Separation and Purification of Sinigrin and Gluconapin from Defatted Indian Mustard Seed Meals by Macroporous Anion Exchange Resin and Medium Pressure Liquid Chromatography

Defatted seed meals, the byproducts of Indian mustard seed oil industry, which are just used as animal feedstock and nitrogen fertilizers, possess various nutrients and phytochemical compounds, such as sinigrin and gluconapin. The present study demonstrated a novel, low-cost, and efficient method for the co-production of sinigrin and gluconapin from defatted Indian mustard seed meals by D261 strong basic anion-exchange macroporous resin and medium pressure liquid chromatography (MPLC). The adsorption/desorption capacities of eight adsorbents characterized by BET, IR, and EDS were screened. 26.4 g defatted Indian mustard seed meals, which contained about 1210.3 mg sinigrin and 696.6 mg gluconapin, could produce 2059.6 mg of glucosinolate-rich extract of 53.67% sinigrin and 31.30% gluconapin after separation by D261 resin. Then, 949.40 mg of 97% sinigrin and 568.40 mg of 95% gluconapin could be obtained after the extracts were purified by MPLC, with the recovery of 76.03% of sinigrin and 77.38% of gluconapin. The products were assessed by analytical HPLC and characterized by UV, MS, ¹H NMR, and ¹³C NMR. In conclusion, this method is practical and environmentally friendly, and it is a low-cost process to make full use of Indian mustard seeds.     

Preparative Separation of Glabridin from Glycyrrhiza glabra L. Extracts with Macroporous Resins

Abstract

In the present study, the performance and adsorption characteristics of five macroporous resins for the separation of glabridin from Glycyrrhiza glabra L. have been evaluated. The adsorption and desorption properties of glabridin on macroporous resins including HPD100, HPD300, HPD800, NKA and H103 were compared. HPD100 resin offered the best adsorption and desorption capacities based on the research results. Both Langmuir and Freundlich isotherms were used to describe the interactions between the solutes and resins at different initial concentrations. Dynamic adsorption and desorption experiments on HPD100 resin packed column were conducted to optimize the separation process of glabridin from licorice extracts. After the treatment with stepwise elution on HPD100 resin, the content of glabridin in the product increased from 0.21% to 32.2% which is 153-fold higher than it in G. glabra L. roots and the recovery yield was 79.7%. The results indicated the good ability of HPD100 resin for separation glabridin and the study may provide scientific references for the large-scale glabridin production from G. glabra L. or other plants extracts.     

Preparation of high purity squalene from soybean oil deodorizer distillate with the combination of macroporous resin and thin-film evaporation coupling distillation

ABSTRACT

In this study, the combination of macroporous adsorbent resins (MARs) and thin-film evaporation coupling distillation (TFECD) was studied systematically, with aim to obtain high value-added squalene from soybean oil deodorizer distillate (SODD). Five types of resins (X-5, D-101, D4020, DM301, and AB-8) were first used to evaluate the adsorption/desorption properties of squalene. D101 resin exhibited higher adsorption/desorption capacities and desorption ratios for squalene based on static adsorption results among the tested resins. We further investigated the adsorption kinetics, isotherms, and thermodynamics with D101 resin as adsorbent. The adsorption of squalene on D101 was best fitted to pseudo-second-order kinetic model and the Langmuir equation. The dynamic adsorption and desorption indicated that similar results were observed in the static adsorption test. The purity of squalene was increased from 7.5 to 82.5% with the recovery up to 88.5% after separation on D101 column. The resin-refined sample was directly subjected to TFECD purification. Under optimized process parameters, the final product with purity of 98.5% and recovery yield of 76.5% was confirmed by high-performance liquid chromatography (HPLC) analysis. The present study provided an effective method for large-scale production of high purity squalene.     

