Applications of liquid chromatography in the quality control of traditional Chinese medicines: An overview

CHAPTER

Applications of liquid chromatography in the quality control of traditional Chinese medicines: An overview

20

Shing-Chung Lama,a, Zong-Lin Yanga, Jing Zhao, Shao-Ping Li University of Macau, Macao, China

­C HAPTER OUTLINE 20.1 Introduction..................................................................................................... 618 20.2  Separation Modes of Liquid Chromatography..................................................... 619 20.2.1  Reversed-Phase Liquid Chromatography........................................ 619 20.2.2  Hydrophilic Interaction Liquid Chromatography.............................. 631 20.2.3  Ion-Exchange Chromatography..................................................... 632 20.2.4  Size Exclusion Chromatography.................................................... 632 20.2.5  Two-Dimensional Liquid Chromatography (2DLC)........................... 636 20.2.6 Miscellaneous............................................................................. 639 20.3 Detections....................................................................................................... 639 20.3.1  UV-Vis Detection......................................................................... 639 20.3.2  Nonspectrometry Detection (RID, ELSD, CAD, and ECD)................. 643 20.3.3  Multiangle Laser Light Scattering (MALLS) Detection..................... 644 20.3.4  Mass Spectrometry (MS) Detection............................................... 644 20.3.5  Biochemical Detection (BCD)....................................................... 646 20.3.6 Miscellaneous............................................................................. 646 20.4  Concluding Remarks........................................................................................ 648 References............................................................................................................... 648

­ABBREVIATIONS 1DLC 2DLC AEC a

one-dimensional liquid chromatography two-dimensional liquid chromatography anion-exchange chromatography

The authors contributed this work equally.

Liquid Chromatography. http://dx.doi.org/10.1016/B978-0-12-805392-8.00020-7 © 2017 Elsevier Inc. All rights reserved.

617

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CHAPTER 20  Quality control of traditional Chinese medicines

APCI atmospheric pressure chemical ionization BCD biochemical detection CAD charged aerosol detector DAD diode array detector dn/dc differential refractive index increment ECD electrochemical detection ELSD evaporative light scattering detector FLD fluorescence detector FPLC fast protein liquid chromatography FT Fourier transform HILIC hydrophilic interaction liquid chromatography HPLC high-performance liquid chromatography HSCCC high-speed countercurrent chromatography IEC ion-exchange chromatography IT ion trap LC liquid chromatography LTQ linear triple quadrupole MALDI matrix-assisted laser desorption ionization MALLS multiangle laser light scattering MS mass spectrometry NMR nucleic magnetic resonance NPLC normal-phase liquid chromatography PAD pulsed amperometric detector PDA photodiode array detector pMRM predictive multiple reaction monitoring Q quadrupole RID refractive index detector RPLC reversed-phase liquid chromatography SEC size exclusion chromatography SFC supercritical fluid chromatography TCM traditional Chinese medicine TOF time of flight UHPLC ultrahigh-performance liquid chromatography UPLC ultra-performance liquid chromatography UV-Vis ultraviolet-visible VIS viscometer VWD variable wavelength detector

20.1 ­INTRODUCTION Traditional Chinese medicines (TCMs) have been widely used to prevent and treat human diseases under the guidance of traditional Chinese medical theories [1]. Known for its long medicinal history and reliable curative effects, TCMs are attracting increasing attention and gradually acquiring acceptance worldwide [2]. However, TCMs are complicated systems reflected in hundreds of components in one herb and having various therapeutic uses, unlike the single-component and definite efficacy of Western medicines. As the demands and uses for TCM products are growing rapidly, there are also

20.2 ­ Separation modes of liquid chromatography

increasing reports on their adverse effects, which mainly comes from uncertain recognitions of the amounts and types of phytochemicals in TCMs [3]. Therefore, identification and determination of the type and amount of active and/or toxic compounds is beneficial for an understanding of the bioactivity and the potential adverse effects of TCMs. Indeed, the quality control of TCMs is crucial for ensuring their safety and efficacy [4]. Liquid chromatography (LC) is considered as the most powerful analytical technique for the quality control of TCMs [5,6]. Several LC techniques according to their separation mechanisms, such as reversed-phase liquid chromatography (RPLC), normal-phase liquid chromatography (NPLC), hydrophilic interaction liquid chromatography (HILIC), ion-exchange chromatography (IEC), size exclusion chromatography (SEC), and other related techniques, have been widely applied to the quality control of multiple components in TCMs [5–8]. After LC separation, the detector is the basic unit for the whole analysis procedure because it can convert the components to chromatographic peaks [9]. The innovation of detectors or newly applied hyphenations shortens the analysis time, improves the resolution, and enhances the sensitivity for the analysis of TCM constituents. LC applications in this field between 2010 and 2016 have been summarized and discussed in this chapter, which will help to illustrate the current status and perspectives of LC in the quality control of TCMs.

20.2 ­SEPARATION MODES OF LIQUID CHROMATOGRAPHY 20.2.1 ­ REVERSED-PHASE LIQUID CHROMATOGRAPHY RPLC is one of the most adoptive separation modes in the quality control of TCMs, due to its broad adaptability and high separation efficiency toward analytes of diverse polarity, as well as superior compatibility with hyphenated detectors [10,11]. At present, octadecyl-bonded silica (C18) is considered as the most used stationary phase for the separation of small-molecule organic compounds in TCMs, such as alkaloids [12–14], flavonoids [15–22], phenolic acids [23–25], triterpenoids [26–29], and saponins [30–36]. There are several factors that contribute to separation efficiency, such as the particle size and the mobile phase. The application of smaller particle size columns can significantly reduce the analysis time as well as improve the resolution and peak capacity [37]. The general particle sizes of commercial chromatographic columns range from 1.7 to 5 μm, in which 5 μm are classified as HPLC, whereas those smaller than 5 μm are considered as UHPLC (usually 2–3.5 μm i.d.) and UPLC (usually < 2 μm i.d.) [38]. Different particle size columns (1.8 and 5 μm) were compared by separating 5 different bile acid derivatives in Calculus bovis, and the results have shown that retention times of the analytes passed through the 1.8 μm column were reduced by 2–3 times [39]. Besides, 1.8-μm and 3.5-μm chromatographic columns were compared in the quality control of 5 different mogrosides in Siraitia grosvenorii, and the results demonstrated that smaller particle size columns (1.8 μm) showed better resolutions and higher capacities [33]. The elementary mobile phases for RPLC consist of organic (mostly methanol or acetonitrile) and aqueous (AQ) phases (water). However, aiming at better separation efficiency for some specific

619

620

CHAPTER 20  Quality control of traditional Chinese medicines

compounds, such as phenolic acids and alkaloids, the conditioning agents are added to adjust the peak shapes and improve the resolutions of chromatograms, mainly including weak organic acids (such as formic acid), weak alkali (such as ammonia), or weak-acid weak-base salts (such as ammonium formate) [40]. For the analysis of organic acids, such as phenolic acids, flavonoids and triterpenic acids, acidic mobile phases are mostly applied in chromatographic conditions [24,27,41,42]. In addition, acidic phases with buffer salt solutions were prevalent in the analysis of alkaloids, such as ammonium formate, monobasic potassium phosphate, and so on [43]. It was reported that AQ phase with monobasic potassium phosphate and sodium dodecyl sulfate was used for the analysis of several alkaloids in Coptis chinensis [12]. Besides, ion-pairing reagents also served as the conditioning agents to retain the hydrophilic components in RP18 columns [44]. Previously, pentadecafluorooctanoic acid (PDFOA), which was a long n-alkyl chain perfluorinated carboxylic acid, was used to serve as ion-pairing agents for the qualitative and quantitative analyses of nucleosides and nucleotides in Cordyceps species [45]. PDFOA is volatile so that it could be compatibly used for mass spectrometry (MS) detection and for diminishing background interferences [45]. RPLC coupled to several detectors, such as the ultraviolet-visible detector (UV-Vis), diode array detector (DAD), evaporative light scattering detector (ELSD), charged aerosol detector (CAD), and MS, have been widely applied to the quality control of TCMs [46–48]. However, conventional C18 stationary phases are not suitable when a highly AQ condition is required for the analysis of high-polar components. The AQ ratio of the mobile phase of less than 5% might lead to column collapse or dewetting of C18 stationary phases. Therefore, in order to solve this problem, the surfaces of stationary phases are polar modified using the endcapping reagents or embedded with polar functional groups (such as amide and carbamate groups) [49]. Among the commercially available columns, AQ columns are widely used for the analysis of hydrophilic components in TCMs [45,50,51]. Compared with conventional C18 chromatographic columns, AQ columns are broadening separation ranges of high-polar components, and improve retention of hydrophilic components, as well as protect the columns against highly AQ conditions [49]. Han et  al. established an HPLC fingerprint to separate 22 marker compounds and quantify 8 hydrophilic components of them in Polygonum multiflorum [50]. The SB-Aq column (50 × 4.6 mm, 3.5 μm) was selected to perform HPLC fingerprint and was proved to retain hydrophilic compounds better than the SB-C18 column. Another example is the quality control of 5 bioactive saponins in Panax notoginseng, which compared a Zorbax SB-Aq column (50 × 4.6 mm, 3.5 μm i.d.) with other commercial C18 columns [51]. It is easier to reach baseline separation for SB-Aq column than other commercial C18 columns under optimized conditions. Besides, RP-HPLC separation based on SB-Aq column (250 × 4.6 mm, 5 μm i.d.) and volatile ion-pairing reagent as AQ mobile phase was applied in the quality assessment of nucleotides, nucleosides, and their transformation products in Cordyceps [45]. It requires high AQ conditions maintained for a long time so that the hydrophilic components could retain in the AQ column. The applications of RPLC in the quality control of TCMs are listed in Table 20.1.

Table 20.1  Applications of 1DLC (RPLC, HILIC, and IEC) in the Analysis of Small Molecules From TCMs Source

Analytes

Columns

Detections

References

6 TCMs Acorus tatarinowii Schott (Shi Chang Pu) Alismatis Rhizoma (Ze Xie)

6 Triterpenic acids 7 Methoxybenzene types

FLD; MS: APCI (+) MS: ESI (+, −)

[28] [52]

MS: ESI (+)

[29]

Allii Macrostemonis Bulbus (Xie Bai)

15 Components, including steroid glycosides, saponins, and nucleotides 4 Cholic acids

Hypersil C18 column (4.6 × 200 mm, 5 μm) Poroshell 120 EC C18 column (2.1 × 100 mm, 2.7 μm) 1.  Cortecs C18 column (2.1 × 100 mm, 1.6 μm); 2.  Ultimate XB-C18 column (4.6 × 150 mm, 5 μm) BEH C18 column (2.1 × 50 mm, 1.7 μm)

MS: ESI (+, −)

[53]

Zorbax SB-C18 column (4.6 × 250 mm, 5 μm) HSS C18 column (2.1 × 100 mm, 1.8 μm)

DAD

[34]

MS: ESI (+)

[54]

BEH C18 column (2.1 × 100 mm, 1.7 μm)

MS: ESI (+, −)

[55]

Phenomenex C18 column (2.1 × 100 mm, 2.6 μm) Spursil C18 column (4.6 × 250 mm, 5 μm)

PAD

[56]

PDA; ELSD

[57]

BEH C18 column (2.1 × 50 mm, 1.7 μm)

ELSD

[39]

RPLC

Bu Fei Granule

Bupeuri radix (Chai Hu), Chaihu-Shugan-San Bu-Zhong-Yi-Qi-Wan

Calculus Bovis (Niu Huang)

18 Components, including aristolochic acids, alkamides, essential oils, flavonoids, and lignans 10 Components, including flavonoids, phenylpropanoids, and triterpenoids 4 Saikosaponins 10 Components, including flavonoids, lactones, and triterpenoids 7 Bile acids

20.2 ­ Separation modes of liquid chromatography

Artificial Calculus bovis (Ren Gong Niu Huang) Asari Radix et Rhizoma (Xi Xin)

14 Triterpenoids

Continued

621

622

Source

Analytes

Columns

Detections

References

Caulis Trachelospermi (Luo Shi Teng) Codonopsis Radix (Dang Shen)

14 Components, including flavonoids and lignans 7 Components, including polyacetylenes, phenylpropanoids, and pyrrolidine alkaloids 4 Alkaloids

Eclipse plus C18 column (4.6 × 150 mm, 5 μm) YMC-Pack Pro-C18 column (4.6 × 250 mm, 5 μm)

UV, MS: ESI (+)

[58]

DAD

[59]

Extend C18 column (4.6 × 250 mm, 5 μm)

UV

[12]

16 Components, including nucleotides, nucleosides, and Cordycepin 12 Components, including alkaloids, lactones, and phenolic acids, 3 Flavonoids

Zorbax SB-Aq column (4.6 × 250 mm, 5 μm)

DAD, MS: ESI (+)

[45]

BEH C18 column (2.1 × 100 mm, 1.7 μm)

PDA; MS: ESI (+, −)

[60]

Diamonsil C18 column (4.6 × 250 mm, 5 μm) BEH C18 column (2.1 × 100 mm, 1.7 μm) C18HCE column (2.1 × 150 mm, 3 μm) HSS C18 column (2.1 × 50 mm, 1.8 μm)

DAD

[21]

MS: ESI (+, −) CAD MS: ESI (+, −)

[25] [48] [61]

Zorbax SB-Aq column (4.6 × 250 mm, 5 μm)

DAD, TOF-MS

[62]

Sepax amethyst C18-P column (4.6 × 250 mm, 5 μm)

ECD

[19]

Coptis chinensis (Huang Lian) Cordyceps

Danmu preparations

Flos Sophorae Immaturus (Huai Mi) Frankincenses (Ru Xiang) Fritillaria (Bei Mu) Fructus Alpinia oxyphylla (Yi Zhi)

Fructus Corni

Fructus Psoraleae (Bu Gu Zhi)

6 Phenolic acids 5 Isosteroidal alkaloids 18 Components, including flavonoids, phenylpropanoids and triterpenoids 7 Components, including phenolic acids and iridoids glycosides 2 Flavonoids, including bavachin and isobavachalcone

CHAPTER 20  Quality control of traditional Chinese medicines

Table 20.1  Applications of 1DLC (RPLC, HILIC, and IEC) in the Analysis of Small Molecules From TCMs—cont’d

Ganoderma lucidum (Ling Zhi) Gardenia jasminoides Ellis (Zhi Zi) Gardeniae Fructus (Shan Zhi Zi)

Gegen-Qinlian Decoction

Gentiana (Long Dan)

Gentianae macrophyllae (Qin Jiao) Gua-Lou-Gui-Zhi decoction

Guanjiekang Preparation

Gui-Zhi Fu Ling Wan

14 Components, including flavonoids and phenylpropanoids 7 Components, including carotenoids, chlorogenic acid, flavonoids, and iridoids 5 Components, including phenols and nucleoside 12 Components, including alkaloids and flavonoids 50 Components, including alkaloids, coumarins, flavonoids and saponins 6 Components, including iridoids, lactones, phenolic acids, and xanthonoid 5 Iridoids 24 Components, including flavonoids, galloyl glucosides, gingerols, phenolic acids, monoterpene glycosides 13 Components, including alkaloids, triterpenoids, and flavonoids 4 Paeoniflorins

Knauer Eurospher C18 column (4.6 × 250 mm, 5 μm) Unitary C18 column (4.6 × 250 mm, 5 μm)

DAD

[26]

DAD; MS: ESI (+, −)

[63]

SinoChrom ODS-BP C18 column (4.6 × 250 mm, 5 μm)

PDA, MS: ESI (+, −)

[64]

Purospher STAR column (4.6 × 250 mm, 5 μm) BEH C18 column (2.1 × 100 mm, 1.7 μm)

DAD

[14]

MS: ESI (+, −)

[65]

CSH C18 column (2.1 × 100 mm, 1.7 μm)

MS: ESI (+)

[66]

Shin-pack XR-ODS III column (2.0 × 150 mm, 2.2 μm)

UV; MS: ESI (+, −)

[67]

Kinetex XB-C18 column (4.6 × 150 mm, 2.6 μm) Cortest C18 column (2.1 × 100 mm, 1.6 μm)

PDA

[68]

MS: MRM

[41]

Acquity C18 column (2.1 × 100 mm, 1.7 μm)

MS: ESI (+)

[69]

SunFire C18 column (4.6 × 250 mm, 5 μm)

DAD

[36] Continued

20.2 ­ Separation modes of liquid chromatography

Gastrodia Rhizoma (Tian Ma) Ge Gen Qin Lian Decoction

4 Genodenic acids

623

Analytes

Columns

Detections

References

Hibisci mutabilis Folium (Fu Rong Ye) Huangqi decoction

Rutin and isoquercetin

Hypersil C18 column (4.6 × 250 mm, 5 μm)

DAD

[70]

18 Components, including flavonoids and triterpenic saponins 7 Components, including sesquiterpene lactones and flavonoids 28 Components, including diterpenoids, flavonoids, and phenolic acids 4 Components, including iridoids and phenylpropanoids 15 Components, including terpenoids and n-pentyl benzoate 6 Alkaloids

