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文章编号: 2020020308  

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人参皂苷类化合物样品前处理及分析检测

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  • 1. 烟台大学药学院 分子药理和药物评价教育部重点实验室 新型制剂与生物技术药物研究山东省高校协同创新中心 烟台 264005
  • 2. 中国科学院烟台海岸带研究所 中国科学院海岸带环境过程与生态修复重点实验室 山东省海岸带环境过程重点实验室 烟台 264003
  • 3. 中国科学院大连化学物理研究所 中国科学院分离分析化学重点实验室 大连 116023

收稿日期:2019-08-13

  要求修回日期:2019-09-13

  网络出版日期:2019-11-18

基金资助

烟台市重点研发计划(2018ZHGY085)

药学国家级实验教学示范中心(烟台大学)()

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版权所有,未经授权,不得转载、摘编本刊文章,不得使用本刊的版式设计。

Sample Pretreatment, Analysis and Detection of Ginsenosides

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  • 1. Key Laboratory of Molecular Pharmacology and Drug Evaluation, Ministry of Education, Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of Shandong, School of Pharmacy, Yantai University, Yantai 264005, China
  • 2. CAS Key Laboratory of Coastal Environmental Processes and Ecological Remediation, Yantai Institute of Coastal Zone Research(YIC), Chinese Academy of Sciences(CAS), Shandong Provincial Key Laboratory of Coastal Environmental Processes, Yantai 264003, China
  • 3. CAS Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
** e-mail: (Zhihua Song);
(Lingxin Chen)

Received:13 Aug. 2019

  rev-requestrev-request:13 Sept. 2019

  Online:18 Nov. 2019

Fund

Key Research Project of Yantai City(2018ZHGY085)

National Demonstration Center for Experimental Pharmacy Education(Yantai University)()

Copyright

Copyright reserved © 2020.

摘要

人参皂苷类化合物是参属类植物中的一类重要活性成分,主要包括原人参二醇、原人参三醇、齐墩果酸型、奥克梯隆型四类。最近,已发现的人参皂苷类化合物化学结构多达620余种,它们具有相似的化学结构,但药理活性具有明显差异。该类化合物所在基质复杂多样,选择简便高效的样品前处理方法及检测技术对于有效检测样品中人参皂苷含量至关重要。本文综述了测定各类样品中多种人参皂苷含量的样品前处理技术(溶剂提取、固相萃取等)及常用检测方法(高效液相色谱法、超高效液相色谱法、薄层色谱法、气相色谱法等),对各种方法的灵敏度及回收率等参数进行了总结,并评述了每种方法的优缺点及研究进展。

关键词: 人参皂苷 ; 样品前处理 ; 分析方法

中图分类号: O652 ()  

本文引用格式

宋志花, 李盛红, 杨刚强, 周娜, 陈令新. 人参皂苷类化合物样品前处理及分析检测[J]. 化学进展, 2020, 32(2/3): 239-248. DOI: 10.7536/PC190814

Zhihua Song, Shenghong Li, Gangqiang Yang, Na Zhou, Lingxin Chen. Sample Pretreatment, Analysis and Detection of Ginsenosides[J]. Progress in Chemistry, 2020, 32(2/3): 239-248. DOI: 10.7536/PC190814

Abstract

Ginsenosides are an important kind of active ingredients in Panax genus and can be classified into four main types: the protopanaxadiols(PPD), protopanaxatriols(PPT), oleanolic acids(OA) and ocotillol type(OT). Recently, more than 620 kinds of these compounds have been isolated. Their chemical structures are similar but have quite different medicinal functions. It is of vital importance to develop simple and facile sample pretreatment methods and detection techniques to detect the content of ginsenosides in complex matrix. This review includes many kinds of sample pretreatment methods(such as liquid phase extraction, and solid phase extraction) and detection methods(such as high performance liquid chromatography, ultra-performance liquid chromatography, thin layer chromatography, and gas chromatography), etc., summarizes the sensitivity and recovery of various methods, and reviews the advantages and disadvantages of each method and its research progress.

Contents

1 Introduction
2 Sample pretreatment technologies
2.1 Solvent extraction methods
2.2 Solid phase extraction(SPE)
3 Detection methods
3.1 High performance liquid chromatography(HPLC)
3.2 Ultra-high performance liquid chromatography(UPLC)
3.3 Other detection methods
4 Conclusion and outlook

1 引言

人参皂苷(ginsenoside)主要存在于人参属植物的根、茎、叶、花等器官中[1],属于固醇类三萜化合物,是人参属中药材的主要活性成分[2],一般不超过人参属药用植物干重的2%~5%[3]。该类化合物具有抗癌[4,5,6]、增强机体免疫力[7]、消肿抗炎[8]、抗菌[4]、抗心肌缺血再灌注损伤[9]、抗氧化[10]、增强记忆力[10]等良好的药理活性,成为近期科研工作者研究的热点[2, 11]
目前,人们从人参属植物中分离出的人参皂苷类化合物多达620余种[12],其化学结构主要分为:原人参二醇(protopanaxadiol(PPD),包括Rb1、Rb2、Rb3、Rd、Rg3、Rh2等)、原人参三醇(protopanaxatriol(PPT),包括Re、Rf、Rg1、Rg2、Rh1等)、齐墩果酸型(oleanolic acid(OA),包括Ro等)、奥克梯隆型(Octillol(OT))四大类[12,13,14,15,16](如表1 [13])。人参皂苷类化合物不同的化学结构决定了其具有不同的药理活性[17, 18],为了进一步研究其作用机理,对该类化合物进行分析检测将会促进中药事业的发展。人参皂苷类化合物通常存在于复杂的植物样品基质中,其代谢过程及作用机理非常复杂,因此,分析检测各种人参皂苷的单体成分面临巨大的挑战。本文对最近五年来人参皂苷类化合物的样品预处理技术及检测方法进行文献综述,以期为解决该难题提供一定的理论参考。
表1 四类常见人参皂苷化合物:(a)20(s)-原人参二醇、(b)20(s)-原人参三醇、(c)齐墩果酸型、(d)24(R)-奥克梯隆型[13]

Table 1 Four types of common ginsenosides:(a) 20(s)-PPD,(b) 20(s)-PPT,(c) OA,(d) 24(R)-OT[13]. Copyright 2000 American Chemical Society

(a) ginsenoside R1 R2 formula M.W.
Rb1 -glc[2→1]glc -glc[6→1]glc C54H92O23 1108
Rb2 -glc[2→1]glc -glc[6→1]ara(p) C53H90O22 1078
Rc -glc[2→1]glc -glc[6→1]ara(f) C53H90O22 1078
Rd -glc[2→1]glc -glc C48H82O18 946
(b) ginsenoside R1 R2 formula M.W.
Rg1 -glc -glc C42H72O14 800
Re -glc[2→1]rha -glc C48H82O18 946
Rf -glc[2→1]glc -H C42H72O14 800
(c) ginsenoside R1 R2 formula M.W.
Ro -glcUA[2→1]glc -glc C48H76O19 956
(d) pseudoginsenoside R formula M.W.
F11 -glc[2→1]rha C42H72O14 800

Sugar units: glc: β-D-glucose; ara(p): α-L-arabinofuranose; rha: α-L-rhamnose; glcUA: β-D-glucuronide.

2 样品前处理技术

2.1 溶剂提取法

人参皂苷类化合物的提取非常关键,溶剂提取法是最为常用的提取方法,该方法主要依据溶剂对人参皂苷类化合物相似相溶的原理,通过选取不同极性的溶剂实现目标成分的有效提取,尽量去除杂质。传统的溶剂提取方法主要包括:煎煮[19]、浸渍[20]、索氏提取等方法。传统技术所需仪器简单、操作方便,Xiong等[21]以60%的乙醇水溶液作为提取溶剂,加热至80 ℃回流萃取三次,每次1.51 h,对三七中三七皂苷R1,人参皂苷Rg1、Re、Rb1及Rd的萃取收率分别可达1.12~5.63、10.52~23.35、1.13~3.43、14.09~18.85及5.15~6.76 mg·g-1。但传统提取技术仍存在一些不足之处,如操作温度过高易破坏热稳定低的活性成分、提取效率低、有机溶剂消耗量大造成环境污染(萃取1 g干燥样品需要几十毫升有机溶剂)、提取时间长、萃取效率低、能耗高等[22, 23]。为了克服以上不足,一些新兴的快速提取方法应运而生,主要包括:微波辅助提取法、超声辅助提取法、加速溶剂萃取、加压溶剂提取、亚/超临界流体萃取和酶解法等(如表2 [24,25,26,27,28,29]所示)。
表2 人参皂苷类化合物的样品前处理方法

