ABSTRACT
In commercial herbal markets, Polygoni Multiflori Radix (PMR, the tuberous roots of Polygonum multiflorum Thunb.), a commonly-used Chinese medicinal material, is divided into different grades based on morphological features of size and weight. While more weight and larger size command a higher price, there is no scientific data confirming that the more expensive roots are in fact of better quality. To assess the inherent quality of various grades and of various tissues in PMR and to find reliable morphological indicators of quality, a method combining laser microdissection (LMD) and ultra-performance liquid chromatography triple-quadrupole mass spectrometry (UPLC-QqQ-MS/MS) was applied. Twelve major chemical components were quantitatively determined in both whole material and different tissues of PMR. Determination of the whole material revealed that traditional commercial grades based on size and weight of PRM did not correspond to any significant differences in chemical content. Instead, tissue-specific analysis indicated that the morphological features could be linked with quality in a new way. That is, PMR with broader cork and phloem, as seen in a transverse section, were typically of better quality as these parts are where the bioactive components accumulate. The tissue-specific analysis of secondary metabolites creates a reliable morphological criterion for quality grading of PMR.
Subject(s)
Drugs, Chinese Herbal/metabolism , Fallopia multiflora/metabolism , Plant Roots/metabolism , Secondary Metabolism , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Fallopia multiflora/chemistry , Organ Specificity , Plant Roots/chemistry , Tandem Mass SpectrometryABSTRACT
BACKGROUND: Bupleuri Radix (Chaihu) represents one of the most successful and widely used herbal medicines in Asia for the treatment of many diseases such as inflammatory disorders and infectious diseases over the past 2000 years. In the Chinese Pharmacopoeia, Chaihu is recorded as the dried roots of Bupleurum chinense DC. and B. scorzonerifolium Willd. (Umbelliferae). However, the widespread demand for the herb has tended to far outstrip the supply. Whether the aerial parts, which account for 70 ~ 85% of the dry weights of Bupleurum species, could be used as an alternative for the root has become an important scientific issue for the sustainable utilization of Bupleurum species. On the other hand, in some areas including the southeast of China as well as in Spain, the aerial parts of Bupleurum species have already been used in the folk medications. Therefore, to clarify whether the root and aerial parts of Bupleurum species are "equivalent" in the types and quantities of chemical constituents which subsequently influence their biological activities and therapeutic effects is of great importance for both the rational and sustainable use of this herb. METHODS: In the present study, the chemical profiles between the root and aerial parts of Bupleurum species from different species and collected from various locations were analyzed and compared by the ultra-high performance liquid chromatography quadrupole/time of flight-mass spectrometry (UHPLC-QTOF-MS). RESULTS: A total of 56 peaks were identified in the root and/or aerial parts from different batches of Bupleurum species, by comparison of references standards or with those reported in the literature. Principal Component Analysis (PCA) was conducted for displaying the differentiating clustering between these two parts. CONCLUSION: The results disclosed the distinct variations between them, which indicated that the aerial parts could not be used as an alternative of root from a chemodiversity perspective. The differentiating markers resulted from the PCA analysis could also be utilized for the differentiation between them. Further validation of their biological differences is anticipated in the future study.
Subject(s)
Bupleurum/chemistry , Drugs, Chinese Herbal/chemistry , Plant Components, Aerial/chemistry , China , Chromatography, High Pressure Liquid , Mass Spectrometry , Metabolomics , Plant Roots/chemistryABSTRACT
Based on a systematic review of morphology and distribution of plants, alternate names, actions, and properties of herbs recorded in ancient and modern literatures, in combination of field investigation, 18 Chinese herbal medicines recorded in ancient bencao literature were regarded to be derived from 7 species in the Ardisia genus. Among them, the variety Ardisia crenata f. hortensis was identified as the source of Zhushagen and Zijinniu. A. hanceana is referenced as Tiesan in the illustrated atlas of Botanical Nomenclature (Zhiwu Mingshi Tukao). The name Pingdimu refers to a different substance in the illustrated atlas of Botanical Nomenclature and the Flower Mirror (Huajing). The medicinals named Yedihong, Aicha, and Duanjiao sanlangare all derived from A. japonica. The origin of the herb Xiaoqing referenced in the Illustrated Classic of the Materia Medica (Bencao Tujing) is A. pusilla. The medicinals Bailiangjin, Jiuguanxue and Zoumatai are derived from A. crispa, A. brevicaulis, and A. gigantifolia, respectively. This investigation clarifies the botanical sources and actions of related Chinese medicinal materials in the genus Ardisia, and provides clues and evidence for utilizing and developing their medicinal plant resources.
