An Economical Method for Preparative Purification of Five Alkaloids from Coptis Chinensis Franch by High-Speed Counter-Current Chromatography Using Singled Prepared Solvent System by GC

Coptis chinensis Franch, a widely used Traditional Chinese Medicine, shows various kinds of bioactivity. The major active components of the herb are considered to be alkaloids. Thus, preparative separation of these alkaloids is critical important for further pharmacology and mechanism studies. In the paper, five alkaloids from C. chinensis were purified by HSCCC using the solvent system composed of chloro-form-metha- nol-water (2:1:1, v/v/v) single prepared. The content of each solvent in solvent system were determined by gas chromatography (GC), then according the ratios of solvents in each phase to prepare the mobile and stationary phase respectively. And a comparative study was carried out between together preparation and single preparation of the solvent system. The purities and recoveries of all the products were over 98.5% and 92%. However, 134 mL chloroform, 336 mL methanol and 452 mL water were saved when the two phase were singled by GC. Our research showed an economical method for separating alkaloids from C. chinensis by HSCCC using the solvent system single prepared by GC.

Separation and Purification of Acetophenones from Cynanchum bengei Decne Root Bark by Combination of Silica Gel and High-speed Counter-current Chromatography

[Objectives] To develop a method for separation and purification of acetophenones from Cynanchum bengei Decne root bark by combination of silica gel and high-speed counter-current chromatography( HSCCC). [Methods]The crude extract of Cynanchum bengei Decne root bark was separated by silica gel column chromatography,and parts A and B containing acetophenones were obtained. Then,parts A and B were separated by HSCCC with a two-phase solvent system composed of petroleum ether-ethyl acetate-methanol-water( 4∶ 6∶ 4. 5∶ 5. 5 and4∶ 6 ∶ 3 ∶ 7, V/V), respectively. [Results] From 260 mg of part A, four compounds with p-dihydroxybenzene 3. 9 mg(Ⅰ),4-hydroxyacetophenone 17. 1 mg( Ⅱ),2,5-di-hydroxyacetophenone 13. 3 mg(Ⅲ) and 2,4-dihydroxyaceto-phenone 21. 0 mg(Ⅳ) were obtained. And from 300 mg of part B,136 mg of Radix Cynanchi Bungei benzophenone(Ⅴ) was obtained. The purity of compounds determined by HPLC was 97. 0%,96. 6%,99. 2%,99. 7%,99. 5%,respectively. [Conclusions] The established method is simple and efficient. It can be used for separation of acetophenones from Cynanchum bengei Decne root bark and has better practical value,which could provide a reference basis for development and utilization of Cynanchum bengei Decne root bark.

Isolation and Purification of Isoaloeresin D and Aloin from Aloe vera by High-speed Counter-current Chromatography

Objective To develop an efficient method to isolate and purify the main components isoaloeresin D and aloin from Aloe vera for its industrial production.Methods High-speed counter-current chromatography was used to isolate isoaloeresin D and aloin in a one-step separation from dried crude extract of A.vera.The biphasic solvent system composed of hexane-ethyl acetate-acetone-water(0.2:5:1.5:5) was used at a flow rate of 1.0 mL/min,while the lipophilic phase was selected as the mobile phase and the apparatus was rotated at 840 r/min.The effluent was detected at 254 nm.Results Isoaloeresin D(53.1 mg) and aloin(106.9 mg) were separated from the crude extract(384.7 mg) with the purities of 98.6% and 99.5%,respectively.Conclusion HSCCC is a powerful technique for isolation and separation of chemical composition from aloe.

