基于近红外漫反射光谱快速测定淫羊藿蛋白质含量

采用近红外漫反射光谱技术对淫羊藿(Epimedium)的蛋白质含量进行快速且无损检测。近红外漫反射光谱经二阶导数处理、标准多元离散校正及主成分分析聚类处理后, 采用改进最小二乘法回归得到的定标模型预测效果最佳, 定标决定系数、交互验证标准差及交互验证相关系数分别为0.923、0.554和0.717。近红外光谱分析法的测定结果与用凯氏定氮法所得结果无显著差异, 两种方法测定值的相关性较高(R²=0.933 9)。重复性实验表明, 近红外光谱分析法的相对标准偏差为0.937%。该研究首次采用近红外光谱分析法测定了8种淫羊藿的蛋白质含量。该方法简便、精确, 在淫羊藿资源开发利用和药材质量控制方面具有参考意义。     

近红外漫反射光谱法快速测定药用植物淫羊藿总黄酮含量

利用一种基于近红外漫反射光谱技术的淫羊藿总黄酮快速定量分析方法, 测定了淫羊藿属(Epimedium)7个种中总黄酮的含量。研究结果表明, 近红外光谱经二阶导数处理、主成分分析聚类及加权多元离散校正处理后, 采用改进最小二乘法回归得到的定标模型的预测效果最佳; 定标样品集预测方程的定标决定系数、交互验证标准差及交互验证相关系数分别为0.985 5、0.023 0和0.044 3; 检验样品集方程的定标决定系数、检验标准差及偏差分别为0.965 1、0.018和0.006, 达到了快速测定淫羊藿总黄酮含量的要求。该方法为淫羊藿总黄酮的快速定量分析提供了新的依据。     

栽培灵芝不同生长阶段多糖和 三萜类化合物的含量测定

将灵芝（Ganoderma lucidum）生长过程分为6个阶段,分别对其不同体积分数乙醇沉淀的多糖和三萜类化合物的含量进行了测定.结果表明:灵芝总多糖、40％和80％体积分数乙醇沉淀多耱含量均在菌蕾期最高;65％乙醇沉淀多耱含量以子实体成熟期最高;三萜类化合物的含量在菌盖分化期最高.     

灵芝多糖的研究进展

灵芝古称瑞草,在我国作为药物有2000多年的历史。目前,已知灵芝属约有100多种,分布最广的为赤芝（Ganodermelucidum）,其次为紧芝（Ga,;oderl;lcisinensis）。经过大量临床和药理分析表明,灵芝对慢性支气管炎、冠心病、心绞痛、慢性肝炎、神经衰弱、心悸头晕等均有不同程度的疗效。目前已知灵芝属的一些种具有明显的抗癌活性和抗衰老活性［’－‘］。l灵芝多糖的发现和结构组成目前,已分离到200多种灵芝多糖,其中大部分为卜型的葡聚糖,大多存在于灵芝细胞壁内壁。液体培养的发酵液和固体培养的培养基中有灵芝菌丝体分泌的胞外多…     

灵芝多糖的生物活性及开发利用

灵芝为最新纳入“药食同源”目录的名贵中药材,具有很高的营养价值。灵芝多糖是灵芝中最主要的成分之一,具有抗氧化、免疫调节、降血糖、抗肿瘤、调节肠道菌群等多种生物活性,在保健食品和药用资源开发方面具有很大的利用价值。本文对近年来灵芝多糖生物活性的研究进展进行总结,梳理了其开发应用的现状,对后续的开发和利用进行了展望,以期为灵芝多糖相关健康产品的开发利用提供参考和借鉴。     

灵芝多糖发酵工艺条件的研究

通过对灵芝菌As5.504发酵,研究不同糖源、氮源、生长因子及接种量、装液量对灵芝细胞生长和灵芝多糖生产的影响。结果表明当以葡萄糖,蛋白胨,VB5分别为碳源、氮源、生长因子及接种量、装液量分别为7.5mL、150mL时,为As5.504产多糖最佳发酵工艺条件,产多糖量最高为0.940g·L-1。     

