Research Progress And Application in Fingerprint Technology On Chinese Materia Medica Ⅱ

1.2 Electrochemical fingerprint

Electrochemistry is a simple analytical method based on the electrochemical properties and principles of the substance to be tested, which is low-cost, highly sensitive, and requires few consumables [19]. However, this method has certain limitations and is currently mainly applicable to the analysis of Chinese medicines that contain chemical components such as terpenes (such as Angelica sinensis), flavonoids (such as Scutellaria baicalensis), alkaloids (such as Aconite), and anthraquinones (such as Rhubarb) that can undergo electrochemical reactions.

Wu Di et al. [20] used an electrochemical fingerprint to establish the optimal extraction process of Scutellaria baicalensis Georgi, providing a good method for rationally using Chinese medicinal materials. Shi Huihui et al. [21] used the Belousov-Chabotinsky (B-Z) reaction chemical oscillation technology to investigate the effects of temperature, rotation speed, and the number of medicinal materials added to the experimental results and established reasonable experimental conditions to obtain the electrochemical fingerprint of Magnolia biondii Pamp., providing a new method for the quality control of Magnolia biondii. Zou Guihua et al. [22] established the nonlinear electrochemical fingerprint of Codonopsis pilosula (Franch.) Nannf. And Gentiana macrophylla Pall. The electrochemical fingerprints of the two medicinal materials are visually different, which can be used as a simple and effective method to distinguish Codonopsis pilosula from Gentiana macrophylla. Dai Hongxia et al. [23] conducted an electrochemical analysis on 20 batches of Angelica sinensis (Oliv.) Diels with a growth period of 1, 2, and 3 years. Combined with principal component analysis, it was found that the electrochemical fingerprints and oscillation parameters (including oscillation induction time, maximum potential, oscillation life, maximum amplitude, stop potential, and number of cycles) of Angelica sinensis at different growth periods were significantly different, which can be used as a basis for identifying Angelica sinensis at various periods.

CHINESE MATERIAL MEDICAL HERB CISTANCHE HPLC FINGERPRINT

1.3 Chromatographic fingerprint

1.3.1 Thin layer chromatography 

Thin-layer chromatography is a chromatographic method that achieves separation by relying on the different adsorption capacities of the same adsorbent for different chemical components. It is the most commonly used method for identifying traditional Chinese medicines, with the advantages of easy operation, simple equipment, good sensitivity, high comparability, strong specificity, and easy color development. The authenticity of the medicinal material can be determined by the presence or absence of spots on the thin layer plate. The depth and size of the color of the spots can reflect the quality of the medicinal material to a certain extent[24]. Currently, high-performance thin-layer chromatography (HPTLC) is often used to replace high-performance liquid chromatography (HPLC) to qualitatively and quantitatively identify traditional Chinese medicine products due to its simplicity, accuracy, low cost, high efficiency, and rapidity. HPTLC fingerprints have better resolution and can reasonably and accurately estimate the active ingredients in the drug in a shorter time[25]. However, due to the large number of influencing factors (humidity, temperature, etc.), the reproducibility of the results is poor compared to HPLC. Loescher et al. [26] used HPLC and HPTLC to analyze the extract of Calendula officinalis and found that HPLC was more suitable for quantitative analysis than HPTLC.

In response to this effect, Li Xiangjun et al. [27] made a series of corresponding solutions, including conducting the development experiment in a refrigerator or a sealed temperature-controlled device to reduce the influence of temperature and adding different proportions of sulfuric acid at one end of the development cylinder to control humidity.

CHINESE MATERIAL MEDICAL HERB CISTANCHE HPLC LABLE

Patil et al. [28] established the HPTLC fingerprint of the petroleum ether extract of Catsia tora Linn. The results showed that the leaves, seeds, and flowers contained 10, 7, and 11 types of ingredients, respectively. This method can be used to identify the authenticity of Catsia tora Linn. Xiao Honghua et al. [29] used thin-layer chromatography combined with mass spectrometry to establish a thin-layer chromatographic fingerprint of chromones, organic acids, and triterpenoid saponins in Cimicifuga foetida L., and performed similarity analysis, cluster analysis and principal component analysis on 16 batches of Cimicifuga samples from five sources, providing a basis for the quality control of Cimicifuga.