Preparative Separation of Four Alkaloids from Gelsemium elegans by High-speed Counter-current Chromatography

Objective To develop an efficient preparative method for the separation of Gelsemium alkaloids from Gelsemium elegans. Methods High-speed counter-current chromatography （HSCCC） with several two-phase solvent systems was investigated for the separation of Gelsemium alkaloids. The purity and structure identification of the purified compounds were performed with HPLC and NMR spectra, respectively. Results In a single operation, 206.6 mg of crude alkaloid sample was separated to yield 28.7 mg of koumine, 24.9 mg of gelsemine, 26.9 mg of humantenine, and 7.2 mg of gelsevirine, with the purities of 97.8%, 95.4%, 97.4%, and 93.5%, respectively. Conclusion A preparative HSCCC method is successfully established for the separation of four Gelsemium alkaloids from G. elegans with a modified two-phase solvent system com posed of n-hexane-ethyl acetate-ethanol-O. 5% triethylamine-H2O （3：5：3：4）.

Application of high-speed counter-current chromatography coupled with high performance liquid chromatography for the separation and purification of Quercetin-3-O-sambubioside from the leaves of Nelumbo nucifera

Quercetin-3-O-sambubioside[Quercetin-3-O-β-D-xylopyranosyl(1→2)-β-D-glucopyranoside]was separated and purified by semi-preparative high-speed counter-current chromatography(HSCCC)with a twophase-solvent system composed of ethyl acetate-nbutanol-water(4∶1∶5,v/v)from the leaves of Nelumbo nucifera(Lotus).A total of 5.0 mg of the targeted compound with a purity of 98.6%as determined by high performance liquid chromatography(HPLC)was obtained from 100 mg of the crude extract cleaned up by AB-8 macroporous resin in a one-step separation.Quercetin-3-O-sambubioside was a novel flavonoid glycoside from the leaves of Nelumbo nucifera,and its chemical structure was identified by means of ESI-MS,1D NMR and 2D NMR.

AN EFFICIENT AND TARGET-ORIENTED SAMPLE ENRICHMENT METHOD FOR PREPARATIVE SEPARATION OF MINOR ALKALOIDS BY PH-ZONE-REFINING COUNTER-CURRENT CHROMATOGRAPHY

A sample enrichment method focusing on the minor targeted components was established to help them to be successfully separated by pH-zone refining CCC.Seven minor indole alkaloids in Uncaria rhynchophylla(Miq.)Miq.ex Havil(UR)were chosen to show the advantage of this method.The sample enrichment and separation were

Comparison of Co-Current and Counter-Current Flow Fields on Extraction Performance in Micro-Channels

Several applications such as liquid-liquid extraction in micro-fluidic devices are concerned with the flow of two immiscible liquid phases. The commonly observed flow regimes in these systems are slug-flow and stratified flow. The latter regime in micro-channels has the inherent advantage that separation of the two liquids at the exit is efficient. Recently extraction in a stratified counter-current flow has been studied experimentally and it has been shown to be more efficient than co-current flow. An analytical as well as a numerical method to determine the steady-state solution of the corresponding convection-diffusion equation for the two flow-fields is presented. It is shown that the counter-current process is superior to the co-current process for the same set of parameters and operating conditions. A simplified model is proposed to analyse the process when diffusion in the transverse direction is not rate limiting. Different approaches to determining mass transfer coefficient are compared. The concept of log mean temperature difference used in design of heat exchangers is extended to describe mass transfer in the system.

Optimization of a Method for the Simultaneous Determination of Phloroglucinol and Trimethylphloroglucinol by High Performance Liquid Chromatography in Routine Use in a Pharmaceutical Industry in Abidjan