Table 2 Sample pretreatment methods for ginsenosides

Sample pretreatment methods Principles Advantages Disadvantages
Solvent extraction method Microwave assisted extraction(MAE) Cell walls of the Chinese traditional medicine are disrupted by the microwave with energy of 300 MHz~300 GHz, and then the extracting rate is increased. Convenient operation, short extraction time, high extraction yield, less solvent consumption, and low cost[24, 25]. Some heat-sensitive
compounds are easy to be destroyed.
Ultrasonic assisted extraction(UAE) The disruption of cell walls, reduction of particle-size and enhancing mass transfer of the cell contents are caused by cavitation bubble collapse, mechanical and thermal effects. Highly efficient, low consumption of solvent, fast(tens minutes), mild conditions(20~30 ℃), simple and low cost[26]. Low degree of automation and difficult to be used on-line.
Accelerated solvent extraction(ASE) ASE is carried out in closed container with a high pressure and a temperature above the boiling point of organic solvent. The extraction rate increases with the increase of pressure and temperature. Fast(a few minutes to ten minutes), save solvent, realize automation easily. Some heat-sensitive
compounds are easy to be destroyed; the instruments are expensive; and operation skills are hard to master.
Sub-and supercritical fluid extraction The extraction process proceeds by using sub-and supercritical fluids near the critical point of temperature and pressure with the good properties of high-density, low-viscosity, and high-permeability. Non-toxic and safe, no organic solvent residual, environment friendly, low energy cost [27]. The instrument is expensive
due to high pressure
resistant.
Enzymatic dissociation method Active ingredients can beextracted from plant tissues by using suitable enzymes(cellulase, amylase, etc.) under mild conditions[28]. Furthermore, the fragmentation of cell walls will be accelerated with the increase of the pressure(100~600 MPa) [29]. Efficient and has great potential in extraction of active ingredients from traditional Chinese medicine. Susceptible to external
conditions.
Solid phase extraction (SPE) The ginsenoside compounds and impurities are separated by adsorbents according to the difference in adsorption. Simple, low cost and wide
range ofapplication.
Large amount of organic reagents consumption, only suitable for pretreatment of small batches of samples.
(1)微波辅助提取法(MAE)
MAE借助于微波能(频率介于300 MHz~300 GHz)加速提取速率,最早于1986年被应用于天然产物中有效成分的提取[30]。在提取过程中,微波可有效破坏药材细胞壁,使待提取成分有效析出,是一种操作简便、快速、节能高效的提取方法[24, 25],易于实现在线联用。
Jin等[31]以水作为萃取溶剂,在145 ℃条件下,采用1600 W微波对西洋参中的人参皂苷类化合物提取15 min,该方法对9种人参皂苷类化合物的总提取收率分别是加热回流提取法及加压升温提取法的1.67倍和2.57倍。为了避免持续高温引起人参皂苷类化合物降解,Zhang等[23]发展了动态微波辅助提取法提取人参根须中的人参总皂苷,相比于普通微波辅助提取法,其产率提升了15%。MAE虽然有效提高了萃取效率,但极易破坏某些热敏性活性化合物。
(2)超声辅助提取法(UAE)
超声波的声波频率范围是20~50 MHz,其工作过程主要涉及到三种作用机制:① 空化效应:植物细胞在超声波的作用下,细胞内产生的气泡声压达到最大耐受值而瞬间破裂,使得细胞被破坏,待测物质被有效释放;② 机械效应与热效应:超声波使得体系吸收机械能,内能增加,分子运动速度有效增加,加速了物质溶出速度。在三种机制的作用下,植物细胞被破坏,活性成分有效扩散到提取溶剂中,从而有效提高了回收率[26, 32]
Mathur等[33]系统考察了超声辅助提取过程中萃取剂种类及温度对中药材中各类人参皂苷类化合物提取效率的影响,结果发现100%甲醇利于西洋参中Rb(0.189 mg·g-1)和Rg(3.163 mg·g-1)的提取;70%甲醇利于Rg1(1.812 mg·g-1)的提取。Liu等[34]以70%甲醇水溶液为萃取剂,对人参根粉末在室温(25 ℃)超声萃取30 min,将所得溶液在40 ℃减压蒸馏后转移至容量瓶中,采用甲醇定容,经HPLC检测出新鲜人参中11种人参皂苷类化合物,其中,Rg1、Re、Rb1及Rd含量分别为2.96、2.58、2.38、0.70 mg·g-1。Fu等[35]发展了水溶液中添加表面活性剂的方式,采用超声辅助有效提取了三七根中三七皂苷R1,人参皂苷Rg1、Re、Rb1及Rd,样品收率分别可达28.66、46.52、12.91、29.31及9.26 mg·g-1;相比于传统溶剂提取过程,该过程无须使用有机溶剂,更加绿色环保。
总之,在人参皂苷类化合物提取过程中,超声辅助提取法的优点主要包括[26]:提取效率高,可以做到短时间内(几十分钟)提取活性成分;操作条件温和(20~30 ℃),有效避免了高温对活性成分的破坏;过程简单、易操作,设备运行成本低。目前,该方法已被广泛应用于中药材中有效成分提取、环境分析、化学化工等多个领域的样品前处理过程中。
(3)加速溶剂萃取(ASE)
ASE是一种在密闭容器内,借助高温(100~200 ℃)、高压(6~20 MPa)条件提取固体基质中化合物的技术。其主要原理是:随着体系压力升高,溶剂的沸点逐渐升高,从而有效增加待提取物的溶解度;待提取物扩散速度随温度升高而加快,从而加速提取过程。在人参皂苷类化合物的提取过程中,ASE的优势主要是:快速(几分钟至十几分钟)、节约溶剂、借助于仪器易实现自动化。
Yu等[36]采用ASE联用离心分配色谱(CPC)及UPLC技术对西洋参叶中20种弱极性人参皂苷类化合物进行了在线提取检测,最佳提取条件为:时间5 min,温度130 ℃。相比于离线分离检测,该联用技术将过滤、浓缩、离心、降解等步骤进行合并,节省人力,改善实验结果的重现性。该技术的不足之处在于热敏性活性成分易被破坏,仪器价格昂贵,要求操作者掌握仪器操作技能等。
(4)亚/超临界流体萃取
亚/超临界流体萃取是采用温度和压力处于临界点附近的流体对目标物进行提取的技术。最常用的流体为二氧化碳和水,相比于以上几种提取方式,该技术具有低毒、无溶剂残留、环境友好、低能耗等优点[27],适用于人参皂苷等一系列具有生物活性化合物的提取[22]。Han等[37]在30 kPa、50 ℃条件下,采用超临界CO2提取了理冲生髓饮药物中的抗卵巢癌活性成分,经HPLC测出人参皂苷Rd、Rc等19种活性成分。
亚临界水的压力介于20~150 bar,温度介于100~374 ℃,具有无毒性、安全、环境友好等优点[22, 27]。Chung等[27]发现,亚临界水对红参中人参皂苷Rg3的萃取效率((1.59 ± 0.05) mg·g-1)是热水、甲醇、乙醇萃取效率的3.6~6.9倍。亚/超临界流体萃取技术的弊端在于实验条件要求高,需要专用设备。
(5)酶解法
人们选取合适的酶(纤维素酶、淀粉酶等),在一定压力(100~600 MPa)及温度条件下分解植物组织,加速人参属植物中人参皂苷类化合物等有效成分的释放,即酶解法[28, 29]。加压酶解法,即酶解过程中,增加体系压力(100~600 MPa)可以加快细胞壁的破碎,增加酶的活性,利于提高萃取效率,在中药活性成分提取中显现出较大潜力[37]
近年来,酶解法在中药活性成分提取中显现出较大潜力[28]。Sunwoo等[38]采用加压酶解法从新鲜人参根中提取了人参皂苷类化合物,在同等条件下(100 MPa,50 ℃),纤维素酶酶解法提取总皂苷产量为40.2 mg·mL-1,明显高于β-淀粉酶酶解法提取收率(36.1 mg·mL-1)。另外,酶解法还可以将某些含量丰富的人参皂苷类化合物成分转变为某些含量低并且药用价值高的稀缺种类[28, 39]。Wan等[40]采用酶解法结合加压溶剂萃取技术在140 ℃、6.89 × 103 kPa条件下,取22 mL甲醇提取三七叶中的人参皂苷,静态萃取时间15 min,经UPLC/Q-TOF MS共检测出57种皂苷类化合物。

2.2 固相萃取(SPE)

SPE通过改变萃取材料种类实现人参皂苷等样品的富集纯化,是一种较为成熟的样品前处理技术。大孔树脂是分离纯化天然产物中人参皂苷类化合物常用的材料,其具有价格低廉,萃取效果较好等优势。近年来,一批新型的商品化SPE材料发展迅猛。Waters Oasis HLB SPE小柱(1 mL,30 mg)对人参皂苷类化合物具有很好的富集能力,Qi等[41]将其用于人血浆、尿液、粪便中西洋参代谢产物的富集,经LC-Q-TOF-MS分别测出42种、44种、45种人参皂苷类化合物。Ju等[42]对患有慢性心力衰竭的小鼠静脉注射益气复脉制剂,采用Waters Oasis HLB SPE小柱吸附该小鼠的血浆,经超快速液相色谱质谱联用技术(UFLC-MS/MS)测出10种人参皂苷类化合物。为了进一步提高人参皂苷类化合物的富集效率,科研工作者不断尝试发展新型的SPE材料,如分子印迹材料、碳纳米材料、金属氧化物纳米材料等。
(1)分子印迹聚合物(MIPs)
分子印迹技术(MIT)是制备空间结构结合位点与模板分子完全匹配的聚合物基质的合成技术[43],是人工模拟抗原-抗体、生物素-抗生素及酶-底物等之间的特异性作用[44],并以此为基础发展起来的一种制备特异性分离富集材料的技术[45,46,47,48]。最近,张伟等[49]制备了人参皂苷Re印迹整体柱,该整体柱对Re具有特异性吸附作用,吸附容量达0.247 mg·g-1
Hou等[50]以Rb1、Rg1和R1为模板分子,丙烯酰胺(AM)为单体,乙二醇二甲基丙烯酸酯(EGDMA)为交联剂,发展了多模板印迹SBA-15材料,将其填充SPE空柱管后用于雄鼠血浆中Rb1、Rg1及R1的富集,最大吸附容量可达123.11 μmol·g-1。Yin等[51]研究发现,表面分子印迹技术制备的Rg1印迹材料对于Rg1的吸附容量(46.80 mg·g-1)高于沉淀聚合技术所得Rg1印迹聚合物(27.74 mg·g-1)。Zhou等[52]发展了Rb1印迹硅胶包覆Fe3O4材料(Rb1-MMIPs),该材料对小鼠粪便中的58种人参皂苷类化合物具有较好的富集效果,Rb1-MMIPs材料对Rb1、Rg1、R1的萃取收率分别是非印迹材料(MNIPs)的5.66、5.58、4.75倍。此外,该材料对人参皂苷类化合物的富集能力优于有机溶剂(34种)和商品化Waters Oasis HLB SPE小柱(30种)。
总之,分子印迹材料对人参皂苷类化合物具有较高的富集效率,可有效减少有机溶剂消耗量,成本低廉,环境友好。但是,该材料的制备过程重现性并不理想。因此,寻找制备分子印迹材料的新方法及新型的富集材料仍然是亟待解决的关键科学问题。
(2)其他材料
多壁碳纳米管(MWCNTs)由石墨片层构成,是一类多孔碳纳米材料,具有独特的物理化学性质(如比表面积高达2600 m2·g-1等)[53],被广泛应用于样品前处理[54]。修洋等[55]发现MWCNTs对人参皂苷类化合物具有特异性吸附作用,对人参提取物中人参皂苷的吸附回收率在3 min内可达100%,而大孔树脂在10 min内对人参皂苷的吸附回收率不足45%,MWCNTs材料在中药材中有效成分的提取中具有潜在的应用价值。
Wei等[56]将氧化铝纳米颗粒涂覆的聚醚砜纤维中空膜材料用于口服液中人参皂苷类化合物的萃取,回收率可达88.61%~94.29%。

3 检测方法

3.1 高效液相色谱法(HPLC)

在人参皂苷类化合物的分离分析中,HPLC是最为常用的分析检测技术(如表3 [12, 24, 40, 42, 55, 57~62]所示),其具有分析速度快(几到几十分钟)、分离效果好(柱效可达几万到几十万理论塔板数每米)、灵敏度高(取决于检测器)、重现性好等优点[63]。在该类化合物的分析中,与HPLC联用的检测器包括:紫外检测器(UV)、二极管阵列检测器(DAD)、蒸发光散射检测器(ELSD)、质谱检测器(MS)等。
表3 液相色谱法检测人参皂苷类化合物

Table 3 The application of liquid chromatography for analysis of active compounds in Chinese herbs