Subject(s)
Ardisia/chemistry , Drugs, Chinese Herbal/chemistry , Plants, Medicinal/chemistry , Materia MedicaABSTRACT
To explore the processing mechanism of Aurantii Fructus decoction pieces used in Guangdong province and Hong Kong by analysing the chemical variation between raw and processed Aurantii Fructus with different methods based on UHPLC-Q-TOF-MS. The total ion chromatograms detected in positive and negative ion modes, and ion peak area ratio before and after processing were taken as variation indexes in the comparison. The results indicated that fermented Aurantii Fructus could produce three new ingredients, namely eriodictyol-7-glucoside, hesperetin-7-O-glucoside and 5-demethylnobiletin. At the same time, it could significantly increase the content of naringenin and hesperetin components, and could increase the content of such limonin derivatives as sudachinoid A, obacunoic acid and limoninand nomilinic acid. This suggests that the fermentation processing method of Aurantii Fructus decoction pieces used in Guangdong province and Hong Kong is of important significance for enhancing biological activity and bioavailability, and improving the clinical efficacy of Aurantii Fructus decoction pieces, and so is worth further protection and promotion.
Subject(s)
Citrus/chemistry , Drugs, Chinese Herbal/chemistry , Flavones/analysis , Glucosides/analysis , Chromatography, High Pressure Liquid , Fruit/chemistry , Mass SpectrometryABSTRACT
Chinese medicinal authentication is fundamental for the standardization and globalization of Chinese medicine. The discipline of authentication addresses difficult issues that have remained unresolved for thousands of years, and is essential for preserving safety. Chinese medicinal authentication has both scientific and traditional cultural connotations; the use of scientific methods to elucidate traditional experience-based differentiation carries the legacy of Chinese medicine forward, and offers immediate practical significance and long-term scientific value. In this paper, a path of inheritance and innovation is explored through the scientific exposition of Chinese medicinal authentication, featuring a review of specialized publications, the establishment of a Chinese medicine specimen center and Chinese medicinal image databases, the expansion of authentication technologies, and the formation of a cultural project dedicated to the Compedium of Materia Medica.
Subject(s)
Drugs, Chinese Herbal/chemistry , Materia Medica/standards , Medicine, Chinese Traditional/standards , Drug Contamination/prevention & control , Drugs, Chinese Herbal/standards , Humans , Materia Medica/chemistry , Reference StandardsABSTRACT
Astragali Radix (AR), prepared from the roots of Astragalus membranaceus (FISCH. ex LINK) BUNGE or its variey, A. membranaceus (FISCH. ex LINK) BUNGE var. mongholicus (BUNGE) HSIAO., is one of the most used and valuable traditional Chinese medicines (TCMs). Historically, Hunyuan, Shanxi Province in China is the geo-authentic producing area of AR and crude AR from here called "geo-authentic." According to tradition, geo-authentic TCMs define both authenticity and quality. However, no scientific investigation has ever determined whether the superior quality of Hunyuan AR is due to the genetic characteristics or to the local environment. In our study, seeds of 30 AR samples representing the two varieties from different regions were cultivated in Hunyuan under the same conditions. A method, using ultra-performance liquid chromatography coupled with photodiode array detector and evaporative light scattering detectors, was developed to evaluate the quality through a simultaneous determination of four major isoflavonoids and four major saponins. The two AR varieties were successfully distinguished by principal component analysis while samples of the same species with different seeds origins could not be distinguished. A genetic study demonstrated that the internal transcribed spacer sequences of the nuclear ribosomal DNA in A. membranaceus var. mongholicus samples from different geographical regions were highly conservative. These results indicate that the content of active components in AR depends on the interaction of genotype and environment. At the varietal level, genetic properties appear to be more important for pharmaceutical quality than environmental factors, while on the intraspecific level environmental factors might be more important than genetic properties.