In order to provide the population with safe, effective and good quality medicines, the pharmaceutical industries, before releasing batches of their products into the pharmaceutical circuit, put in place internal dosage methods to control the quality of these products. The present study consisted in optimizing a method for the simultaneous determination of Phloroglucinol (PHG) and Trimethylphloroglucinol (TPH) by high performance liquid chromatography (HPLC) routinely used in a pharmaceutical industry located in a township in Abidjan (Ivory Coast). The basic chromatographic conditions were those routinely used for the determination of these two molecules: mobile phase: acetonitrile/water (60/40), stationary phase (C18 BDS Hypersil 250 mm * 4.6 mm - 5 μm), detection wavelength (265 nm), flow rate, injection volume and run time configured at the equipment level were respectively 1 mL/min, 10 μL and 8 min. The method of preparation of the analytes (PHG and TPHG) was also applied by the pharmaceutical industry. The application of these different parameters at the equipment level made it possible to determine a chromatogram which highlights three chromatographic peaks with respective retention times (RT) of 0.773 min (unidentified compound), 2.275 min (PHG) and 7.269 min for an analysis time of 8 min with a better resolution of the peaks and baseline. The progressive optimization of different parameters such as the stationary phase (C18 YMC 150 mm * 4.6 mm - 3 μm), the proportion of the mobile phase: acetonitrile/water (80/20), the flow rate impelled by the pump (0.8 mL/min) and the modification of the analyte preparation mode (same amount of PHG and TPHG in a 50 mL volumetric flask) resulted in a final chromatogram that highlighted two chromatographic peaks at the respective RT of 2.391 min (PHG) and 3.735 min (TPHG) at a run time of 6 min. The chromatographic conditions that led to the final chromatogram can be used routinely by the pharmaceutical industry for the determination of several PHG and TPHG drug matrices after prior validation of the determination method.

离子色谱指纹图谱分析在烟用香精香料品质评价中的应用

为保证烟用香精香料品质的一致性,采用离子色谱法对烟用香精香料样品中的9种有机酸及7种无机阴离子进行分析。1.0 g样品中加入10 mL去离子水,振荡萃取30 min后,水相溶液用0.45μm微孔滤膜过滤,滤液进一步采用RP前处理柱除去基质,经离子色谱仪分离测定。采用Dionex IonPac AS11-HC阴离子交换柱及淋洗液梯度洗脱,9种有机酸和7种常规无机阴离子标准工作溶液的保留时间和峰面积日内精密度(RSD)分别为0.01%~0.69%和1.34%~2.98%,日间RSD分别为0.03%~0.68%和3.54%~5.16%。同时对烟用香精香料标准样品A~D(各5个批次)中有机酸和无机酸的保留时间和峰面积RSD进行考察,得到的保留时间和峰面积日内RSD分别为0.01%~0.71%和2.39%~3.22%,日间RSD分别0.05%~0.81%和3.61%~6.02%。建立了4种烟用香精香料(各5个批次)的指纹谱图库,并采用系统聚类分析和相似度分析法对其他生产厂家样品进行品质评价。结果表明,聚类分析法能有效地从不同生产厂家的产品中筛选出与标准样品质量最相近的样品;实际样品AY3、BY2、CY2及DY1与香精香料标准品性质最接近,品质较好。相似度分析可以对不同生产厂家的产品质量进行更加具体、量化的评价,实际样品AY3、BY2、CY2及DY1与香精香料标准品的化学成分相似度值均大于97.7%,符合相似度评价要求。相较于超声辅助液液萃取-气相色谱法,对于个别特殊样品,离子色谱法能更有效地区分香精香料样品间的品质差异。

高等农业院校现代色谱分析课程思政探索与实践

课程思政建设是当前加强高校思想政治工作和专业课程教学改革的重要内容,是构建全方位育人格局,将其他课程与思政课同向同行,协同育人,以最终实现“立德树人”根本教育任务的一种综合教育理念。现代色谱分析是高等农业院校化学、农学、食品科学、动物药学、动物医学等专业研究生的必修课程。该课程开课专业广、修读人数多,是开展课程思政教育的重要载体。作者以高等农业院校现代色谱分析课程为例,介绍了实施课程思政的重要意义,课程思政元素的挖掘及思政内容与专业内容的融合,希望能为相关学科课程思政教学提供有益参考。