Analyte Matrix Sample
pretreatment
Column and temperature Mobile phases and detection Recovery(%) LOD Ref
Re, Rh1, Rg2,
Rg1, Rf
White ginseng, red ginseng, American ginseng, and ginseng. Extracted by methanol Shiseido UG 80 Capcell Pak
NH2 column
(250 × 4.6 mm
i.d., 5 μm), 25 ℃
0~3 min, 89% A; 3~25 min, 89%→84% A; 25~30 min, 84%→82% A; 30~35 min, 82%→76% A; 35~40 min,89% A.
A:acetonitrile; B water. UV:203 nm
95.31%~103.85% 0.0047 ~
0.225
(mg·L-1)
12
20 (S)-Rh1, 20
(R)-Rh1, Rg6,
F4, Rk3, 20(S)-Rg3, 20(R)-Rg3, Rk1, Rg5
Roots of P. quinquefolius L. Reflux extraction by water and
methanol
Acchrom Technologies ODS-C18 type column(250 ×4.6 mm i.d., 5 μm), 30 ℃. 0~10 min, 33% B; 10~15 min, 33%→40% B; 15~40 min, 40%→60% B; 40~70 min, 60% B.
A: water; B: acetonitrile. HPLC-UV: 203 nm.
HPLC-ESI-MS
97.97%~103.24% 0.18~
0.45
(μg·mL-1)
24
ginsenosides Rb1, Rc, Rb2, Rb3;
notoginsenosides Fc, Fe, Fd
Panax notoginseng leaves Rxtracted by methanol Agilent Zorbax ODS C8 column(250 × 4.6 mm
i.d.,5 μm),
35 ℃
0~5 min, 15%→30% B; 5~15 min, 30%→32% B; 15~35 min, 32%→32% B; 35~45 min, 32%→45% B; 45~60 min, 45%→50% B. A: water; B: acetonitrile.
HPLC-UV: 203 nm
98.7%~106.1% 98 (ng) 40
Ginsenoside Rg1,
Ginsenoside Rb1,
Ginsenoside Rc,
Ginsenoside Rd,
Ginsenoside Re,
Ginsenoside Rf,
Ginsenoside Rg3,
Ginsenoside Rh1,
Ginsenoside Rb2,
Ginsenoside Rb3
Ginsenosides in chronic
heart failure(CHF) rats
Solid-phase extraction(SPE) ACQUITY UPLC􀳏 HSS T3 column(100 × 2.1 mm i.d.,
1.8 μm),
40 ℃
0~1 min, 30%→35% B; 1.0~5.0 min, 35%→38% B; 5.0~5.5 min, 38%→45% B; 5.5~6.0 min, 45%→80% B; 6.0~7.0 min, 80%→90% B; 7.0~7.5 min, 90%→30% B; 7.5~8.0 min, 30%→30% B. A: water with 0.1% formic acid; B: acetonitrile.
UFLC-MS/MS
60%~105% - 42
Ginsenoside Rg1,
Ginsenoside Re,
Ginsenoside Rb1
Panax quinquefolii Radix Reflux extraction by water-saturated
n-butanol
Venusil XBP
C18 column
(250 × 4.6 mm i.d., 5 μm),
30 ℃.
0.01~25 min, 19%→20% A;
25.01~60 min, 20%→40% A;
60.01~80 min, 40.1%→100% A.
A: acetonitrile; B: 0.1%
phosphoric acid solution.
UV: 203 nm
88.61%~94.29% 0.066~
0.400
(μg·mL-1)
55
notoginsenoside R1,
ginsenoside Rg1,
ginsenoside Rb1,
astragaloside Ⅳ,
ginsenoside Rd
Qishen Yiqi
Dripping Pills(QYDP)
liquid-liquid Extraction using water-
saturated n-butanol
Acquity UPLC HSS T3 column (100 × 2.1 mm i.d., 5 μm), 30 ℃ 0~1 min, 83% A; 1~14 min, 83%→56%; 14~15 min, 56% A. A: water containing 0.1% formic acid; B: acetonitrile. UPLC-ELSD 96.87%~99.97% 2.36~
7.68
(μg·mL-1)
57
Analyte Matrix Sample
pretreatment
Column and temperature Mobile phases and detection Recovery(%) LOD Ref
G-Ra1, G-Ra2,
G-Rb1, G-Rb2,
G-Rb3, G-Rc,
G-Rd, G-Re,
G-Re4, G-Rf,
G-Rg1, G-Rg2,
G-Ro, G-Rs2,
G-RoMe, 20-Glc-
G-Rf, Ma-G-Rb2,
NG-R1, NG-R2
Roots and
rhizomes of
Panax ginseng samples.
Extracted by 70% aqueous MeOH solutions Diamonsil ODS C18 column
(250 × 4.6 mm i.d., 5 μm), room temperature
0~20 min, 10%→20% A; 20~30 min, 20%→22% A; 30~40 min, 22%→31%A; 40~75 min, 31%→33% A; 75~80 min, 33%→40% A; 80~90 min, 40%→50% A; 90~100 min, 50%→60% A; 100~110 min, 60%→70% A. Flow rate: 0~32 min, 0.8 mL/min; 32.1~110 min, 0.5 mL/min.
A: MeCN; B: MeCN: H2O: 0.1% formic acid aqueous solution(5∶90∶8; v/v/v)
HPLC-ESI-MS
94.87%~102.45% 0.159~
9.052 (ng)
58
N-R1, G-Rg1,
G-Re, G-Rf,
G-F3, G-Rg2,
G-Rh1, G-Rb1,
G-Ro, G-Rc,
G-Rb2, G-Rb3,
CS-IV, CS-Iva,
G-Rd, G-Rg3
Panax japonicas(PJ), Panax japonicus var. major(PM), and Panax zingiberensis (PZ) Extracted by 60% aqueous methanol solutions Waters C18
column(150×3.9 mm i.d., 4.6 μm), room
temperature
0~3 min, 20%→23% A; 3~8 min, 30%→35% A; 8~15 min, 35% A; 15~20 min, 35%→60% A; 20~22 min, 60%→80% A; 22~24 min, 80%→95% A; 24~25 min, 95%→20% A.
A: acetonitrile; B: 0.05% formic acid aqueous solution.
HPLC-ESI-MS/MS
99.25%~104.10% 0.13~
2.22
(ng·mL-1)
59
ginsenosides Rg1, 20(S)-Rg2, Re, 20(S)-Rh, Rb1, Rb2, Rd Tissue extracts from the root and rhizome of Panax ginseng C.A. Mey. Extracted by methanol Waters C18
column(100 mm ×2.1 mm i.d., 1.7 μm), room temperature ~20 ℃.
0~3 min, 10%→20% B; 3~25 min, 20%→38% B; 25~30 min, 38%→85% B; 30~30.1 min, 85%→100% B; 30.1~32 min, 100% B; 32~32.1 min 100%→10% B. A: formic acid aqueous solution; B: acetonitrile containing 0.1% formic acid.
UPLC-QTOF-MS
- 6.08~
108.72
(ng·mL-1)
60
Rg1 and
its metabolites
Sprague-
Dawley rat
bile, urine,
and feces
Extracted by methanol Shim-Pack XR-ODS Ⅱ(75 × 2 mm, 2.3 μm) column, 40 ℃. 0~7 min, 22%→80% B; 7~7.01 min, 80%→22%; 7.01~10 min, 22%→22% B.
A: 0.05% formic acid aqueous solution; B: 0.05% formic acid in acetonitrile.
HPLC-MS/MS
Rg1, ginsenoside Rh1(Rh1), and protopanaxatriol(Ppt) in bile, urine, and feces ≥70%. The fecal excretion recoveries of Rg1, Rh1, and Ppt, 22.19%~ 40.11%. Rg1 in bile, 6.88%; Rh1 and Rg1in Urinary excretion 0.04%~0.09%. - 61
Rg1, Re, Rf,
Rg2, Rb1, Rc,
Rb2, Rd
Kang’ai
injection
Aqueous two-
phase system
based Deep
eutectic solvent
and K2HPO4
solution
Agilent Zorbax SB-C18 column(250×4.6 mm i.d., 5 μm),
30 ℃
0~34 min, 19.2% A; 34~35 min, 19.2%→28.0% A; 35~48 min, 28.0% A; 48~56 min, 28.5% A; 56~72 min, 36.0% A. A: acetonitrile; B: 0.1% phosphoric acid aqueous solution. HPLC-DAD: 203 nm 92.7%~110.8% 0.3~1.5
(μg·mL-1)
62
由于UV价格低廉、操作简便、检测灵敏度较高,通常被应用于简单基质中样品的检测,是测定人参皂苷类化合物的主力检测器[64]。Xu等[65]采用HPLC在203 nm波长条件下对人参主根的周皮、表皮及中柱中8种人参皂苷进行分离检测,发现周皮中人参皂苷含量(≥ 25 mg·g-1)分别为表皮及中柱中含量的4.6及74倍。在人参皂苷的检测中,UV的不足之处在于,由于待测化合物的分子结构中缺少共轭双键等生色团,检测波长通常介于200~205 nm,检测结果受溶剂干扰严重。
ELSD属于通用型检测器,适用于紫外吸收很弱的人参皂苷类化合物的检测。Kwon等[66]采用HPLC-ELSD测定越南野生人参中紫外响应较弱的OT类化合物,共检出17种以OT为主的人参皂苷类化合物。Park等[67]采用HPLC-ELSD对9种人参皂苷类化合物进行有效分离检测,并考察了流动相组成、洗脱条件等因素对ELSD检测器的影响。Yu等[68]以FeCl3作为催化剂,促进高含量低活性的Rb1转化为低含量高活性的珍贵人参皂苷类化合物;并采用HPLC-ELSD对产物进行分离检测,除Rb1外,共测得6种Rk1等人参皂苷类化合物;证实催化反应成功完成,为低含量珍贵人参皂苷类化合物的合成提供了有力指导,但是,其不足之处在于检测灵敏度低,检测时间较长。
相比于上述两种检测器,MS或MS/MS具有更高的检测灵敏度(ng以下)[69, 70]、高通量、高分辨率、强选择性等优势[69],可鉴定人参的分子结构,在近期的中药分析应用中日趋广泛[15, 71, 72]。Popovich等[14]采用LC-QTOF-MS/MS在40 min内实现了102种人参皂苷类化合物的分离检测;并测出人参主根、根须、根茎、茎、叶部位的人参皂苷种类数量分别是76、69、74、44、57,发现生长13年的人参根部人参皂苷含量最丰富,生长14年的人参茎叶中人参皂苷含量最丰富。

3.2 超高效液相色谱法(UPLC)