Zorbax SB-C18 column (4.6 × 250 mm, 5 μm)

MS: ESI (+, −)

[71]

Luna C18 column (4.6 × 250 mm, 5 μm)

DAD

[46]

Diamonsil C18 column (4.6 × 250 mm, 5 μm)

MS: ESI (+, −)

[72]

Luna Phenyl-Hexyl column (2.0 × 250 mm, 5 μm)

CAD

[73]

Phenomenex C18 column (2.1 × 150 mm, 2.6 μm)

DAD

[30]

Eclipse XDB C18 column (4.6 × 250 mm, 5 μm) Zorbax SB C18 column (2.1 × 100 mm, 1.8 μm) Grace Vydac 201TP54 C18 column (4.6 × 250 mm, 5 μm)

DAD; TOF-MS

[74]

DAD

[75]

DAD; MS: ESI (−)

[76]

TC-C18 column (4.6 × 250 mm, 5 μm)

DAD

[77]

Zorbax SB-C18 column (4.6 × 250 mm, 5 μm)

PDA

[13]

BEH C18 column (2.1 × 100 mm, 1.7 μm)

UV; MS: ESI (−)

[78]

Inula salsoloides (Xuan Fu Hua) Isodon rubescens (Dong Ling Cao) Leonurus sibiricus (Yi Mu Cao) Liriope muscari (Mai Dong)

Litsea cubeba (Dou Chi Jiang) Liu Wei Di Huang preparation Lycium barbarum (Ning Xia Gou Qi) Magnoliae Cortex (Hou Po) Magnoliae Officinalis Cortex (Long Xu Cai) Mahuang Fuzi Xixin

11 Components, including iridoids and phenolic acids 6 Components, including flavonoids and phenolic acids 2 Phenols, including honokiol and magnolol 4 Components, including alkaloid and phenylpropanoids 6 Components, including flavonoids and lignans

CHAPTER 20  Quality control of traditional Chinese medicines

Source

624

Table 20.1  Applications of 1DLC (RPLC, HILIC, and IEC) in the Analysis of Small Molecules From TCMs—cont’d

4 Components, including lactones and phenol

XBridge C18 column (4.6 × 250 mm, 5 μm)

UV

[79]

16 Ginsenosides

DAD

[32]

Panax notoginseng

5 Saponins

DAD

[51]

Perilla frutescens seeds and pomaces (Zi Su)

4 Components, including flavonoids and phenolic acids 6 Components, including alkaloids and phenolic acids 12 Flavonoids

Zorbax SB-C18 column (4.6 × 250 mm, 5 μm) Zorbax SB-Aq column (4.6 × 50 mm, 3.5 μm) Atlantis T3 C18 column (4.6 × 250 mm, 5 μm)

PDA; MS: ESI (+, −)

[80]

Zorbax SB-C18 column (4.6 × 250 mm, 5 μm)

DAD, MS: ESI (+)

[81]

Phenomenex C18 column (2.1 × 100 mm, 1.7 μm) Zorbax SB-Aq column (4.6 × 50 mm, 3.5 μm)

CAD

[82]

DAD

[50]

SD column (1.8 × 100 mm, 2.1 μm)

[27]

Athena C18 column (4.6 × 250 mm, 5 μm)

DAD; FT-MS; MS: ESI (+, −) DAD

[83]

Extend C18 column (2.1 × 150 mm, 5 μm)

MS: ESI (+, −)

[42]

Eclipse XDB C18 column (4.6 × 150 mm, 5 μm)

MS: MRM, ESI (+, −)

[84]

Phellodendron amurense Rupr (Guan Huang Bo) Polygoni avicularis herba (Bian Xu) Polygonum multiflorum (He Shou Wu)

Poria cocos (Fu Ling) Qi-Fang-Xi-Bi-Granules Qi-Fu-Yin

Radix Glehniae (Sha Shen)

8 Components, including anthraquinone, alkaloids, flavonoids, and phenolic acid 9 Triterpenic acids 2 Alkaloids, fangchinoline and tetrandrine 26 Components, including alkaloids, flavonoids, lactones, phenolic acids, triterpene saponins, oligosaccharide esters, and xanthone 15 Components, including lactones, nucleosides, and phenolic acids

20.2 ­ Separation modes of liquid chromatography

Notopterygium incisum and Notopterygium franchetii (Qiang Huo) Panax ginseng (Ren Shen)

625

Continued

626

Source

Analytes

Columns

Detections

References

Radix polygoni multiflori (He Shou Wu)

14 Components, including anthraquinones, flavonoids, and phenolic acids 10 Flavonoids

Phenomenex Hydro-RP-C18 column (2.0 × 150 mm, 4 μm)

MS: ESI (−)

[85]

BEH C18 column (2.1 × 100 mm, 1.7 μm)

UV

[16]

9 Saponins

Kromasil C18 column (4.6 × 250 mm, 5 μm) YMC-Pack ODS column (4.6 × 150 mm, 5 μm) Phecda C18 column (4.6 × 250 mm, 5 μm)

ELSD

[31]

PDA; MS: ESI (+, −) DAD, MS: ESI (+, −)

[17]

Radix scutellariae (Huang Qin) Rhizoma Paridis (Chong Lou) Rhizoma Smilacis Glabrae (Tu Fu Ling) Rhododendron dauricum (Du Juan Hua) Salvia miltiorrhiza Bge. (Dan Shen) Salvia miltiorrhiza Bge. (Dan Shen)

Semen Ziziphi Spinosae (Suan Zao Ren) Shexiang Tongxin dropping pill Shuang-Huang-Lian oral liquid

6 Flavonoids 7 Components, including flavonoids, lactones, and phenolic acids 11 Phenolic acids 5 Components, including danshensu, protocatechualdehyde, and phenolic acids 6 Components, including flavonoids, triterpenic acids, and saponins 13 Components, including cholic acids, phenolic acids, and saponins 3 Components, including baicalin, chlorogenic acid, and forsythin

[86]

BEH C18 column (2.1 × 50 mm, 1.7 μm)

MS: ESI (−)

[23]

BEH C18 column (2.1 × 100 mm, 1.7 μm)

PDA

[87]

BEH C18 column (2.1 × 100 mm, 1.7 μm)

ELSD

[47]

Cortecs C18 column (2.1 × 100 mm, 1.6 μm)

MS: ESI (+)

[88]

Reversed-phase C18 column (4.6 × 250 mm, 5 μm)

DAD

[18]

CHAPTER 20  Quality control of traditional Chinese medicines

Table 20.1  Applications of 1DLC (RPLC, HILIC, and IEC) in the Analysis of Small Molecules From TCMs—cont’d

5 Saponins

Tong-Xie-Yao-Fang

7 Components, including flavonoids, phenolic acid, paeoniflorin, cimifugin, and its derivatives 4 Lignans

Vitex negundo seeds (Huang Jing) Wu Ji Pill Xiao Bu Xin Tang Xiao Chai Hu Tang Xiao Yan Li Dan Tablet Xiaoer Chaigui Tuire granules

Xiao-Qing-Long-Tang

7 Phenolic compounds 16 Flavonoids Astragaloside IV

11 Components, including alkaloids and lactones 13 Flavonoids 7 Components, including flavonoids and saponins 5 Components, including lactones and phenolic acid 8 Components, including flavonoids, cinnamaldehyde, and paeoniflorin 16 Components, including alkaloids, flavonoids, nucleosides, and phenolic acids

HSS T3 C18 column (2.1 × 150 mm, 3.5 μm) BEH C18 column (2.1 × 100 mm, 1.7 μm)

MS: MRM, ESI (−)

[33]

MS: ESI (+)

[24]

MS: MRM, ESI (+)

[22]

Eclipse plus C18 column (4.6 × 50 mm, 1.8 μm) Hypersil Gold C18 column (2.1 × 100 mm, 1.8 μm) Kromasil C18 column (6 × 250 mm, 5 μm)

Q-Orbitrap MS

[35]

DAD

[89]

Extend C18 column (4.6 × 250 mm, 5 μm)

DAD

[90]

BEH C18 column (2.1 × 100 mm, 1.7 μm)

MS: ESI (+)

[91]

Inertsil C8-3 column (2.1 × 250 mm, 5 μm) BEH C18 column (2.1 × 100 mm, 1.7 μm)

MS: ESI (−) PDA; MS: ESI (+, −) UV; MS: ESI (+)

[15] [92]

Zorbax SB-C18 column (4.6 × 250 mm, 5 μm)

DAD

[20]

HSS C18 column (2.1 × 100 mm, 1.8 μm)

MS: ESI (+, −)

[94]

BEH C18 column (2.1 × 100 mm, 1.7 μm)

[93]

Continued

20.2 ­ Separation modes of liquid chromatography

Siraitia grosvenorii (Luo Han Guo) Sparganii Rhizoma (San Leng) Spatholobi Caulis (Ji Xue Teng) TCM functional spirit

627

628

Source

Analytes

Columns

Detections

References

Yinhuang Drop Pill

26 Components, including flavonoids, iridoids, phenolic acids, and saponins 12 Components, including anthraquinones, alkaloids, and flavonoids 20 Components, including anthraquinones, flavonoids, iridoids, and phenolic acids 6 Phenols

BEH C18 column (2.1 × 100 mm, 1.7 μm)

MS: ESI (−)

[95]

BEH C18 column (2.1 × 50 mm, 1.7 μm)

PDA

[96]

Phenomenex Kinetex C18 column (4.6 × 150 mm, 2.6 μm)

MS: ESI (−)

[97]

Ultimate XB-C18 column (4.6 × 250 mm, 5 μm) 1.  UPLC: BEH C18 column (2.1 × 100 mm, 1.7 μm); 2.  HPLC: Asahipak NH2P-504E column (4.6 × 250 mm, 5 μm)

DAD; MS: ESI (+, −) 1.  MS: ESI (−); 2. ELSD

[98]

XBridge Amide column (4.6 × 250 mm, 3.5 μm) BEH Amide column (2.1 × 100 mm, 1.7 μm)

CAD

[100]

MS: ESI (+)

[101]

XBridge Amide column (4.6 × 250 mm, 3.5 μm)

ELSD

[102]

Yiqing granule

Zhi-Zi-Da-Huang decoction

Zingiberis Rhizome (Ginger) Shuang-Huang-Lian oral liquid

1.  15 Components, including flavonoids, quinic acids, saponins, and phenylethanoid glycosides; 2.  3 Saccharides

[99]

HILIC

12 TCMs

11 Oligosaccharides

Animal horns

14 Components, including nucleosides and nucleobases 10 Components, including monosaccharides and oligosaccharides

Burdock

CHAPTER 20  Quality control of traditional Chinese medicines

Table 20.1  Applications of 1DLC (RPLC, HILIC, and IEC) in the Analysis of Small Molecules From TCMs—cont’d

Eclipta prostrasta

8 Monosaccharides

Elaphuri Davidiani Cornu and Cervi Cornu

17 Components, including nucleosides and nucleobases 16 Components, including nucleosides and nucleobases Atractyloside

Euryale ferox

Fructus Xanthii (Cang Er Zi)

Geosaurus (Di Long) and Leech (Shui Zhi) Ginkgo biloba leaves

Ginkgo seeds Ginkgo seeds

Mulberry leaf (Sang Ye)

Ginsenoside Rb1, Astragaloside IV, and dulcitol 14 Components, including nucleosides and nucleobases 11 Components, including nucleosides and nucleobases 24 Amino acids 20 Components, including nucleosides and nucleobases 40 Components, including amino acids and alkaloids, nucleosides and nucleobases

CAD

[103]

MS: ESI (+)

[104]

XBridge Amide column (4.6 × 150 mm, 3.5 μm)

MS: ESI (+)

[105]

Zorbax RX-SIL column (150 × 2.1 mm, 5 um) Luna HILIC column (4.6 × 250 mm, 5 μm)

MS: ESI (−)

[106]

ELSD

[107]

TSK-Gel Amide-80 column (2.0 × 150 mm, 3 μm)

PDA

[108]

BEH Amide column (2.1 × 100 mm, 1.7 μm)

MS: ESI (+)

[109]

BEH Amide column (2.1 × 100 mm, 1.7 μm) BEH Amide column (2.1 × 100 mm, 1.7 μm)

MS: ESI (+)

[110]

MS: ESI (+)

[111]

BEH Amide column (2.1 × 100 mm, 1.7 μm)

MS: ESI (+)

[112]

Continued

20.2 ­ Separation modes of liquid chromatography

Fufangfufangteng Heji

BEH Amide column (3.0 × 100 mm, 2.5 μm) BEH Amide column (2.1 × 100 mm, 1.7 μm)

629

630

Source

Analytes

Columns

Detections

References

Radix Isatidis (Ban Lan Gen)

13 Fingerprints, including amino acids, nucleosides, and phenolic acids 20 Components, including nucleosides, nucleotides, and nucleobases

BEH Amide column (2.1 × 50 mm, 1.7 μm)

MS: ESI (+)

[113]

BEH Amide column (2.1 × 100 mm, 1.7 μm)

MS: ESI (+)

[114]

Collagen-derived peptide 8 Monosaccharides 1-Deoxynojirimycin Dencichine

Ion-exchange column (100 × 10 mm) CarboPac PA10 column (2 × 250 mm) CarboPac MA-1 column (4 × 250 mm) Eprogen Synchropak WAX column (4.6 × 250 mm, 6 μm)

MS PAD PAD DAD

[115] [116] [117] [118]

3 Alkaloids

SCX column (4.6 × 250 mm, 5 μm)

PAD

[119]

2 Components, including albiflorin and paeoniflorin 29 Free amino acids and their metabolites

CarboPac PA 20 column (3 × 150 mm)

PAD

[120]

Hitachi custom ion-exchange resin 2622 column (4.6 × 60 mm, 5 μm)

Amino acid analyzer

[121]

Ziziphus

IEC Colla Corii Asini Cyclocarya paliurus Mulberry leaf (Sang Ye) Panax (P. ginseng, P. notoginseng, P. quinquefolium) Si-Mo-Tang oral liquid preparation Si-Ni-San Ziziphus jujube (Hong Zao)

APCI, atmospheric pressure chemical ionization; CAD, charged aerosol detector; DAD, diode array detector; ECD, electrochemical detector; ELSD, evaporative light scattering detector; ESI, electrospray ionization; FLD, fluorescence detector; FT, Fourier transform; HILIC, hydrophilic interaction liquid chromatography; IEC, ion-exchange chromatography; MRM, multiple reaction monitoring; MS, mass spectrometry; PAD, pulsed amperometric detector; PDA, photodiode array detector; Q, quadrupole; RPLC, reversed-phase liquid chromatography; TCM, traditional Chinese medicine; TOF, time of flight; UV, ultraviolet.

CHAPTER 20  Quality control of traditional Chinese medicines

Table 20.1  Applications of 1DLC (RPLC, HILIC, and IEC) in the Analysis of Small Molecules From TCMs—cont’d

20.2 ­ Separation modes of liquid chromatography

20.2.2 ­ HYDROPHILIC INTERACTION LIQUID CHROMATOGRAPHY Nevertheless, AQ columns could not satisfy the requirements (such as good retention behavior and high resolution) of some high-polar components, like carbohydrate analysis. In these cases, HILIC is usually necessary for the analysis of these highly hydrophilic components. As introduced by Alpert in 1990 for the first time [122], HILIC is considered as the variant of NPLC, applying unmodified silica gel stationary phase, or polar chemically bonded stationary phases (amine-, amide-, cyano-, zwitterionic-, and diol-bonded) [123], and reflecting in partitioning as its retention mechanism and applying mobile phases similar to RPLC modes [124]. Compared with RPLC, HILIC is used to separate compounds with higher polarity and hydrophilicity, such as amino acids [110,112], nucleic compounds [101,104,109,111,114], and saccharides [100,102,103]. In addition, HILIC separation also serves as IEC to separate polar ionic molecules [125]. The HILIC applications in component analysis were systematically reviewed and discussed [7,126,127]. For the determination of saccharides, a Waters XBridge Amide column (4.6 × 250 mm, 3.5 μm i.d.) was successfully applied for the determination of 11 inulin-type fructooligosaccharides (DP3-DP13) in 12 TCMs, such as Atractylodes lancea, Codonopsis pilosila, Atractylodes macrocephala, Carthamus tinctorius, Arctium lappa, and so on [100]. For determining monosaccharide composition, a Waters BEH Amide column (3.0 × 100 mm, 2.5 μm) was applied to separate 8 monosaccharides degraded from polysaccharides of Eclipta prostrasta L. [100]. Besides, Zhang et al. have reported on the simultaneous determination of 40 compounds (including 14 nucleosides and nucleobases, 24 amino acids and 2 alkaloids) in the leaves of Morus alba L. using a Waters BEH Amide column (100 × 2.1 mm, 1.7 μm i.d.) [112]. The separation was maintained for 18 min and the HILIC column was confirmed to obtain a relatively stronger retention for analytes than the RP-18 column. Moreover, Qin et al. have reported that a Phenomenex Luna HILIC column (250 × 4.6 mm, 5 μm i.d.) was applied for HILIC separation of 2 saponins (ginsenoside Rb1 and astragaloside IV) and 1 alkanol in a TCMs formula “Fufangfufangteng Heji” [107]. The applications of HILIC in the quality control of phytochemical components in TCMs are listed in Table 20.1. Higher ratio of organic solvents (e.g. acetonitrile and methanol) in mobile phases is required for HILIC, resulting in reduced viscosity of mobile phases and decrease of column pressure [128]. There are several advantages for HILIC compared with RPLC. Compared with the RPLC mode, lower column pressures allow fast componential analysis with high resolution and separation efficiency. Owing to similar mobile phase systems, HILIC has excellent orthogonality and high compatibility with RPLC when the component is analyzed, which has extensions in two-dimensional analytical systems [129]. Besides, higher organic solvents make it more compatible to aerosol-based detectors, such as ELSD, CAD, and MS with ESI interfaces, which could highly increase the sensitivity of detections [130]. Likewise, higher organic solvents also lead to some challenges. When highly hydrophilic components are analyzed, it will be less soluble and further precipitated. The consumptions of highproportion organic solvents for lengthened analysis time are expensive and also unfriendly to the environment. Besides, it requires longer equilibration time prior to the

631

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CHAPTER 20  Quality control of traditional Chinese medicines

analysis [131]. Therefore, IEC, which applies polar ion-exchange stationary phases and adapts to AQ mobile phases, could serve as an extended substitute for HILIC, especially in the analysis of highly hydrophilic and large-molecule compounds, such as saccharides, nucleotides, peptides, and proteins.