Subject(s)
Astragalus Plant/genetics , Drugs, Chinese Herbal/chemistry , Geography , Base Sequence , Calibration , Chromatography, Liquid , DNA Primers , Medicine, Chinese Traditional , Plant Leaves/chemistry , Principal Component Analysis , Reproducibility of Results , Spectrophotometry, UltravioletABSTRACT
OBJECTIVE: To study the efficacy of different anti-adhesion agents used in preventing tubal obstruction after recanalization. METHODS: Five hundred and eight patients with tubal obstruction were divided into 245 cases in control group, 108 cases in chitosan group; 113 cases in sodium hyaluronate group and 42 cases in lipiodol group. The patients in control group were injected with anti-inflammation agents after recanalization, while other groups were injected with chitosan, sodium hyaluronate or lipiodol at dose of 2 - 3 ml in every therapeutic group. The rate of location of tubal obstruction and tubal recanalization were recorded during operation. Then patients in every group were followed up on tubal patency after 3 months, and pregnancy rate after 12 months. RESULTS: Among 1016 fallopian tubes in 508 patients, there were 330 tubes occlusion at isthmus portion and 563 tubes occlusion at interstitial portion of fallopian tube. Thirty-seven fallopian tubes were ablated because of ectopic pregnancy, 86 fallopian tubes were unobstructed. (1) The recanalization rate were 95.7% (179/187) in chitosan group, 97.9% (191/195) in sodium hyaluronate group, 98.7% (75/76) in lipiodol group and 97.7% (425/435) in control group, which did not show statistical difference (P > 0.05). (2) The rates of tubal patency after 3 months of 91.7% (99/108) in chitosan group and 88.5% (100/113) in sodium hyaluronate group were significantly higher than 71.4% (30/42) in lipiodol group and 74.3% (182/245) in control group (P < 0.05). (3) The rates of intrauterine pregnancy after 12 months were 48.1% (52/108) in chitosan group and 41.6% (47/113) in sodium hyaluronate group, which were significantly higher than 23.8% (10/42) in lipiodol group and 24.1% (59/245) in control group (P < 0.05). CONCLUSION: Chitosan and sodium hyaluronate could be effective to prevent tubal obstruction after interventional recanalization and increase pregnancy rate.
Subject(s)
Fallopian Tube Diseases , Hysterosalpingography , Animals , Fallopian Tube Diseases/surgery , Fallopian Tubes/surgery , Humans , Pregnancy Rate , Sterilization, TubalABSTRACT
This study demonstrates the feasibility of feature extraction and similarity measurement for the identification of starch grains, which are often found in microscopic images of Chinese Materia Medica (CMM) and as such is an important feature for use in the authentication of CMM with abundance of starch grains and otherwise without other special microscopic characteristics. Our proposed image signature, namely, the size distribution, indicates that it is effective to characterize starch grains in microscopic images of those CMM. Experimental results in a small scale study show that the recognition rate for the proposed signature using different measures of discrepancy is around 90%, which is considered very good while the benchmark wavelet-based energy signature has only about 60% recognition. Moreover, the extraction of the size distribution only requires moderate level of computational complexity. In this study, our proposed image signature provides greater accuracy and flexibility in capturing starch grains information for the authentication of CMM, compared with other existing preliminary identification approach in which manual image segmentation is necessary.
Subject(s)
Drugs, Chinese Herbal/chemistry , Image Processing, Computer-Assisted/methods , Starch/classification , Drugs, Chinese Herbal/standards , Particle Size , Starch/chemistry , Starch/ultrastructureABSTRACT
Microscopic authentication is an effective method for quality control of Chinese Materia Medica (CMM) because of its speed, convenience and low cost. However, the application of modern microscopic technique in quality evaluation of Toxic and Potent Chinese Materia Medica (T/PCMM) of animal origin is seldom reported. This gap in published knowledge is increasingly serious because confusion in T/PCMM has led to serious medical problems in China and other countries in recent years. To ensure the safe and effective use of T/PCMM, an accurate and convenient method, based on macroscopic and microscopic techniques, was developed for the authentication of animal T/PCMM. The color microscopic photos of the crude drug were acquired with the light microscope, and from these their morphological and microscopic characters were described. The present method was successfully applied in the analysis of 31 T/PCMM including 17 samples originating from plants, 3 from animals, 9 from minerals, and the remaining 2 from secreta. The macro- and microscopic characters of three animal T/PCMM, namely Mylabris (Mylabris phalerata or Mylabris cichorii), Huechys (Huechys sanguinea), and Lytta (Lytta caraganae) were conclusively determined and are here presented. The results demonstrated that it was feasible to use the present microscopic characters for the authentication of the above three animal T/PCMM.