离子交换树脂对白扦中莽草酸的分离纯化

建立了离子交换树脂分离纯化白扦中莽草酸的工艺条件和参数。通过研究D261、D296、D301-R、D301-G、D290、201*7(717)、D201和D280共8种离子交换树脂对莽草酸的吸附和解吸附能力,筛选出最佳树脂为D290,确定了最佳的吸附与解吸附工艺参数,吸附条件为pH=6、25 ℃、流速为3 mL/min;脱附条件为：洗脱液为2.5%NaOH水溶液,洗脱流速为3 mL/min。莽草酸样品溶液经D290树脂吸附与脱附后回收率为92.53%,纯度由2.97%提高到46.76%,提高了15.74倍。实验结果表明,D290树脂对莽草酸的吸附量大,脱附容易,可以应用于莽草酸的分离纯化。     

大孔树脂法分离纯化阿维菌素工艺研究

研究了大孔树脂分离纯化阿维菌素的工艺。采用HPLC检测方法,从7种大孔树脂中筛选出吸附阿维菌素性能最好的树脂并优化其吸附和洗脱参数。结果表明,采用大孔树脂HZ816吸附阿维菌素的效果最佳,其动态吸附量为62mg·mL^-1,在吸附流速为1．5～2BV·h^-1、90％乙醇作为洗脱剂的优化条件下,解吸收率大于90％、阿维菌素中B1a含量大于91％、总收率大于65％。该阿维菌素分离纯化方法工艺简单。分离效果好,适于工业化生产。     

Process research of macroporous resin chromotography for separation of N-(p-coumaroyl)serotonin and N-feruloylserotonin from Chinese safflower seed extracts

N-(p-Coumaroyl)serotonin (CS) and N-feruloylserotonin (FS) are two bioactive serotonin derivatives in safflower (Carthamus tinctorius L.) seeds with many biological effects. In the present study, the sorption and desorption characteristics of six widely used macroporous resins, coded D312, 860021, DM131, HZ801, AB-8 and XDA-1, respectively, is critically evaluated and compared for enrichment and preparative separation of CS and FS. Static adsorption and desorption experiments on these resins showed that XDA-1 had the best adsorption and desorption equilibrium for CS and FS, and its adsorption equilibrium fits the best to the Langmuir isotherm. Dynamic adsorption and desorption experiments on XDA-1 resin packed column were conducted to establish the optimum parameters as: CS and FS concentration in sample solution 0.346 and 0.276 mg/mL, respectively, sample size 5-bed volumes (BV), sample flow rate 2 mL/min, temperature 25 °C (for adsorption); eluent 80% ethanol, 11 BV, flow rate 1 mL/min (for desorption). After one run of adsorption and desorption, the contents of CS and FS were increased from 2.01%, 1.67% to 24.1% and 22.4%, and the recoveries were 77.2% and 68.5%, respectively. The chromatographic process optimized in the present work is a promising basis for large scale preparation of CS and FS upon further scaling up tests.     

Preparative Purification of Solanesol from Tobacco Leaf Extracts by Macroporous Resins

In the present study, the preparative purification of solanesol by macroporous resins was studied. The adsorption and desorption properties of solanesol on macroporous resins including SP207, SP700, SP825, SP850 and HP20 were compared. SP850 resin offered the best adsorption and desorption capacities based on the research results. Both Langmuir and Freundlich isotherms were used to describe the interactions between the solutes and resin at different temperatures, and the experimental data fits best to the Freundlich isotherms. Adsorption and desorption kinetic curves were obtained for solanesol adsorption on SP850 resin in the temperature range of 298–303 K. The results showed that the mobility of solanesol adsorption increased and the proportion of adsorbed solanesol decreased with increasing temperature. The adsorption kinetic curve can be classified into three portions, i.e., instantaneous adsorption stage, intraparticle diffusion stage and the final equilibrium stage. Dynamic adsorption and desorption experiments were carried out with the column packed by SP850 resin to optimize the separation process. The isocratic and stepwise operations were designed for the separation of solanesol. The use of stepwise elution allowed a reduction in the process time and solvent consumption, but also led to some loss in purity. The corresponding optimal parameters for adsorption and desorption were determined. According to the research results, it can be concluded that SP850 resin possesses a good ability to separate solanesol and the results in this study may provide scientific references for large‐scale solanesol production from tobacco leaf extracts.     