HPLC测定灵芝子实体水提物及相关产品中三萜的含量

灵芝药品大多以灵芝子实体水提物为原料,为快速准确测定灵芝子实体水提物及相关产品中三萜的含量,建立了具有较好分离效果的HPLC分析测定方法。通过优化色谱柱和洗脱条件,优选出Agilent Zorbax SB-Aq C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-乙酸水溶液(0.01%)为流动相梯度洗脱,流速为1.0 mL/min,检测波长252 nm,柱温30℃,该条件下灵芝酸A、灵芝酸F等10种灵芝酸得到较好的分离。方法学考察显示该分析方法精密度、重复性、稳定性和加样回收率的RSD值均小于5%,可以用于灵芝酸C2、灵芝酸G、灵芝烯酸B、灵芝酸B等10种灵芝酸的定量检测。通过对灵芝子实体原料、水提物和市售灵芝产品中10种三萜类成分分析发现,灵芝子实体水提物中均含有这10种三萜,含量为2.52%–6.83%,较子实体原料大幅提高,市售的灵芝产品中的三萜含量为0.27%–0.84%。该方法的建立为灵芝水提物及其产品质量标准的建立奠定基础。     

灵芝子实体多糖对小鼠急性酒精肝损伤预防的代谢组分析

灵芝多糖是灵芝的主要药理活性成分之一。本研究通过检测血清指标分析灵芝子实体多糖（Ganoderma lingzhi fruitbody polysaccharides,GLFPS）对小鼠急性酒精性肝损伤的预防作用并结合代谢组学探究作用机制。结果显示,GLFPS显著抑制因酒精作用而升高的小鼠血清中ALT、AST、TG、TC和ADH水平。通过代谢组分析,在模型组与对照组中得到85个差异代谢物,其中三磷酸腺苷（adenosine triphosphate）、L-天门冬氨酸（L-aspartic acid）以及赖氨酸（L-lysine）等在相互作用网络中起重要作用,说明酒精能引起小鼠肝脏腺苷和氨基酸代谢的改变。GLFPS组与模型组有58个差异代谢物,主要包括脂质和有机氧化物,说明GLFPS可以通过调节小鼠肝脏中脂质与有机化合物代谢来预防急性酒精肝损伤。对差异代谢物进行KEGG富集分析发现主要涉及胆碱代谢、甘油磷脂代谢和ABC转运蛋白。GLFPS能够有效缓解这3个代谢通路中因酒精作用发生明显改变的代谢。综上可见,灵芝子实体多糖能够通过调节小鼠胆碱代谢、甘油磷脂代谢以及部分ABC转运蛋白有效预防酒精性肝损伤。     

拱状灵芝多糖的分离与鉴定

由拱状灵芝Ganoderma Fornicolum提取的水溶性多糖,经甲醇分级与DEAE-纤维素柱层析获拱状灵芝葡聚糖,平均分子量为50,000。该葡聚糖经酶试验、高碘酸氧化、红外光谱、Smith降解,乙酰化产物质谱分析,证明为多枝结构。由β(1→6)(1→2)-D-葡萄糖组成。     

Process analytical technology case study,part III: Calibration monitoring and transfer

This is the third of a series of articles detailing the development of near-infrared spectroscopy methods for solid dosage form analysis. Experiments were conducted at the Duquesne University Center for Pharmaceutical Technology to develop a system for continuous calibration monitoring and formulate an appropriate strategy for calibration transfer. Indcators of high-flux noise (noise factor level) and wave-length uncertainty were developed. These measurements, in combination with Hotelling’s T² and Q residual, are used to continuously monitor instrument performance and model relevance. Four calibration transfer techniques were compared. Three established techniques, finite impulse response filtering, generalized least squares weighting, and piecewise direct standardization were evaluated. A fourth technique, baseline subtraction, was the most effective for calibration transfer. Using as few as 15 transfer samples, predictive capability of the analytical method was maintained across multiple instruments and major instrument maintenance.     

吗啡的精神神经免疫学作用及灵芝多糖肽对吗啡依赖小鼠的免疫保护效应

通过多种急慢性吗啡给药动物模型,从整体,细胞和分子水平,系统研究了吗啡的精神神经免疫学作用及灵芝多糖肽对吗啡依赖小鼠的免疫保护效应,首次发现：反复吗啡处理小鼠脾细胞内原癌基因ｃ－ｍｙｂ和ｃ－ｍｙｃ的表达比正常动物降低;ＧＰＰ可以明显恢复吗啡处置小鼠降低的各项免疫学实验指标达到甚至超出对照水平,从而为应用ＧＰＰ等免疫反应修饰剂控制阿片滥用所致的免疫功能缺陷提供了动物实验依据。     