Subject(s)
Drugs, Chinese Herbal/chemistry , Materia Medica/chemistry , Minerals/chemistry , Animals , Drugs, Chinese Herbal/classification , Drugs, Chinese Herbal/toxicity , Insecta/classification , Insecta/ultrastructure , Microscopy , Minerals/classificationABSTRACT
Aristolochiae Fructus, a Chinese herbal medicine derived from the fruit of Aristolochia contorta Bge., contains nephrotoxic aristolochic acid analogues (AAAs). According to ancient medical texts, various medicinal parts of the fruit of A. contorta were ever used. In order to reveal which part could be safely and effectively used, it is necessary to analyze the chemical profiles of different medicinal parts. Herein we compared the chemical compositions and determined aristolochic acid I (AA-I) and aristolochic acid II (AA-II) in the four parts viz. outer pericarp, inner pericarp, septum, and seed. Ultra-high performance liquid chromatography equipped with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) was applied for chemical profiling. Ultra-high performance liquid coupled with triple quadrupole mass spectrometry (UHPLC-QqQ-MS) was employed to quantify AA-I and AA-II in different parts. It was found that the chemical compositions of the four parts varied both qualitatively and quantitatively. A total of 10 AAAs, including 5 aristolochic acids and 5 aristolactams, together with 3 alkaloids, were unambiguously or tentatively identified by UHPLC-QTOF-MS. The quantitatively analytical results obtained by UHPLC-QqQ-MS showed that AA-I and AA-II exclusively accumulate in the seeds of A. contorta. These findings provide supporting data for the rational selection of medicinal parts.
Subject(s)
Aristolochia/chemistry , Aristolochic Acids/chemistry , Drugs, Chinese Herbal/chemistry , Chromatography, High Pressure Liquid , Fruit/chemistry , Molecular Structure , Tandem Mass SpectrometryABSTRACT
The light microscope is routinely used for microscopic identification of Chinese Materia Medica (CMM). However, the fluorescence microscope has not been used for this purpose until now. The CMM, Baihuasheshecao, mostly used as a component of herb tea, is a well known folk-medicine in China. According to the Chinese Pharmacopoeia, its source is the species Oldenlandia diffusa (Willd.) Roxb. Two other species of the same genus, namely O. corymbosa (L.) Lam and O. tenelliflora Bl., had been found in use as Baihuasheshecao. To find a quick and easy method to distinguish O. diffusa from these similar species of the same genus, the fluorescence microscope was used to investigate the fluorescence emission characteristics of the three tissues, which were compared with light microscopy images of the same material. The results showed that some tissues of the three herbs emit autofluorescence. Specially, the wall of endoderm cells of O. diffusa and O. tenelliflora emit autofluorescence, while similar tissue of O. corymbosa does not. Hence, fluorescence microscopy can be helpful in the identification of CMM.
Subject(s)
Oldenlandia/classification , Plants, Medicinal/classification , Microscopy, Fluorescence , Oldenlandia/cytology , Plants, Medicinal/cytologyABSTRACT
Ku-Ding-Cha, a kind of herbal tea, has been widely used in China for a long time to support cardiovascular health. Confusion arises because in different parts of China different plants are used to produce the commercial teas. It is important to determine the identity of the used species in order to authenticate genuine Ku-Ding-Cha. However, once the plants have been processed, it is difficult to identify them through macroscopic identification; other means are necessary. A comparative study was made on 24 samples of Ku-Ding-Cha, including five standard identified and authenticated plants and 19 commercial samples, by microscopic techniques. The results showed that the shapes of leaf blades, xylem cells, stone cells, and calcium oxalate crystals could be used for the identification of the plants from which the commercial products were made. The method was proven to be quick, handy, simple, and convenient.