20.2.3 ­ION-EXCHANGE CHROMATOGRAPHY IEC is used to separate charged and polar molecules based on their binding capacities toward ion exchangers [132]. On the basis of the functional groups of ion-exchange resins, IEC includes two separation modes, the cation and the anion-exchange chromatography (AEC) [133]. Cation-exchange chromatography resin has the negatively charged functional groups that are used to separate cations, while those of AEC are positively charged to separate anions [134]. By changing pH of the eluents, the molecules that are bound to ion-exchange resins could be subsequently eluted from the column. IEC has been applied to the purification of macromolecules, such as carbohydrates, peptides, and proteins [135–137]. Moreover, IEC has been applied to the analysis of charged substances, such as inorganic metal ions, alkaloids, and amino acids [138,139]. IEC was applied in the quality control of bioactive polar molecules in TCMs. A nonprotein amino acid, dencichine, was quantitatively determined by an established HPAEC-DAD method in 3 Panax species [118]. Dencichine was separated on an Eprogen SynChropak WAX analytical column (4.6 × 250 mm, 6 μm, Darien, USA) with an isocratic elution of 50 mM sodium dihydrogen phosphate AQ solution (pH 4.0 adjusted with phosphoric acid). Besides, 1-deoxynojirimycin, which was an azasugar derived from mulberry leaf, was quantified by an analytical HPAEC column coupled to pulsed amperometric detection (PAD), with mobile phases of 0.2 M and 1 M sodium hydroxide solutions [117]. Another HPAEC-PAD example was the determination of neutral monosaccharide composition of Cyclocarya paliurus. Eight neutral monosaccharides were quantified using a CarboPac PA-10 column (2.0 × 250 mm) and sodium hydroxide solutions as the mobile phase [116]. In addition, an Agilent strong cation-exchange (SCX) column (4.6 × 250 mm, 5 μm) and an isocratic elution (methanol-0.17% phosphoric acid solution with pH adjusted to 3.8 using ammonia water, 65:35, v/v) were applied to the quantitative analysis of 3 alkaloids from a Chinese patent medicine, Si-Mo-Tang oral liquid preparation [119]. The cation-exchange column obtained better resolutions and higher theoretical plates in separating those 3 alkaloids as compared with those for another RP18 column (4.6 × 250 mm, 5 μm).

20.2.4 ­SIZE EXCLUSION CHROMATOGRAPHY SEC has a predominant status on molecular characterization and analysis of polymeric compounds [140,141]. SEC analysis is established based on gel filtration chromatography (GFC) columns [142]. Compared with a single SEC column, a serial combination of multiple SEC columns allows broader molecular weight

20.2 ­ Separation modes of liquid chromatography

(Mw) distribution and higher resolution for the analysis of macromolecules, such as water-soluble polysaccharides, proteins, and nucleotides [143]. The data obtained from HPSEC coupled to several detectors, such as refractive index (RI) detectors, multiangle laser light scattering (MALLS) detectors, viscometers (VIS), and mass spectrometers (MS), can provide sufficient information for the investigation of macromolecules [141]. In our previous studies, HPSEC has been proved to be a powerful tool for the characterization and quality evaluation of polysaccharides in different TCMs, such as the Cordyceps species, Dendrobium species, Panax species and the Gymnadenia conopsea [144–151]. The applications of HPSEC in the analysis of TCMs are listed in Table 20.2. HPSEC coupled to a MALLS detector (HPSEC-MALLS) can provide sufficient data for the investigation of molecular parameters, such as the polydispersity index, absolute molecular mass, radius of gyration, and chain conformation [153]. Moreover, HPSEC-MALLS coupled with RID realize the determination of differential RI increment (dn/dc), which could be further utilized for direct calculation of their content based on the concentration-specific RI increment equation [148]. The quantification accuracy of HPSEC-MALLS-RID had been compared with that of HPSEC-ELSD in 3 Panax species, such as P. ginseng, P. notoginseng, and P. quinquefolium. The results demonstrated that HPSEC-MALLS-RID with the dn/dc method had better recoveries and did not require any polysaccharide standards, while HPSEC-ELSD and phenol-sulfuric acid assay had much lower recoveries and required a large amount of polysaccharide standards as references. Therefore, it was demonstrated in previous studies that HPSEC-MALLS-RID based on the dn/dc method could be a simple and reliable approach for the quantification of polysaccharides in TCMs [146,148,154]. Lycium barbarum polysaccharide (LBP) is considered as one of the bioactive constituents in L. barbarum species, which exhibit immunomodulatory activities [159,160]. In previous studies from our laboratory, Mw and distributions of LBPs from different origins of China have been quantified using HPSEC-MALLS-RID with the dn/dc method [154]. Monosaccharide compositions of LBPs from different origins of China have been previously investigated by high-performance thin-layer chromatography and polysaccharide analysis by using carbohydrate gel electrophoresis analysis [161]. The results based on the established methods demonstrated that HPSEC chromatograms and Mw distributions for LBPs from different origins were in similar patterns (Fig. 20.1). It is considered that no significant differences of total polysaccharide fractions are found in Ningxia and other origins (Inner Mongolia, Xinjiang and Gansu), whereas average amounts of total polysaccharide fractions in Qinghai were significantly lower than in Ningxia. Therefore, the established HPSEC-MALLS-RID with the dn/dc method has been proved to be the routine approach for quality control of polysaccharides from natural products. Besides, SEC-MALDI-TOF-MS had also been applied for the characterization of bioactive peptides in cinobufacini injection, which was a TCM injection derived from AQ extracts from toad skin [158].

633

634

Source

Analyte

Stationary Phase

Detections

References

Gymnadenia conopsea tubers (Shou Zhang Shen) Panax species (Ren Shen, San Qi, Xi Yang Shen)

Polysaccharides

TSK-Gel G5000PWXL (300 mm × 7.8 mm, 10 μm)

MALLS, RID

[146]

Polysaccharides

1.  TSK-Gel G4000PWXL (300 mm × 7.8 mm, 10 μm); 2.  TSK-Gel G6000PWXL (300 mm × 7.8 mm, 13 μm) and G3000PWXL (300 mm × 7.8 mm, 7 μm) in series TSK-Gel G5000PWXL (300 mm × 7.8 mm, 10 μm) and G3000PWXL (300 mm × 7.8 mm, 7 μm) in series TSK-Gel G6000PWXL (300 mm × 7.8 mm, 13 μm) and G3000PWXL (300 mm × 7.8 mm, 7 μm) in series 1.  TSK-Gel G3000PWXL (300 mm × 7.8 mm, 7 μm); 2.  TSK-Gel G4000PWXL (300 mm × 7.8 mm, 10 μm); 3.  TSK-Gel G4000PWXL (300 mm × 7.8 mm, 10 μm) TSK-Gel G6000PWXL (300 mm × 7.8 mm, 13 μm) and G3000PWXL (300 mm × 7.8 mm, 7 μm) in series TSK-Gel G4000PWXL (300 mm × 7.8 mm, 10 μm)

MALLS, RID; ELSD

[148]

DAD, MALLS, RID

[149]

MALLS, RID

[147]

DAD, ELSD; MALLS; TDA

[145]

MALLS, RID

[152]

DAD, ELSD

[150]

Astragalus membranaceus (Huang Qi)

Polysaccharides

Cultured Cordyceps sinensis fungus (Dong Chong Xia Cao)

Polysaccharides

Cordyceps sinensis (Dong Chong Xia Cao)

Hyperbranched polysaccharides

Hericium erinaceus fruiting body (Hou Tou Gu)

Polysaccharides

Panax (P. ginseng, P. notoginseng, P. quinquefolium), Cordyceps (C. sinensis, C. militaris), Ganoderma (G. lucidum, G. sinense), Astragalus membranaceus, Angelica sinensis

Polysaccharides

CHAPTER 20  Quality control of traditional Chinese medicines

Table 20.2  Applications of SEC in Analysis of Macromolecules From TCMs

Dendrobium (D. huoshanense, D. officinale, D. fimbriatum, D. chrysanthum, D. nobile, and D. officinale) Lentinan injection

Lycium barbarum (Ning Xia Gou Qi)

Cinobufacini injection

TSK-Gel G3000PWXL (300 mm × 7.8 mm, 10 μm)

DAD, ELSD

[151]

Polysaccharides (β-(1 → 6) branched-(1 → 3)-glucan) Water-soluble polysaccharides

MALLS, RID

[153]

MALLS, RID, UV

[154]

Glucogalactomannan

TSK-Gel G6000PWXL (300 mm × 7.8 mm, 13 μm) and G4000PWXL (300 mm × 7.8 mm, 10 μm) in series TSK-Gel G5000PWXL (300 mm × 7.8 mm, 10 μm) and G3000PWXL (300 mm × 7.8 mm, 7 μm) in series TSK-Gel G4000PWXL (300 mm × 7.8 mm, 10 μm)

TDA, RID

[155]

Polysaccharides

TSK-Gel G4000PWXL (300 mm × 7.8 mm, 10 μm)

ELSD

[144]

Water-soluble polysaccharides Polysaccharides

TSK-Gel G3000PWXL (300 mm × 7.8 mm, 7 μm)

RID

[156]

1.  TSK-Gel G3000PWXL (300 mm × 7.8 mm, 10 μm); 2.  TSK-Gel G6000PWXL (300 mm × 7.8 mm, 13 μm) and TSK-Gel G3000PWXL (300 × 7.8 mm, 10 μm) in series TSK-Gel G2000SWXL (300 mm × 7.8 mm, 5 μm)

ELSD; MALLS, RID

[157]

UV

[158]

Peptides

DAD, diode array detector; ELSD, evaporative light scattering detector; MALLS, multiangle laser light scattering detector; RID, refractive index detector; TDA, triple diode array detector; UV, ultraviolet.

20.2 ­ Separation modes of liquid chromatography

Cultured Cordyceps sinensis fungus (Dong Chong Xia Cao) Natural and cultured Cordyceps Euphorbia fischeriana (Lang Du) Ganoderma (G. lucidum and G. sinense)

Polysaccharides

635

CHAPTER 20  Quality control of traditional Chinese medicines

0.2

5.0x10−7

0.1 20.0 30.0 Time (min)

40.0

0.4

1

2 3 Mw<3kDa

0.3

−6

5.0x10

4.0x10−6 3.0x10−6

0.2

2.0x10−6

0.1 0.0

10.0

20.0 30.0 Time (min)

1

2 3

50.0

dRI −6 UV 4.0x10 Mw<3kDa 3.0x10−6

0.3

2.0x10−6

0.2 1.0x10 0.1 0.0

10.0

20.0 30.0 40.0 Time (min)

50.0

−6

1

2 3

dRI 5.0x10−6 UV Mw<3kDa 4.0x10−6

0.3

3.0x10−6

0.2

2.0x10−6

0.1

1.0x10−6 0.0

0.5

10.0

20.0 30.0 40.0 Time (min)

XJ 1

0.4

2 3

50.0

dRI −6 UV 5.0x10 Mw<3kDa 4.0x10−6

0.3

3.0x10−6

0.2

2.0x10−6

0.1 0.0

0.5

10.0

20.0 30.0 Time (min)

NX

0.4

1

2 3

40.0

50.0

dRI UV Mw<3kDa

0.3

4.0x10−6 3.0x10−6 2.0x10−6

0.2

1.0x10−6

0.1 0.0

10.0

20.0 30.0 40.0 Time (min)

50.0

Differential refractive index (RIU)

GS

0.4

40.0

Differential refractive index (RIU)

0.5

6.0x10−6

QH

0.4

Differential refractive index (RIU)

dRI UV

IM

Differential refractive index (RIU)

0.5

10.0

0.0 50.0

Detector voltage (V)

1.0x10−6

Detector voltage (V)

0.3

0.0

Detector voltage (V)

1.5x10

0.5

Detector voltage (V)

Detector voltage (V)

0.4

−6

Differential refractive index (RIU)

2.0x10−6

dRI UV

Blank

Differential refractive index (RIU)

0.5

Detector voltage (V)

636

FIG. 20.1 HPSEC-MALLS-RID chromatograms of polysaccharides of Lycium barbarum in different regions of China (QH, Qinghai; IM, Inner Mongolia; XJ, Xinjiang; GS, Gansu; NX, Ningxia). From Wu DT, Lam SC, Cheong KL, Wei F, Lin PC, Long ZR, et al. Simultaneous determination of molecular weights and contents of water-soluble polysaccharides and their fractions from Lycium barbarum collected in China. J Pharm Biomed Anal 2016;129:210–8 with permission of Elsevier.

20.2.5 ­TWO-DIMENSIONAL LIQUID CHROMATOGRAPHY (2DLC) As introduced above, different LC modes, such as NPLC, RPLC, HILIC, IEC, SEC, and so on, are classified based on their separation mechanisms. However, each onedimensional LC (1DLC) mode has its limitations, such as beyond applicable separation ranges or insufficient for complex analytes. Two-dimensional LC (2DLC) combines two of these separation modes in order to broaden the separation range, enhance the resolution, and to improve the selectivity [162]. The applications of 2DLC in the quality control of TCMs have been systematically reviewed [163,164]. On the basis of the transferring patterns of fractions from first-dimension (1D) to second-dimension (2D) chromatography, 2DLC is classified into offline and online

20.2 ­ Separation modes of liquid chromatography

modes [165]. Eluted fractions of interest from the 1D column are manually collected and subsequently reinjected to the 2D column in the offline mode, while the fractions could be automatically transferred from 1D to 2D columns through special interfaces (column switching) in the online mode. Online 2DLC is further divided into comprehensive and heart-cutting 2DLC on account of whether the fractions from 1D are entirely transferred to 2D columns. Comprehensive 2DLC enables two-step separation of the whole sample while the heart-cutting mode separates selected fractions in the 2D chromatography [166]. To date, 2DLC approaches of different mixed modes and of different combinations have been widely and successfully applied to the quality control of TCMs. Li et al. reported an offline (RP × RP) LC method to characterize coumarins from the roots of Angelica dahurica [167]. The 1D separation collected the fractions of interest monitored by DAD and further concentrated for subsequent 2D separation. The 2 D separation coupled to DAD-ESI/MSn was successfully applied to characterize 50 coumarins in the tested samples. Yao et al. reported an online multiple heart-­cutting 2DLC method for quality control of five saponins in P. notoginseng and related samples [168]. The established method was a combination of RP18 column and AQ column coupled to DAD, by using a 10-port 2-position interface with a sample loop. The analytical results by the established (RP × RP) LC-DAD method were compared with those by the existing 1D-RPLC method from Chinese Pharmacopoeia, which demonstrated that the 2DLC method possessed a more desirable resolution, better specificity, and higher analytical efficiency. Apart from RP × RP combination, LC with different separation mechanisms had been hyphenated in order to enlarge the separation ranges and improve the peak capacity. Song et al. reported an HILIC × RPLC method to qualitatively characterize the components in Shenfu injection, in which 154 compounds were characterized by ESI-MS/MS with pMRM and sMIM modes [169]. Further, the established method was also used for simultaneous quantification of 40 compounds with different polarities in Shenfu injection and related TCM extracts (aconite and ginseng extracts). The results demonstrated that the combination of HILIC and RPLC could achieve simultaneous analysis of hydrophilic and hydrophobic components, while the conventional 1D RPLC could not satisfy the quantification of hydrophilic components. Ma et al. reported an HPSEC × UPLC (RPLC) method coupled to TOF-MS for analysis of Qingkailing injection [170]. A total of 398 peaks were detected based on the fourdimensional data generated by HPSEC × UPLC coupled to TOF-MS, including 2D retention times, peak intensity, and m/z ratio. In our previous study, an LC system consisting of multiple columns and detectors has been designed and conducted for the analysis of global components in the Ganoderma species [171]. Two precolumns (PCs), PC1 and PC2, as well as three LC column systems, including SEC, RPLC, and HILIC, have been connected in series by four 6-port 2-position column switching valves, followed by three detections, which are DAD, ELSD, and MS (Fig. 20.2). The designed systems could be applied to the analysis of different types of compounds. First, macromolecular components could be analyzed by the coupling information shown in Fig. 20.2A and B.

