47.25%。小鼠溶血素含量最高为水洗脱组的0.4。IgM含量最高为水提液组的34.19 mg·L-1,而IgG最高则是水浸液组的44.50 mg·L-1。淋巴细胞增殖指数最高为40%乙醇洗脱液组的1.23。 结论 芝芪菌质水提液、芝芪菌质水浸液和40%乙醇洗脱液具有显著的免疫增强功效,为芝芪菌质制成增强人体抵抗力的功能性食品及药品提供实验数据和理论参考。     

灵芝三萜成分分析

目的建立灵芝子实体、孢子粉、发酵菌丝体三萜成分的HPLC含量测定方法.方法以赤芝孢子酸A(ganosporeric acid A,1),赤芝酸A(lucidenic acid A,2),灵芝酸B(ganoderic acid B,3)和灵芝酸C(ganoderic acid C,4)为对照品;色谱柱:反相C18;流动相:乙腈-水(37∶63);流速:1.0 mL*min-1;检测波长:UV 254 nm.结果进样量在0.2～1 μg有良好的线性关系,(1),(2),(3)和(4)的加样回收率分别为100.9%,101.2%,101.3%和101.7%.结论本法快速、简便、灵敏和分离度好,适用于灵芝子实体、孢子粉、发酵菌丝体及相关制剂的三萜类物质检查和含量测定.     

三种灵芝不同部位的活性成分含量差异性分析

目的比较三种灵芝（霍芝I号、韩芝、日芝）不同药用部位的总多糖、总三萜、灵芝酸A及灵芝酸B的含量差异,为全面评价灵芝药材及孢子粉的质量及其药用成分的合理利用提供依据。方法采用苯酚一硫酸法测定总多糖含量;香草醛一冰乙酸一高氯酸显色法测定总三萜的含量;HPLC法测定灵芝酸A、B的含量。结果每种灵芝不同药用部位的总多糖含量由高到低依次为：孢子粉〉菌盖〉菌柄;每种灵芝不同药用部位的总三萜含量由高到低依次为：菌盖〉菌柄〉孢子粉;每种灵芝不同药用部位的灵芝酸A和灵芝酸B含量由高到低依次为：菌盖〉菌柄〉孢子粉,且灵芝酸A的含量均明显高于灵芝酸B。结论该研究为全面评价灵芝子实体及孢子粉的质量及其药用成分的合理利用提供依据。,     

灵芝发酵终点的确定及高产灵芝多糖菌种的筛选

目的确定灵芝发酵终点,筛选高产灵芝多糖的菌种,用于产业化生产。方法对六株来源于不同地区的灵芝菌种进行同条件发酵测其生物量、灵芝胞内多糖及pH值,革兰染色镜检菌丝。结果与讨论灵芝发酵终点应控制在pH值3.5~4.5,镜检菌丝出现少量自溶点,此时灵芝胞内多糖含量最高;BLGL32(福州)所产灵芝多糖最高,胞内多糖1.53g.L-1,胞外多糖7.02g.L-1。     

Sensitive and selective liquid chromatography-tandem mass spectrometry method for the determination of five ganoderic acids in Ganoderma lucidum and its related species

The present paper describes a novel, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous analysis of ganoderic acids C(2), B, A, H, D in Ganoderma lucidum and its related species. Ganoderma samples were prepared using simple ultrasonic extraction. Chromatographic separation was carried out on an Agilent Zorbax XDB C(18) column (250 mm × 4.6 mm i.d., 5μm) with an isocratic mobile phase consisting of acetonitrile, water and formic acid (42:58:0.5, v/v/v). Mass spectrometric detection was achieved by a triple-quadrupole mass spectrometer equipped with an atmospheric pressure chemical ionization (APCI) interface operating in negative and positive ionization mode via a single within-run polarity switching. Quantitation of five ganoderic acids was performed using selective reaction monitoring (SRM) mode. The intra- and inter-day precision was less than 6.2% and the accuracy ranged from 90.0% to 105.7%. The limit of quantification (LOQ) was 20.0-40.0 ng/mL and the limit of detection (LOD) was 3.0-25.0 ng/mL. With this method, low levels of ganoderic acids in the fruiting bodies of Ganoderma sinense and Ganoderma applanatum were accurately quantified for the first time. Importantly, the method allows unequivocal quantification of the five ganoderic acids in the spores and fruiting bodies of Ganoderma lucidum, whereas the previously published methods have lacked the capability. The method presented will be a powerful tool for quality control of Ganoderma lucidum and its related species.     

