大孔吸附树脂法柚皮果胶脱色工艺研究

比较了AB-8、S-8、X-5、聚酰胺、NKA-Ⅱ、NPD-600等6种大孔吸附树脂对柚皮果胶的脱色效果.在静态吸附试验研究的基础上,筛选出效果较好的树脂进行动态试验研究.实验表明:AB-8型大孔吸附树脂对柚皮果胶提取液具有较好的吸附脱色效果和较低的果胶损失率,该树脂对柚皮果胶脱色的适宜工艺参数为:室温(约20℃),流速3BV/h,溶液处理量为5BV,上柱液pH值为4～6.此工艺对果胶提取液中色素的吸附率约80%.树脂吸附后可用70%乙醇进行洗涤再生,果胶提取液脱色后经喷雾干燥所得的果胶成品质量符合QB2484-2000标准.     

油樟叶多糖的大孔树脂纯化工艺研究

为研究大孔树脂纯化油樟叶多糖的最佳工艺,以脱色率、多糖保留率和蛋白质脱除率的加权综合评分为指标,首先通过静态及动态实验考察8种不同型号大孔树脂的纯化效果,优选出效果较好的树脂,再通过单因素和正交试验优化油樟叶多糖的纯化工艺参数。研究结果表明,AB-8型大孔树脂对油樟叶多糖有良好的综合纯化效果,其最佳纯化工艺参数为：油樟叶多糖的上样浓度为0.3mg/mL,洗脱剂为40%乙醇,洗脱剂流速为1.0mL/min,洗脱剂体积为2.2BV;在此工艺下纯化,油樟叶多糖保留率为86.35%,脱色率为73.03%,蛋白质去除率达81.18%。AB-8型大孔树脂对油樟叶多糖的纯化工艺稳定可靠、效果良好,为油樟叶多糖的开发利用提供了参考。     

响应面法优化大孔树脂对地参多糖脱色工艺及抗氧化活性分析

采用响应面法优化大孔树脂对地参多糖的脱色工艺,并分析其抗氧化性能。利用综合评分法,从5种大孔树脂中筛选出对地参多糖脱色较好的树脂,根据前期单因素试验结果,利用响应面法对多糖脱色率影响较大的NKA-9树脂脱色条件进行优化,依据回归分析确定其最佳脱色条件,同时测定地参多糖对羟基自由基(·OH)和超氧阴离子自由基(O_2~-·)的清除能力。结果表明,NKA-9树脂是地参多糖脱色的适宜树脂,多糖脱色的最佳工艺条件为:NKA-9树脂用量3.3wt%、脱色温度40℃、脱色时间88min。在此条件下,多糖保留率为96.20%,预测值为96.80%;多糖脱色率为66.54%,预测值为66.71%。地参多糖对·OH和O_2~-·有明显的清除能力,且随多糖质量浓度的增加而逐渐增强;当地参多糖的质量浓度为0.420mg/mL时,对·OH和O_2~-·的清除率分别为74.56%和69.98%。通过响应面试验优化的NKA-9树脂脱色工艺可对地参多糖进行有效脱色;地参多糖具有一定的体外抗氧化能力,可作为食品及医药行业的天然抗氧化剂资源进行有效开发利用。     

聚酰胺-大孔树脂纯化海棠花中总黄酮及其抗肿瘤活性研究

采用聚酰胺-大孔树脂联用技术纯化海棠花总黄酮成分,检测总黄酮对小鼠S-180肉瘤的抑制作用。选择提取液-聚酰胺(g/g,1∶1)拌匀,低温烘干,加入NKA-9大孔树脂柱,树脂-提取液(g/g,10∶2)上柱(柱径高比1∶10),以水洗脱至无色,再以60%乙醇洗脱,洗脱液收集量为10BV,总黄酮纯度达85.96%。海棠花总黄酮体外抗肿瘤活性显示,当剂量为50mg·kg-1时,抑瘤率达44.28%,并可显著提高小鼠的胸腺指数和脾脏指数。     

海地瓜多糖脱色工艺研究

比较了活性炭、H2O2及大孔树脂对海地瓜多糖的脱色效果。从脱色率及多糖保留率两方面进行考察,树脂D392处理优于活性炭和H2O2,对其脱色工艺进行优化,通过响应面实验建立了数学模型,并确定树脂D392脱色的最优工艺为时间:5h,pH 10,温度:60℃,此时,脱色率达(84.2±0.6)%,多糖保留率为(82.9±1.1)%。研究工作有利于海参多糖品质的提高,为海参多糖后续研究及应用奠定了基础。     