A novel method for analyzing thick tablets by near infrared spectroscopy

A near-infrared (NIR) spectroscopic method to determine content uniformily of a large, thick tablet using an approach that could facilitate future validations has been developed. A CT ibuprofen 800-mg tablet weighs about 1150 mg and is about 18.6 mm wide and 7.6 mm thick. The FT NIR spectrometer was optimized for transmission spectra of the tablets by moving it to the sample compartment and placing it immediately behind the tablet. In spite of this dedicated setup, the transmission spectra obtained were very poor, indicating that the NIR radiation was not reaching the detector. The spectra of the tablet improved with use of a simple preparation in which a flat-face die applies pressure of 20 000 psi to the tablet, this reduced the thickness of the tablet from 7.6 mm to 3.6 mm. A calibration model was developed for tablets with drug content ranging from 70% to 130% of label. The calibration model was tested using a validation set of tablets with a drug content of 752, 800, and 848 mg. The results obtained were within 1.5% of the known drug content of the validation set, tablets. Even with the sample preparation, the content uniformity results of 10 tablets could be determined using this method in less than 1 hour. The approach described in this article could also be used to validate NIR content uniformity methods for orther formulations. Published: July 12, 2001.     

Near‐infrared spectroscopic evaluation of lyophilized viral vaccine formulations

This article examines the applicability of near‐infrared spectroscopy (NIRS) to evaluate the virus state in a freeze‐dried live, attenuated vaccine formulation. Therefore, this formulation was freeze‐dried using different virus volumes and after applying different pre‐freeze‐drying virus treatments (resulting in different virus states): (i) as used in the commercial formulation; (ii) without antigen (placebo); (iii) concentrated via a centrifugal filter device; and (iv) stressed by 96 h exposure to room temperature. Each freeze‐dried product was measured directly after freeze‐drying with NIR spectroscopy and the spectra were analyzed using principal component analysis (PCA). Herewith, two NIR spectral regions were evaluated: (i) the 7300–4000 cm^?1 region containing the amide A/II band which might reflect information on the coated proteins of freeze‐dried live, attenuated viruses; and (ii) the C–H vibration overtone regions (10,000–7500 and 6340–5500 cm^?1) which might supply information on the lipid layer surrounding the freeze‐dried live, attenuated viruses. The different pre‐freeze‐drying treated live, attenuated virus formulations (different virus states and virus volumes) resulted in different clusters in the scores plots resulting from the PCA of the collected NIR spectra. Secondly, partial least squares discriminant analysis models (PLS‐DA) were developed and evaluated, allowing classification of the freeze‐dried formulations according to virus pretreatment. The results of this study suggest the applicability of NIR spectroscopy for evaluating live, attenuated vaccine formulations with respect to their virus pretreatment and virus volume. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1573–1586, 2013     

灵芝多糖对小鼠细胞免疫功能调节作用的实验研究

研究灵芝多糖 (GLB7)对小鼠细胞免疫功能调节作用的影响。实验小鼠分成 4组 ,分别经胃灌注不同剂量 (高、中、低 )灵芝多糖 ,每天 1次 ,连续 14d ,对照组小鼠用以等量蒸馏水代替。分别于实验第 15 ,2 8d进行小鼠细胞免疫功能调节作用的测试。灵芝多糖喂药后 2周 ,3个剂量组小鼠脾淋巴细胞转化试验、小鼠腹腔巨噬细胞对鸡血红细胞的吞噬百分率和吞噬指数 ;低剂量组小鼠迟发型变态反应 (DTH)、小鼠碳廓清 ;高剂量组小鼠NK细胞活性和对照组比较均有显著性差异 (P <0 .0 5 )。喂药后 4周 :上述实验结果和对照组比较均无显著性差异 (P >0 .0 5 )。结果提示灵芝多糖能提高小鼠的非特异性和特异性细胞免疫功能。     

灵芝多糖对荷瘤小鼠肿瘤免疫系统的影响

目的:观察灵芝多糖对荷瘤小鼠免疫系统的影响.方法:纯系BALB/c小鼠30只,用U14瘤细胞荷瘤建立动物模型,随机平均分为三组:灵芝多糖治疗组、环磷酰胺治疗组、生理盐水对照组.观察灵芝多糖对荷瘤小鼠NK细胞活性、淋巴细胞转化率、TNF-α等免疫指标的影响.结果:灵芝多糖能够显著提高治疗组小鼠的NK细胞活性、淋巴细胞转化率和血清中TNF-α、IL-2的含量.结论:灵芝多糖能够显著提荷瘤小鼠免疫系统的活性.     