637

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CHAPTER 20  Quality control of traditional Chinese medicines

Pump-1

Pump-1

Injector

Pre-2

Injector

Pre-1

Pre-1

Pump-2

Pump-2 Pre-2

Mixer

Mixer

SEC Pump-3

SEC Pump-3

DAD

DAD

Mixer

Mixer HILIC

RP

ELSD

MS

(A)

HILIC

RP

ELSD

MS

(B)

Pump-1

Pump-1

Injector

Injector Pre-1

Pre-1

Pump-2

Pump-2 Pre-2

Mixer SEC

Pre-2

Mixer SEC

Pump-3

DAD

Pump-3

DAD

Mixer

Mixer HILIC

RP

ELSD

MS

HILIC

RP

ELSD

MS

(D)

(C) Pump-1

Injector Pre-1

Pump-2 Pre-2

Mixer SEC

Pump-3

DAD

Mixer HILIC

RP

ELSD

MS

(E) FIG. 20.2 Schematic diagrams of multiple columns and detectors LC system for loading (A) and analysis (B) of macromolecules, loading (C), and analysis (D) of high-polarity small molecules, and analysis (E) of low-polarity small molecules. From Qian ZM, Zhao J, Li DQ, Hu DJ, Li SP. Analysis of global components in Ganoderma using liquid ­chromatography system with multiple columns and detectors. J Sep Sci 2012;35(20):2725–34 with ­permission of John Wiley and Sons.

20.3 ­ Detections

The ­samples were injected into PC1, and macromolecules were eluted from the PC1 while small molecular compounds could be retained (Fig.  20.2A). After that, the macromolecules were separated in SEC column and further analyzed by multiple detectors (Fig. 20.2B). Then, small molecular compounds were eluted from PC1 to PC2 and low-polarity small molecular compounds were retained on PC2 (Fig.  20.2C). The high-polarity small molecular compounds were eluted through PC2 and further analyzed in the HILIC column coupled to multiple detectors (Fig. 20.2D). Finally, low-polarity compounds were analyzed by the RPLC system after column switching (Fig. 20.2E). The multiple columns and detectors LC systems were successfully applied to the quality control of global components in Ganoderma (G. lucidum and G. sinense), which is considered as an efficient analysis protocol for the quality control of TCMs. The applications of 2DLC in the quality control of TCMs are listed in Table 20.3.

20.2.6 ­ MISCELLANEOUS Apart from several major chromatographic techniques introduced above, other less used approaches were relevant to the quality control of TCMs, such as high-speed countercurrent chromatography (HSCCC) and supercritical fluid chromatography (SFC). HSCCC is a support-free liquid-liquid partition chromatography with two immiscible mobile phases [189]. By the centrifugal forces of HSCCC, the constituents of the samples got separated based on the dissociation constant (K) in either phase. Owing to high sample loading capacity and powerful separation efficiency, HSCCC had been widely used for preparative isolation of different components in TCMs [190–192]. In addition to HSCCC, SFC is a form of normal-phase chromatography that used supercritical fluid as the mobile phase, such as the supercritical CO2, with the advantages of low viscosity, high separation efficiency, and environmental friendliness [193].

20.3 ­DETECTIONS 20.3.1 ­ UV-VIS DETECTION In the present, UV-Vis detectors, such as the DAD and the photodiode array detector (PDA), are considered as nearly universal detectors for the analysis of compounds with chromophores under ultraviolet (λ = 190–400 nm) and visible (λ = 400–800) ranges. Moreover, DAD and PDA could provide contour maps and 3D spectra of the eluting peaks that can be used for peak identification and peak purity monitoring as compared with the old fixed wavelength detector (FWD) and variable wavelength detector (VWD). UV-Vis detectors, as the standard equipment of commercial HPLC instruments, is considered as the most prevailing detectors for the analysis of phytochemicals in TCMs, such as alkaloids [12,13,55,60,81], flavonoids [16,17,20 ,21,50,53,70,76,78,80,86,89,92,96], phenolic acids [18,75,81,87,93], triterpenoids [26,27,30,32,36,51,57], and iridoids [62,64,67,68].

639

640

Parameters Source

Analyte

Mode

1

Angelica dahurica roots (Bai Zhi)

50 Coumarins

Offline

Scutellaria barbata (Ban Zhi Lian)

206 and 543 Detected

Offline

Dracaena cochinchinensis (Long Xue Jie) Salvia miltiorrhiza (Dan Shen) and related preparations Panax notoginseng (San Qi)

55 Detected

Offline

366, 460, 451, and 476, respectively 224 Saponins

Offline

Zorbax SB-C18 column (4.6 × 250 mm, 5 μm) 1.  Atlantis HILIC Silica column (4.6 × 250 mm, 5 μm); 2.  XAmide column (4.6 × 150 mm, 5 μm) Ultimate XB-CN column (4.6 × 250 mm, 5 μm) Unitary XAmide column (4.6 × 250 mm, 5 μm) XAmide column (150 × 4.6 mm, 5 μm)

Stevia Rebaudiana (Tian Ju) Xueshuantong injection

13 Steviol glycosides 143 Components

Offline

Qingkailing injection

398 Detected

Offline

Offline

Online (comprehensive)

D column

XCharge C18 column (20 × 150 mm, 10 μm) XBridge Amide column (4.6 × 150 mm, 3.5 μm) TOSOH Toyopearl HW-40 S column (2.1 × 200 mm)

2

D column

Detections

References

Extend C18 column (4.6 × 250 mm, 5 μm)

DAD; ESI-MS

[167]

1.  XAmide column (4.6 × 150 mm, 5 μm); 2. XUnion C18 column (2.1 × 150 mm, 5 μm) Ultimate XBC18 column (4.6 × 250 mm, 5 μm) Acquity HSS T3 C18 column (2.1 × 50 mm, 1.7 μm) Acquity BEH C18 column (2.1 × 100 mm, 1.7 μm) XAmide column (10 × 150 mm, 5 μm) Acquity HSS T3 C18 column (2.1 × 100 mm, 1.8 μm) Acquity C18 column (2.1 × 100 mm, 1.7 μm)

UV; ESI-MS

[172]

UV

[173]

PDA; IT-TOF-MS

[174]

UV; ESI-TQ-MS

[175]

DAD; QTOF-MS VWD; QTOF-MS

[176]

UV; TOF-MS

[170]

[177]

CHAPTER 20  Quality control of traditional Chinese medicines

Table 20.3  Applications of 2DLC in Analysis of Small Molecules From TCMs

154/157 Identified, 40 quantified

Online (comprehensive)

XBridge BEH Amide column (4.6 × 150 mm, 3.5 μm)

Hdyotis diffusa (Bai Hua She She Cao) and Scutellaria barbata (Ban Zhi Lian)

–

Online (comprehensive)

Huang-Lian-ShangQing tablet

4 Components

Online (comprehensive)

Hdyotis diffusa (Bai Hua She She Cao)

25 Characterized (22 flavonoids and 3 iridoid glycosides) 14/40 Identified

Online (comprehensive)

Two PhenoSphere column (2.0 × 150 mm, 5 μm) and one Luna CN column (3.0 × 50 mm, 3 μm) SCX Poly-SEA capillary column (0.3 × 150 mm, 3 μm) Luna CN column (2.0 × 150 mm, 3 μm)

Online (comprehensive)

Immobilized liposome column (4.6 × 250 mm)

–

Online (comprehensive)

Luna CN column (2.0 × 150 mm, 3 μm)

4 Components

Online (comprehensive)

HepG2/CMC column (2.0 × 10 mm, 5 μm)

Schisandra chinensis (Wu Wei Zi)

Hdyotis diffusa (Bai Hua She She Cao) and Scutellaria barbata (Ban Zhi Lian) Cortex phellodendri amurensis (Huang Bo) and Radix sophorae flavescentis (Ku Shen)

ESI-MS

[169]

DAD

[178]

Magic C18AQ capillary column (0.3 × 50 mm, 5 μm) Kinetex C18 column (3.0 × 50 mm, 2.6 μm)

UV

[179]

DAD; QTOF-MS

[180]

Chromolith Performance RP-18e column (4.6 × 100 mm, 5 μm) Kinetex C18 column (3.0 × 50 mm, 2.6 μm)

DAD; ESI-MS

[181]

DAD

[182]

Chromolith Performance RP-18e column (4.6 × 100 mm, 5 μm)

TOF-MS

[183]

Phenomenex Synergi Polar-RP 100A column (2.0 × 100 mm, 2.5 μm) Kinetex C18 column (3.0 × 50 mm, 2.6 μm)

Continued

20.3 ­ Detections

Shenfu injection

641

642

Parameters Source

Analyte

Mode

1

Radix et Rhizoma Asari (Xi Xin)

Asarinin

Online (comprehensive)

EGFR/CMC column (2.0 × 10 mm, 5 μm)

Salvia miltiorrhiza (Dan Shen)

102/328 Identified

Online (comprehensive)

Gegen-Qinlian Decoction

125/280 Characterized

Pueraria (P. lobata and P. thomsonii) (Ye Ge, Fen Ge) Panax notoginseng (San Qi)

271 and 254 detected

Online (comprehensive +  heart-cutting) Online (comprehensive +  heart-cutting) Online (heart-cutting)

Hypersil gold CN column (1.0 × 150 mm, 3 μm) Acquity CSH C18 column (2.1 × 100 mm, 1.7 μm) Acquity CSH C18 column (2.1 × 100 mm, 1.7 μm) Poroshell SB C18 column (2.1 × 100 mm, 2.7 μm)

Hypericum ascyron (Huang Hai Tang)

5 Saponins, noto-R1, Rg1, Re, Rb1, Rd 54 Detected

Online (heart-cutting)

D column

Zorbax SB-Phenyl column (4.6 × 75 mm, 3.5 μm)

2

Shim-pack VPODS column (2.0 × 150 mm, 5 μm) Accucore C18 column (4.6 × 50 mm, 2.6 μm) Poroshell 120 Phenyl-Hexyl column (3.0 × 50 mm, 2.7 μm) Poroshell 120 Phenyl-Hexyl column (3.0 × 50 mm, 2.7 μm) Zorbax SB-Aq column (4.6 × 100 mm, 3.5 μm) Poroshell 120 C18 column (4.6 × 150 mm, 2.7 μm)

D column

Detections

References

DAD; MS

[184]

LTQ-Orbitrap MS

[185]

DAD; QTOF-MS

[186]

DAD; ESI-MS

[187]

DAD

[168]

DAD

[188]

DAD, diode array detector; ESI, electrospray ionization; IT, ion trap; LTQ, linear triple quadrupole; MS, mass spectrometry; Q, quadrupole; TOF, time of flight; TQ, triple quadrupole; UV, ultraviolet; VWD, variable wavelength detector.

CHAPTER 20  Quality control of traditional Chinese medicines

Table 20.3  Applications of 2DLC in Analysis of Small Molecules From TCMs—cont’d

20.3 ­ Detections

However, the uses of DAD and PDA detectors might be limited when those compounds with lower UV or Vis absorbance are analyzed, which may require a higher concentration of analytes and a more desirable detection method for substitution or hyphenation. Owing to its good compatibility with other instruments, it allows coupling of DAD and other types of detectors, such as ELSD, CAD, or MS, which makes it possible to determine different types of compounds and acquire sufficient data for identification and quantification based on different detectors [57,194–197]. For example, Peng et al. have reported an HPLC-DAD-ELSD method for simultaneous determination of flavonoids, isochlorogenic acids, and triterpenoids in Ilex hainanensis [194]. To obtain a more accurate quantification, 283 nm was selected as an optimized wavelength for the detection of flavonoids and isochlorogenic acids, while ELSDs were applied to the quantification of triterpenoids. Besides, the precolumn derivatization reaction is also applied to those compounds with no UV absorbance, in order to allow the quantification under UV detection. Shi et al. have reported a precolumn derivatization followed by the HPLC-PDA analysis in the quality control of major bile acids in artificial C. bovis samples [34].

20.3.2 ­ NONSPECTROMETRY DETECTION (RID, ELSD, CAD, AND ECD) Universal detectors could offer the analysis of nonspecific components with no UV absorbance [198]. Refractive index detector (RID) is a universal detector that measures the intensity of the analyte through determinations of a RI relative to that of the solvent, which demonstrates that the greater the differences in RIs between the analytes and the mobile phase, the higher the sensitivity of the method. It is currently used for the analysis of saccharides in TCMs, which determine the RI of polysaccharides when combined with HPSEC [147–149,152,154–156]. Besides, fast protein liquid chromatography (FPLC) coupled to RID has been used for preparative isolation of oligosaccharides in TCMs [199,200]. Owing to several disadvantages of RID, including only available in isocratic separations and very sensitive to temperature variations, aerosol-based detectors are offering alternatives for the analysis of nonvolatile compounds [201]. ELSD is a cost-effective and versatile detector that can provide favorable baseline separation and good compatibility with gradient separation as compared with RID [202,203]. HPLC-ELSD has been a prevalent technique for the analysis of components with no or low UV absorption in TCMs, such as saponins [31,107,204] and cholic acids [39,205]. Hu et al. have reported an HPLC-DAD-ELSD method for the quantification of 10 constituents in TCMs, two of which were simultaneously quantified and compared by DAD and ELSD [57]. However, the results have shown that the ELSD has a poorer resolution than UV detectors for flavonoids which had responses in each chromatogram [57]. Another gradually popular aerosol-based detector is CAD, which measures charged particle flux after evaporating the mobile phases, whereas the ELSD measures light scattering by particles. CAD is a powerful mass-selective d­ etector

643

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CHAPTER 20  Quality control of traditional Chinese medicines

with several advantages, such as easier operation and much higher sensitivity (<1 ng), as well as better reproducibility than the ELSD [206,207]. Therefore, a lower concentration of analytes is required for CAD. So far, HPLC-CAD has been applied to the quality control of alkaloids [48], saponins [207], and saccharides [100,103]. Besides, ECD is a sensitive detector for the analysis of electrochemically reactive analytes [208]. Li et  al. have reported an HPLC-ECD method for the determination of bavachin and isobavachalcone in Fructus Psoraleae [19]. The results have demonstrated that the sensitivity of the analytes detected by ECD is 10 times higher than those obtained by DAD. Besides, PAD is the extension of single-potential amperometry for ECD and has widely been used in the detection of carbohydrates after HPAEC separation, but it has extensively been applied to alkaloids, amines, flavonoids, and saponins [116,117,119,120]. Kwon et al. have reported an RPLCPAD method for the determination of 4 saikosaponins in Bupleuri Radix and CaihuShugan-San, which has demonstrated that the determination of saponins by PAD is more sensitive than that by DAD [56].

20.3.3 ­MULTIANGLE LASER LIGHT SCATTERING (MALLS) DETECTION MALLS detector is considered as a powerful detector for the determination of molar mass and average size of macromolecules by measuring scattered light from the samples in solution, in contrast to a light scattering detector that measures the particles suspended in gas [141]. The mass and average size of the analyte could be measured using proper mathematical transformations without reference standards. HPSEC coupled to MALLS allows the measurement of Mw and distribution, and further provide absolute characterization information for macromolecules based on MALLS detection. The principles for absolute characterization of macromolecules by HPSEC-MALLS have been reviewed in the previous study [153]. Chen et  al. have reported an HPSEC-MALLS-RID method for quality control of lentinan injection produced in China, based on several parameters, including Mw, polydispersity index, and triple helical conformation [153]. In the present, HPSEC-MALLS had extensively been applied to the quality control of polysaccharides in TCMs [145,147–149,152,154,157].

20.3.4 ­MASS SPECTROMETRY (MS) DETECTION MS is a powerful analytical technique that could determine the chemical parameters and further elucidate the chemical structures of known and unknown molecules, as well as possesses superior sensitivity and selectivity [209]. Therefore, MS is considered as a more preferable universal detector for the identification of pure or complex analytes and trace amounts of molecules [210–212]. Combined with LC separation, LC-MS could be applied to qualitative and quantitative analyses of multiple components in complex analytes. It is considered that LC-MS is the major technique with more than half of the applications in the analysis of TCMs [5].