纳米晶纤维素组装的手性介孔材料及其对手性药物的分离

纤维素纳米晶体(CNC)是由硫酸催化的纤维素水解制备的,根据纤维素的来源和水解条件,纳米晶体的直径约为5~15 nm,长度约为100~300 nm。CNC在水中可以形成具有手性向列型结构的液晶相。当干燥成固体薄膜时,CNC保持螺旋手性向列顺序并组装成层状结构。在CNC自组装过程中,可以添加溶胶-凝胶前体,如Si(OMe)_(4)。随着溶剂的蒸发,这些前体会发生水解和缩合,从而形成手性向列相二氧化硅/CNC复合材料。复合材料在空气中煅烧破坏纤维素模板,留下高表面积手性向列结构的介孔二氧化硅膜。此外,有机二氧化硅或二氧化钛薄膜也采用类似的方法得到。这些薄膜可用作滤光片、反射器和薄膜。最重要的是,合成的手性介孔二氧化硅可用作手性色谱固定相材料分离手性药物。结果显示,手性分离的效果显著,为CNC开发手性新材料提供了新方法。

不同干燥方式八角果实挥发性化合物代谢组分析

【目的】探明不同干燥方式对八角果实挥发性成分的影响,筛选出适宜八角果实干燥的处理方式,为八角后期深加工、保存及应用提供参考。【方法】以新鲜八角果实为材料,采用自然晾晒、40℃、50℃、60℃、70℃、80℃和90℃烘干7种处理方式进行干燥,通过液相色谱-质谱(LC-MS)技术检测分析八角果实代谢产物,阐明八角果实不同温度干燥与自然晾晒处理的代谢物差异。【结果】与自然晾晒相比,八角果实40℃干燥差异代谢产物有11类17种,其中,萜类、醇类和醛类3类代谢物上调;50℃干燥差异代谢物有10类25种,其中,上调代谢物6类(萜类、醇类、酯类、酮类、卤代烃类、酚类);60℃干燥差异代谢物有8类13种,其中,醇类和烃类代谢物上调;70℃干燥差异代谢物为13类40种,其中,醇类、萜类和酯类3类代谢物上调;80℃干燥差异代谢物有13类38种,仅醇类代谢物上调;90℃干燥差异代谢物有13类52种,其中,上调代谢物4类(萜类、醇类、酮类、卤代烃类)。40℃、50℃和60℃干燥的八角果实与自然晾晒差异代谢物较少,上调代谢物主要是苯甲醛、醋酸植物醇、茴香脑、6-异羟基樟脑、乙酸异丁香酚酯和甲基丁香酚等挥发性化合物,提升了八角浓厚的香味。70℃、80℃和90℃干燥的八角果实与自然晾晒差异代谢物较多,且与八角香味相关的许多代谢物下调表达,主要有香芹酮、茴香脑、E-氧化玫瑰、4-蒈烯、对薄荷脑1,5,8-三烯、苯甲醛、芳樟醇、3-甲基苯并噻吩、2-甲基-2-四氢呋喃和2-乙酰基-5-甲基呋喃等挥发性化合物,减弱了八角香味。【结论】不同干燥处理八角果实的挥发性化合物种类均存在一定差异。40℃、50℃和60℃干燥八角果实,苯甲醛、醋酸植物醇和茴香脑等上调表达的挥发性化合物提升了八角香味。八角果实经50℃干燥后,上调表达的代谢物有萜类、醇类、酯类、酮类、卤代烃类和酚类6类,果实独特香气保持最好,宜在生产上推广应用。