八角莲中黄酮类化合物的分离纯化工艺研究

研究了八角莲中黄酮类化合物的分离纯化工艺。考察各种因素对树脂吸附和洗脱效果的影响,确定了AB-8型大孔树脂分离纯化八角莲中黄酮类化合物的最佳工艺参数。最佳工艺参数为:静态吸附,树脂与样液比为1:20 g/m L、吸附时间为3 h,静态解吸过程解吸液(乙醇)体积分数为70%、树脂与解吸液的比例为1∶50 g/m L;动态吸附中动态流速为0.5 m L/min、静置时间为80 min,动态解吸中洗脱液(乙醇)的体积分数为60%、树脂与洗脱液的比例为1∶40 g/m L。     

Preparative separation and purification of rosavin in Rhodiola rosea by macroporous adsorption resins

Rosavin is one of the main bioactive components in Rhodiola rosea L. with known pharmacological effects. In this study, a simple method for preparative separation and purification of rosavin from R. rosea L. was developed with macroporous adsorption resins. The adsorption isotherms and desorption performances of some resins have been determined and compared. Depending on its hydrophobic force, HPD-200 resin, a styrene-divinylbenzene (SDVB) copolymer, offers the best adsorption and desorption properties for rosavin based on the research results, its adsorption data fit best to the Langmuir adsorption model. Dynamic adsorption and desorption was performed on HPD-200 resin packed in a glass column to obtain optimal parameters for the separation of rosavin. The optimum conditions were as follow. The volume, concentration, bed height and flow rate of loading sample were 4 bed volumes (BV), 0.24 mg/mL, 30 cm and 2 BV/h, respectively, at 25 °C; desorption was performed successively with 30% ethanol of 4 bed volumes, 40% ethanol of 2.5 bed volumes and 60% ethanol of 3 bed volumes. After one run treatment with HPD-200, the rosavin content in the product was increased from 0.69% to 11.02% with a recovery of 82.46%. In this case, the process throughout is 1.34 g rosavin/L adsorbent/day, solvent usage is 7.14 L ethanol/g rosavin. The simple purification scheme avoids toxic organic solvent and thus, increases the safety of the process with a potential industrial application prospect.     

Enrichment and Purification of Syringin, Eleutheroside E and Isofraxidin from Acanthopanax senticosus by Macroporous Resin

In order to screen a suitable resin for the preparative simultaneous separation and purification of syringin, eleutheroside E and isofraxidin from Acanthopanax senticosus, the adsorption and desorption properties of 17 widely used commercial macroporous resins were evaluated. According to our results, HPD100C, which adsorbs by the molecular tiers model, was the best macroporous resin, offering higher adsorption and desorption capacities and higher adsorption speed for syringin, eleutheroside E and isofraxidin than other resins. Dynamic adsorption and desorption tests were carried out to optimize the process parameters. The optimal conditions were as follows: for adsorption, processing volume: 24 BV, flow rate: 2 BV/h; for desorption, ethanol-water solution: 60:40 (v/v), eluent volume: 4 BV, flow rate: 3 BV/h. Under the above conditions, the contents of syringin, eleutheroside E and isofraxidin increased 174-fold, 20-fold and 5-fold and their recoveries were 80.93%, 93.97% and 93.79%, respectively.     

