Rapid characterization and identification of non-volatile constituents in Pogostemonis Herba by UPLC-Q-TOF-MS combined with UNIFI and an in-house library / 中国中药杂志

This study aimed to characterize and identify the non-volatile components in Pogostemonis Herba by using ultra-perfor-mance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS) combined with UNIFI and an in-house library. The chemical components in 50% methanol extract of Pogostemonis Herba were detected by UPLC-Q-TOF-MS in both positive and negative MS~E continuum modes. Then, the MS data were processed in UNIFI combined with an in-house library to automatically characterize the metabolites. Based on the multiple adduct ions, exact mass, diagnostic fragment ions, and peak intensity of compounds and the fragmentation pathways and retention behaviors of reference substances, the structures identified by UNIFI were further verified and those of the unidentified compounds were tentatively elucidated. A total of 120 compound structures were identified or tentatively identified, including flavonoids, phenylpropanoids, phenolic acids, terpenes, fatty acids, alkaloids, and phenylethanoid glycosides. Sixteen of them were accurately identified by comparison with reference substances, and 53 compounds were reported the first time for Pogostemonis Herba. This study systematically characterized and identified the non-volatile compounds in Pogostemonis Herba for the first time. The findings provide a scientific basis for revealing the pharmacodynamic material basis, establishing a quality control system, and developing products of Pogostemonis Herba.

Analysis on Contents of Main Constituents in Prepared Morindae Officinalis Radix，Morindae Officinalis Radix Processed with Steaming and Salt / 中国实验方剂学杂志

Objective:To investigate the differences of main components of prepared Morindae Officinalis Radix，Morindae Officinalis Radix processed with steaming and salt. Method:A total of 83 batches samples were collected in the market， including 41 batches of prepared Morindae Officinalis Radix，32 batches of Morindae Officinalis Radix processed with steaming and 10 batches of Morindae Officinalis Radix processed with salt. The contents of main components were determined with high performance liquid chromatography coupled with four-pole tandem time-of-flight mass spectrometry（UPLC-Q-TOF-MS），and the differences were analyzed. Result:The main components of prepared Morindae Officinalis Radix and Morindae Officinalis Radix processed with salt were fructo-oligosaccharides （GF<italic>n</italic>），monotropein. The main components of Morindae Officinalis Radix processed with steaming were fructose，glucose，sucrose，and monotropein. The main differences of prepared Morindae Officinalis Radix and<italic> </italic>Morindae Officinalis Radix processed with steaming and salt were the contents of fructose，glucose，sucrose and GF2-GF11. The contents of GF2-GF11 in Morindae Officinalis Radix processed with steaming and salt were all lower than those in prepared Morindae Officinalis Radix，with extremely significant differences（<italic>P</italic><0.01）. The contents of fructose，glucose and sucrose in<italic> </italic>Morindae Officinalis Radix processed with steaming were significantly higher than those in prepared Morindae Officinalis Radix. The content of GF3 in each batch was higher than 40.0 mg·g^-1 in prepared Morindae Officinalis Radix and Morindae Officinalis Radix processed with salt，and significantly higher than the limit in<italic> Chinese Pharmacopoeia</italic>. However，there were only a few batches of Morindae Officinalis Radix processed with steaming in line with the requirements of <italic>Chinese Pharmacopoeia</italic>. The contents of monotropein in processing Morindae Officinalis Radix and Morindae Officinalis Radix processed with steaming and salt were 42.6，39.8，32.3 mg·g^-1，respectively. The content of monotropein in prepared Morindae Officinalis Radix was higher than that in Morindae Officinalis Radix processed with steaming. The content of monotropein in Morindae Officinalis Radix processed with steaming was higher than that in Morindae Officinalis Radix<italic> </italic>processed with salt. Compared with the components of GF2-GF11，the effect of processing with steaming process and/or salt on monotropein content was relatively less. Conclusion:The contents of GF2-GF11 components in prepared Morindae Officinalis Radix were converted into fructose，glucose and sucrose after processing with steaming and/or salt. The results showed that the content limit of Morindae Officinalis Radix processed with steaming needs to be revised in line with the requirements of <italic>Chinese Pharmacopoeia </italic>for the quality control of Morindae Officinalis Radix. The results provide a reference basis for revising the quality standards and studying the pharmacodynamic material basis of prepared Morindae Officinalis Radix，Morindae Officinalis Radix processed with steaming and salt.