Subject(s)
Beverages/analysis , Microscopy/methods , Calcium Oxalate/analysis , Plant Leaves/cytology , Xylem/cytologyABSTRACT
Traditional macroscopic and microscopic identification methods of medicinal materials are economical and practical, but usually experience-based due to few chemical supports. Here histochemical evaluation on bioactive components of Coptidis Rhizoma (CR) in anatomic sections using laser microdissection and liquid chromatography-mass spectrometry (LMD-LC-MS) was developed to correlate the inner quality and outer features of materials from different growing areas. Results of a total 33 peaks representing potential different alkaloids were detected and 8 common peaks were identified as the major alkaloids, namely magnoflorine, thalifendine, columbamine, epiberberine, jatrorrhizine, coptisine, palmatine, and berberine. Six major alkaloids were quantified in the top and middle sections of raw materials and in their tissues and cells at the same time. Histochemical analyses showed consistent results with direct determination in raw materials and explained the reason why top sections of all samples contained higher contents of alkaloids by giving out attributions of each alkaloid in different anatomic sections. Besides, results manifested the distribution and accumulation rules of alkaloids in diverse tissues and cells of CR. This study demonstrates an effective and scientific way to correlate bioactive components and morphological features of medicinal materials, which is beneficial to future research, agriculture and application.
Subject(s)
Alkaloids/analysis , Coptis/anatomy & histology , Coptis/chemistry , Histological Techniques/methods , Laser Capture Microdissection , Rhizome/anatomy & histology , Rhizome/chemistry , Chromatography, Liquid , Drugs, Chinese Herbal/chemistry , Mass SpectrometryABSTRACT
BACKGROUND: Rhizoma Chuanxiong (RC) is the dried rhizome of Ligusticum chuanxiong Hort., and various types of processed Rhizoma Chuanxiong (PRC) are widely used in China. However, quality assurance and quality control of these processed medicines remain challenging. This study aims to investigate the chemical compositions of various PRC preparations by a high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD) method. METHODS: A HPLC-DAD method with validation was developed for PRC samples. Seven batches of plant samples from two processing methods, stir-frying and steaming, were analyzed by the HPLC-DAD method. Common peaks in PRC chromatograms were chosen to calculate their relative retention time (RRT) and relative peak area (RPA), and similarity analyses of the chromatographic fingerprints were conducted by Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine software (Version 2004 A). RESULTS: In the 24-h stability test, the relative standard deviation for the RRT and RPA was less than 0.07% and 2.57%, respectively. The precision was less than 0.08% for the RRT and 2.48% for the RPA. The repeatability for the RRT and RPA was less than 0.03% and 2.64%, respectively. The similarities between the seven PRC batches were range from 0.956 to 0.990. After stir-frying or steaming, the amount of ferulic acid in PRC was much higher than that in the raw material. CONCLUSIONS: The fingerprint analysis of PRC by different processing methods was feasible by HPLC-DAD.
ABSTRACT
Rabdosia japonica var. glaucocalyx (Maxim.) Hara, belonging to the Labiatae family, is widely used as an anti-inflammatory and antitumor drug for the treatment of different inflammations and cancers. Aim of the Study. To investigate therapeutic effects and possible mechanism of the flavonoids fraction of Rabdosia japonica var. glaucocalyx (Maxim.) Hara (RJFs) in acute lung injury (ALI) mice induced by lipopolysaccharide (LPS). Materials and Methods. Mice were orally administrated with RJFs (6.4, 12.8, and 25.6 mg/kg) per day for 7 days, consecutively, before LPS challenge. Lung specimens and the bronchoalveolar lavage fluid (BALF) were isolated for histopathological examinations and biochemical analysis. The level of complement 3 (C3) in serum was quantified by a sandwich ELISA kit. Results. RJFs significantly attenuated LPS-induced ALI via reducing productions of the level of inflammatory mediators (TNF- α , IL-6, and IL-1 ß ), and significantly reduced complement deposition with decreasing the level of C3 in serum, which was exhibited together with the lowered myeloperoxidase (MPO) activity and nitric oxide (NO) and protein concentration in BALF. Conclusions. RJFs significantly attenuate LPS-induced ALI via reducing productions of proinflammatory mediators, decreasing the level of complement, and reducing radicals.