Guzelmeric et al. [30] used ethyl acetate-formic acid-acetic acid-water (30:1.5:1.5:3) as the developing solvent to develop the HPTLC fingerprint of apigenin-7-glucoside, an active marker of Matricaria chamomilla L., which can be used as a major guide for the quality assessment of chamomile. Bazylko et al. [31] established the HPTLC fingerprint of five flavonoid compounds, namely tagetoides, quercetin, flavonoids, caffeic acid, and chlorogenic acid, from Galinsoga parviflora Cav. This method can be used to identify Galinsoga parviflora. Ethanolic et al. [32] established the fingerprint of the ethanol extract of Acacia catechu (L. f.) Willd. Using HPTLC performed a simple and rapid quantitative evaluation of rutin and quercetin in the extract. Agatonovic-Kustrin et al. [33] used HPTLC to quantitatively analyze apigenin, Matricaria, and bisabolol in the leaf and flower extracts of Pyrethrum parthenium (L.) Smith, Matricaria chamomilla L., and Calendula officinalis L., combined it with the DPPH method to compare the antioxidant capacity of these components. This method is simple, rapid, reliable, and inexpensive, and can also be used to screen the antioxidant activity of plant extracts.

1.3.2 HPLC method HPLC is a universal analytical method for detecting various chemical components in traditional Chinese medicine. 

It is widely used in the detection of chemical components in traditional Chinese medicine due to its advantages of high pressure, high sensitivity, high efficiency and automation [34]. The UV detector, diode array detector (DAD), electrochemical detector, evaporative light detector, etc. that can be connected to it all show their irreplaceable superiority.

Qiao et al. [35] used reversed-phase × reversed phase comprehensive two-dimensional liquid chromatography (RP × RP 2DLC) to separate the phenolic compounds obtained by ethyl acetate extraction of licorice Glycyrrhiza uralensis Fisch.

311 compounds were detected within 40 minutes, and the structures of 21 unknown compounds were preliminarily characterized by mass spectrometry, and 8 of them were found in licorice for the first time. The results showed that the RP×RP2DLC/MS system has a good effect on the separation of complex traditional Chinese medicine extracts and the qualitative analysis of natural products. Peng Liang et al. [36] analyzed the flavonoid components of 10 batches of Gynostemma pentaphyllum (Thunb.) Makino established the HPLC fingerprint of Gynostemma pentaphyllum and calibrated 11 common characteristic peaks. The similarity analysis results showed that the highest similarity of the 10 batches of Gynostemma pentaphyllum fingerprints relative to the reference spectrum was 0.989, and the lowest was 0.128, indicating that the flavonoid components of Gynostemma pentaphyllum from different origins were similar in type but had significant differences in content. Sun et al. [37] used the HPLC-DAD method to analyze the similarity of the chemical components of Bupleurum chinense DC. and B. scorzonerifolium Willd. For the first time, and found that the two had 12 common flavonoid peaks and 4 saponin peaks, respectively, and at the same time, each had 6 unique flavonoid peaks and 5 saponin peaks, respectively. The results showed that this method can reasonably and effectively reflect the difference in chemical components between Bupleurum and Bupleurum chinense. The correlation between the two is poor. Therefore, Bupleurum chinense should be used with caution in clinical practice. He et al. [38] quantitatively analyzed 10 kinds of bioactive gallic acid, chlorogenic acid, caffeine, and catechins contained in 10 batches of Ziyang green tea. The HPLC fingerprint developed was simple and reliable. Li et al. [39] established the HPLC fingerprint of pomegranate peel. They selected 15 characteristic peaks and evaluated the similarity of 10 batches of pomegranate peel to be 0.968. They also separated and quantitatively analyzed the punicalagin, gallic acid, catechin, chlorogenic acid, caffeic acid, epicatechin, rutin, and ellagic acid contained in it, providing a reliable and effective method for identifying the authenticity and quality control of pomegranate peel. Sumathy et al. [1] used HPLC combined with HPTLC and gas chromatography-mass spectrometry (GC-MS) to determine the chemical composition of the methanol extract of Ixora chinensis Lam. HPLC analysis showed the presence of biochin A, myricetin, quercetin, rutin, daidzein, and formononetin; ursolic acid was found in the HPTLC fingerprint; 24 phytochemical components were found by GC-MS; this combined qualitative and quantitative method can easily evaluate the quality and stability of Ixora chinensis Lam. Ge et al. [40] established the UHPLC-DAD fingerprint of Radix Angelicae Pubescentis, quantitatively analyzed the coumarins and phenolic acids contained in Radix Angelicae Pubescentis, and qualitatively identified its active ingredients using quadrupole-time of flight mass spectrometry (Q-TOF-MS). They found 9 phenolic acids, 30 coumarins, and 41 compounds of myrrh and adenosine. They combined the discriminant analysis method to effectively and quickly distinguish 32 batches of Radix Angelicae Pubescentis from different provinces. This method provides a new idea for the quality control of Radix Angelicae Pubescentis.