Dry matter production and boron concentrations of vegetative and reproductive tissues of canola and sunflower plants grown in nutrient solution

Prediction of nitrogen (N) mineralization is important for specifying the optimum rate of N fertilizer for flooded rice at the time of sowing. To develop a predictive test, soils (0–0.1 m) were sampled from 22 farms throughout the rice-growing region of southern Australia over a 4-year period. Near infrared reflectance (NIR) spectra of the soils were compared with sixteen biological and chemical soil tests for the prediction of N-uptake by rice plants from these soils in the field and glasshouse. The aim of the study was to develop a soil-NIR calibration as an accurate, rapid and economical mineralization test. Nitrogen uptake by field-grown and glasshouse-grown plants was poorly correlated (r = 0.30), even though significant NIR calibrations were developed with both. Since N uptake by rice in the field was affected by varying weather and management, the field calibration is probably spurious. The calibration of soil NIR spectra with N uptake by glasshouse plants was satisfactory, with a standard error (SE) of 13 kg ha^–1 over a range of 11 – 95 kg ha^–1, and a correlation between calculated and measured N uptake (r = 0.87, P<0.001). An even better soil-NIR calibration was found with N-mineralization after 21 days of anaerobic incubation (SE 16 mg kg^–1, range 52–175 mg kg^–1). Analysis of the soil spectra showed that similar wavelengths were correlated with both plant-N uptake and mineralization. NIR spectroscopy shows considerable potential to predict soil N mineralization, and may assist future fertiliser decision support.     

Effects of oven- or freeze-drying on chemical composition and NIR spectra of pasture silage

In order to assess effects of drying method on chemical composition and NIR spectra of pasture silage, 20 samples of silages from permanent pastures of different qualities were subsampled and dried either by forced draught oven at 65°C or freeze-dried. Samples were analysed for crude protein (CP), crude fibre (CF), neutral detergent fibre (NDF), acid detergent fibre (ADF), gross energy (GE) and in vitro digestible organic matter in the dry matter (DOMD) and composition expressed on a dry-matter (DM) basis, either obtained as oven 105°C DM (DM_105°) or toluene DM (DM_tol). Effects of the drying method on chemical composition were estimated by paired comparisons. Near-infrared reflectance (NIR) spectra were taken and the difference spectra between treatments for each sample were plotted in order to visually inspect effects of treatments, either as log 1/R or transformed by their first derivative. Principal components explaining spectral variability were computed and samples graphically displayed according to the eigenvalues of the first (1 and 2), or second (2 and 3) pairs of principal components. Oven-drying resulted in a reduction in CP (p<0.02) content and an increase in CF (p<0.1), NDF (p<0.001) and ADF (p<0.06) content of silages, when expressed on a DM_105° basis. When expressed on a DM_tol basis, a reduction in CP (p<0.02) and DOMD (p<0.06) content was observed with oven drying, but fibre fractions were not affected (p>0.1). Effects of the drying treatment were visually apparent on NIR spectra when plotted as log 1/R against wavelengths, mainly as a baseline shift. Difference spectra (OD − FD), both as log 1/R and its first derivative, showed consistent absorption bands, indicative of different molecular responses of OD and FD silages to incident light. Plotting samples according to eigenvalues of the first two or second two principal components suggests that drying methods affect the distribution of samples. This is also indicative of an effect of drying treatment on spectral features and reinforces the general advice that sample processing should be consistent in order to avoid adding errors to NIR analysis.     

灵芝多糖对AD模型大鼠海马组织Caspase-3和FasL表达的影响

目的研究灵芝多糖（GLP）对阿尔茨海默病（AD）大鼠海马组织半胱氨酰天冬氨酸特异性蛋白静3（caspase-3）和FasL基因以及空间学习记忆能力的影响。方法双侧海马内一次性注射淀粉样多肽25-35片段（Aβ25—35）制作大鼠AD模型,治疗组24h后腹腔注射灵芝多糖水溶液,1周后进行Morris水迷宫检测大鼠空间学习记忆能力变化。采用免疫组织化学法和逆转录-聚合酶链式反应法（RT-PCR）分别检测大鼠海马组织caspase-3蛋白的表达量和FasL基因的表达,以及灵芝多糖对上述各指标的影响。结果灵芝多糖能明显改善AD模型大鼠低下的空间学习记忆能力。显著降低模型大鼠海马组织caspase-3和FasL的表达,而且灵芝多糖能明显改善模型大鼠脑组织海马CAI区神经元的退行性变化。结论灵芝多糖能降低海马组织内FasL和caspase-3表达量,改善海马CAI区神经元的退行性变化。对老年性痴呆大鼠学习记忆能力可能有增强和提高作用。