半枝莲多糖脱色及清除羟基自由基作用的研究

研究了半枝莲多糖的脱色的最佳条件及多糖体外清除羟基自由基的作用,结果表明,从活性炭、中性氧化铝、大孔吸附树脂AB-8、LSA-21和NKA-9等5种脱色剂中筛选出的最佳脱色剂为AB-8,AB-8脱色的最佳条件是pH7,温度40℃,溶液中多糖含量1046.5mg/L,此时多糖保留率为88.446%,脱色率为90.65%;体外清除羟基自由基作用结果表明,半枝莲粗多糖、SBP、SBP1随浓度的升高,其清除作用逐渐增强,SPB2几乎没有清除羟基自由基的能力.     

香菇多糖脱色工艺研究

研究了6种大孔吸附树脂和2种离子交换树脂对香菇多糖提取液中色素脱除的影响,在静态吸附试验研究的基础上,筛选出效果较好的树脂进行动态试验研究.研究发现,采用树脂DA201-C,在pH值4～6,温度40℃,流速3BV/h的工艺条件下,香菇多糖提取液中的色素脱除率达到87.8%,多糖保留率为85%,蛋白质的去除率为42%,树脂吸附后可先用1%NaOH和5%NaCl溶液浸泡,再用80%乙醇浸泡淋洗再生.     

超滤-吸附法制备高纯度L-辛弗林的优化工艺

以枳实提物液为原料,探讨了膜过滤和吸附树脂分离杂质,制备纯化L-辛弗林的新方法并获得纯度高达90%的冻干粉。先用不同截留分子量的超滤膜对枳实提取液进行超滤分级分离,脱除分子量差异大的杂质,再用筛选出的大孔吸附树脂吸附分离超滤透过液中的色素,确定其优化工艺条件,然后用反渗透浓缩及冷冻干燥,获得高纯度L-辛弗林冻干粉。超滤分级富集枳实粗提液中的L-辛弗林的优化条件为:以截留分子量为30kDa和5kDa的超滤膜顺序进行分级分离L-辛弗林浓度为4.5mg/mL的枳实提取液,压力为0.20MPa,获得的枳实超滤液中L-辛弗林透过率达89.7%,多糖、蛋白质和色素截留率分别为95.2%、93.8%和75.7%;D3520大孔吸附树脂脱除超滤透过液中色素的优化工艺条件为:上柱液L-辛弗林浓度为3mg/mL,上柱流速为1BV/h,上柱体积为3BV,获得的脱色液中色素仅残留3.82%,L-辛弗林损失率仅为6.54%;该脱色液经反渗透浓缩及冷冻干燥,制备出L-辛弗林冻干粉,其纯度为89.61%,总收率为77.34%。通过系列超滤膜顺序分级分离枳实提取液中分子量差异大的杂质及采用D3520大孔吸附树脂脱除色素的集成创新方法,制备出高纯度的L-辛弗林,丰富了天然产物分离纯化领域的应用理论和实证,并对分离枳实中L-辛弗林提供指导和参考。     

人参茎叶中人参皂甙的树脂脱色研究

选择了8种不同类型的脱色树脂,通过比较脱色液中色素脱色率的差异和脱色后人参皂甙纯度、收率的不同,筛选出较优的人参皂甙脱色树脂,并研究了其动态脱色实验和动态脱色曲线。实验结果表明,人参皂甙脱色母液经LX-TS4大孔强碱型阴离子交换树脂进行脱色,其脱色率达到85%以上,人参皂甙纯度达到80%以上。静态、动态的脱色实验表明,LX-TS4大孔强碱型阴离子交换树脂可作为人参茎叶中人参皂甙较为理想的脱色材料。     

蚕砂总生物碱提取物的脱色研究

通过动态吸附实验,考察了D296R(-Cl型)、D296R(-OH型)、D2040、X-5、D380(-Cl型)和D380(-OH型)6种树脂对蚕砂提取液的脱色作用,筛选出分离效果较好的树脂并确定了其分离条件,在此基础上使用正交设计法对它进行树脂再生条件的探究。通过对比这6种树脂对色素及活性物质的分离效果,发现D296-R(-Cl型)大孔阴离子交换树脂脱色效果最为理想,以95%乙醇溶液稀释的蚕砂提取液,经该树脂上样分离后,用3BV70%的乙醇溶液洗脱,所得洗脱液吸光度0.1,活性成分保留较多。正交设计法得出该树脂的适宜再生条件:乙醇浓度为80%,流速为3BV/h,NaOH溶液浓度为3%。     