随着液相色谱泵耐压性能的改善,UPLC的应用得到推广,相比于HPLC,UPLC的分析速度更快[20]。最近,Yang等[58]采用超高效液相色谱串联二极管阵列检测器及飞行时间质谱检测器 (UPLC-DAD-QTOF-MS/MS)在10 min内实现了131种人参皂苷类化合物有效分离检测。串联质谱(MS)分析复杂中药优点主要是:分析速度快,分辨率高,灵敏度高[5]
Kuang等[20]采用超高效液相色谱-电喷雾二级质谱联用技术(UPLC-ESI-FT-MS),首次检测出红毛七粉末中23种丙二酰三萜烯皂苷类化合物。Yu等[57]采用UPLC-ELSD在15 min内实现芪参益气滴丸中5种人参皂苷类化合物的分离检测。

3.3 气相色谱法(GC)

GC适用于分离检测挥发性和半挥发性的小分子化合物[73],Tang等[74]采用GC-MS对人参及西洋参根、茎、叶组织的初级代谢产物进行分离分析,最终检测出149种化合物,主要包括14种糖类、39种酸类、14种醇类、29种氨基酸和48种未知及未分类化合物。Wang等[75]采用GC-MS从人参中测出91种以挥发性萜烯类和醇类芳香类化合物为主的物质,并发现人参芳香类化合物含量与其生长年限相关。Qi等[76]采用GC-MS对比研究了高丽参和西洋参的代谢产物。Tang等[77]采用GC-MS对人参的叶、茎、叶柄、侧根、主根的代谢产物进行检测,测出代谢产物种类数目分别是30、16、20、36、31。
在人参皂苷类化合物的分离检测中,GC对待测样品具有一定挥发性的要求并且要求检测前衍生化反应[78, 79],GC多应用于人参皂苷类化合物代谢产物的检测。

3.4 其他检测方法

(1)薄层色谱法(TLC)
TLC是一种非常简便快捷的分离及定性检测方式,该技术对人参皂苷类化合物的分析始于80年代初期。Paik等[80] VCHCl3 : VCH3OH : VH2O =65∶35∶10的混合溶液为展开剂,将Rb1酶解液中Rb1、Rd、Rg3、Rh2等化合物在60F254硅胶薄层色谱板上展开,以10%硫酸溶液为显色剂,通过颜色变化定性描述酶解反应程度。在人参皂苷类化合物的检测中,TLC的不足之处:(1)难以实现成分复杂的样品(十几至几十种组分)的分离;(2)难以实现精确定量。
(2)酶联免疫分析法(ELISA)
ELISA基于抗原-抗体特异性识别作用,将人参皂苷类化合物与对应抗体进行结合,从而测定该类化合物含量,Qu等[81]制备了人参皂苷Rh1的单克隆抗体,并将其应用于中药中人参皂苷Rh1和Rg2的测定。相比于上述几种分析方法,ELISA的优势在于样品前处理过程简单、易操作、特异性强、环保、样品用量少(几微升)、适用于大批量痕量样品的快速检测。该方法的弊端在于复杂样品多种成分之间相互干扰,易出现假阳性结果。
(3)质谱成像技术(MSI)
在人参皂苷类化合物的分析测定中,传统的检测方法通常需要复杂的样品预处理过程,该过程占据分析过程用时的60%以上,因此,寻求简单快速的分析方法引发科研工作者的广泛关注。质谱成像技术无需复杂的样品预处理步骤,有利于研究物质在复杂基质中的空间分布。
最近,Liu等[82]采用基质辅助激光解吸电离飞行时间质谱成像技术(MALDI-TOF-MSI)对不同生长年限的人参根切片进行分析,检出人参皂苷类化合物总计31种,PPD、PPT、OA及其他种类人参皂苷类化合物的数量分别是20、7、1、3种,可有效鉴定不同生长年限的人参根。质谱成像技术具有一系列优点[83]:快速(几十秒)、空间分辨率高(微米级)、灵敏度高(与MS相当)、节省样品(切片厚度几十微米)等。因此,质谱成像技术在珍贵中药材检测中展现出巨大的开发潜力。
综上所述,在人参皂苷类化合物的分析检测中,HPLC及UPLC联用各类检测器是最为常用的方式。虽然TLC难以实现准确定量,但具有操作过程简单、成本低廉的优势,目前仍被广泛使用。GC检测灵敏度高,但该技术对分析对象有一定挥发性的要求限制了其在人参皂苷类化合物分析检测中的应用,多应用于人参皂苷类化合物代谢产物的分析检测。ELISA适合微量样品的快速检测,但检测结果易受结构类似化合物干扰。质谱成像技术具有较高的时空分辨率,作为一种新兴技术吸引着人们不断探索。

4 结论与展望

人参皂苷类化合物具有繁多的化学分子结构,结构差异导致其药理活性差异明显。近年来,科研工作者发展了多种样品前处理及检测方法,该类方法对实现人参皂苷类化合物的定性定量分析具有重要意义。
虽然现有的样品前处理技术及分析方法在人参皂苷类化合物的分析测定方面取得了很好的结果,但仍然存在一些问题,如溶剂提取过程所用试剂污染环境、提取不充分,检测过程耗时等。因此,在未来我们应当向以下几个方向不断努力:(1)在样品前处理方面,发展切实可行的处理方法,尽量减少样品前处理过程引入新的杂质;(2)发展集成化、易操作的样品前处理-分析测试在线联用技术,节省能源及人力;(3)寻求绿色环保无毒的溶剂,提升食用安全性及减少环境污染。
[1]
Gurung B, Bhardwaj P K, Rai A K, Sahoo D . Nat. Prod. Res., 2018,32:234. https://www.ncbi.nlm.nih.gov/pubmed/28649854

DOI: 10.1080/14786419.2017.1343322   PMID: 28649854

This study compared eight major ginsenosides (Rg1, Rg2, Rf, Re, Rd, Rc, Rb1 and Rb2) between Panax sokpayensis and Panax bipinnatifidus collected from Sikkim Himalaya, India. High-performance liquid chromatographic analysis revealed that all major ginsenosides were present in the rhizomes of P. sokpayensis except ginsenoside Rc, whereas ginsenoside Rf, Rc and Rb2 were not detected in P. bipinnatifidus.

[2]
Gao Y L, Wang T, Wang G F, Li G S, Sun C F, Jiang Z M, Yang J R, Li Y S, You Y L, Wu X R, Sun L Q, Wang H B, Li C M, Tian J W, Zhu J, Wang K Z . Cho S. Food Chem. Toxicol., 2019,131:110578. https://www.ncbi.nlm.nih.gov/pubmed/31201900

DOI: 10.1016/j.fct.2019.110578   PMID: 31201900

Ginsenoside compound K (CK) is a hydrolysate of ginsenosides in the soil bacteria. This study evaluated the toxicity of CK as acute and the 26-week repeated-dose. The results of acute toxicity show that CK administered orally to rats and mice did not cause mortality or toxicity at the maximum dosage of 8 g/kg and 10 g/kg, respectively. In the toxicity study for 26-week, rats were administered with CK at doses of 13, 40, or 120 mg/kg, and were observed for 26 weeks and recovery periods of four weeks. Under the conditions, asthenia, hypoactivity, loss of fur and body weight reduction were transiently noticed in males of 120 mg/kg group. Hepatotoxicity and nephrotoxicity also were evident including the elevation of liver and kidney relative weight, along with focal liver necrosis as well as the increase in plasma enzymes (ALT and ALP) in male rats receiving CK (120 mg/kg), but this toxicity might be reversible. For 13 and 40 mg/kg CK groups, there was no significant variation in food habits, clinical signs, urine analysis, body weight, biochemical and hematological values, organ coefficient and histopathology examination. The NOAEL for male and female rats were observed to be 40 and 120 mg/kg, respectively.

[3]
Wang W Y, Ni Y Y, Wang L, Che X, Liu W H, Meng Q G . Xenobiotica, 2015,45:385. https://www.ncbi.nlm.nih.gov/pubmed/25430797

DOI: 10.3109/00498254.2014.986562   PMID: 25430797

1. In this study, the oxidative metabolites of 20(S)-protopanaxatriol (PPT) were identified in human liver microsomes (HLMs) and in rats using liquid chromatography-electrospray ionization tandem mass spectrometry. 2. Twelve oxidative metabolites were found in HLM, eight of which have not been previously reported. Twenty-four oxidative metabolites were found in rat feces after oral administration and 20 of these, including six found in HLM, were first reported. The results indicated PPT was more extensively metabolized in rats than in HLM. C20-24 epoxides, a pair of epimers (namely, M1-1 and M1-2) were the major metabolites, and other metabolites were derived from their further metabolism. 3. Enzyme kinetics experiments showed that the apparent formation Vmax of M1-1 was 10.4 folds and 2.4 folds higher than that of M1-2 in HLM and in rat liver microsomes (RLMs), respectively. The depletion rate of M1-2 was 11.0 folds faster than M1-1 in HLM, and was similar in RLM. Hence, the remarkable species differences of PPT metabolism mainly resulted from the stereoselective formation and further metabolic elimination of M1-1 and M1-2. 4. Chemical inhibition study and recombinant human P450 isoforms analysis showed that CYP3A4 was the predominant isoform involved in the oxidative metabolism of M1-1 and M1-2.

[4]
Ren Q W, Yang G Q, Guo M Q, Guo J W, Li Y, Lu J, Yang Q, Tang H H, Li Y, Fang X J, Sun Y X, Qi J G, Tian J W, Wang H B . Eur. J. Med. Chem., 2019,161:118. https://www.ncbi.nlm.nih.gov/pubmed/30347326

DOI: 10.1016/j.ejmech.2018.10.038   PMID: 30347326

Multidrug resistance (MDR) is a major cause of failure in cancer treatment, in which the overexpression of P-glycoprotein (Pgp) plays a crucial role. Herein, a novel class of ocotillol-type amide derivatives has been designed, synthesized, and evaluated for their ability to reverse MDR. The structure-activity relationship of the reversal activity was analyzed. Ten compounds showed promising chemo-reversal ability, among which the 24R-ocotillol-type amide derivative 6c with an N-Boc-hexanoyl unit exhibited the most potency in reversing paclitaxel resistance in KBV cells. Compound 6c could inhibit Pgp-mediated rhodamine123 efflux function via stimulating Pgp-ATPase activity and exhibited high binding affinity with Pgp in molecular docking studies. Importantly, compound 6c enhanced the efficacy of paclitaxel against KBV cancer cell-derived xenograft tumors in nude mice after oral administration. These results indicate that ocotillol-type amide derivatives are promising lead compounds for overcoming MDR in cancer.

[5]
Xu X F, Gao Y, Xu S Y, Liu H, Xue X, Zhang Y, Zhang H, Liu M N, Xiong H, Lin R C, Li X R . J. Ginseng Res., 2018,42:277. https://www.ncbi.nlm.nih.gov/pubmed/29983609

DOI: 10.1016/j.jgr.2017.02.003   PMID: 29983609

Temperature is an essential condition in red ginseng processing. The pharmacological activities of red ginseng under different steam temperatures are significantly different.