Angiogenesis Activity of Morindae Officinalis Radix from Different Origins Based on Zebrafish Model / 中国实验方剂学杂志

Objective: To study the effect of Morindae Officinalis Radixe extract from different origins on the vascular growth of damaged internodes of zebra fish, so as to discuss regional differences of Morindae Officinalis Radix from different origins. Method: A zebra fish model of internode vascular injury induced by PTK787 was established to study the effect of Morindae Officinalis Radix extract from Guangdong, Guangxi and Fujian on the growth of internode vascular in zebra fish, and detect the number of internode angiogenesis in zebra fish treated with PTK787.On the basis of principal component analysis, relative angiogenesis rate was taken as the index, and the experimental data was analyzed by SPSS 17.0 statistical software. Result: The extract of Morindae Officinalis Radix had a repairing effect on the growth of damaged internode blood vessels of zebra fish, and the extracts of Morindae Officinalis Radix from Guangdong Gaoyao, and Guangdong Yunan, were better than those from other producing areas. The average number of internode angiogenesis was 9, the average length of angiogenesis was 448-504 μm, and the relative rate of angiogenesis was between 7.9% and 8.86%. The results of cluster analysis showed that Morindae Officinalis Radix of 12 different origins were classified into four categories, Guangdong Deqing, and Guangdong Gaoyao, Guangxi Cangwu and Guangxi Cenxi, and Guangdong Yunan, and Guangxi Cangwu could be better clustered. Conclusion: There are obvious regional differences in the pro-angiogenic effect of Morindae Officinalis Radix from different origins. Relevant studies provide basic data for the quality assessment of Morindae Officinalis Radix medicinal materials.

Comparison of volatile components in Schisandra propinqua from different habitats by HS-SPME-GC-MS / 中国中药杂志

A headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) method was developed and optimized for qualitative and semi quantitative analysis of volatile components from Schisandra propinqua, a kind of Yi Nationality herb medicine. This method was used for analysis and evaluation of volatile components from S.propinqua from four different geometrical origins (Yunnan Wuding, Yunnan Luoping, Guizhou Qingzhen and Hubei Shennongjia). 51, 53, 52， 50 compounds were identified from the above four kinds of samples respectively, in which 46 volatile components were contained in all of these four origins. The volatile components in these samples from different geographical origins were then compared by using principal component analysis, cluster analysis and other chemometrics methods. The results suggested that the analysis of volatile components can be used to distinguish the S.propinqua from four origins, and those samples from close geographical origins were more similar in volatile components. The developed method was stable, reliable, and suitable for rapid analysis of volatile components from S.propinqua, providing reference for quality control, drug development and scientific utilization of the herb.

Rapidly identify oligosaccharides in Morinda officinalis by UPLC-Q-TOF-MSE / 中国中药杂志

In this paper, an approach was applied for separation and identification of oligosaccharides in Morinda officinalis How by Ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) with collision energy. The separation was carried out on an ACQUITY UPLC BEH Amide C₁₈(2.1mm×100 mm，1.7 μm) with gradient elution using acetonitrile(A) and water(B) containing 0.1% ammonia as mobile phase at a flow rate of 0.2 mL·min⁻¹. The column temperature was maintained at 40 °C. The information of accurate mass and characteristic fragment ion were acquired by MSE in ESI negative mode in low and high collision energy. The chemical structures and formula of oligosaccharides were obtained and identified by the software of UNIFI and Masslynx 4.1 based on the accurate mass, fragment ions, neutral losses, mass error, reference substance, isotope information, the intensity of fragments, and retention time. A total of 19 inulin oligosaccharide structures were identified including D(+)-sucrose, 1-kestose, nystose, 1F-fructofuranosyl nystose and other inulin oligosaccharides (DP 5-18). This research provided important information about the inulin oligosaccharides in M. officinalis. The results would provide scientific basis for innovative utilization of M. officinalis.

Climate change impacts on yield of Cordyceps sinensis and research on yield prediction model of C. sinensis / 中国中药杂志

Cordyceps sinensis is a Chinese unique precious herbal material, its genuine producing areas covering Naqu, Changdu in Qinghai Tibet Plateau, Yushu in Qinghai province and other regions. In recent 10 years, C. sinensis resources is decreasing as a result of the blindly and excessively perennial dug. How to rationally protect, develop and utilize of the valuable resources of C. sinensis has been referred to an important field of research on C. sinensis. The ecological environment and climate change trend of Qinghai Tibet plateau happens prior to other regions, which means that the distribution and evolution of C. sinensis are more obvious and intense than those of the other populations. Based on RS (remote sensing)/GIS(geographic information system) technology, this paper utilized the relationship between the snowline elevation, the average temperature, precipitation and sunshine hours in harvest period (April and may) of C. sinensis and the actual production of C. sinensis to establish a weighted geometric mean model. The model's prediction accuracy can reach 82.16% at least in forecasting C. sinensis year yield in Naqu area in every early June. This study can provide basic datum and information for supporting the C. sinensis industry healthful, sustainable development.