固相萃取-超高效液相色谱-三重四极杆/线性离子阱质谱法检测水中大环内酯类抗生素

采用固相萃取-超高效液相色谱-三重四极杆线性离子阱质谱技术(SPE-UPLC-QTRAP MS)建立了环境水样中阿奇霉素、林可霉素、克林霉素、红霉素及替米考星5种大环内酯类抗生素的检测方法。调节水样pH为10后经HLB固相萃取柱净化、富集,然后用Kinetex F5色谱柱进行分离,用0.1%甲酸水溶液和0.1%甲酸乙腈溶液进行洗脱,采用电喷雾正离子源(ESI+),多反应监测(MRM)-信息依赖采集(IDA)-增强子离子(EPI)扫描模式对样品进行检测,用多反应监测(MRM)进行定量,增强子离子(EPI)谱图库辅助定性。结果表明:5种大环内酯类抗生素线性关系良好,相关系数均大于0.9992;检出限为0.01~0.30μg·L^(-1),定量限为0.10~0.50μg·L^(-1);3种不同浓度的水样加标回收率为71.60%~111.05%,相对标准偏差均在10%以内,EPI谱库比对纯度值均大于90%。该方法将传统的MRM扫描模式结合EPI谱图库检索,实现了同时定性和定量分析,对未知物准确定性定量检测提供了重要参考。

色谱技术在纺织服装化学残留物检测中的应用

纺织服装生产过程中使用的各种染料、助剂等会形成化学残留物,不仅会对人体健康产生风险,也会影响生态系统的健康发展。色谱技术具有高分离效率和准确的定量能力等优势,可以有效检测纺织服装中的化学残留物,有利于保护消费者健康,减少环境污染。探讨色谱技术在纺织服装化学残留物检测中的应用,可以为纺织行业提供理论参考。

科研成果转化为教学资源对提升教学效果的作用:以药物分析学课程中色谱方法的教学为例

药物分析学课程是将多个课程联系起来形成新的综合知识节点的三维知识网,涉及知识体系庞大,知识结构灵活。其中,色谱分析法在这门课程中涉及面极广,要准确掌握其内容难度很大,这会导致教学效果不好。本着立德树人的教育宗旨和培养复合型高素质专业人才的目标,本文以药物分析学课程中色谱方法的教学为例,将与色谱方法相关的科研成果转化为教学资源,融入教学过程,将创新理念和科研思路渗透给学生,这不仅有利于学生理解并掌握知识,而且能够锻炼学生提出问题和解决问题的能力。本文进一步阐述了如何通过课程设计引入科学发展前沿并渗透科学问题,从而阐明将科研成果转化为教学资源对达成教学目标的促进作用。针对药物分析学课程知识更新速度快、理论与实践并重的特点,我们通过引入色谱科学发展前沿、渗透科学问题、采用问题式与任务驱动式相结合的教学方法对课程进行设计,并采用多元化考核、学生反馈、自我评价3个方面对教学效果进行评估。结果表明,将科研成果转化为教学资源对激发学生学习兴趣、提升学生解决问题的能力,以及达成课程目标具有显著作用,在很大程度上提升了教学效果,实现了学生知识、能力和素质的共同提高。

采用自主研发的在线前处理系统同时分析脱硫废水中阴阳离子

电厂专用的全自动脱硫废水分析仪可以实现对超高浓度脱硫废水的自动化前处理以及分析,样品通过自动二步稀释,并与双系统离子色谱仪联用,对脱硫废水中的阴、阳离子同时进行检测。脱硫废水中阳离子主要有Na+、NH4+、K+、Mg2+、Ca2+,阴离子主要有F-、Cl-、NO3-、SO42-。本研究对仪器相关参数进行了验证,测试结果为,4种阴离子和5种阳离子的混合标准溶液在其线性关系内相关系数均大于0.999;4种阴离子的峰高和峰面积重复性RSD小于2.29%,5种阳离子的峰高和峰面积重复性RSD小于1.78%,实际样品中4种加标阴离子回收率在93.5%~100.2%之间;5种阳离子加标回收率在95.4%~103.8%之间。结果证明,该系统大大提高了分析效率,节省了人工成本,填补了市场空白。