[6]
Liu Z Y, Zhang H Y, Bi Y, Liu X X, Lu J, Zhang X C, Xu J Y, Wang C Z, Yuan C S . Nat. Prod. Res., 2017,31:1523. https://www.ncbi.nlm.nih.gov/pubmed/28107791

DOI: 10.1080/14786419.2017.1280488   PMID: 28107791

To explore the antitumour mechanism of 20(S)-protopanaxadiol (PPD) while maintaining its uncovered pharmacological active site 3-hydroxyl, 28-hydroxy protopanaxadiol (17), a small molecular probe template of PPD was first designed and synthesised based on the Baldwin's reaction. Thus, 28-hydroxyl of 17 was built successfully as a derivatized site of molecular probe's functional and report groups. The important intermediates and final product were confirmed by ESI-MS and nuclear magnetic resonance spectra with good yield. These studies provided a valuable basis for probe research of PPD.

[7]
Liu F, Ma N, Xia F B, Li P, He C W, Wu Z Q, Wan J B . J. Ginseng Res., 2019,43:105. https://www.ncbi.nlm.nih.gov/pubmed/30662299

DOI: 10.1016/j.jgr.2017.09.003   PMID: 30662299

Ginsenosides with less sugar moieties may exhibit the better adsorptive capacity and more pharmacological activities.

[8]
Song Y Q, Zhao F, Zhang L M, Du Y, Wang T, Fu F H . Fitoterapia, 2013,91:173. https://www.ncbi.nlm.nih.gov/pubmed/24035860

DOI: 10.1016/j.fitote.2013.09.001   PMID: 24035860

Glucocorticoids (GCs) are usually used to treat inflammatory diseases. However, they cause severe and irreversible side effects, which limit the use of these compounds. Ginsenoside Rg1 had been demonstrated to possess anti-inflammatory and anti-cancer effects. The present study was designed to investigate whether Rg1 exhibits synergistic anti-inflammatory effects when combined with glucocorticoids. After stimulated by lipopolysaccharide (LPS), murine macrophagic RAW264.7 cells were treated with Rg1, corticosterone (Cort) or Rg1 and Cort. Then nitric oxide (NO), tumor necrosis factor-α (TNF-α) and glucocorticoid receptor (GR) expression were measured. The results showed that Rg1 or Cort could reduce the production of NO and TNF-α, and Rg1 dose-dependently up-regulated GR expression, while Cort dose-dependently down-regulated GR expression. The combination of low concentrations of Rg1 with Cort, which alone could not markedly inhibit the release of inflammatory factors, inhibited the secretion of NO and TNF-α in LPS-stimulated RAW264.7 macrophage cells, and up-regulated the expression of GR. The findings suggested Rg1 can synergize with glucocorticoid to enhance its anti-inflammatory effect.

[9]
杨刚强(Yang G Q), 李阳(Li Y), 杨青(Yang Q), 岳馨(Yue X), 姚雷(Yao L), 姜永涛(Jiang Y T) . 有机化学 (Chinese Journal of Organic Chemistry), 2017,37:1530.
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An K S, Choi Y O, Lee S M, Ryu H Y, Kang S J, Yeon Y, Kim Y R, Lee J G, Kim C J, Lee Y J, Kang B J, Choi J E, Song K S . Nutrients, 2019,11:1120.
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Zhang J Q, Zhang Q, Xu Y R, Li H X, Zhao F L, Wang C M, Liu Z, Liu P, Liu Y N, Meng Q G, Zhao F . Planta Med., 2019,85:292. https://www.ncbi.nlm.nih.gov/pubmed/30380571

DOI: 10.1055/a-0770-0994   PMID: 30380571

in vitro anti-inflammatory activity of C20 epimeric ocotillol-type triterpenes (2, 3, 9: , and 10: ) and protopanaxadiol [20(S/R)-protopanaxadiol] were investigated. Epimers 2: and 3: were prepared starting from 20(S)-protopanaxadiol. Epimers 9: and 10: were synthesized from 20(R)-3-acetylprotopanaxadiol (7: ). The anti-inflammatory activity of 2, 3, 9, 10: , 20(S)-protopanaxadiol, and 20(R)-protopanaxadiol was evaluated in cultured mouse macrophage RAW 264.7 cells. The MTT assay was used to measure the cytotoxicity. RAW 264.7 cells were stimulated by lipopolysaccharide to release the inflammatory mediators nitric oxide, prostaglandin E2, TNF-α, and interleukin-6 and anti-inflammatory mediator interleukin-10. The effect of the compounds on the overproduction of nitric oxide, prostaglandin E2, TNF-α, interleukin-6, and interleukin-10 was determined using Griess and ELISA methods. The results demonstrated that the in vitro anti-inflammatory activities of C20 epimeric ocotillol-type triterpenes and protopanaxadiol were different. Both the 20S-epimers (2: and 3: ) and 20R-epimers (9: and 10: ) inhibited the release of inflammatory mediator nitric oxide, while mainly the 20S-epimers inhibited the release of inflammatory mediator prostaglandin E2, and the 20R-epimers inhibited the release of inflammatory cytokine TNF-α. Both the 20S-epimers [2, 3: , and 20(S)-protopanaxadiol] and 20R-epimers [9, 10: , and 20(R)-protopanaxadiol] inhibited the release of inflammatory cytokine interleukin-6, but mainly the 20S-epimers [2, 3: , and 20(S)-protopanaxadiol] increased the release of anti-inflammatory mediator interleukin-10.]]>

[12]
Zhang L N, Wang S Y, Qu B Q, Chi H J, Quan Y L, Wu X H . J. Pharmaceut. Biomed., 2019,170:48.
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Chan T W D, But P P H, Cheng S W, Kwok M Y I, Lau F W, Xu H X . Anal. Chem., 2000,72:1281. https://www.ncbi.nlm.nih.gov/pubmed/10740871

DOI: 10.1021/ac990819z   PMID: 10740871

An LC/MS-based method is established for the differentiation and authentication of specimens and commercial samples of Panax ginseng (Oriental ginseng) and Panax quinquefolius (American ginseng). This method is based on the separation of ginsenosides present in the ginseng methanolic extracts using high-performance liquid chromatography (HPLC), followed by detection with electrospray mass spectrometry. Differentiation of ginsenosides is achieved through simultaneous detection of intact ginsenoside molecular ions and the ions of their characteristic thermal degradation products. An important parameter used for differentiating P. ginseng and P. quinquefolius is the presence of ginsenoside Rf and 24-(R)-pseudoginsenoside F11 in the RICs of Oriental and American ginsengs, respectively. It is important to stress that ginsenoside Rf and 24(R)-pseudoginsenoside F11, which possess the same molecular weight and were found to have similar retention times under most LC conditions, can be unambiguously distinguished in the present HPLC/MS method. The method developed is robust, reliable, reproducible, and highly sensitive down to the nanogram level.

[14]
Chen W, Balan P, Popovich D. G ., J. Ginseng Res., 2019, DOI: 10.1016/j.jgr.2019.04.007.
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Huang X, Liu Y, Zhang Y, Li S P, Yue H, Chen C B, Liu S Y . J. Ginseng Res., 2019,43:27. https://www.ncbi.nlm.nih.gov/pubmed/30662291

DOI: 10.1016/j.jgr.2017.08.001   PMID: 30662291

Panax quinquefolium L. under steaming were investigated, and the possible mechanisms were discussed.]]>

[16]
Liu J, Xu Y R, Yang J J, Wang W Z, Zhang J Q, Zhang R M, Meng Q G . J. Ginseng Res., 2017,41:373. https://www.ncbi.nlm.nih.gov/pubmed/28701880

DOI: 10.1016/j.jgr.2017.01.001   PMID: 28701880

Panax quinquefolius L., Panax japonicus, Hana mina, and Vietnamese ginseng. In recent years, the semisynthesis of 20(S/R)-ocotillol-type saponins has been reported. The biological activities of ocotillol-type saponins include neuroprotective effect, antimyocardial ischemia, antiinflammatory, antibacterial, and antitumor activities. Owing to their chemical structure, pharmacological actions, and the stereoselective activity on antimyocardial ischemia, ocotillol-type saponins are subjected to extensive consideration. In this review, we sum up the discovery, semisynthesis, biological activities, and metabolism of ocotillol-type saponins.]]>

[17]
Bi Y, Ma C, Zhou Z W, Zhang T T, Zhang H Y, Zhang X C, Lu J, Meng Q G, Lewis P J, Xu J Y . Rec. Nat. Prod., 2015,9:356.
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Bi Y, Yang J, Ma C, Liu Z Y, Zhang T T, Zhang X C, Lu J, Meng Q G . Pharmazie, 2015,70:213. https://www.ncbi.nlm.nih.gov/pubmed/26012249

PMID: 26012249

A series of novel ocotillol-type furoxan derivatives was synthesized by coupling various furoxans to 3-OH of 6-deoxy ocotillol, and their in vitro nitric oxide (NO) releasing capability was studied. The discharge of NO was examined after 30 min at two different concentrations, the results showed that all of the compounds tested could release NO in a dose-dependent manner. All of the synthesized compounds released similar amounts of NO at 100 μM, whereas at 500 μM these compounds showed more difference, in which compound II1, II3, II4, III2 displayed higher potency in releasing NO at this concentration. Analysis of the in vitro data showed that the derivatives bearing the same furoxan group on different ocotillol cores possessed various NO releasing capacity, suggesting that the structure of carrier of NO releasing groups may affect the NO release. Indeed, except compound II2, 24(S)-6-deoxy ocotillol derivatives from compound 6 with different furoxan substitutions at 3-OH and III2 displayed enhanced NO releasing capacity, compared to other compounds derived from compounds 5 and 9. The results illustrated that the functional group and the stereochemistry on the ocotillol structure may affect the NO release of furoxans.

[19]
Kim S J, Shin J Y, Ko S K . J. Ginseng Res., 2016,40:86. https://www.ncbi.nlm.nih.gov/pubmed/26843826

DOI: 10.1016/j.jgr.2015.04.008   PMID: 26843826

This study compared the contents of prosapogenin depending on the extracting conditions of Red ginseng to provide basic information for developing Red ginseng-based functional foods. The content of ginsenoside Rg3 reached their maximum value at 24 h of extraction, followed by 36 h and 72 h of extraction at 100°C.