大孔吸附树脂对沙棘叶总黄酮的吸附性能考察

以吸附值(mg/g)和解吸附率(%)为评价指标,通过静态吸附/解吸附分别考察LSA-40、LSA-21、DM-130、HPD450、D101和XDA-1六种大孔吸附树脂对沙棘叶总黄酮的吸附/解吸附性能。通过动态吸附/解吸附优选沙棘叶总黄酮最佳分离工艺条件。结果表明中极性树脂LSA-21在吸附/解吸附方面显示出最佳的综合性能,对沙棘叶总黄酮的静态吸附值/解吸附率分别为131.78mg/g和55.53%,经静态和动态吸附后得到的总黄酮纯度分别为19.4%和20.3%。     

Enrichment and Purification of Total Chlorogenic Acids from Tobacco Waste Extract with Macroporous Resins

In the present study, an evaluation was conducted on the performance and separation characteristics of nine macroporous resins for the enrichment and purification of total chlorogenic acids from tobacco (Nicotiana tobaccum L.) waste extracts. Based on the results, XAD-4 offered higher adsorption and desorption capacities for total chlorogenic acids than other resins. To optimize the separation process of total chlorogenic acids, a column packed with XAD-4 resin was used to perform dynamic adsorption and desorption tests. The results show that the highest purity of the total chlorogenic acids product was 89.27% when optimum parameters for the adsorption process packed with the XAD-4 resin were as follows: flow rate 3.6 BV/h, pH 3.0; for desorption: ethanol–water (40:60, v/v) used as eluent, flow rate 3.6 BV/h, respectively. Therefore, the XAD-4 resin revealed a good ability to enrich and purify total chlorogenic acids. The method developed will provide a potential approach for the large-scale separation and purification of chlorogenic acid in pharmaceutical applications as a medical intermediate or a material for traditional Chinese medicine.     

大孔吸附树脂分离纯化追风伞总黄酮的研究

以总黄酮的吸附量、回收率及解吸率为考察指标,研究了大孔吸附树脂分离纯化追风伞总黄酮的工艺条件。通过静态吸附实验比较了7种不同类型大孔吸附树脂的吸附特性,确定了D101型大孔吸附树脂用于追风伞总黄酮的纯化富集。通过动态吸附实验,确定了D101型大孔吸附树脂分离纯化追风伞总黄酮的最佳工艺条件为:上样液浓度1.839 mg.mL-1,上样流速为2.0 mL.min-1,洗脱剂为70%乙醇,洗脱剂用量为6 BV。在此条件下,D101型大孔吸附树脂对追风伞总黄酮的动态饱和吸附量为80.05 mg.g-1,纯化后追风伞总黄酮的纯度达到86.2%。D101型大孔吸附树脂可以较好地分离纯化追风伞总黄酮。     

大孔树脂法分离纯化申嗪霉素的工艺研究

通过大孔吸附树脂对申嗪霉素发酵滤液静态吸附和解吸试验,从6种大孔吸附树脂中筛选出分离纯化申嗪霉素最优的树脂,考察了该树脂对申嗪霉素的静态、动态吸附与解吸性能并对吸附与洗脱的最佳条件进行了研究。结果表明:AB-8树脂对申嗪霉素有很好的吸附和解吸性能,其最优的动态吸附工艺条件为:上样液浓度3 000μg/mL,上样量4 BV,上样流速2 BV/h;最优的解吸条件为:洗脱剂为80%乙醇溶液,洗脱液用量3 BV,洗脱流速1 BV/h。在此优化条件下,申嗪霉素的吸附率、解吸率、收率、纯度的平均值分别达到(90.33±0.14)%、(90.87±0.12)%、(82.1±0.1)%和(90.74±0.14)%(n=5)。     

Preparative enrichment and purification of nevadensin from Lysionotus pauciflorus using macroporous resins

An efficient method using macroporous adsorption resins for preparative enrichment and purification of nevadensin from Lysionotus pauciflorus Maxim was developed. The results indicated that non-polar HPD-100 resin offered the best adsorption and desorption performance, its adsorption data were well-fitted to the Freundlich isotherm and the pseudo-second-order kinetics model. Dynamic adsorption and desorption experiments have been investigated for optimization of chromatographic parameters. Through one cycle of dynamic adsorption/desorption, the purity of nevadensin in the extract, increased about 8.82-fold from 8.58% to 75.70%, with a recovery yield of 69.90%. The results suggested that HPD-100 resin can separate nevadensin effectively from plant material.     