20.3 ­ Detections

MS has three elementary units, i.e. ion source, mass analyzer, and detector. The ion source is to induce molecules into ions through ionization. Soft ionizations impart less energy to molecules and result in less fragmentation which could be applied for liquid and solid samples, as compared with hard ionizations which are used for gases and vapors [213]. The interfaces for soft ionization include several applications, such as electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI) and matrix-assisted laser desorption ionization (MALDI) [5,6,214]. Among them, ESI interface with positive and/or negative modes has been the most applied interface and is widely used in the identification and quantification of phytochemicals in TCMs [215–217]. After the ionization, charged molecules are transported by magnetic or electric fields to a mass analyzer. The mass analyzer determines the mass resolving power and accuracy of the mass spectrometer. Several mass analyzers, such as quadrupole (Q), ion trap (IT), time of flight (TOF), and their hybrid forms, such as triple Q (QQQ), Q-TOF and LTQ-Orbitrap, have been widely used in LC-MS applications [218,219]. Among these analyzers, MS with TOF mass analyzers or hybrid mass analyzers, such as Q-TOF and LTQ-Orbitrap, are considered as high resolution MS, which allows fast and large-scale analysis, as well as provides accurate elucidation of mass data and structural information for the analytes [220]. LC-MS has a wide application for the quality control of bioactive components in TCMs, such as alkaloids [60,65,66,69,74,91], phenolic compounds [22– 25,58,63,64,67,72,76,78,80,85], terpenoids [16,27–29,61], saponins [33,35,42,53,87,88], nucleosides, and nucleobases [45,84,94,105,109,112,113]. Shen et al. have reported an HPLC-QTrap-MS/MS method for the identification and delicate determination of 5 major mogrosides in S. grosvenorii, followed by further confirmation of the UPLCQTOF-MS method [33]. Zhao et  al. have reported a quality control example for identifying and quantifying major triterpenoids in Alismatis Rhizoma based on HPLC-DAD-Q-TOF-MS and UPLC-QQQ-MS methods, respectively [29]. In all, 25 components have been characterized by Q-TOF-MS in 60 min, and have also been classified into 7 types based on the elucidation results from DAD and Q-TOF-MS. In the following steps, 14 triterpenoids have been simultaneously determined by UPLCQQQ-MS with an ESI interface and positive ion MRM mode for 8.5 min. Wang et al. have established an efficient, sensitive, and accurate method based on UPLC-QOrbitrap-MS/MS, for the determination of astragaloside IV that exists in Chinese functional spirits [35]. The combination of Q and Orbitrap allows high sensitivity and accurate mass orbitrap detection. Zhang et al. have developed a hybrid IT-TOF-MS method for the characterization of 6 aporphine alkaloids from Litsea cubeba, and the results demonstrate that the combination of IT and TOF has high resolving capacity [74]. Wang et  al. have reported a UHPLC-DAD coupled to the Fourier transform (FT) MS method for the identification of lanostane-type triterpene acids in different parts of Poria cocos, and were further quantified by UHPLC-QQQ-MS [27]. Results have shown that the established UHPLC-DAD-FT-MSn method have implemented accurate characterization with high sensitivity and high resolution.

645

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CHAPTER 20  Quality control of traditional Chinese medicines

20.3.5 ­BIOCHEMICAL DETECTION (BCD) The essential step for ensuring pharmacological bases of TCMs is to discover and identify bioactive substances. The conventional methods for the discovery of bioactive phytochemicals from natural products include the bioassay-guided fractionation and the screening of pure compounds, which are very laborious and time consuming, as well as hard to obtain pure compounds [221]. Therefore, HPLC coupled to online bioassays, or BCD, is considered as a robust and efficient method for screening of bioactive compounds in complex analytes. Combined with structure elucidation approaches, such as MS and nucleic magnetic resonance (NMR), screened compounds could be further elucidated. The applications for online bioassay in bioactivity-based quality control of medicinal herbs have been systematically reviewed [4,5,221]. HPLC-BCD methods have been applied to the discovery of antioxidants [222,223], enzyme inhibitors [224–226], and receptor binding agents [227–230] in TCMs. Li et al. have reported an online HPLC-DAD-MS/MS-BCD method for screening and characterization of α-glucosidase inhibitor from herbal tea extracts [195]. The instrumental setup of the established method is shown in Fig. 20.3A. The herbal extracts were first injected to a C18 column (4.6 × 250 mm, 5 μm) by a binary pump (consisting of 0.1% formic acid and methanol for two phases) for separation, and were followed by a makeup procedure to maintain the methanol at 30% constantly using another binary pump (consisting of water and methanol for two phases) in case of protein denaturation. Then a splitter was used to obtain a continuous flow rate of 0.1 mL/min to BCD, while the others were distributed to DAD and MS at 1 mL/min by a 1:1 ratio, as well as to waste at 0.9 mL/min. For BCD, 0.1 mL/min eluent, 0.05 mL/min α-glucosidase (0.4 U/mL), and 0.05 mL/min PNPG (2.5 mM, p-nitrophenyl α-d-glucopyranoside) on ice were continuously entered and mixed in a reaction coil. The products of inhibition, p-nitrophenol, were measured at 405 nm, while the measurement of the analytes were at 270 nm. As a result, (-)-epigallocatechin gallate (EGCG) and (-)-epicatechin gallate (ECG) were selected as potent α-glucosidase inhibitors from Pu-er tea (Fig. 20.3B). The established platform could be used for rapid screening of enzyme inhibitors.

20.3.6 ­MISCELLANEOUS Apart from the common detectors mentioned above, there are other superior detectors with great characteristics. Fluorescence detector (FLD) is used to measure the compounds that have natural fluorescence or those made to fluorescence through derivatization [231]. After the analytes have been excited at a higher energy excitation wavelength (λex), the fluorescence of the analytes could be detected under lower energy emission wavelength (λem). FLD can be 100 times more sensitive than UV detectors, and thus can be applied to the analysis of trace components. Wu et al. have reported a precolumn derivatization coupled to the HPLC-FLD method for quality control of six triterpenic acids in 6 different TCMs [28]. The synthesized derivatization reagent (DDCETS) was bound to triterpenic acids and further analyzed

Make-up pumps Water

LC Pumps 0.1% Formic acid MeOH

MeOH

Waste

Waste

MS

DAD

Column

Injector Enzyme

Substrate

Reaction coil 37°C

(A) BCD mAU

2.51 min

50

2.55 min

0

DAD

−50

BCD

−100 −150 5.06 min

−200 2

(B) mAU

4

6

8 30.33 min

24.77 min

ECG

EGCG

400

min

200

2.54 min

DAD BCD

0 2.52 min

−200 −400

(C)

27.29 min 32.87 min 10

mAU 500 400 300 200 100 0 −100 −200 −300

(D)

20

30

24.86 min

40

min

30.44 min

EGCG

ECG

2.49 min

DAD BCD

2.51 min 27.37 min 32.93 min 10

20

30

40

min

FIG. 20.3 Schematic diagram (A) of online HPLC-DAD-MS/MS-BCD platform for the rapid screening and identification of α-glucosidase inhibitors in complex mixtures, and typical UV chromatograms before (DAD, 270 nm) and after (BCD, 405 nm) online bioassays of acarbose (B), Pu-er tea (C), and mixed solution of EGCG and ECG (D). Modified from Li DQ, Qian ZM, Li SP. Inhibition of three selected beverage extracts on α-glucosidase and rapid identification of their active compounds using HPLC-DAD-MS/MS and biochemical detection. J Agric Food Chem 2010;58(11):6608–13 with permission of American Chemical Society.

648

CHAPTER 20  Quality control of traditional Chinese medicines

by HPLC-FLD under λex of 292 nm and λex of 402 nm. The established method has been proved to be a highly sensitive method with low detection limits.

20.4 ­CONCLUDING REMARKS LC is suitable for phytochemical components in TCM extracts and possesses distinctive separation capability. Moreover, LC separation modes based on different isolation mechanisms have been developed and continuously applied to the quality control of specific components of TCMs. The innovations of LC stationary phases and the extension of separation ranges are contributing to the developing trends of LC. First, transformations from HPLC to UPLC are considered as the substantial leap for LC in the quality control of TCMs, reflecting in smaller particle sizes and stronger separation capacity. The shortened analysis time and high resolution for UPLC allow fast detection of multiple compounds, which were highly suitable to complicated systems especially TCMs. Second, 2DLC incorporates separation ranges and efficiency of different LC modes, which is capable of analyzing global components in two or more LCs connected by miscellaneous interfaces. 2DLC exhibits high resolving powers and can be used for pattern recognition and identification of large-scale components in TCMs. Third, LC coupled to BCD or biochemical assays is another developing trend for the quality control of TCMs. The identification of effective components is essential to recognize the pharmacological bases of TCMs and online bioassays can boost up isolation and identification procedures. Overall, efficient and large-scale LC separation is the significant tendency for the quality control of TCMs.

­REFERENCES [1] Zhang  NL, Yuan  S, Chen  T, Wang  Y. Statistical validation of Traditional Chinese Medicine theories. J Altern Complement Med 2008;14(5):583–7. [2] Hsiao WL, Liu L. The role of traditional Chinese herbal medicines in cancer therapy— from TCM theory to mechanistic insights. Planta Med 2010;76(11):1118–31. [3] Lai  JN, Tang  JL, Wang  JD. Observational studies on evaluating the safety and adverse effects of traditional Chinese medicine. Evid Based Complement Alternat Med 2013;2013(6):1–9. [4] Li SP, Zhao J, Yang B. Strategies for quality control of Chinese medicines. J Pharm Biomed Anal 2011;55(4):802–9. [5] Zhao J, Ge LY, Xiong W, Leong F, Huang LQ, Li SP. Advanced development in phytochemicals analysis of medicine and food dual purposes plants used in China (2011– 2014). J Chromatogr A 2016;1428:39–54. [6] Zhao  J, Lv  GP, Chen  YW, Li  SP. Advanced development in analysis of phytochemicals from medicine and food dual purposes plants used in China. J Chromatogr A 2011;1218(42):7453–75. [7] Jin  HL, Liu  YF, Guo  ZM, Wang  JX, Zhang  XL, Wang  CR, et  al. Recent development in liquid chromatography stationary phases for separation of Traditional Chinese Medicine components. J Pharm Biomed Anal 2016;130:336–46.

­References

[8] Zhao J, Deng JW, Chen YW, Li SP. Advanced phytochemical analysis of herbal tea in China. J Chromatogr A 2013;1313(19):2–23. [9] Michael  S. HPLC detectors: a brief review. J Liq Chromatogr Relat Technol 2010;33(9):1130–50. [10] Claessens HA, Straten MAV. Review on the chemical and thermal stability of stationary phases for reversed-phase liquid chromatography. J Chromatogr A 2004;1060(1-2):23–41. [11] Huang HL, Liu M, Chen P. Recent advances in ultra-high performance liquid chromatography for the analysis of traditional Chinese Medicine. Anal Lett 2014;47(11):1835–51. [12] Chen X, Wang J, Hu S, Bai X. Hollow-fiber double-solvent synergistic microextraction with high-performance liquid chromatography for the determination of antitumor alkaloids in Coptis chinensis. J Sep Sci 2016;39(5):827–34. [13] Yan R, Yu S, Liu H, Xue Z, Yang B. An HPLC-DAD method for simultaneous quantitative determination of four active hydrophilic compounds in Magnoliae Officinalis cortex. J Chromatogr Sci 2015;53(4):598–602. [14] Zhang X, Ning Z, Ji D, Chen Y, Mao C, Lu T. Approach based on high-performance liquid chromatography fingerprint coupled with multivariate statistical analysis for the quality evaluation of Gastrodia Rhizoma. J Sep Sci 2015;38(22):3825–31. [15] Cen M, Ruan J, Huang L, Zhang Z, Yu N, Zhang Y, et al. Simultaneous determination of thirteen flavonoids from Xiaobuxin-Tang extract using high-performance liquid chromatography coupled with electrospray ionization mass spectrometry. J Pharm Biomed Anal 2015;115:214–24. [16] Cui X, Cai H, Li H, Tao Y, Huang P, Qian X, et al. Simultaneous determination of 10 flavonoids in crude and wine-processed radix scutellariae by UHPLC. J Chromatogr Sci 2016;54(3):312–7. [17] Dai W, Zhao W, Gao F, Shen J, Lv D, Qi Y, et al. Simultaneous chemical fingerprint and quantitative analysis of Rhizoma Smilacis Glabrae by accelerated solvent extraction and high-performance liquid chromatography with tandem mass spectrometry. J Sep Sci 2015;38(9):1466–75. [18] Li BQ, Chen J, Li JJ, Wang X, Zhai HL, Zhang XY. High-performance liquid chromatography with photodiode array detection and chemometrics method for the analysis of multiple components in the traditional Chinese medicine Shuanghuanglian oral liquid. J Sep Sci 2015;38(24):4187–95. [19] Li  Y, Wang  F, Chen  Z. Determination of bavachin and isobavachalcone in Fructus Psoraleae by high-performance liquid chromatography with electrochemical detection. J Sep Sci 2011;34(5):514–9. [20] Liu  HM, Nie  L. Quantitative analysis combined with chromatographic fingerprint for comprehensive evaluation of Xiaoer Chaigui Tuire granules by HPLC-DAD. J Chromatogr Sci 2015;53(5):749–56. [21] Xie  ZS, Lam  SC, Wu  JW, Yang  DP, Xu  XJ. Chemical fingerprint and simultaneous determination of flavonoids in Flos Sophorae Immaturus by HPLC-DAD and HPLC-DAD-ESI-MS/MS combined with chemometrics analysis. Anal Methods 2014;6(12):4328–35. [22] Zhang Y, Guo L, Duan L, Dong X, Zhou P, Liu EH, et al. Simultaneous determination of 16 phenolic constituents in Spatholobi Caulis by high performance liquid chromatography/electrospray ionization triple quadrupole mass spectrometry. J Pharm Biomed Anal 2015;102:110–8. [23] Chen Y, Zhang N, Ma J, Zhu Y, Wang M, Wang X, et al. A Platelet/CMC coupled with offline UPLC-QTOF-MS/MS for screening antiplatelet activity components from aqueous extract of Danshen. J Pharm Biomed Anal 2016;117:178–83.

649

650

CHAPTER 20  Quality control of traditional Chinese medicines

[24] Wang  X, Wu  Y, Wu  Q, Qian  Y, Yue  W, Liang  Q. Ultra-high performance liquid ­chromatography-tandem mass spectrometry for rapid analysis of seven phenolic compounds of Sparganii rhizoma. Acta Chromatogr 2015;27(4):755–66. [25] Zhang C, Sun L, Tian RT, Jin HY, Ma SC, Gu BR. Combination of quantitative analysis and chemometric analysis for the quality evaluation of three different frankincenses by ultra high performance liquid chromatography and quadrupole time of flight mass spectrometry. J Sep Sci 2015;38(19):3324–30. [26] Keypour S, Rafati H, Riahi H, Mirzajani F, Moradali MF. Qualitative analysis of ganoderic acids in Ganoderma lucidum from Iran and China by RP-HPLC and electrospray ionisation-mass spectrometry (ESI-MS). Food Chem 2010;119(4):1704–8. [27] Wang W, Dong H, Yan R, Li H, Li P, Chen P, et al. Comparative study of lanostane-type triterpene acids in different parts of Poria cocos (Schw.) Wolf by UHPLC-Fourier transform MS and UHPLC-triple quadruple MS. J Pharm Biomed Anal 2015;102:203–14. [28] Wu H, Li G, Liu S, Liu D, Chen G, Hu N, et al. Simultaneous determination of six triterpenic acids in some Chinese medicinal herbs using ultrasound-assisted dispersive liquid-liquid microextraction and high-performance liquid chromatography with fluorescence detection. J Pharm Biomed Anal 2015;107:98–107. [29] Zhao W, Huang X, Li X, Zhang F, Chen S, Ye M, et al. Qualitative and quantitative analysis of major triterpenoids in Alismatis Rhizoma by high performance liquid chromatography/diode-array detector/quadrupole-time-of-flight mass spectrometry and ultra-performance liquid chromatography/triple quadrupole mass spectrometry. Molecules 2015;20(8):13958–81. [30] Li YW, Qi J, Zhang W, Zhou SP, Wu Y, Yu BY. Determination and Fingerprint Analysis of Steroidal Saponins in roots of Liriope muscari (Decne.) L. H. Bailey by ultra high performance liquid chromatography coupled with ion trap time-of-flight mass spectrometry. J Sep Sci 2014;37(14):1762–72. [31] Man S, Gao W, Zhang Y, Wang J, Zhao W, Huang L, et al. Qualitative and quantitative determination of major saponins in Paris and Trillium by HPLC-ELSD and HPLC-MS/ MS. J Chromatogr B 2010;878(29):2943–8. [32] Shan SM, Luo JG, Huang F, Kong LY. Chemical characteristics combined with bioactivity for comprehensive evaluation of Panax ginseng C.A. Meyer in different ages and seasons based on HPLC-DAD and chemometric methods. J Pharm Biomed Anal 2014;89:76–82. [33] Shen Y, Lin S, Han C, Zhu Z, Hou X, Long Z, et al. Rapid identification and quantification of five major mogrosides in Siraitia grosvenorii (Luo-Han-Guo) by high performance liquid chromatography-triple quadrupole linear ion trap tandem mass spectrometry combined with microwave-assisted extraction. Microchem J 2014;116:142–50. [34] Shi Y, Xiong J, Sun D, Liu W, Wei F, Ma S, et al. Simultaneous quantification of the major bile acids in Artificial Calculus bovis by high-performance liquid chromatography with precolumn derivatization and its application in quality control. J Sep Sci 2015;38(16):2753–62. [35] Wang  L, Zhang  Z, Yu  X, Zhao  H, Wu  X, Chen  X, et  al. An efficient, sensitive and ­accurate method for the detection of astragaloside IV in Chinese functional spirit by ultra-performance liquid chromatography-quadrupole-orbitrap mass spectrometry. ­ Anal Methods 2015;7(24):10199–206. [36] Wang S, Huang J, Mao H, Wang Y, Kasimu R, Xiao W, et al. A novel method HPLC-DAD analysis of the contents of Moutan cortex and Paeoniae radix alba with similar constituentsmonoterpene glycosides in Guizhi Fuling Wan. Molecules 2014;19(11):17957–67.