RP-HPLC和UV-VIS法测定灵芝不同收获期的多糖肽和灵芝酸

目的研究灵芝不同栽培方式和不同收获期中多糖肽和灵芝酸A、B的变化,为灵芝规范化栽培及制定产品质量标准提供依据。方法 RP-HPLC测定菌草灵芝和仿野生栽培的灵芝提取物的灵芝酸A、B的变化。色谱柱为XB-C18柱(150mm×4.6mm,5μm),流动相为0.05%磷酸-乙腈(65:35),检测波长254nm,常温,进样量50μL。以灵芝多糖肽为对照品,采用Folin-酚试剂法,通过紫外-可见吸收光谱法检测菌草灵芝和仿野生栽培灵芝不同收获期提取物的多糖肽。结果灵芝酸A、B线性范围分别为14.8～371.0和16.4～409.0μg/mL,平均回收率为95.0%、100.5%,RSD分别为1.18%、3.37%(n=6)。灵芝多糖肽在100～600μg/mL与吸光度值呈线性关系,r=0.9995,回收率95%～100%。结论该方法简便、快速,可用于灵芝及其产品中多糖肽和灵芝酸A、B的质量控制。菌草灵芝沸水提取能将多糖肽和灵芝酸分离,既简化了灵芝水、醇提取工艺,又提高经济效益。     

大鼠灌胃灵芝总三萜后血浆中灵芝酸类成分的检测

研究灵芝中总三萜在大鼠体内的吸收情况。大鼠灌胃给予灵芝总三萜后,利用HPLC-DAD和LC-MS方法鉴定大鼠血浆中的灵芝酸类成分。在大鼠血浆中检测到5种灵芝酸类成分,即灵芝酸C2、灵芝酸C6、灵芝酸G、灵芝酸B和灵芝酸A。大鼠口服灵芝总三萜后灵芝中的灵芝酸类成分能被大鼠吸收。     

A new lanostane-type triterpene from the fruiting bodies of Ganoderma lucidum

A new lanostane-type triterpene, named ganoderic acid LM2 (5), was isolated from the fruiting bodies of Ganoderma lucidum. Its structure was characterized as (23S) 7beta, -dihydroxy-3, 11, 15-trioxo-5alpha-lanosta-8, 24-dien-26-oic acid by 1D- and 2D-NMR spectra. In addition, a known triterpene, ganoderic acid epsilon (4), was obtained. Both of them exhibited potent enhancement of ConA-induced mice splenocytes proliferation in vitro.     

灵芝多糖的研究进展

灵芝多糖是灵芝的主要活性成分之一,具有抗肿瘤、抗氧化、抗衰老和降血糖等生物活性,对近年来研究灵芝多糖的超声法、微波法、复合酶法等提取方法和径向流色谱、柱色谱联用等分离纯化方法,含量测定及其在抗肿瘤和抗氧化等方面的药理活性进行综述,为灵芝多糖的进一步开发利用提供依据.     

Ganoderic acid Me inhibits tumor invasion through down-regulating matrix metalloproteinases 2/9 gene expression

The effect of ganoderic acid Me (GA-Me), which was purified from the fermentation mycelia of the traditional Chinese medicinal mushroom Ganoderma lucidum as reported (Tang W, Gu TY, Zhong JJ. Biochem Eng J. 2006;32:205-210), on anti-invasion was investigated. Wound healing assay indicated that GA-Me inhibited cell migration of 95-D, a human highly metastatic lung tumor cell line, in dose- and time-dependent manners. Results of cell aggregation and adhesion assays showed that GA-Me promoted cell homotypic aggregation and inhibited cell adherence to extracellular matrix (ECM). In addition, GA-Me suppressed matrix metalloproteinases 2/9 (MMP2/9) gene expressions at both mRNA and protein levels in 95-D cells according to qRT-PCR and Western blotting, respectively. The results demonstrated that GA-Me effectively inhibited tumor invasion, and it might act as a new MMP2/9 inhibitor for anti-metastasis treatment of carcinoma cells.     

A new lanostane-type triterpene from the fruiting bodies of Ganoderma lucidum

A new lanostane-type triterpene, named ganoderic acid LM 2 ( 5 ), was isolated from the fruiting bodies of Ganoderma lucidum . Its structure was characterized as (23S) 7 g , -dihydroxy-3, 11, 15-trioxo-5 f -lanosta-8, 24-dien-26-oic acid by 1D- and 2D-NMR spectra. In addition, a known triterpene, ganoderic acid l ( 4 ), was obtained. Both of them exhibited potent enhancement of ConA-induced mice splenocytes proliferation in vitro .