亚硫酸氢镁预处理麦秆废液中同步分离木质素磺酸盐和低聚木糖的方法研究

分别采用有机溶剂萃取法、超滤法、大孔树脂吸附法、离子交换法分离提取亚硫酸氢镁预处理麦秆废液中的木质素磺酸盐和低聚木糖。研究结果表明超滤法不能达到分离目的,有机溶剂沉淀和大孔树脂吸附可实现木质素磺酸镁的纯化,采用D380离子交换树脂进行离子交换层析可将废液中低聚木糖和木质素磺酸镁完全分离,回收所得低聚木糖和木质素磺酸盐纯度分别可达63.95%和91.28%。因此,D380树脂固定床离子交换法是一种简单有效的提取废液中高附加值产品的方法,可实现亚硫酸氢镁预处理麦秆废液的高值化利用,具有强劲的市场应用潜力。     

大孔树脂分离纯化核桃青皮总黄酮的研究

以总黄酮回收率为考察指标,研究了大孔树脂分离纯化核桃青皮总黄酮的工艺。结果表明:D101型树脂对核桃青皮总黄酮有较好的吸附分离性能,是分离纯化核桃青皮总黄酮的适宜大孔树脂;最佳工艺条件为:上柱总黄酮与干树脂质量比为1:12,上样液质量浓度可在3.0875～6.175 g/L 范围内,pH值为5, 6BV(1BV=23.7 mL)的水洗后用5BV的70%乙醇洗脱。经D101处理后的核桃青皮总黄酮回收率在60%,纯度可达80%以上。该工艺简便,能有效分离纯化核桃青皮黄酮类化合物。     

大孔树脂吸附法吸附分离黄酮的研究

研究了D101、LD601、LS-303B、LX-28、LX-38型大孔树脂对黄酮的吸附及分离性能。结果表明,LD601型大孔树脂对黄酮的吸附效果较好,pH 2～4的盐酸溶液中,静态吸附量为5 849.312μg/g,对流速2 mL/min黄酮溶液的吸附率可达96.27%,负载1 500μg黄酮的LD601树脂,用40%乙醇50 mL以3 mL/min流速进行解吸,解吸率可达97.51%。     

H103大孔吸附树脂分离纯化苦参碱与氧化苦参碱的研究

通过对比苦参总碱在九种大孔树脂上的静态吸附,筛选出对苦参总碱吸附能力较强的H103树脂;研究了H103树脂对上述两种生物碱的吸附等温曲线及吸附动力学行为;考察了苦参总碱在H103树脂上的动态吸脱附过程,用薄层色谱法(TLC)对洗脱液跟踪分析,用高效液相色谱法(HPLC)检测苦参碱、氧化苦参碱含量,确定了动态吸脱附的过程参数。结果表明,H103树脂对苦参总碱的等温吸附可采用Freundlich方程描述;由吸附动力学曲线得H103树脂在100 min内达到平衡,具有较快吸附速度,吸附动力学规律可用二级速率方程表示;动态吸脱附过程的洗脱条件为：用30%乙醇-25%氨水（115:1,v/v）,80%乙醇梯度洗脱,氧化苦参碱先被洗脱,苦参碱随后,两种生物碱可达到很好的分离,经计算苦参碱收率为90.1%,氧化苦参碱收率为85.3%。     

大孔树脂对小叶榕叶的总黄酮吸附分离特性

以大孔吸附树脂为吸附剂,研究了其对小叶榕黄酮(FFLF)的吸附分离特性,选择10种大孔吸附树脂,比较了其对FFLF的吸附率和解吸率,筛选出最佳树脂种类,并对其动力学曲线和动态吸附性能进行了考察。结果表明,D101树脂对FFLF有较好的吸附和解吸效果,适合于FFLF的分离纯化。当上样液质量浓度为25 g/L,pH=4,上样液流速为2.5 mL/min时,D101树脂对FFLF吸附量大;以w(EtOH)=50%为洗脱剂,洗脱液流速为3mL/min时,对FFLF解吸率达92%。