­References

[37] Fekete S, Schappler J, Veuthey JL, Guillarme D. Current and future trends in UHPLC. TrAC Trends Anal Chem 2014;63:2–13. [38] Zotou A. An overview of recent advances in HPLC instrumentation. Cent Eur J Chem 2012;10(3):554–69. [39] Kong W, Jin C, Liu W, Xiao X, Zhao Y, Li Z, et al. Development and validation of a UPLC-ELSD method for fast simultaneous determination of five bile acid derivatives in Calculus Bovis and its medicinal preparations. Food Chem 2010;120(4):1193–200. [40] Jemal  M, Hawthorne  DJ. Effect of high performance liquid chromatography mobile phase (methanol versus acetonitrile) on the positive and negative ion electrospray response of a compound that contains both an unsaturated lactone and a methyl sulfone group. Rapid Commun Mass Spectrom 1999;13(1):61–6. [41] Xu W, Huang M, Li H, Chen X, Zhang Y, Liu J, et al. Chemical profiling and quantification of Gua-Lou-Gui-Zhi decoction by high performance liquid chromatography/ quadrupole-time-of-flight mass spectrometry and ultra-performance liquid chromatography/triple quadrupole mass spectrometry. J Chromatogr B 2015;986-987:69–84. [42] Li MN, Dong X, Gao W, Liu XG, Wang R, Li P, et al. Global identification and quantitative analysis of chemical constituents in traditional Chinese medicinal formula Qi-FuYin by ultra-high performance liquid chromatography coupled with mass spectrometry. J Pharm Biomed Anal 2015;114:376–89. [43] Tindall GW, Dolan JW. Mobile phase buffers, part II—buffer selection and capacity. LC GC Eur 2003;16(3):329–44. [44] Franey T. A volatile ion-pairing chomatography reagent for an LC-MS mobile phase. LCGC N Am 2003;21(1):54–8. [45] Yang  FQ, Li  DQ, Feng  K, Hu  DJ, Li  SP. Determination of nucleotides, nucleosides and their transformation products in Cordyceps by ion-pairing reversed-phase liquid chromatography-mass spectrometry. J Chromatogr A 2010;1217(34):5501–10. [46] Bai L, Jiang M, Guo S, Liu Q, Zhang X, Tian X, et al. Simultaneous quantification of six sesquiterpene lactones and a flavonoid in the whole life stage of: Inula salsoloides by high performance liquid chromatography. Anal Methods 2016;8(17):3587–91. [47] Zhao  X, Xie  L, Wu  H, Kong  W, Yang  M. Analysis of six bioactive components in Semen Ziziphi Spinosae by UPLC-ELSD and UPLC-Q/TOF-MS. Anal Methods 2014;6(15):5856–64. [48] Long  Z, Guo  Z, Acworth  IN, Liu  X, Jin  Y, Liu  X, et  al. A non-derivative method for the quantitative analysis of isosteroidal alkaloids from Fritillaria by high performance liquid chromatography combined with charged aerosol detection. Talanta 2016;151:239–44. [49] Majors RE, Przybyciel M. Columns for reversed-phase LC separations in highly aqueous mobile phases. LCGC N Am 2002;20(7):584. [50] Han DQ, Zhao J, Xu J, Peng HS, Chen XJ, Li SP. Quality evaluation of Polygonum multiflorum in China based on HPLC analysis of hydrophilic bioactive compounds and chemometrics. J Pharm Biomed Anal 2013;72(2):223–30. [51] Li SP, Qiao CF, Chen YW, Zhao J, Cui XM, Zhang QW, et al. A novel strategy with standardized reference extract qualification and single compound quantitative evaluation for quality control of Panax notoginseng used as a functional food. J Chromatogr A 2013;1313(19):302–7. [52] Zhang F, Qi P, Xue R, Li Z, Zhu K, Wan P, et al. Qualitative and quantitative analysis of the major constituents in Acorus tatarinowii Schott by HPLC/ESI-QTOF-MS/MS. Biomed Chromatogr 2015;29(6):890–901.

651

652

CHAPTER 20  Quality control of traditional Chinese medicines

[53] Qin  Z, Lin  P, Dai  Y, Yao  Z, Wang  L, Yao  X, et  al. Quantification and semiquantification of multiple representative components for the holistic quality control of Allii Macrostemonis Bulbus by ultra high performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry. J Sep Sci 2016;39(10):1834–41. [54] Wen  H, Gao  HY, Qi  W, Xiao  F, Wang  LL, Wang  D, et  al. Simultaneous determination of twenty-two components in Asari Radix et Rhizoma by ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Planta Med 2014;80(18):1753–62. [55] Cui Q, Fu M, Zhou M, Chang N, Ye P, Jiang M, et al. Bioactivity-based ultra-­performance liquid chromatography-coupled quadrupole time-of-flight mass spectrometry for NFκB inhibitors identification in Chinese Medicinal Preparation Bufei Granule. Biomed Chromatogr 2016;30:1184–9. [56] Kwon HJ, Sim HJ, Lee SI, Lee YM, Lee JH, Park YD, et al. Analysis of saikosaponins in Bupleuri Radix and Caihu-shugan-san using reversed-phase HPLC with pulsed amperometric detection. J Sep Sci 2011;34(6):651–8. [57] Hu F, Zhu R, Liu X, Yang Y, Li C, Feng S, et al. Simultaneous determination of 10 components in Bu-Zhong-Yi-Qi Wan by solid phase extraction-high performance liquid chromatography with diode array detection and evaporative light scattering detection. J Chromatogr Sci 2015;53(5):736–41. [58] Liu XT, Wang XG, Yang Y, Xu R, Meng FH, Yu NJ, et al. Qualitative and quantitative analysis of lignan constituents in Caulis Trachelospermi by HPLC-QTOF-MS and HPLC-UV. Molecules 2015;20(5):8107–24. [59] He  JY, Zhu  S, Komatsu  K. HPLC/UV analysis of polyacetylenes, phenylpropanoid and pyrrolidine alkaloids in medicinally used Codonopsis species. Phytochem Anal 2014;25(3):213–9. [60] Zhu  F, Chen  J, Wang  J, Yin  R, Li  X, Jia  X. Qualitative and quantitative analysis of the constituents in Danmu preparations by UPLC-PDA-TOF-MS. J Chromatogr Sci 2014;52(8):862–71. [61] Hou  L, Zuo  L, Sun  Z, Sun  Z, Zhu  Z, Kang  J, et  al. A new approach to rapid determination of 18 bioactive constituents in Alpinia oxyphylla using UPLC-MS/MS with positive-negative conversion multiple reaction monitor (+/-MRM) technology. Anal Methods 2016;8(15):3163–70. [62] Liu Z, Zhu Z, Zhang H, Tan G, Chen X, Chai Y. Qualitative and quantitative analysis of Fructus Corni using ultrasound assisted microwave extraction and high performance liquid chromatography coupled with diode array UV detection and time-of-flight mass spectrometry. J Pharm Biomed Anal 2011;55(3):557–62. [63] Han  Y, Wen  J, Zhou  T, Fan  G. Chemical fingerprinting of Gardenia jasminoides Ellis by HPLC-DAD-ESI-MS combined with chemometrics methods. Food Chem 2015;188:648–57. [64] Yin F, Wu X, Li L, Chen Y, Lu T, Li W, et al. Quality control of Gardeniae Fructus by HPLCPDA fingerprint coupled with chemometric methods. J Chromatogr Sci 2015;53(10):1685–94. [65] Shi Z, Li Z, Zhang S, Fu H, Zhang H. Subzero-temperature liquid-liquid extraction coupled with UPLC-MS-MS for the simultaneous determination of 12 bioactive ­components in traditional Chinese medicine Gegen-Qinlian Decoction. J Chromatogr Sci 2015;53(8):1407–13. [66] Wang Q, Song W, Qiao X, Ji S, Kuang Y, Zhang ZX, et al. Simultaneous quantification of 50 bioactive compounds of the traditional Chinese medicine formula Gegen-Qinlian decoction using ultra-high performance liquid chromatography coupled with tandem mass spectrometry. J Chromatogr A 2016;1454:15–25.

­References

[67] Pan Y, Shen T, Zhang J, Zhao YL, Wang YZ, Li WY. Simultaneous determination of six index constituents and comparative analysis of four ethnomedicines from genus Gentiana using a UPLC-UVMS method. Biomed Chromatogr 2015;29(1):87–96. [68] Zeng R, Hu H, Ren G, Liu H, Qu Y, Hua W, et al. Chemical profiling assisted quality assessment of Gentianae macrophyllae by high-performance liquid chromatography using a fused-core column. J Chromatogr Sci 2015;53(8):1274–9. [69] Wang C, Xie Y, Xiang Z, Zhou H, Liu L. Simultaneous determination of thirteen major active compounds in Guanjiekang preparation by UHPLC-QQQ-MS/MS. J Pharm Biomed Anal 2016;118:315–21. [70] Liu  D, Mei  Q, Wan  X, Que  H, Li  L, Wan  D. Determination of rutin and isoquercetin contents in Hibisci mutabilis Folium in different collection periods by HPLC. J Chromatogr Sci 2015;53(10):1680–4. [71] Luo HL, Zhong J, Ye FY, Wang Q, Ma YM, Liu P, et al. A systematic quality control method of Huangqi decoction: simultaneous determination of eleven flavonoids and seven triterpenoid saponins by UHPLC-MS. Anal Methods 2014;6(13):4593–601. [72] Du Y, Liu P, Yuan Z, Jin Y, Zhang X, Sheng X, et al. Simultaneous qualitative and quantitative analysis of 28 components in Isodon rubescens by HPLC-ESI-MS/MS. J Sep Sci 2010;33(4-5):545–57. [73] Pitschmann A, Zehl M, Heiss E, Purevsuren S, Urban E, Dirsch VM, et al. Quantitation of phenylpropanoids and iridoids in insulin‐sensitising extracts of Leonurus sibiricus L. (Lamiaceae). Phytochem Anal 2016;27(1):23–31. [74] Zhang S, Zhang Q, Guo Q, Zhao Y, Gao X, Chai X, et al. Characterization and simultaneous quantification of biological aporphine alkaloids in Litsea cubeba by HPLC with hybrid ion trap time-of-flight mass spectrometry and HPLC with diode array detection. J Sep Sci 2015;38(15):2614–24. [75] Qiu Y, Huang J, Jiang X, Chen Y, Liu Y, Zeng R, et al. Quantitative and qualitative determination of LiuweiDihuang preparations by ultra high performance liquid chromatography in dual-wavelength fingerprinting mode and random forest. J Sep Sci 2015;38(21):3720–6. [76] Inbaraj  BS, Lu  H, Kao  TH, Chen  BH. Simultaneous determination of phenolic acids and flavonoids in Lycium barbarum Linnaeus by HPLC-DAD-ESI-MS. J Pharm Biomed Anal 2010;51(3):549–56. [77] Zhang Q, Hong B, Liu J, Mu G, Cong H, Li G, et al. Multiwalled-carbon-nanotubesbased matrix solid-phase dispersion extraction coupled with high-performance liquid chromatography for the determination of honokiol and magnolol in Magnoliae Cortex. J Sep Sci 2014;37(11):1330–6. [78] Zhang L, Wang C, Miao D, Yuan D, Bi K, Yang J, et al. Simultaneous determination of six constituents in Mahuang Fuzi Xixin by UPLC-PDA-MS/MS. Nat Prod Res 2016;29(8):772–5. [79] Wang Y, Huang L. Comparison of two species of Notopterygium by GC-MS and HPLC. Molecules 2015;20(3):5062–73. [80] Guan Z, Li S, Lin Z, Yang R, Zhao Y, Liu J, et al. Identification and quantitation of phenolic compounds from the seed and pomace of Perilla frutescens using HPLC/PDA and HPLC-ESI/QTOF/MS/MS. Phytochem Anal 2014;25(6):508–13. [81] Zhang Z, Liu H, Zhang B, Liao Y, Zhang Y, Zhang Z. Quantitative and chemical fingerprint analysis for quality evaluation of the dried bark of wild Phellodendron amurense Rupr. based on HPLC-DAD-MS combined with chemometrics methods. Anal Methods 2015;7(5):2041–9.

653

654

CHAPTER 20  Quality control of traditional Chinese medicines

[82] Granica S. Quantitative and qualitative investigations of pharmacopoeial plant material polygoni avicularis herba by UHPLC-CAD and UHPLC-ESI-MS methods. Phytochem Anal 2015;26(5):374–82. [83] Lu X, Zhang R, Fu F, Shen J, Nian H, Wu T. Simultaneous determination of fangchinoline and tetrandrine in Qi-Fang-Xi-Bi-Granules by RP-HPLC. J Chromatogr Sci 2015;53(8):1328–32. [84] Yang  W, Feng  C, Kong  D, Shi  X, Zheng  X, Cui  Y, et  al. Simultaneous determination of 15 components in Radix Glehniae by high performance liquid chromatography-­ electrospray ionization tandem mass spectrometry. Food Chem 2010;120(3):886–94. [85] Lin L, Ni B, Lin H, Zhang M, Yan L, Qu C, et al. Simultaneous determination of 14 constituents of Radix polygoni multiflori from different geographical areas by liquid ­chromatography-tandem mass spectrometry. Biomed Chromatogr 2015;29(7):1048–55. [86] Wang L, Zhu X, Lou X, Zheng F, Feng Y, Liu W, et al. Systematic characterization and simultaneous quantification of the multiple components of Rhododendron dauricum based on high-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry. J Sep Sci 2015;38(18):3161–9. [87] Luo H, Kong W, Hu Y, Chen P, Wu X, Wan L, et al. Quality evaluation of Salvia miltiorrhiza Bge. by ultra high performance liquid chromatography with photodiode array detection and chemical fingerprinting coupled with chemometric analysis. J Sep Sci 2015;38(9):1544–51. [88] Chen D, Lin S, Xu W, Huang M, Chu J, Xiao F, et al. Qualitative and quantitative analysis of the major constituents in Shexiang Tongxin dropping pill by HPLC-Q-TOF-MS/ MS and UPLC-QqQ-MS/MS. Molecules 2015;20(10):18597–619. [89] Yan Z, Yang X, Wu J, Su H, Chen C, Chen Y. Qualitative and quantitative analysis of chemical constituents in traditional Chinese medicinal formula Tong-Xie-Yao-Fang by high-performance liquid chromatography/diode array detection/electrospray ionization tandem mass spectrometry. Anal Chim Acta 2011;691(1-2):110–8. [90] Shu Z, Li X, Rahman K, Qin L, Zheng C. Chemical fingerprint and quantitative analysis for the quality evaluation of Vitex negundo seeds by reversed-phase high-­performance liquid chromatography coupled with hierarchical clustering analysis. J Sep Sci 2016;39(2):279–86. [91] Tian T, Jin Y, Ma Y, Xie W, Xu H, Du Y. Simultaneous quantification of 11 constituents in Wuji Pill using ultra performance liquid chromatography coupled with a triple quadrupole electrospray tandem mass spectrometry. J Chromatogr Sci 2016;54(2):237–45. [92] Wang L, Wu C, Zhao L, Lu X, Wang F, Yang J, et al. An ultra-performance liquid chromatography photodiode array detection tandem mass spectrometric method for simultaneous determination of seven major bioactive constituents in xiaochaihutang and its application to fourteen compatibilities study. J Chromatogr Sci 2015;53(9):1570–6. [93] Yang N, Xiong A, Wang R, Yang L, Wang Z. Quality evaluation of traditional Chinese medicine compounds in xiaoyan lidan tablets: fingerprint and quantitative analysis using UPLC-MS. Molecules 2016;21(2):83. [94] Zhou L, Qi W, Xu C, Makino T, Yuan D. A rapid method for simultaneous determination of 52 marker compounds in Xiao-Qing-Long-Tang by ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. J Sep Sci 2014;37(22):3260–7. [95] Wong TL, An YQ, Yan BC, Yue RQ, Zhang TB, Ho HM, et al. Comprehensive quantitative analysis of Chinese patent drug YinHuang drop pill by ultra high-performance liquid chromatography quadrupole time of flight mass spectrometry. J Pharm Biomed Anal 2016;125:415–26.