ABSTRACT
The rhizome of Belamcanda chinensis (L.) DC. is a traditionally used medicinal material in China. Due to increasing demand, B. chinensis has been cultivated widely, and thus the study on its rational utilization of medicinal part and guidelines for the optimal cultivation and harvest is an important issue. Considering flavonoids were the main bioactive secondary metabolites of B. chinensis, fluorescence microscopy, laser microdissection (LMD), ultra-high performance liquid chromatography-quadrupole/time-of-flight-mass spectrometry (UHPLC-Q/TOF-MS), and UHPLC coupled with triple quadrupole mass spectrometer (UHPLC-QqQ-MS) were applied to profile and determine flavonoids in various tissues in this study. Consequently, 43 peaks were detected by UHPLC-Q/TOF-MS, and 26 flavonoid compounds combined with seven triterpene compounds were identified or tentatively identified in the tissue extractions. The results indicated that the hydrophobic compounds, especially flavonoid or isoflavonoid aglycones and xanthone mainly accumulated in the cork, whereas the hydrophilic compounds, namely the flavonoid and isoflavonoid glycosides were usually found in the cortex or center (the part inside of endodermis). Samples of rhizomes from different growth ages and origins were simultaneously analyzed. It was shown that the bulb or lateral part of the rhizome generally possessed more total flavonoids than the vertical part or the primordium. The present study established a new practical method to evaluate the quality of the rhizome of B. chinensis and to explore the relationship between distribution patterns of secondary metabolites and growth years of plants, thus important information for cultivation and processing was provided.
Subject(s)
Chromatography, High Pressure Liquid/methods , Flavonoids/analysis , Iridaceae/chemistry , Laser Capture Microdissection/methods , Metabolomics , Rhizome/chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , China , Organ Specificity , Triterpenes/analysisABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Saussurea laniceps Hand.-Mazz. (SL) has long been used under the herbal name Tibetan "Snow Lotus" for the treatment of rheumatoid arthritis, stomachache and dysmenorrhea in Tibetan folk medicine. Since herbal medicine (HM) is a synergistical system with multiple components, both of the metabolism and pharmacokinetic studies of HM are interdependent. This study aimed to develop an integrated strategy based on the UPLC-DAD-QTOF-MS technique for metabolism and pharmacokinetic studies of HM. MATERIAL AND METHODS: SL was used here as a test herb to verify the feasibility of the proposed strategy. SL was administered to rats, then, the blood plasma, urine and feces were analyzed to determine the metabolic profiles. Using our strategy, umbelliferone and scopoletin were evaluated to be the key bioactive components. Their pharmacokinetic parameters were measured and biotransformation pathways were elucidated. RESULTS: After oral administration of SL to rats, 17 components in blood, 10 components in urine and 2 components in feces were identified and characterized using our UPLC-DAD-QTOF-MS method. Umbelliferone, scopoletin and their metabolites were found to be the major components involved in the metabolism process. Literature reports also suggest that umbelliferone and scopoletin are responsible for the therapeutic effects of SL, thus these two components were selected as the active markers for pharmacokinetic study. In the test of validity, the established method presented good linearity with R(2)>0.99. The relative standard deviation value was below 13.9% for precision, and recovery studies for accuracy were found to be within the range 91.8-112.5%. CONCLUSION: The present strategy offers, simultaneously, precision in quantitative analysis (metabolism study) and accuracy in quantitative analysis (pharmacokinetic study) with greater efficiency and less costs, which is therefore reliably used for integrated metabolism and pharmacokinetic studies of HM.
Subject(s)
Asteraceae , Plant Extracts/pharmacokinetics , Animals , Chromatography, High Pressure Liquid , Feces/chemistry , Male , Mass Spectrometry/methods , Medicine, Traditional , Phytochemicals/pharmacokinetics , Plant Extracts/blood , Plant Extracts/urine , Rats, Sprague-Dawley , Scopoletin/blood , Scopoletin/pharmacokinetics , Umbelliferones/blood , Umbelliferones/pharmacokineticsABSTRACT
Secondary metabolites accumulated in different tissues and cells of herbs are usually bioactive components of herbal medicines. Thus, tissue- and cell-specific phytochemical profiling should be useful for indicating relationship between herbal tissues and chemicals, and evaluating the quality of a medicinal herb. Here, a method that combining laser microdissection and ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (LMD with UPLC-Q/TOF-MS) was established to achieve simultaneous localization and determination of bioactive components in herbal medicines. Sinomenii Caulis, sourced from the stems of Sinomenium acutum (Thunb.) Rehd. et Wils., was set as an illustrative case, and its phytochemicals were profiled by the present method through analyses of different microdissected tissues and cells, involving epidermis, cortex, stone cells, pericycle, vascular bundles and pith. Results revealed that different tissues and cells contained varied alkaloids, among which six alkaloids, i.e. 6-Me-ether-12-O-ß-D-glucopyranoside-laudanosoline (peak 4), sinomenine (peak 6), N-norsinoacutine (peak 7), magnoflorine (peak 11), laurifoline (peak 16) and menisperine (peak 17) were detected in all microdissected parts, and sinomenine and magnoflorine were the two most abundant components. By further quantitative determination, alkaloids were generally demonstrated to distribute in the outer part of the cortex, phloem and xylem. According to the relationship between alkaloids and tissues revealed in our study, Sinomenii Caulis of larger diameter has proportionately more bioactive components, and is therefore of higher quality for medicinal use. The method of LMD with UPLC-Q/TOF-MS developed in this study was initially applied to the research of medicinal herbs, and proved to be high sensitive, low cost, convenient and practical.