气相色谱-质谱法测定酒店空气中7种苯系物

试验运用气相色谱-质谱法测定扩建装修酒店空气中7种苯系物,采用活性炭管采样和二硫化碳解吸,同时对质谱参数和色谱柱进行优化。结果表明,色谱柱能有效分离苯系物中7种待测组分,实现各组分峰完全分离。苯系物具有良好线性关系,相关系数R^(2)>0.999,检出限介于0.60~2.20μg/L,加标回收率为93.9%~98.2%,相对标准偏差为1.8%~4.0%,准确度和精密度良好。苯系物质量控制样品测定表明,该方法符合质控范围。气相色谱-质谱法可用于检测酒店空气中7种苯系物,方法准确可靠,灵敏度高。

基于离子色谱-电感耦合等离子体质谱法分析木耳、香菇、松茸和茶树菇中砷形态分布

为分析木耳、香菇、松茸和茶树菇中砷形态分布,采用离子色谱-电感耦合等离子体质谱(Ion Chromatography-Inductive Coupled Plasma Mass Spectrometer,IC-ICP-MS)法对其砷甜菜碱、二甲基砷、亚砷酸、砷胆碱、一甲基砷、砷酸进行测定,并进行方法学考察和含量测定。结果表明:方法在5 min内能将6种砷形态全部分离开,且峰型好;方法线性关系良好(标线质量浓度0.5~20μg/L,r>0.999),6种砷形态检出限和定量限分别不超过0.005、0.017 mg/kg。木耳、茶树菇和香菇中6种砷形态加标回收率在80%~120%范围;对于松茸,适宜加标量(0.05 mg/kg二甲基砷、砷胆碱和砷酸;0.2 mg/kg亚砷酸和一甲基砷;5 mg/kg砷甜菜碱),加标回收率也在80%~120%范围。结合干制品脱水率,所测样品无机砷含量都符合GB 2762-2022要求;松茸总砷含量最高,但是无机砷(亚砷酸+砷酸)占总砷比例最低为3.7%~6.8%,其占比最高为砷甜菜碱(75.8%~87.3%);木耳、茶树菇和香菇中砷主要形式为无机砷,无机砷占总砷比例分别可达58.4%~66.1%、60.0%~66.7%、81.2%~91.7%,提示其总砷高时有食用安全风险。

水蛭素的HPLC色谱分析条件优化研究

为研究水蛭素的HPLC最佳色谱分析条件,通过对比重组水蛭素的HPLC分析方法的洗脱条件、检测波长、色谱柱的分离效果,拟合水蛭素浓度与峰面积的线性方程,优选最佳水蛭素色谱分析条件。结果显示,ODSC18柱分析最佳条件为:采用梯度洗脱,流动相A和B洗脱程序:0~60 min,B:0%~100%,进样量20μL,柱温35℃,流速为1 mL/min,波长205 nm;TSKgelG2000SW柱最佳分析条件为:流动相磷酸缓冲液(0.02 mol/L,pH 7.0),流速0.5 mL/min,检测波长205 nm,进样量20μL。研究表明,水蛭素浓度与峰面积具有良好的直线线性相关关系,采用TSKgelG2000SW色谱柱的分析效果优于ODSC18柱,可为动物活性多肽的分析提供依据。

啶氧菌酯在香蕉及土壤中的残留分析方法研究

为研究啶氧菌酯在香蕉及土壤中的残留分析方法,采用外标法定量,试验样品经乙腈提取,固相萃取净化后经气相色谱仪-微电子捕获检测器检测,在0.02~0.10 mg/kg和0.01~0.10 mg/kg添加水平范围内,香蕉和土壤中啶氧菌酯的回收率分别为101.4%~109.2%和93.7%~105.0%,相对标准偏差(RSD)分别为2.5%~4.0%和2.8%~8.4%,最小检出量均为0.01 ng,最低检出浓度分别为0.02 mg/kg和0.01 mg/kg。该分析方法灵敏度、准确度、精密度均满足农药残留分析要求,可广泛用于啶氧菌酯残留检测。