Simultaneous qualitative and quantitative analyses of the major constituents in the rhizome of Ligusticum Chuanxiong using HPLC-DAD-MS

An HPLC-DAD-MS method was developed for the qualitative and quantitative analysis of the major constituents in Chuanxiong (the dried rhizome of Ligusticum chuanxiong Hort). Twenty compounds including phenolic constituents, alkylphthalides and phthalide dimers were identified using online ESI-MS and comparisons with literature data and standard compounds, and six of them were quantified by HPLC-DAD simultaneously. A comprehensive validation of the method including sensitivity, linearity, repeatability and recovery was conducted. The linear regressions were acquired with R(2) > 0.99 and limit of detection (LOD, S/N = 3) values were between 1.5 and 2.5 ng. The repeatability was evaluated by intra- and inter-day assays, and relative standard deviation (RSD) values were reported within 1.87%. The recovery studies for the quantified compounds were observed in the range of 96.36-102.37% with RSD values less than 2.63%. These phenolic constituents and alkylphthalides, the major constituents in Chuanxiong, are generally regarded as the index for the quality assessment of this herb. The overall procedure is accurate and reproducible, which is considered suitable for the qualitative and quantitative analyses of a large number of Chuanxiong samples.     

Exploratory analysis of chromatographic fingerprints to distinguish rhizoma Chuanxiong and rhizoma Ligustici

Identification and quality control of products of natural origin, used for preventive and therapeutical goals, is required by regulating authorities, as the World Health Organization. This study focuses on the identification and distinction of the rhizomes from two Chinese herbs, rhizoma Chuanxiong (from Ligusticum chuanxiong Hort.) and rhizoma Ligustici (from Ligusticum jeholense Nakai et Kitag), by chromatographic fingerprints. A second goal is using the fingerprints to assay ferulic acid, as its concentration provides an additional differentiation feature. Several extraction methods were tested, to obtain the highest number of peaks in the fingerprints. The best results were found using 76:19:5 (v/v/v) methanol/water/formic acid as solvent and extracting the pulverized material on a shaking bath for 15 min. Then fingerprint optimization was done. Most information about the herbs, i.e. the highest number of peaks, was observed on a Hypersil ODS column (250 mm × 4.6 mm ID, 5 μm), 1.0% acetic acid in the mobile phase and employing within 50 min linear gradient elution from 5:95 (v/v) to 95:5 (v/v) acetonitrile/water. The final fingerprints were able to distinguish rhizoma Chuanxiong and Ligustici, based on correlation coefficients combined with exploratory data analysis. The distinction was visualized using Principal Component Analysis, Projection Pursuit and Hierarchical Clustering Analysis techniques. Quantification of ferulic acid was possible in the fingerprints of both rhizomes. The time-different intermediate precisions of the fingerprints and of the ferulic acid quantification were shown to be acceptable.     

大孔吸附树脂对天麻素的吸附与分离特性的研究

研究了AB 8、NKA 9和S 83种大孔吸附树脂对中药天麻提取液中有效成分天麻素的吸附与分离特性。结果表明,NKA 9和S 8树脂对天麻素具有较好的吸附和解吸特性。其中经NKA-9树脂纯化的天麻素纯度为16.4%,比粗提物的天麻素纯度提高了1倍多。     

国产大孔吸附树脂浓集分离赤霉素

研究了７种国产大孔吸附树脂对赤霉素的吸附能力,其中Ｓ－８吸附效果较好,赤霉素固／液分配系数为１２．４４,动态吸附容量为４．０６ｍｇ／ｍｌ。８０％丙酮水地赤霉素的洗脱能力较强。用工业赤霉素发酵液进行吸附－解吸实验,分配系数可提高至２５．８８（ＰＨ＝２）,动态吸附容量可提高至１０．０２ｍｇ／ｍｌ,赤霉素收率可达９０％以上,且经吸附－解吸循环,可将发酵液中赤霉素浓缩７倍以上。     

Analysis of Chuanxiong Rhizoma substrate on production of ligustrazine in endophytic Bacillus subtilis by ultra high performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry

Ligustrazine was the active ingredient of the traditional Chinese medicine Chuanxiong Rhizoma. However, the content of ligustrazine is very low. We proposed a hypothesis that ligustrazine was produced by the mutual effects between endophytic Bacillus subtilis and the Ligusticum chuanxiong Hort. This study aimed to explore whether the endophytic B. subtilis LB5 could make use of Chuanxiong Rhizoma fermentation matrix to produce ligustrazine and clarify the mechanisms of action preliminarily. Ultra high performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry analysis showed the content of ligustrazine in Chuanxiong Rhizoma was below the detection limit (0.1 ng/mL), while B. subtilis LB5 produced ligustrazine at the yield of 1.0268 mg/mL in the Chuanxiong Rhizoma‐ammonium sulfate fermentation medium. In the fermented matrix, the reducing sugar had a significant reduction from 12.034 to 2.424 mg/mL, and rough protein content increased from 2.239 to 4.361 mg/mL. Acetoin, the biosynthetic precursor of ligustrazine, was generated in the Chuanxiong Rhizoma‐Ammonium sulfate (151.2 mg/mL) fermentation medium. This result showed that the endophytic bacteria B. subtilis LB5 metabolized Chuanxiong Rhizoma via secreted protein to consume the sugar in Chuanxiong Rhizoma to produce a considerable amount of ligustrazine. Collectively, our preliminary research suggested that ligustrazine was the interaction product of endophyte, but not the secondary metabolite of Chuanxiong Rhizoma itself.     

大孔吸附树脂分离纯化金银花中黄酮类物质的研究

比较了AB-8、S-8、NKA-9和D-101 4种大孔吸附树脂对金银花提取液中黄酮类物质的吸附及解吸附性能.在静态吸附试验基础上,筛选出效果较好的D-101树脂进行动态试验研究,结果表明,D-101树脂在30℃下对金银花黄酮类物质的静态吸附-动态解吸较优的工艺参数为:上样液pH值2.46,解吸液为95%乙醇,解吸液的流速为3mL/min,pH值11,4.5BV解吸液即可完全洗脱被树脂吸附的黄酮类物质,其解吸率高达98.00%.在试验研究范围内,树脂吸附金银花黄酮是自发性放热过程,并且符合Langmuir方程,此外树脂对黄酮的吸附动力学可用Pseudo-second-order模型较好地拟合,其表观吸附速率常数为Kso℃=3.43×10-2g/(mg·min).     

A型吸附树脂分离纯化杜仲中京尼平甙酸

比较了NKA-9、D311、S-8、HPD600、NKA-2、A型、D140和聚酰胺等8种树脂对杜仲中降血压活性成分京尼平甙酸的吸附及脱附性能,从中筛选出吸附率及脱附率均较高的A型树脂进行试验.确定了最佳工艺条件:杜仲皮粉末用50%乙醇水溶液提取后,95%乙醇沉淀,上清液回收乙醇后,用蒸馏水稀释,调节pH=6～9后用A型树脂吸附,15%乙醇溶液洗脱,流速为1.5ml/min,洗脱液浓缩后冷冻干燥得产品.     

大孔树脂精制6种含黄连复方水提液体系的吸附动力学比较研究

研究含黄连的不同复方水提液体系经大孔吸附树脂精制后,黄连中主要指标性成分生物碱的变化,阐明不同复方组合-配伍对药方中组成药味指标性成分的影响,为大孔树脂精制技术在中药复方中的应用提供实验依据.选用黄连用量和复方总用量均相同,组方药味不同的复方1～6.通过考察5种树脂AB-8、S-8、X-5、NKA-II、NKA-9对黄连中盐酸小檗碱的静态吸附率,确定AB-8树脂作为复方1～6吸附动力学比较研究的树脂;通过比较复方1～6水提液体系在相同树脂环境下的静态吸附过程,考察6种不同复方组成药味对复方中指标性成分分离纯化的影响.实验结果表明,由于体系本身溶液环境的差异,复方1～6各指标性成分达到吸附平衡的时问均有差异.AB～8树脂对复方1～6水提液体系的静态吸附在开始阶段吸附量的增加比较明显,在40min后,由于吸附竞争点饱和的原因,吸附量的增加均比较平稳;黄连与不同药材配伍时,虽然配伍药材所含的化合物类型差异较大,但黄连中生物碱类成分均可以在树脂上被选择性吸附,可知在复方水提液复杂体系中可以选择性地分离纯化生物碱类有效成分.     

大孔吸附树脂富集板蓝根中(R,S)-告依春工艺研究

采用静态吸附法考察了D101、AB-8、NKA-2、NKA-9、HPD 100、HPD600等6种大孔吸附树脂对(R,S)-告依春的吸附及解吸性能,筛选出效果最佳的AB-8树脂,并对其进行动态考察.最佳富集条件为:上样液pH 6,生药质量-体积浓度为0.200g/mL,解吸液为2BV量70％乙醇,在优化条件下(R,S)-告依春在浸膏中含量可从0.76％提高到12.48％.结果表明,AB-8型大孔吸附树脂可用来从板蓝根水提取液中富集(R,S)-告依春.