­References

[96] Chen J, Yang Y, Shi YP. Simultaneous quantification of twelve active components in yiqing granule by ultra-performance liquid chromatography: application to quality control study. Biomed Chromatogr 2011;25(9):1045–53. [97] Tang Z, Yin R, Bi K, Zhu H, Han F, Chen K, et al. Simultaneous quantitative determination of 20 active components in the traditional Chinese medicine formula ZhiZi-Da-Huang decoction by liquid chromatography coupled with mass spectrometry: application to study the chemical composition variations in different combinations. Biomed Chromatogr 2015;29(9):1406–14. [98] Deng  X, Yu  J, Zhao  M, Zhao  B, Xue  X, Che  C, et  al. Quality assessment of crude and processed ginger by high-performance liquid chromatography with diode array detection and mass spectrometry combined with chemometrics. J Sep Sci 2015;38(17):2945–52. [99] Zhang TB, Yue RQ, Xu J, Ho HM, Ma DL, Leung CH, et al. Comprehensive quantitative analysis of Shuang-Huang-Lian oral liquid using UHPLC-Q-TOF-MS and HPLCELSD. J Pharm Biomed Anal 2015;102:1–8. [100] Li  J, Hu  D, Zong  W, Lv  G, Zhao  J, Li  S. Determination of inulin-type fructooligosaccharides in edible plants by high-performance liquid chromatography with charged aerosol detector. J Agric Food Chem 2014;62(31):7707–13. [101] Liu  R, Duan  JA, Chai  C, Wen  HM, Guo S, Wang XZ, et al. Hydrophilic interaction ultra-high performance liquid chromatography coupled with triple-quadrupole mass spectrometry for determination of nucleosides and nucleobases in animal horns. J Liq Chromatogr Relat Technol 2015;38(12):1185–93. [102] Li  J, Liu  XM, Zhou  B, Zhao  J, Li  SP. Determination of fructooligosaccharides in burdock using HPLC and microwave-assisted extraction. J Agric Food Chem 2013;61(24):5888–92. [103] Yan J, Shi S, Wang H, Liu R, Ning L, Chen Y, et al. Neutral monosaccharide composition analysis of plant-derived oligo- and polysaccharides by high performance liquid chromatography. Carbohydr Polym 2016;136:1273–80. [104] Li FT, Duan JA, Qian DW, Guo S, Ding YH, Liu XH, et al. Comparative analysis of nucleosides and nucleobases from different sections of Elaphuri Davidiani Cornu and Cervi Cornu by UHPLC-MS/MS. J Pharm Biomed Anal 2013;83:10–8. [105] Wang  H, Wu  Q, Wu  C, Jiang  Z. Simultaneous determination of 16 nucleosides and nucleobases in Euryale ferox Salisb. by liquid chromatography coupled with electro spray ionization tandem triple quadrupole mass spectrometry (HPLC-ESITQ-MS/MS) in multiple reaction monitoring (MRM) mode. J Chromatogr Sci 2015;53(8):1386–94. [106] Yang  L, Chen  LL, Xu  SJ, Zeng  X, Feng  Y, Xie  PS. RRLC-MS/MS method for the quantitation of atractyloside in Fructus Xanthii (Xanthium sibiricum). Anal Methods 2013;5(8):2093–7. [107] Qin  J, Feng  J, Li  Y, Mo  K, Lu  S. Ultrasonic-assisted liquid-liquid extraction and HILIC-ELSD analysis of ginsenoside Rb1, astragaloside IV and dulcitol in sugar-free "Fufangfufangteng Heji". J Pharm Biomed Anal 2011;56(4):836–40. [108] Chen  P, Li  W, Li  Q, Wang  Y, Li  Z, Ni  Y, et  al. Identification and quantification of nucleosides and nucleobases in Geosaurus and Leech by hydrophilic-interaction chromatography. Talanta 2011;85(3):1634–41. [109] Yao X, Zhou G, Tang Y, Guo S, Qian D, Duan JA. HILIC-UPLC-MS/MS combined with hierarchical clustering analysis to rapidly analyze and evaluate nucleobases and nucleosides in Ginkgo biloba leaves. Drug Test Anal 2015;7(2):150–7.

655

656

CHAPTER 20  Quality control of traditional Chinese medicines

[110] Zhou GS, Pang HQ, Tang YP, Yao X, Mo X, Zhu SQ, et al. Hydrophilic interaction ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry for highly rapid and sensitive analysis of underivatized amino acids in functional foods. Amino Acids 2013;44(5):1293–305. [111] Zhou GS, Pang HQ, Tang YP, Yao X, Ding YH, Zhu SQ, et al. Hydrophilic interaction ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (HILIC-UPLC-TQ-MS/MS) in multiple-reaction monitoring (MRM) for the determination of nucleobases and nucleosides in ginkgo seeds. Food Chem 2014;150:260–6. [112] Zhang LL, Bai YL, Shu SL, Qian DW, Zhen OY, Liu L, et al. Simultaneous quantitation of nucleosides, nucleobases, amino acids, and alkaloids in mulberry leaf by ultra high performance liquid chromatography with triple quadrupole tandem mass spectrometry. J Sep Sci 2014;37(11):1265–75. [113] Zhang C, Xiong Y, Dong Q, Gao D, Zhang LL, Ma LN, et al. Comparison of reversedphase liquid chromatography and hydrophilic interaction chromatography for the fingerprint analysis of Radix isatidis. J Sep Sci 2014;37(9-10):1141–7. [114] Guo S, Duan JA, Qian DW, Wang HQ, Tang YP, Qian YF, et al. Hydrophilic interaction ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry for determination of nucleotides, nucleosides and nucleobases in Ziziphus plants. J Chromatogr A 2013;1301:147–55. [115] Wu HZ, Ren CY, Yang F, Qin YF, Zhang YX, Liu JW. Extraction and identification of collagen-derived peptides with hematopoietic activity from Colla Corii Asini. J Ethnopharmacol 2016;182:129–36. [116] Xie JH, Shen MY, Nie SP, Liu X, Zhang H, Xie MY. Analysis of monosaccharide composition of Cyclocarya paliurus polysaccharide with anion exchange chromatography. Carbohydr Polym 2013;98(1):976–81. [117] Yoshihashi T, Do HTT, Tungtrakul P, Boonbumrung S, Yamaki K. Simple, selective, and rapid quantification of 1-deoxynojirimycin in mulberry leaf products by high-­ performance anion-exchange chromatography with pulsed amperometric detection. J Food Sci 2010;75(3):C246–50. [118] Qiao CF, Liu XM, Cui XM, Hu DJ, Chen YW, Jing Z, et al. High-performance anionexchange chromatography coupled with diode array detection for the determination of dencichine in Panax notoginseng and related species. J Sep Sci 2013;36(15):2401–6. [119] Yi YN, Cheng XM, Liu LA, Hu GY, Wang ZT, Deng YD, et al. Simultaneous determination of synephrine, arecoline, and norisoboldine in Chinese patent medicine SiMo-Tang oral liquid preparation by strong cation exchange high performance liquid chromatography. Pharm Biol 2012;50(7):832–8. [120] Sim  H-J, Jeong  J-S, Kwon  H-J, Lee  Y-M, Hong  S-P. Sensitive high-performance anion-exchange chromatographic determination of paeoniflorin and albiflorin by pulsed amperometric detection after solid-phase extraction. J Chromatogr A 2010;1217(32):5302–5. [121] Choi SH, Ahn JB, Kim HJ, Im NK, Kozukue N, Levin CE, et al. Changes in free amino acid, protein, and flavonoid content in jujube (Ziziphus jujube) fruit during eight stages of growth and antioxidative and cancer cell inhibitory effects by extracts. J Agric Food Chem 2012;60(41):10245–55. [122] Alpert AJ. Hydrophilic-interaction chromatography for the separation of peptides, nucleic acids and other polar compounds. J Chromatogr A 1990;499(2):177–96.

­References

[123] Jandera P. Stationary and mobile phases in hydrophilic interaction chromatography: a review. Anal Chim Acta 2011;692(1-2):1–25. [124] Hemström  P, Irgum  K. Hydrophilic interaction chromatography. J Sep Sci 2006;29(12):1784–821. [125] Bogusław  B, Sylwia  N. Hydrophilic interaction liquid chromatography (HILIC)—a powerful separation technique. Anal Bioanal Chem 2012;402(1):231–47. [126] Aurélie P, Krull IS, Davy G. Applications of hydrophilic interaction chromatography to amino acids, peptides, and proteins. J Sep Sci 2015;38(3):357–67. [127] Ares AM, Bernal J. Hydrophilic interaction chromatography in drug analysis. Cent Eur J Chem 2012;10(3):534–53. [128] Zauner  G, Deelder  AM, Wuhrer  M. Recent advances in hydrophilic interaction liquid chromatography (HILIC) for structural glycomics. Electrophoresis 2011;32(24):3456–66. [129] Liu Y, Xue X, Guo Z, Xu Q, Zhang F, Liang X. Novel two-dimensional reversed-phase liquid chromatography/hydrophilic interaction chromatography, an excellent orthogonal system for practical analysis. J Chromatogr A 2008;1208(1):133–40. [130] Buszewski  B, Noga  S. Hydrophilic interaction liquid chromatography (HILIC)—a powerful separation technique. Anal Bioanal Chem 2012;402(1):231–47. [131] Fei F, Bowdish DME, McCarry BE. Comprehensive and simultaneous coverage of lipid and polar metabolites for endogenous cellular metabolomics using HILIC-TOF-MS. Anal Bioanal Chem 2014;406(15):3723–33. [132] Jandera  P, Churáček  J. Ion-exchange chromatography of nitrogen compounds. J Chromatogr A 1974;98(1):1–54. [133] Nesterenko EP, Nesterenko PN, Paull B. Zwitterionic ion-exchangers in ion chromatography: a review of recent developments. Anal Chim Acta 2009;652(1-2):3–21. [134] Janoš  P. Retention models in ion chromatography: the role of side equilibria in ion-exchange chromatography of inorganic cations and anions. J Chromatogr A 1997;789(1–2):3–19. [135] Fekete  S, Beck  A, Veuthey  JL, Guillarme  D. Ion-exchange chromatography for the characterization of biopharmaceuticals. J Pharm Biomed Anal 2015;113(4):111–5. [136] Lenhoff  AM. Protein adsorption and transport in polymer-functionalized ion-­ exchangers. J Chromatogr A 2011;1218(49):8748–59. [137] Zhang  Z, Khan  NM, Nunez  KM, Chess  EK, Szabo  CM. Complete monosaccharide analysis by high-performance anion-exchange chromatography with pulsed amperometric detection. Anal Chem 2012;84(9):4104–10. [138] Paull B, Haddad PR. Chelation ion chromatography of trace metal ions using metallochromic ligands. TrAC Trends Anal Chem 1999;18(2):107–14. [139] Petruczynik A, Waksmundzka-Hajnos M. High performance liquid chromatography of selected alkaloids in ion-exchange systems. J Chromatogr A 2013;1311(18):48–54. [140] Gaborieau M, Castignolles P. Size-exclusion chromatography (SEC) of branched polymers and polysaccharides. Anal Bioanal Chem 2011;399(4):1413–23. [141] Li SP, Wu DT, Lv GP, Zhao J. Carbohydrates analysis in herbal glycomics. TrAC Trends Anal Chem 2013;52(12):155–69. [142] Duong-Ly KC, Gabelli SB. Gel filtration chromatography (size exclusion chromatography) of proteins. Methods Enzymol 2014;541:105–14. [143] Eremeeva T. Size-exclusion chromatography of enzymatically treated cellulose and related polysaccharides: a review. J Biochem Biophys Methods 2003;56(1-3):253–64.

657

658

CHAPTER 20  Quality control of traditional Chinese medicines

[144] Guan J, Zhao J, Feng K, Hu DJ, Li SP. Comparison and characterization of polysaccharides from natural and cultured Cordyceps using saccharide mapping. Anal Bioanal Chem 2011;399(10):3465–74. [145] Wu DT, Meng LZ, Wang LY, Lv GP, Cheong KL, Hu DJ, et al. Chain conformation and immunomodulatory activity of a hyperbranched polysaccharide from Cordyceps sinensis. Carbohydr Polym 2014;110:405–14. [146] Lin  PC, Wu  DT, Xie  J, Zhao  J, Li  SP. Characterization and comparison of bioactive polysaccharides from the tubers of Gymnadenia conopsea. Food Hydrocoll 2015;43:199–206. [147] Wang LY, Cheong KL, Wu DT, Meng LZ, Zhao J, Li SP. Fermentation optimization for the production of bioactive polysaccharides from Cordyceps sinensis fungus UM01. Int J Biol Macromol 2015;79:180–5. [148] Cheong KL, Wu DT, Zhao J, Li SP. A rapid and accurate method for the quantitative estimation of natural polysaccharides and their fractions using high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector. J Chromatogr A 2015;1400:98–106. [149] Lv  GP, Hu  DJ, Cheong  KL, Li  ZY, Qing  XM, Zhao  J, et  al. Decoding glycome of Astragalus membranaceus based on pressurized liquid extraction, microwave-assisted hydrolysis and chromatographic analysis. J Chromatogr A 2015;1409:19–29. [150] Guan J, Li SP. Discrimination of polysaccharides from traditional Chinese medicines using saccharide mapping-Enzymatic digestion followed by chromatographic analysis. J Pharm Biomed Anal 2010;51(3):590–8. [151] Xu J, Guan J, Chen XJ, Zhao J, Li SP. Comparison of polysaccharides from different Dendrobium using saccharide mapping. J Pharm Biomed Anal 2011;55(5):977–83. [152] Wu  DT, Li  WZ, Chen  J, Zhong  QX, Ju  YJ, Zhao  J, et  al. An evaluation system for characterization of polysaccharides from the fruiting body of Hericium erinaceus and identification of its commercial product. Carbohydr Polym 2015;124(2):201–7. [153] Chen YW, Hu DJ, Cheong KL, Li J, Xie J, Zhao J, et al. Quality evaluation of lentinan injection produced in China. J Pharm Biomed Anal 2013;78-79(9):176–82. [154] Wu DT, Lam SC, Cheong KL, Wei F, Lin PC, Long ZR, et al. Simultaneous determination of molecular weights and contents of water-soluble polysaccharides and their fractions from Lycium barbarum collected in China. J Pharm Biomed Anal 2016;129:210–8. [155] Cheong  KL, Meng  LZ, Chen  XQ, Wang LY, Wu DT, Zhao J, et al. Structural elucidation, chain conformation and immuno-modulatory activity of glucogalactomannan from cultured Cordyceps sinensis fungus UM01. J Funct Foods 2016;25:174–85. [156] Liu J, Sun Y, Liu L, Yu C. A water-soluble polysaccharide (EFP-AW1) from the alkaline extract of the roots of a traditional Chinese medicine, Euphorbia fischeriana: fraction and characterization. Carbohydr Polym 2012;88(4):1299–303. [157] Xie J, Zhao J, Hu DJ, Duan JA, Tang YP, Li SP. Comparison of polysaccharides from two species of Ganoderma. Molecules 2012;17(1):740–52. [158] Wu X, Si N, Bo G, Hu H, Yang J, Bian B, et al. Characterization and quantitative amino acids analysis of analgesic peptides in cinobufacini injection by size exclusion chromatography, matrix-assisted laser desorption/ionization time of flight mass spectrometry and gas chromatography mass spectrometry. Biomed Chromatogr 2015;29(1):138–47. [159] Bo RN, Zheng SS, Xing J, Luo L, Niu YL, Huang Y, et al. The immunological activity of Lycium barbarum polysaccharides liposome in vitro and adjuvanticity against PCV2 in vivo. Int J Biol Macromol 2016;85:294–301.