[20]
Xia Y G, Liang J, Li G Y, Yang B Y, Kuang H X . J. Mass Spectrom., 2016,51:947. https://www.ncbi.nlm.nih.gov/pubmed/27383264

DOI: 10.1002/jms.3806   PMID: 27383264

- /ESI+ -FT-MS(1, 1) to MS(1, 4) spectra was constructed for the identification of structural elements in the MTSs. As a result, a total of 23 MTSs were discovered and tentatively identified, which had not been reported from Caulophyllum species before. All of these were potentially new compounds. This study provides an excellent example for discovery and identification of MTSs in herb medicines. Copyright © 2016 John Wiley & Sons, Ltd.]]>

[21]
Hu Y P, Cui X M, Zhang Z J, Chen L J, Zhang Y M, Wang C X, Yang X Y, Qu Y, Xiong Y . Molecules, 2018,23:1206.
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Zhong Z X, Li G K, Luo Z B, Zhu B H . Talanta, 2019,194:46. https://www.ncbi.nlm.nih.gov/pubmed/30609558

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-1 and 0.85-11.0 mg kg-1, respectively. The recoveries ranged from 90.2% to 106.1% with relative standard deviations of 0.30-3.1%. The method was successfully applied to the analysis of cosmetics, in which all the colorants could be quantified, and their median values ranged from 4.94 to 591 mg kg-1 for seventy-two lipsticks, and from 7.70 to 1.73 × 103 mg kg-1 for fifty eye shadows, respectively. The proposed protocol could achieve batch preparation of samples and avoid measurement errors from the obvious volume reduction of the recovered extract, and it could serve as a powerful tool for high-throughput analysis of multiple colorants in complex samples.]]>

[25]
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Ganzler K, Salgo A, Valko K . J. Chromatogr., 1986,371:299. https://www.ncbi.nlm.nih.gov/pubmed/3558551

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The applicability of microwave irradiation to the extraction of various types of compounds from soil, seeds, foods and feeds as a novel sample preparation method for chromatography was investigated. Samples were ground and mixed with an appropriate solvent, methanol or methanol-water for polar compounds and hexane for non-polar compounds. The suspensions were irradiated for 30 s, but they were not allowed to boil. After cooling, the irradiation was repeated several times. The samples were then centrifuged, and aliquots of the supernatant were injected into a chromatographic column. The yields of the extracted compounds obtained by microwave irradiation were compared with those obtained by the traditional Soxhlet or shake-flask extraction methods. The microwave extraction method was more effective than the conventional methods. Due to the considerable savings in time and energy, this novel method is suitable for fast extractions of large sample series.

[31]
Yao H, Li X, Liu Y, Wu Q, Jin Y . J. Ginseng Res., 2016,40:415. https://www.ncbi.nlm.nih.gov/pubmed/27746695

DOI: 10.1016/j.jgr.2016.06.007   PMID: 27746695

Panax quinquefolius L. have strong bioactivities. The fact that it is hard to obtain large amounts of rare ginsenosides seriously restricts further research on these compounds. An easy, fast, and efficient method to obtain different kinds of rare ginsenosides simultaneously and to quantify each one precisely is urgently needed.]]>

[32]
Yoon S H, Nam Y M, Hong J T, Kim S J, Ko S K . J. Ginseng Res., 2016,40:300. https://www.ncbi.nlm.nih.gov/pubmed/27616907

DOI: 10.1016/j.jgr.2015.09.001   PMID: 27616907

The result of USRG-12 indicated that ultrasonication-processed (100°C, 12 h) red ginseng extracts had the highest amount of ginsenosides Rg3 (0.803%), Rg5 (0.167%), and Rk1 (0.175%).

[33]
Biswas T, Ajayakumar P V, Mathur A K, Mathur A Nat . Prod. Res., 2015,29:1256. https://www.ncbi.nlm.nih.gov/pubmed/25813381

DOI: 10.1080/14786419.2015.1024119   PMID: 25813381

The present study aims at developing an extraction protocol for efficient ginsenoside recovery from cell suspensions of Panax quinquefolius and P. sikkimensis. Methanol (100%, 70% and 30%), water (40°C, 90°C), water-saturated butanol and butanol-saturated water were compared for their ultrasonication-assisted ginsenoside retrieval efficacy. HPLC and HP-TLC analysis revealed 100% methanol as the best solvent for maximum retrieval of Rb (diol) and Rg (triol) ginsenosides (P. quinquefolius: Rb: 0.189, Rg: 3.163 mg/g DW; P. sikkimensis: Rb: 0.245, Rg: 4.073 mg/g DW), followed by water (90°C). Methanolic solutions, especially 70%, proved to be significant retrievers of Rg1 (1.812 and 1.327 mg/g DW in P. quinquefolius and P. sikkimensis), with poor Re recovery (0.328 and 0.342 mg/g DW). Water-saturated butanol also led to significant ginsenoside extraction (72.4% of content extracted by methanol), selectively in P. quinquefolius, with a less than 50% of total content extracted by methanol, in P. sikkimensis.

[34]
Liu Z, Xia J, Wang C Z, Zhang J Q, Ruan C C, Sun G Z, Yuan C S . J. Agr. Food Chem., 2016,64:5389. https://www.ncbi.nlm.nih.gov/pubmed/27295137

DOI: 10.1021/acs.jafc.6b00963   PMID: 27295137

Panax ginseng contains many chemical components, including acidic ginsenosides and organic acids. However, whether these acidic substances play a role in ginsenoside transformation during steaming treatment has not yet been explored. In this paper, the content of neutral ginsenosides, acidic ginsenosides, and their degradation products in unsteamed and steamed P. ginseng were simultaneously quantified by high-performance liquid chromatography. We observed that neutral ginsenosides were converted to rare ginsenosides during the root steaming but not during the individual ginsenoside steaming. In contrast, acidic malonyl ginsenosides released malonic acid and acetic acid through demalonylation, decarboxylation, deacetylation reactions during the steaming at 120 °C. These malonyl ginsenosides not only were converted to rare ginsenosides but also promoted the degradation of neutral ginsenosides. Further studies indicated that a low concentration of organic acid was the determining factor for the ginsenoside conversion. The related mechanisms were deduced to be mainly acidic hydrolysis and dehydration. In summary, acidic ginsenosides and organic acids remarkably affected ginsenoside transformation during the steaming process. Our results provide useful information for precisely understanding the ginsenoside conversion pathways and mechanisms underlying the steaming process.

[35]
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[36]
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DOI: 10.1016/j.chroma.2016.12.068   PMID: 28027838

1, Rg5, Rs5, 20R-Rg3, and Rs4 exceeded 50.00%; indicating that the aforementioned chemical compounds have potential for further development. The results were validated by comparison with authentic standards, indicating that the method used in this research was accurate and advantageous for matrix analysis.]]>

[37]
Wang Y H, Li Y, Zhang Y, Feng G, Yang Z X, Guan Q X, Wang R, Han F J . Molecules, 2017,22:17.
[38]
Sunwoo H H, Gujral N, Huebl A C, Kim C T . Food Bioprocess Tech., 2014,7:1246. http://link.springer.com/10.1007/s11947-013-1234-1

DOI: 10.1007/s11947-013-1234-1

[39]
Zhong F L, Ma R, Jiang M L, Dong W W, Jiang J, Wu S Q, Li D H, Quan L H . J. Microbiol. Biotechnol., 2016,26:1661. https://www.ncbi.nlm.nih.gov/pubmed/27435543

DOI: 10.4014/jmb.1605.05052   PMID: 27435543

bgy2) was cloned from Lactobacillus brevis. We expressed this gene in Escherichia coli BL21(DE3), isolated the resulting protein, and then utilized the enzyme for the biotransformation of ginsenosides. The bgy2 gene contains 2,223 bp, and encodes a protein of 741 amino acids that is a member of glycosyl hydrolase family 3. β-Glucosidase (Bgy2) cleaved the outer glucose moieties of ginsenosides at the C-20 position, and the inner glucose at the C-3 position. Under optimal conditions (pH 7.0, 30°C), we used 0.1 mg/ml Bgy2 in 20 mM sodium phosphate buffer (PBS) for enzymatic studies. In these conditions, 1.0 mg/ml ginsenoside Rb1 and ginsenoside F2 were converted into 0.59 mg/ml ginsenoside Rd and 0.72mg/ml compound K, with molar conversion productivities of 69% and 91%, respectively. In pharmaceutical and commercial industries, this recombinant Bgy2 would be suitable for producting ginsenoside Rd and compound K.]]>

[40]
Liu F, Ma N, He C W, Hu Y J, Li P, Chen M W, Su H X, Wan J B . J. Ginseng Res., 2018,42:149. https://www.ncbi.nlm.nih.gov/pubmed/29719461

DOI: 10.1016/j.jgr.2017.01.007   PMID: 29719461

Panax notoginseng leaves (PNL) exhibit extensive activities, but few analytical methods have been established to exclusively determine the dammarane triterpene saponins in PNL.]]>

[41]
Wan J Y, Wang C Z, Liu Z, Zhang Q H, Musch M W, Bissonnette M, Chang E B, Li P, Qi L W, Yuan C S . J. Chromatogr. B, 2016,1015:62.
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Zheng H R, Chu Y, Zhou D Z, Ju A C, Li W, Li X, Xia Y, Polachi N, Li D K, Zhou S P, Sun H, Liu C X . J. Chromatogr. B, 2018,1072:282. https://www.ncbi.nlm.nih.gov/pubmed/29202359

DOI: 10.1016/j.jchromb.2017.10.056   PMID: 29202359

1, Rd, Re and Rh1, 2.5ng/mL for ginsenoside Rf, Rg3, Rb2 and Rb3 and 5.0ng/mL for ginsenoside Rb1 and Rc, respectively. All the precision (RSD) data ranged from 1.7-14.5% and the accuracy (RE) data was within ±13.73%. Moreover, the validated method has been applied to investigate the integrated pharmacokinetic profiles of ginsenosides in CHF rats following intravenous administration of YQFM successfully.]]>

[43]
Yang Q, Li J H, Wang X Y, Xiong H, Chen L X . Anal. Chem., 2019,91:6561. https://www.ncbi.nlm.nih.gov/pubmed/31010290

DOI: 10.1021/acs.analchem.9b00082   PMID: 31010290

A novel ternary-emission fluorescence sensor was proposed by post-imprinting mixing blue-/green-/red-emission bovine hemoglobin (BHb) imprinted polymers (b-MIPs, g-MIPs, and r-MIPs) at a proper ratio and realized the multiplexed and visual detection of BHb. The three MIPs were individually embedded with blue-emission 7-hydroxycoumarin, green-emission CdTe quantum dots (QDs), and red-emission CdTe/ZnS QDs. Upon interaction with BHb within 8 min, the fluorescence of CdTe and CdTe/ZnS QDs were simultaneously turned off, whereas the 7-hydroxycoumarin turned on the fluorescence intensity. Thereupon, the ratiometric fluorescence intensity of the ternary emission linearly varied within 0.025-3 μM BHb, accompanying the profuse fluorescence color evolution from yellowish green to yellow to salmon to plum to purple to finally blue. In comparison with the dual- or single-emission sensor, the ternary-emission fluorescence MIPs sensor provided a wider color variation covering the green-red-blue window for accurate naked-eye determination of BHb, as well as a lower detection limit down to 7.8 nM and a higher imprinting factor of 15.2. Moreover, the satisfactory recoveries of 99.25-111.7% in determining the spiked BHb in bovine urine samples, as well as the optical stability and post-imprinting construction convenience, indicated that the developed tricolor-emission fluorescence MIPs sensor provided an ideal alternative for rapid, sensitive, and visual determination of proteins in complicated samples.