CISTANCHE SPECS LIST

Yang et al. [41] used ultra-high performance liquid chromatography (UPLC)-DAD detector and ESI-MS technology to obtain the HPLC fingerprint and mass spectrometry fingerprint of Xiaoyanlidan Tablets, respectively. They analyzed 113 samples of Xiaoyanlidan Tablets and obtained 39 peaks, of which 26 were from Quassia, 9 from Andrographis paniculata, and 4 from Rhizoma Cynoglossi. Using the chemometric methods of similarity and PCA, they determined that the five significant components were 4-methoxy-5-hydroxyferrocephalostomone, andrographolide, dehydroandrographolide, neoandrographolide, and rosmarinic acid. They established a systematic and comprehensive fingerprint to improve and evaluate the quality of Xiaoyanlidan Tablets. Fan et al. [42] established HPLC and UPLC fingerprints of flavonoid glycosides in Lophatherum gracile Brongn., and isolated a new flavonoid C-glycoside luteolin 6-C-β-D-glucuronic acid-(1→2)-β-D-pyranoglucoside for the first time. They also discovered the optical isomers of two compounds, flavonoid flavonoid and flavonoid hydroxyflavonoid, for the first time. They used one-dimensional and two-dimensional nuclear magnetic resonance and mass spectrometry to distinguish and determine their structures, providing a new method for quality control of Lophatherum gracile Brongn. Tshibangu et al. [43] used HPLC-DAD combined with thin layer chromatography, nuclear magnetic resonance, and mass spectrometry to effectively separate the main biflavonoid compounds GB1, GB2, GB-1a and flavonoids from Garcinia kola Linn., and confirmed their structures, which can be used as a routine analysis method for pharmaceutical preparations. Zhao et al. [44] established the HPLC-UV-MS fingerprint of Andrographis paniculata (Burm. f.) Nees, quantitatively determined andrographolide and dehydroandrographolide contained in Andrographis paniculata, and evaluated the differences of 10 batches of Andrographis paniculata by combining similarity analysis and PCA. This method provides a basis for the quality control of Andrographis paniculata.

Ahmed et al. [45] used HPLC to obtain the fingerprint of Commiphora wightii (Arn.) Bhandari resin, and quantitatively analyzed the trans- and cis-sterolone contained in it. At the same time, this method can effectively distinguish 22 batches of Commiphora wightii resin and 9 batches of adulterated resin, which can be used as a method to identify the authenticity of Commiphora wightii resin.

2 Biological fingerprints

Biological fingerprints are the use of gene sequence analysis methods at the molecular level to distinguish and classify the geographical distribution and changes in Chinese medicinal materials, including genomic fingerprints, proteomic fingerprints[60], and DNA fingerprints[61]. At present, there are fewer studies on biological fingerprints of Chinese medicines than chemical fingerprints. The most commonly used methods are polyacrylamide gel electrophoresis (PAGE), random amplified polymorphic DNA molecular marker technology (RAPD), polymerase chain reaction.

(PCR) and other methods.

Liu Li et al.[62] used RAPD markers, combined with PCR and cluster analysis, to effectively identify Panax notoginseng (Burk.) f. H. Chen, Panax ginseng C. A. Mey., and American ginseng Panaxquinquefolium L. Fan Wei et al. [63] first established the SDS-PAGE protein fingerprint of Eupolyphaga sinensis Walker, Bombyx mori Linnaeus, and Scolopendra subsidies mutilans L. Koch, providing a reliable and effective method for the identification of these three medicinal materials. Wei Yicong et al. [64] designed dual-site specific primers and constructed a multiplex PCR The rapid identification of two medicinal materials, Houttuynia cordata Thunb. And Gymnotheca chinensis Decne., can simultaneously identify the two species positively and determine whether they are mixed. Wang Fu et al. [65] used ITS2 (internal transcribed spacer 2) sequence as a DNA barcode to accurately identify the Chinese medicine, Zanthoxylumbungeanum Maxim. And its spice Zanthoxylumbungeanum Maxim., providing a basis for the identification of Chinese medicine Zanthoxylumbungeanum and its spice Zanthoxylumbungeanum.