­References

[160] Xie JH, Tang W, Jin ML, Li JE, Xie MY. Recent advances in bioactive polysaccharides from Lycium barbarum L., Zizyphus jujuba Mill, Plantago spp., and Morus spp.: structures and functionalities. Food Hydrocoll 2016;60:148–60. [161] Wu DT, Cheong KL, Deng Y, Lin PC, Wei F, Lv XJ, et al. Characterization and comparison of polysaccharides from Lycium barbarum in China using saccharide mapping based on PACE and HPTLC. Carbohydr Polym 2015;134:12–9. [162] Shellie RA, Haddad PR. Comprehensive two-dimensional liquid chromatography. Anal Bioanal Chem 2006;386(3):405–15. [163] Cao  JL, Wei  JC, Chen  MW, Su  HX, Wan  JB, Wang  YT, et  al. Application of two-­ dimensional chromatography in the analysis of Chinese herbal medicines. J Chromatogr A 2014;1371:1–14. [164] Li Z, Chen K, Guo MZ, Tang DQ. Two-dimensional liquid chromatography and its application in traditional Chinese medicine analysis and metabonomic investigation. J Sep Sci 2015;39(1):21–37. [165] Jandera P. Programmed elution in comprehensive two-dimensional liquid chromatography. J Chromatogr A 2012;1255(17):112–29. [166] Filippo  B, Schoenmakers  PJ, Hans-Gerd  J. Theories to support method development in comprehensive two-dimensional liquid chromatography—a review. J Sep Sci 2012;35(14):1697–711. [167] Li B, Zhang X, Wang J, Zhang L, Gao B, Shi S, et al. Simultaneous characterisation of fifty coumarins from the roots of Angelica dahurica by off-line two-dimensional highperformance liquid chromatography coupled with electrospray ionisation tandem mass spectrometry. Phytochem Anal 2014;25(3):229–40. [168] Yao  CL, Yang  WZ, Wu  WY, Da  J, Hou  JJ, Zhang  JX, et  al. Simultaneous quantitation of five Panax notoginseng saponins by multi heart-cutting two-dimensional liquid chromatography: method development and application to the quality control of eight Notoginseng containing Chinese patent medicines. J Chromatogr A 2015;1402:71–81. [169] Song Y, Zhang N, Shi S, Li J, Zhang Q, Zhao Y, et al. Large-scale qualitative and quantitative characterization of components in Shenfu injection by integrating hydrophilic interaction chromatography, reversed phase liquid chromatography, and tandem mass spectrometry. J Chromatogr A 2015;1407:106–18. [170] Ma  S, Liang  Q, Jiang  Z, Wang  Y, Luo  G. A simple way to configure on-line twodimensional liquid chromatography for complex sample analysis: acquisition of fourdimensional data. Talanta 2012;97:150–6. [171] Qian ZM, Zhao J, Li DQ, Hu DJ, Li SP. Analysis of global components in Ganoderma using liquid chromatography system with multiple columns and detectors. J Sep Sci 2012;35(20):2725–34. [172] Liang Z, Li K, Wang X, Ke Y, Jin Y, Liang X. Combination of off-line two-dimensional hydrophilic interaction liquid chromatography for polar fraction and two-dimensional hydrophilic interaction liquid chromatography × reversed-phase liquid chromatography for medium-polar fraction in a traditional Chinese medicine. J Chromatogr A 2012;1224:61–9. [173] Teng ZQ, Dai RJ, Meng WW, Chen Y, Deng YL. Offline two-dimensional RP/RPLC method to separate components in Dracaena cochinchinensis (Lour.) S.C.Chen xylem containing resin. Chromatographia 2011;74(3-4):313–7. [174] Sun W, Tong L, Miao J, Huang J, Li D, Li Y, et al. Separation and analysis of phenolic acids from Salvia miltiorrhiza and its related preparations by off-line two-dimensional hydrophilic interaction chromatography × reversed-phase liquid chromatography coupled with ion trap time-of-flight mass spectrometry. J Chromatogr A 2015;1431:79–88.

659

660

CHAPTER 20  Quality control of traditional Chinese medicines

[175] Xing QQ, Liang T, Shen GB, Wang XL, Jin Y, Liang XM. Comprehensive HILIC × RPLC with mass spectrometry detection for the analysis of saponins in Panax notoginseng. Analyst 2012;137(9):2239–49. [176] Fu  Q, Guo  ZM, Zhang  XL, Liu  YF, Liang  XM. Comprehensive characterization of Stevia Rebaudiana using two-dimensional reversed-phase liquid chromatography/hydrophilic interaction liquid chromatography. J Sep Sci 2012;35(14):1821–7. [177] Yang W, Zhang J, Yao C, Qiu S, Chen M, Pan H, et al. Method development and application of offline two-dimensional liquid chromatography/quadrupole time-of-flight mass spectrometry-fast data directed analysis for comprehensive characterization of the saponins from Xueshuantong Injection. J Pharm Biomed Anal 2016;128:322–32. [178] Li D, Dück R, Schmitz OJ. The advantage of mixed-mode separation in the first dimension of comprehensive two-dimensional liquid-chromatography. J Chromatogr A 2014;1358:128–35. [179] Zeng Y, Shao D, Fang Y. On-line two-dimension liquid chromatography for the analysis of ingredients in the medicinal preparation of Coptis Chinensis Franch. Anal Lett 2011;44(9):1663–73. [180] Li D, Schmitz OJ. Comprehensive two-dimensional liquid chromatography tandem diode array detector (DAD) and accurate mass QTOF-MS for the analysis of flavonoids and iridoid glycosides in Hedyotis diffusa. Anal Bioanal Chem 2015;407(1):231–40. [181] Wang S, Wang C, Zhao X, Mao S, Wu Y, Fan G. Comprehensive two-dimensional high performance liquid chromatography system with immobilized liposome chromatography column and monolithic column for separation of the traditional Chinese medicine Schisandra chinensis. Anal Chim Acta 2012;713:121–9. [182] Li D, Schmitz OJ. Use of shift gradient in the second dimension to improve the separation space in comprehensive two-dimensional liquid chromatography. Anal Bioanal Chem 2013;405(20):6511–7. [183] Chen X, Cao Y, Lv D, Zhu Z, Zhang J, Chai Y. Comprehensive two-dimensional HepG2/ cell membrane chromatography/monolithic column/time-of-flight mass spectrometry system for screening anti-tumor components from herbal medicines. J Chromatogr A 2012;1242:67–74. [184] Han S, Huang J, Hou J, Wang S. Screening epidermal growth factor receptor antagonists from Radix et Rhizoma Asari by two-dimensional liquid chromatography. J Sep Sci 2014;37(13):1525–32. [185] Cao JL, Wei JC, Hu YJ, He CW, Chen MW, Wan JB, et al. Qualitative and quantitative characterization of phenolic and diterpenoid constituents in Danshen (Salvia miltiorrhiza) by comprehensive two-dimensional liquid chromatography coupled with hybrid linear ion trap Orbitrap mass. J Chromatogr A 2015;1427:79–89. [186] Qiao X, Wang Q, Song W, Qian Y, Xiao Y, An R, et al. A chemical profiling solution for Chinese medicine formulas using comprehensive and loop-based multiple heart-cutting two-dimensional liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. J Chromatogr A 2015;1438:198–204. [187] Qiao X, Song W, Ji S, Li YJ, Wang Y, Li R, et al. Separation and detection of minor constituents in herbal medicines using a combination of heart-cutting and comprehensive two-dimensional liquid chromatography. J Chromatogr A 2014;1362:157–67. [188] Li  XM, Luo  XG, Zhang  CZ, Wang  N, Zhang  TC. Quality evaluation of Hypericum ascyron extract by two-dimensional high-performance liquid chromatography coupled with the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. J Sep Sci 2015;38(4):576–84.

­References

[189] Oka F, Oka H, Ito Y. Systematic search for suitable two-phase solvent systems for highspeed counter-current chromatography. J Chromatogr A 1991;538(1):99–108. [190] Hou ZG, Luo JG. Recent progress of high-speed counter-current chromatography coupling with other relative technologies in natural product. Chin J Nat Med 2010;8(1):62–7. [191] Xie ZS, Sun YJ, Lam SC, Zhao MQ, Liang ZK, Yu XX, et al. Extraction and isolation of flavonoid glycosides from Flos Sophorae Immaturus using ultrasonic-­assisted extraction followed by high-speed countercurrent chromatography. J Sep Sci 2014;37(8):957–65. [192] Xie ZS, Huang JY, Xie ZY, Yu XX, Yang M, Yang DP, et al. Isolation and purification of isoquercitrin and quercitrin from Hypericum Japonicum Thunb. ex Murray by countercurrent chromatography. Sep Sci Technol 2014;49(5):778–82. [193] Hartmann A, Ganzera M. Supercritical fluid chromatography—theoretical background and applications on natural products. Planta Med 2015;81(17):1570–81. [194] Peng B, Qiao CF, Zhao J, Huang WH, Hu DJ, Liu HG, et al. Simultaneous determination of flavonoids, isochlorogenic acids and triterpenoids in Ilex hainanensis using high performance liquid chromatography coupled with diode array and evaporative light scattering detection. Molecules 2013;18(3):2934–41. [195] Li DQ, Qian ZM, Li SP. Inhibition of three selected beverage extracts on α-glucosidase and rapid identification of their active compounds using HPLC-DAD-MS/MS and biochemical detection. J Agric Food Chem 2010;58(11):6608–13. [196] He  X, Yang  W, Ye  M, Wang  Q, Guo  D. Differentiation of Cuscuta Chinensis and Cuscuta Australis by HPLC-DAD-MS analysis and HPLC-UV quantitation. Planta Med 2011;77(17):1950–7. [197] Liao LJ, Won TH, Kang SS, Shin JH. Simultaneous analysis of bioactive metabolites from Ziziphus jujuba by HPLC-DAD-ELSD-MS/MS. J Pharm Investig 2012;42(1):21–31. [198] Zhang  B, Li  XF, Yan  B. Advances in HPLC detection—towards universal detection. Anal Bioanal Chem 2008;390(1):299–301. [199] Li J, Cheong KL, Zhao J, Hu DJ, Chen XQ, Qiao CF, et al. Preparation of inulin-type fructooligosaccharides using fast protein liquid chromatography coupled with refractive index detection. J Chromatogr A 2013;1308:52–7. [200] Zong WR, Cheong KL, Wu DT, Li J, Zhao J, Li SP. Preparation and purification of raffinose family oligosaccharides from Rehmannia glutinosa Libosch. by fast protein liquid chromatography coupled with refractive index detection. Sep Purif Technol 2014;138:98–103. [201] Dolan  JW. Avoiding refractive index detector problems. LCGC N Am 2012;30(12): 1032–7. [202] Young CS, Dolan JW. Success with evaporative light-scattering detection. LC GC Eur 2003;16(3):132. [203] Hopia  AI, Ollilainen  VM. Comparison of the evaporative light scattering detector (ELSD) and refractive index detector (RID) in lipid analysis. J Liq Chromatogr Relat Technol 1993;16(12):2469–82. [204] Zhu  LL, Zhao  Y, Xu  YW, Sun  QL, Sun  XG, Kang  LP, et  al. Comparison of ultrahigh performance supercritical fluid chromatography and ultra-high performance liquid chromatography for the separation of spirostanol saponins. J Pharm Biomed Anal 2016;120:72–8. [205] Kong  W, Jin  C, Xiao  X, Zhao  Y, Liu  W, Li  Z, et  al. Determination of multicomponent contents in Calculus bovis by ultra-performance liquid chromatography-­ evaporative light scattering detection and its application for quality control. J Sep Sci 2010;33(10):1518–27.

661

662

CHAPTER 20  Quality control of traditional Chinese medicines

[206] Vehovec T, Obreza A. Review of operating principle and applications of the charged aerosol detector. J Chromatogr A 2010;1217(10):1549–56. [207] Eom  HY, Park  S, Kim  MK, Suh  JH, Yeom  H, Min  JW, et  al. Comparison between evaporative light scattering detection and charged aerosol detection for the analysis of saikosaponins. J Chromatogr A 2010;1217(26):4347–54. [208] Wang J. Electrochemical detection for microscale analytical systems: a review. Talanta 2002;56(2):223–31. [209] van Hove ERA, Smith DF, Heeren RMA. A concise review of mass spectrometry imaging. J Chromatogr A 2010;1217(25):3946–54. [210] Motilva M-J, Serra A, Macià A. Analysis of food polyphenols by ultra high-performance liquid chromatography coupled to mass spectrometry: an overview. J Chromatogr A 2013;1292:66–82. [211] Petrović M, Hernando MD, Díaz-Cruz MS, Barceló D. Liquid chromatography—tandem mass spectrometry for the analysis of pharmaceutical residues in environmental samples: a review. J Chromatogr A 2005;1067(1):1–14. [212] Mann  M, Hendrickson  RC, Pandey  A. Analysis of proteins and proteomes by mass spectrometry. Annu Rev Biochem 2001;70(1):437–73. [213] Daniel JM, Friess SD, Rajagopalan S, Wendt S, Zenobi R. Quantitative determination of noncovalent binding interactions using soft ionization mass spectrometry. Int J Mass Spectrom 2002;216(1):1–27. [214] Raffaelli A, Saba A. Atmospheric pressure photoionization mass spectrometry. Mass Spectrom Rev 2003;22(5):318–31. [215] Zhang FX, Li M, Qiao LR, Yao ZH, Li C, Shen XY, et al. Rapid characterization of Ziziphi Spinosae Semen by UPLC/Qtof MS with novel informatics platform and its application in evaluation of two seeds from Ziziphus species. J Pharm Biomed Anal 2016;122:59–80. [216] Wang Z, Qu Y, Wang L, Zhang X, Xiao H. Ultra-high performance liquid chromatography with linear ion trap-Orbitrap hybrid mass spectrometry combined with a systematic strategy based on fragment ions for the rapid separation and characterization of components in Stellera chamaejasme extracts. J Sep Sci 2016;39(7):1379–88. [217] Dong P, Zhang L, Zhan L, Liu Y. Ultra high performance liquid chromatography with mass spectrometry for the rapid analysis and global characterization of multiple constituents from Zibu Piyin Recipe. J Sep Sci 2016;39(3):595–602. [218] Yang M, Sun J, Lu Z, Chen G, Guan S, Liu X, et al. Phytochemical analysis of traditional Chinese medicine using liquid chromatography coupled with mass spectrometry. J Chromatogr A 2009;1216(11):2045–62. [219] Tolstikov VV, Fiehn O. Analysis of highly polar compounds of plant origin: combination of hydrophilic interaction chromatography and electrospray ion trap mass spectrometry. Anal Biochem 2002;301(2):298–307. [220] Zhou  JL, Qi  LW, Li  P. Herbal medicine analysis by liquid chromatography/time-offlight mass spectrometry. J Chromatogr A 2009;1216(44):7582–94. [221] Li DQ, Zhao J, Wu D, Li SP. Discovery of active components in herbs using chromatographic separation coupled with online bioassay. J Chromatogr B 2016;1021:81–90. [222] Chen  LX, Hu  DJ, Lam  SC, Ge  L, Wu  D, Zhao  J, et  al. Comparison of antioxidant activities of different parts from snow chrysanthemum (Coreopsis tinctoria Nutt.) and identification of their natural antioxidants using high performance liquid chromatography coupled with diode array detection and mass spectrometry and 2, 2’-azinobis (3-ethylbenzthiazoline-sulfonic acid) diammonium salt-based assay. J Chromatogr A 2016;1428:134–42.

­References

[223] Meng Q, Qian Z, Li X, Li D, Huang W, Zhao J, et al. Free radical scavenging activity of Eagle tea and their flavonoids. Acta Pharm Sin B 2012;2(3):246–9. [224] Li  DQ, Zhao  J, Li  SP. High-performance liquid chromatography coupled with post-­ column dual-bioactivity assay for simultaneous screening of xanthine oxidase inhibitors and free radical scavengers from complex mixture. J Chromatogr A 2014;1345:50–6. [225] Li  DQ, Zhao  J, Li  SP, Zhang  QW. Discovery of xanthine oxidase inhibitors from a complex mixture using an online, restricted-access material coupled with column-­ switching liquid chromatography with a diode-array detection system. Anal Bioanal Chem 2014;406(7):1975–84. [226] Li DQ, Li SP, Zhao J. Screening of xanthine oxidase inhibitors in complex mixtures using online HPLC coupled with postcolumn fluorescence-based biochemical detection. J Sep Sci 2014;37(4):338–44. [227] Han SL, Lv YN, Xue WJ, Cao J, Cui RH, Zhang T. Screening anaphylactic components of MaiLuoNing injection by using rat basophilic leukemia-2H3 cell membrane chromatography coupled with HPLC-ESI-TOF-MS. J Sep Sci 2016;39(3):466–72. [228] Fan J, Wei F, Zhang Y, Su H, Ji Z, He J, et al. Combining Sprague—Dawley rat uterus cell membrane chromatography with HPLC/MS to screen active components from Leonurus artemisia. Pharm Biol 2016;54(2):279–84. [229] Han SL, Huang J, Cui RH, Zhang T. Screening antiallergic components from Carthamus tinctorius using rat basophilic leukemia 2H3 cell membrane chromatography combined with high-performance liquid chromatography and tandem mass spectrometry. J Sep Sci 2015;38(4):585–91. [230] Xue H, Cheng YJ, Wang X, Yue Y, Zhang WF, Li XN. Rutaecarpine and evodiamine selected as β1-AR inhibitor candidates using β1-AR/CMC-offline-UPLC/MS prevent cardiac ischemia—reperfusion injury via energy modulation. J Pharm Biomed Anal 2015;115:307–14. [231] Lingeman  H, Underberg  WJM, Takadate  A, Hulshoff  A. Fluorescence detection in high performance liquid chromatography. J Liq Chromatogr Relat Technol 1985;8(5):789–874.

663