[44]
明魏娜(Ming W N), 王晓艳(Wang X Y), 明永飞(Ming Y F), 李金花(Li J H), 陈令新(Chen L X) . 化学进展 (Progress in Chemistry), 2016,28:552.
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DOI: 10.1016/j.jcis.2019.01.081   PMID: 30710820

Green ion imprinted polymers (IIPs) were prepared in aqueous phase via the synergy of three functional monomers of low-cost eco-friendly gelatin (G), 8-hydroxyquinoline (HQ) and chitosan (C), namely G-HQ-C IIPs, and were applied as an effective and recyclable adsorbent to remove Cu(II) from aqueous solution. The as-prepared G-HQ-C IIPs were systematically characterized, and several major factors affecting adsorption capacity including solution pH, temperature and contact time were investigated in detail. The adsorption of Cu(II) on G-HQ-C IIPs followed the pseudo-second-order kinetic and Langmuir isotherm models, and the adsorption capacity increased with temperature increase. Moreover, the maximum adsorption capacities of G-HQ-C IIPs toward Cu(II) reached up to 111.81 mg/g at room temperature, much higher than those of most of the reported adsorbents for Cu(II). The G-HQ-C IIPs displayed excellent selectivity against seven common divalent ions with selectivity coefficients above 18.71, as well as high anti-interference ability. Additionally, a good reusability was demonstrated without significant loss in adsorption capacity after at least ten cycles. The IIPs were applied to environmental water samples for selective removal of Cu(II) with satisfactory results. By replacing Cu(II) template by Cd(II), Hg(II) and Pb(II), respectively, the obtained three kinds of IIPs based on G-HQ-C presented convincing imprinting properties, and therefore the work could provide a simple and general imprinting strategy toward various concerned heavy metal ions through multi-point interactions from multiple functional monomers.

[46]
Xing R R, Wen Y R, Dong Y R, Wang Y J, Zhang Q, Liu Z . Anal. Chem., 2019,91:9993. https://www.ncbi.nlm.nih.gov/pubmed/31347834

DOI: 10.1021/acs.analchem.9b01826   PMID: 31347834

2 nanoparticles were prepared as nanotags for the specific labeling of captured protein. The formed MIP-protein-MIP sandwich-like complexes could produce a significantly enhanced SERS signal. The dual MIP-based recognitions ensured high specificity of the assay, while SERS detection provided ultrahigh sensitivity. The duMIP-PISA of neuron-specific enolase (NSE) in human serums was demonstrated, which permitted the differentiation of small cell lung cancer patients from healthy individuals. As compared to regular ELISA, the duMIP-PISA exhibited multiple merits including a simpler procedure, faster speed, lower sample volume requirement, and wider linear range. The approach well demonstrated the great potentials of MIPs and can be easily modified and extended to other protein biomarkers. Therefore, the duMIP-PISA approach holds great promise in many important applications such as disease diagnosis.]]>

[47]
BelBruno J J . Chem. Rev., 2019,119:94. https://www.ncbi.nlm.nih.gov/pubmed/30246529

DOI: 10.1021/acs.chemrev.8b00171   PMID: 30246529

Molecularly imprinted polymers are synthetic receptors for a targeted molecule. As such, they are analogues of the natural antibody-antigen systems. In this review, after a recounting of the early history of the general field, we specifically focus on the application of these polymers as sensors. In these applications, the polymers are paired with a reporting system, which may be electrical, electrochemical, optical, or gravimetric. The presence of the targeted molecule effects a change in the reporting agent, and a calibrated quantity of the target is recorded. In this review, we describe the imprinted polymer production processes, the techniques used for reporting, and the applications of the reported sensors. A brief survey of recent applications to gas-phase sensing is included, but the focus is primarily on the development of sensors for targets in solution. Included among the applications are those designed to detect toxic chemicals, toxins in foods, drugs, explosives, and pathogens. The application of computational chemistry to the development of new imprinted polymers is included as is a brief assessment of future developments.

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18 solid-phase extraction column. Overall, a new, innovative method was developed to efficiently enrich high-polarity bioactive molecules present at low concentrations in complex matrices.]]>

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In the present work, an advanced pretreatment method magnetic molecular imprinted polymers-dispersive solid phase extraction (MMIPs-DSPE) combined with the high sensitivity LTQ-Orbitrap mass spectrometry was applied to the complicated metabolites analysis of Traditional Chinese Medicines (TCMs) in complex matrices. The ginsenoside Rb1 magnetic molecular imprinted polymers (Rb1-MMIPs) were successfully synthesized for specific recognition and selective enrichment of Panax notoginseng saponin metabolites in rat faeces. The polymers were prepared by using Fe3O4@SiO2 as the supporting material, APTES as the functional monomer and TEOS as the cross-linker. The Rb1-MMIPs showed quick separation (10.8 emu/g), large adsorption capacity (636μmol/g), high selectivity and fast binding kinetics (25min). Dispersion solid-phase extraction using Rb1-MMIPs (Rb1-MMIPs-DSPE) integrated with LTQ-Orbitrap MS was applied to fish out and identify saponin metabolites from rat faeces, and totally 58 related compounds were detected within 20min, including 26 PPD-group and 32 PPT-group notoginsenoside metabolites. Parallel tests showed that Rb1-MMIPs-DSPE obtained the lowest matrix effects of 0.98-14.84% and captured the largest number of structural analogues compared with traditional pretreatment methods organic solvent extraction (OSE) and solid phase extraction (SPE).

[53]
李健(Li J), 官亦标(Guan Y B), 傅凯(Fu K), 苏岳锋(Su Y F), 包丽颖(Bao L Y), 吴锋(Wu F) . 化学进展( Progress in Chemistry), 2014,26:1233. 10b2a9eb-4495-4417-9d72-ab630532eba9 http://www.progchem.ac.cn//CN/abstract/abstract11392.shtml

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当今社会日益增长的能源与环境需求对储能电池技术的发展既是机遇也是严峻的挑战。纳米碳材料如碳纳米管与石墨烯因其优异的导电能力、良好的机械性能以及独特的形貌与结构特征在储能电池技术领域中的应用越来越普遍。本文通过综述近年来碳纳米管与石墨烯分别作为锂离子电池的复合电极材料、负极活性材料、导电添加剂以及新型锂硫电池用复合导电载体的最新应用进展,重点讨论了这两类纳米碳材料的不同应用模式对储能电池容量性能、倍率性能以及循环寿命的影响。同时对目前研究中存在的问题进行了总结,并对未来发展方向,如开发低成本与环境友好的高质量材料合成技术、提升材料的分散能力以有效构筑复合电极结构以及开发新的应用模式等进行了展望。

[54]
韩强(Han Q), 王宗花(Wang Z), 张晓琼(Zhang X), 丁明玉(Ding M) . 化学进展( Progress in Chemistry), 2014,26:820. 758efe2b-7014-4c6e-a098-8e292f8a8cf0 http://www.progchem.ac.cn//CN/abstract/abstract11356.shtml

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样品前处理新技术与方法研究是现代分析化学的重要研究课题与发展方向之一。固相萃取是目前应用最为广泛的样品前处理技术,其技术核心是吸附材料,因此开发新型吸附材料是样品前处理领域的研究热点。石墨烯是一种新型碳纳米材料,由于其良好的物理化学性质,在短短几年内迅速成为众多学科的研究热点。其高比表面积、良好的化学稳定性和热稳定性使之在分离科学领域得到广泛的应用。本文系统综述了石墨烯及其复合材料在样品前处理中的应用研究,主要包括其作为固定相在固相萃取、固相微萃取、磁固相萃取等技术在环境、食品、生物等样品前处理中的应用。

[55]
苗瑞(Miao R), 吴冬雪(Wu D X), 王秋颖(Wang Q Y), 赵幻希(Zhao H X), 李雪(Li X), 修洋(Xiu Y), 刘淑莹(Liu S Y) . 高等学校化学学报 (Chemical Journal of Chinese Universities), 2018,39:2178.
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Panax ginseng (GRR). This study was carried out to qualitatively and quantitatively determine the ginsenosides in the cultivated and forest GRR.]]>

[59]
Du Z X, Li J H, Zhang X, Pei J, Huang L F . Molecules, 2018,23:20.
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摘要

r>0.995) within the determined ranges. Both intra-day and inter-day variances were less than 15%, and the accuracy was within 80-120%. The excretion recoveries of Rg1, ginsenoside Rh1 (Rh1), and protopanaxatriol (Ppt) in bile, urine, and feces combined were all greater than 70%. The fecal excretion recoveries of Rg1, Rh1, and Ppt were 40.11%, 22.19%, and 22.88%, respectively, whereas 6.88% of Rg1 and 0.09% of Rh1 were excreted in bile. Urinary excretion accounted for only 0.04% of Rg1. In conclusion, the observed excretion profiles for Rg1 and its metabolites after oral administration are helpful for understanding the poor oral bioavailability of Rg1 and will aid further investigations of Rg1 as a pharmacologically active component.]]>

[62]
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Cobalt nitrate, nickel sulphate, hydrogen peroxide, sodium nitroprusside, and culture filtrates of Pseudomonas monteili, Bacillus circularans, Trichoderma atroviridae, and Trichoderma harzianum were tested to elicit ginsenoside production in a cell suspension line of Panax quinquefolius. Abiotic elicitors preferentially increased panaxadiols whereas biotic elicitors upregulated the panaxatriol synthesis. Cobalt nitrate (50 μM) increased total ginsenosides content by twofold (54.3 mg/L) within 5 days. It also induced the Rc synthesis that was absent in the control cultures. Elicitation with P. monteili (2.5 % v/v, 5 days) also supported 2.4-fold enhancement in saponin yield. Elicitation by T. atroviridae or hydrogen peroxide induced the synthesis of Rg3 and Rh2 that are absent in ginseng roots. The highest ginsenosides productivity (3.2-fold of control) was noticed in cells exposed to 1.25 % v/v dose of T. atroviridae for 5 days. Treating cells with T. harzianum for 15 days afforded maximum synthesis and leaching (8.1 mg/L) of ginsenoside Rh1.