Subject(s)
Alkaloids/analysis , Laser Capture Microdissection/methods , Mass Spectrometry/methods , Sinomenium/chemistry , Sinomenium/ultrastructure , Alkaloids/metabolism , Chromatography, Liquid/methods , Plant Stems/chemistry , Plant Stems/metabolism , Plant Stems/ultrastructure , Plants, Medicinal/chemistry , Plants, Medicinal/metabolism , Plants, Medicinal/ultrastructure , Sensitivity and Specificity , Sinomenium/metabolismABSTRACT
Authentication is the first priority in quality evaluation of Chinese herbal medicines (CHMs). The most commonly used authentication methods are morphological identification and microscopic identification. Unfortunately, these two methods cannot provide the chemical information needed to assess the quality of CHMs. In this study, a combination of fluorescence microscopy and high performance liquid chromatography-mass spectrometry (HPLC-MS) was used to analyze the chemical profiles of the tissues of the raw root tubers of Polygonum multiflorum Thunb. The results showed that the cork cells, cortex, and vessels in transverse sections of the raw root tuber of P. multiflorum fluoresced differently. Further analysis by HPLC-MS revealed that anthraquinones are mainly distributed in the cortex, and 2,3,5,4'-tetrahydroxystilbene-2-O-ß-D-glucopyranoside could be found in all tissues of the raw root tubers of P. multiflorum with its relative content as cork > cortex > xylem of allotype vascular bundles > xylem of central vascular bundles. Moreover, the fluorescence characteristics of the tissues from the steamed root tuber of P. multiflorum were compared and showed different fluorescence from those of raw material. From these results, it can be deduced that the root tuber of P. multiflorum with a broader cortex and fewer vascular bundles visible in a transverse section should be of better quality. The different fluorescence characteristics can be used to differentiate raw root tubers of P. multiflorum from those that have been steamed.
Subject(s)
Polygonum/chemistry , Polygonum/cytology , Root Nodules, Plant/chemistry , Root Nodules, Plant/cytology , Anthraquinones/analysis , Chromatography, High Pressure Liquid , Glucosides/analysis , Histocytochemistry , Mass Spectrometry , Microscopy, Fluorescence , Stilbenes/analysisABSTRACT
The light microscope has been successfully used in identification of Chinese herbal medicines (CHMs) for more than a century. However, positive identification is not always possible. Given the popularity of fluorescence microscopy in bioanalysis, researchers dedicated to finding new ways to identify CHMs more effectively are now turning to fluorescence microscopy for authentication purposes. Some studies on distinguishing confused species from the same genus and on exploring distributions of chemicals in tissues of CHMs by fluorescence microscopy have been reported; however, no systematic investigations on fluorescent characteristics of powdered CHMs have been reported. Here, 46 samples of 16 CHMs were investigated. Specifically, the mechanical tissues including stone cells and fibers, the conducting tissues including three types of vessels, and ergastic substances including crystals of calcium oxalate and secretions, in various powdered CHMs were investigated by both light microscope and fluorescence microscope. The results showed many microscopic features emit fluorescence that makes them easily observed, even against complex backgrounds. Under the fluorescence microscope, different microscopic features from the same powdered CHM or some same features from different powdered CHMs emitted the different fluorescence, making this information very helpful for the authentication of CHMs in powder form. Moreover, secretions with unique chemical profiles from different powdered CHMs showed different fluorescent characteristics. Hence, fluorescence microscopy could be a useful additional method for the authentication of powdered CHMs if the fluorescent characteristics of specific CHMs are known.