[65]
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3 catalysis, the reaction conditions are similar to enzymatic reaction conditions. In FeCl3 catalysis, the only 20-O-sugar-moiety of ginsenoside Rb1 was decomposed into the minor ginsenosides Rk1 and Rg5 with newly produced C-20 ethylene bands; but also hydrolyzed into 20(S)-Rg3 and 20(R)-Rg3; subsequently the C-24(25) ethylene bands of 20(S)-Rg3 and 20(R)-Rg3 were hydrated to 20(S)-25-OH-Rg3 and 20(R)-25-OH-Rg3. After separation of reaction mixture from 34 g ginsenoside-Rb1 by silica-gel-column, the 3.3 g sample I of TLC top-band consisting of Rg5 and Rk1, 8.7 g sample II of TLC middle-band consisting of 20(S)-Rg3 and 20(R)-Rg3, 3.5 g sample III of TLC bottom-band consisting of unknown product-I and -II including 20(S)-25-OH-Rg3, were obtained. The sample III consisting of unknown product-I and -II was purified by crystallization, and identified to 20(S)-25-OH-Rg3 and 20(R)-25-OH-Rg3 by HPLC-Evaporative Light Scattering Detector (ELSD) and NMR. Therefore, six types of minor-ginsenosides Rk1, Rg5, 20(S)-Rg3, 20(R)-Rg3, 20(S)-25-OH-Rg3 and 20(R)-25-OH-Rg3 were successfully prepared from ginsenoside Rb1 by FeCl3 catalysis. FeCl3 has low toxicity and is inexpensive, and the reaction conditions are similar to enzymatic reaction conditions; thus, this method is applicable to the development of ginseng-based drugs.]]>

[69]
Guo C, Li D M, Liu C M, Guo Z P, Chen Y . Anal. Bioanal. Chem., 2018,410:4293. https://www.ncbi.nlm.nih.gov/pubmed/29748756

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2, cholic acid, and chenodeoxycholic acid in saliva. The method consists of two successive steps: fast and direct labeling of the target analytes with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide followed by ultrahigh-performance liquid chromatography-tandem mass spectrometry analysis. The method exhibited a wide linear range from 2.5 to 2500 pg/mL, with linear coefficients greater than 0.9963 and limits of detection and quantification as low as 0.10 and 0.33 pg/mL, respectively. The method precision was evaluated, with relative standard deviations ranging from 2.12% to 10.63% for intraday assays and from 2.98% to 12.88% for interday assays. The recoveries were measured by our spiking saliva samples with standards at three different levels, and ranged from 72.5% to 98.0%. Real applicability was validated by direct quantification of trace target analytes in human saliva, with simple pretreatment, use of a small sample volume, and a short analysis time. Graphical abstract Sequential steps to extract, label, and determine the ultratrace carboxylic acids in saliva. CDCA chenodeoxycholic acid, γ-CEHC 2-(β-carboxyethyl)-6-hydroxy-2,7,8-trimethylchroman, α-LA α-lipoic acid, PGE2 prostaglandin E2, UHPLC-MS/MS ultrahigh-performance liquid chromatography-tandem mass spectrometry.]]>

[70]
张俊杰(Zhang J J), 贾金萍(Jia J P), 秦雪梅(Qin X M) . 分析测试学报 (Journal of Instrumental Analysis), 2017,36:579.
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Panax ginseng has received much attention as a valuable health supplement with medicinal potential. Its chemical diversity and multiple pharmacological properties call for comprehensive methods to better understand the effects of ginseng and ginsenosides. Liquid chromatography-mass spectrometry (LC-MS) based metabonomic approaches just fit the purpose.

[73]
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Panax quinquefolius (PQ) is one of the best-selling natural health products due to its proposed beneficial anti-aging, anti-cancer, anti-stress, anti-fatigue, and anxiolytic effects. In recent years, the quality of PQ has received considerable attention. Sensitive and accurate methods for qualitative and quantitative analyses of chemical constituents are necessary for the comprehensive quality control to ensure the safety and efficacy of PQ. This article reviews recent progress in the chemical analysis of PQ and its preparations. Numerous analytical techniques, including spectroscopy, thin-layer chromatography (TLC), gas chromatography (GC), high-performance liquid chromatography (HPLC), liquid chromatography/mass spectrometry (LC/MS), high-speed centrifugal partition chromatography (HSCPC), high-performance counter-current chromatography (HPCCC), nuclear magnetic resonance spectroscopy (NMR), and immunoassay, are described. Among these techniques, HPLC coupled with mass spectrometry (MS) is the most promising method for quality control. The challenges encountered in the chemical analysis of PQ are also briefly discussed, and the remaining questions regarding the quality control of PQ that require further investigation are highlighted.

[74]
Liu J, Liu Y, Wang Y, Abozeid A, Zu Y G, Tang Z H . J. Pharmaceut. Biomed., 2017,135:176. https://linkinghub.elsevier.com/retrieve/pii/S0731708516314613

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i = 883 nM was discovered. This compound exhibited low cell toxicity and was able to selectively inhibit shedding of known ADAM10 substrates in several cell-based models. We hypothesize that differential glycosylation of these cognate substrates is the source of selectivity of our novel inhibitor. The data indicate that this novel inhibitor can be used as an in vitro and, potentially, in vivo, probe of ADAM10 activity. Additionally, results of the present and prior studies strongly suggest that glycosylated substrate are applicable as screening agents for discovery of selective ADAM probes and therapeutics.]]>

[77]
Liu J, Liu Y, Wang Y, Abozeid A, Zu Y G, Zhang X N, Tang Z H . Molecules, 2017,22:14.
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Park S E, Seo S H, Lee K I, Na C S and Son H S J . Ginseng Res., 2018,42:57. https://www.ncbi.nlm.nih.gov/pubmed/29348723

DOI: 10.1016/j.jgr.2016.12.010   PMID: 29348723

Ginseng contains many small metabolites such as amino acids, fatty acids, carbohydrates, and ginsenosides. However, little is known about the relationships between microorganisms and metabolites during the entire ginseng fermentation process. We investigated metabolic changes during ginseng fermentation according to the inoculation of food-compatible microorganisms.

[79]
Sun Y F, Chen S Q, Wei R M, Xie X, Wang C C, Fan S H, Zhang X, Su J, Liu J, Jia W, Wang X Y . Food Funct., 2018,9:3547. https://www.ncbi.nlm.nih.gov/pubmed/29896600

DOI: 10.1039/c8fo00025e   PMID: 29896600

Ginseng, a widely used functional food and food additive, has been proven to have promotion effects of health on the body. However, whether the long-term intake of Ginseng is beneficial or has side effects on an organism is still unclear. In this study, untargeted GC-TOFMS metabolomic analysis of serum, cecum and ileum intestinal contents was conducted to understand the effect of the long-term intake of Ginseng extracts. 16S rRNA microbial sequencing technology was applied to investigate the effect of Ginseng extracts on the structure of gut microbiota. Cytokines in spleen were detected to determine the effect of Ginseng extracts on the immune system. Compared to control groups, the metabolites in serum, cecum and ileum, such as amino acids, amines and other metabolites related to carbohydrate metabolism, significantly varied between the C and GS groups. Ginseng extracts affected the structure of gut microbiota with a decreased abundance of TM7, while the abundance of Proteobacteria, Methylobacteriaceae, Parasutterella, Sutterella increased in the GS group. The increased abundance of Bifidobacterium and Lactobacillus demonstrated that Ginseng extracts contribute to probiotic amplification. Highly correlated with Bifidobacterium and Lactobacillus, interleukin 4 (IL4), IL10 and immunoglobulin A (IgA) levels were significantly elevated after the long-term intake of Ginseng extracts. These results indicated that the long-term administration of Ginseng extracts positively affected the host-gut metabolism, immune system, the anti-inflammation process and the gut intestinal microbiota structure.

[80]
Chang K H, Jo M N, Kim K T, Paik H D . J. Ginseng Res., 2014,38:47. c83995f3-6a1e-4331-83a2-5319de937469 https://www.ncbi.nlm.nih.gov/pubmed/24558310

DOI: 10.1016/j.jgr.2013.11.008   PMID: 24558310

The transformation of ginsenoside Rb1 into a specific minor ginsenoside using Aspergillus niger KCCM 11239, as well as the identification of the transformed products and the pathway via thin layer chromatography and high performance liquid chromatography were evaluated to develop a new biologically active material. The conversion of ginsenoside Rb1 generated Rd, Rg3, Rh2, and compound K although the reaction rates were low due to the low concentration. In enzymatic conversion, all of the ginsenoside Rb1 was converted to ginsenoside Rd and ginsenoside Rg3 after 24 h of incubation. The crude enzyme (β-glucosidase) from A. niger KCCM 11239 hydrolyzed the β-(1→6)-glucosidic linkage at the C-20 of ginsenoside Rb1 to generate ginsenoside Rd and ginsenoside Rg3. Our experimental demonstration showing that A. niger KCCM 11239 produces the ginsenoside-hydrolyzing β-glucosidase reflects the feasibility of developing a specific bioconversion process to obtain active minor ginsenosides.

[81]
Yan X, Zhao Y, Zhang Y, Qu H H . Molecules, 2017,22:29.
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Bai H R, Wang S J, Liu J J, Gao D, Jiang Y Y, Liu H X, Cai Z W . J. Chromatogr. B, 2016,1026:263. https://www.ncbi.nlm.nih.gov/pubmed/26520809

DOI: 10.1016/j.jchromb.2015.09.024   PMID: 26520809

The root of Panax ginseng C.A. Mey. (P. ginseng) is one of the most popular traditional Chinese medicines, with ginsenosides as its main bioactive components. Because different ginsenosides have varied pharmacological effects, extraction and separation of ginsenosides are usually required for the investigation of pharmacological effects of different ginsenosides. However, the contents of ginsenosides vary with the ages and tissues of P. ginseng root. In this research, an efficient method to explore the distribution of ginsenosides and differentiate P. ginseng roots with different ages was developed based on matrix assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-TOF-MSI). After a simple sample preparation, there were 18 peaks corresponding to 31 ginsenosides with distinct localization in the mass range of m/z 700-1400 identified by MALDI-TOF-MSI and MALDI-TOF-MS/MS. All the three types of ginsenosides were successfully detected and visualized in images, which could be correlated with anatomical features. The P. ginseng at the ages of 2, 4 and 6 could be differentiated finely through the principal component analysis of data collected from the cork based on the ion images but not data from the whole tissue. The experimental result implies that the established method for the direct analysis of metabolites in plant tissues has high potential for the rapid identification of metabolites and analysis of their localizations in medicinal herbs. Furthermore, this technique also provides valuable information for the component-specific extraction and pharmacological research of herbs.

[83]
Buchberger A R, DeLaney K, Johnson J, Li L J . Anal. Chem., 2018,90:240. https://www.ncbi.nlm.nih.gov/pubmed/29155564

DOI: 10.1021/acs.analchem.7b04733   PMID: 29155564

/