Determination of eight steroidal saponins in 15 kinds of genus Paris / 中国中药杂志

The study was established an HPLC method for simultaneous determination of 8 steroidal saponins (polyphyllins Ⅶ, H, Ⅵ, Ⅱ, Ⅰ, and Ⅴ, dioscin, and gracillin) in Paridis Rhizoma, and made an evaluation by determining steroidal saponins in 15 kinds of genus Paris. The analysis was performed on a Waters Acquity H-ClassTM UPLC ultrafine liquid chromatography system coupled with a PDA detector. The chromatographic separation was achieved through a CAPCELL PAK ADME (4.6 mm× 250 mm, 5 μm) column and the optimal mobile phase consisted of acetonitrile and water. The column was maintained at 21 ℃, and the flow rate was 0.8 mL•min ⁻¹. The UV detection wavelength was 203 nm. The results showed that ① the detected components can be well separated and all with good correlation coefficients. The standard calibration curves were linearly good (R2>0.999 9). The linearity was obtained over 0.041 70-3.812 00 μg. The average recoveries ranged from 95.91% to 103.8%. ② there are significant differences in the content of steroidal saponins from different species. The steroidal saponins are low content or almost none in P. mairei, P. polyphylla var. stenophylla, and P. delavayi have low content or almost did not contain, so these species are not suitable for medicinal use. The contents of steroidal saponins in P. polyphylla var. chinensis are varied from different places. There were high content of steroidal saponins in P. polyphylla var. yunnanensis, P. forrestii, P. daliensis, and P. axialis, even up to 5.0%, which indicated that they had the potential pharmic value of development.

Qualitative and quantitative analyses of primary saponins in Paris forrestii / 中国中药杂志

In order to study whether Paris forrestii could be developed as a substitute of Paridis Rhizome, chemical compositions of P. forrestii and P. polyphylla var. yunnanensis were investigated by UPLC-Q-TOF MS. In addition, the contents of eight primary steroidal saponins in 77 batches of P. forrestii samples from different habitats were simultaneously determined by HPLC-UV. The results showed that P. forrestii and P. polyphylla var. yunnanensis have similar chemical compositions, and all 22 major common peaks were identified as steroid derivatives. Meanwhile, there were some differences in the contents of saponins in P. forrestii samples from different habitats. The contents of 4 steroidal saponins in Chinese Pharmacopoeia ranged from 0.068% to 3.30%, and the highest content of the 8 kinds of steroidal saponins was 6.18%, while the lowest was just 0.71%. Moreover, 78% of P. forrestii samples were in conformity with the requirements of Chinese Pharmacopoeia, indicating that P. forrestii samples had relatively stable quality and could be further studied as a substitute for Paridis Rhizome.

Rapidly identify chemical constituents of Schisandra propinqua by UPLC-Q-Tof/MSE / 中国中药杂志

This study was to identify the chemical constituents of Schisandra propinqua, one herbal medicine of Yi nationality in China by using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF/MSE). Acetonitrile- water containing 0.1% formic acid was used as the mobile phase for gradient elution. Data were collected under ESI negative mode and ESI positive mode, and then screened and verified by the software of UNIFI and Masslynx4.1. Based on the accurate mass, fragment ions, neutral losses, mass error, retention time, reference substance, isotope information, the intensity of fragments, as well as the previous reports, the known compounds were validated and identified. The chemical structures of the unknown components were identified according to exact molecular weight, MS fragment, chromatographic retention behavior, and characteristic fragments of known congener compounds. A total of 68 chemical components were identified from S. propinqua, including 3 flavonoids, 10 flavanols, 34 lignans compounds (including 20 dibenzocyclooctene lignans), 4 triterpenoids, 17 organic acid and other compounds. 37 compounds of them were found in S. propinqua for the first time, and one potential compound needed to be identified.

Study on distribution of five heavy metal elements in different parts of Cordyceps sinensis by microwave digestion ICP-MS / 中国中药杂志

The contents of five heavy metals (Cu, Pb, As, Cd, Hg) in 17 batches of Cordyceps sinensis were determined by microwave digestion-ICP-MS, and their distribution in C. sinensis were analyzed. The results showed that the contents of Cu, Pb, Cd and Hg in all batches were in accordance with the international standards of Chinese Medicine-Chinese Herbal Medicine Heavy Metal Limit, with their contents in the stroma higher than that in the caterpillar body, and the excess rate of As, which mainly concentrated in the caterpillar body part of C. sinensis, was 88.24%, as the content of As in the caterpillar body was 7 to 12 fold of that in the stroma. In this study, the distribution of five heavy metals in C. sinensis was clarified, and the existing problems of arsenic limit of heavy metal in C. sinensis were analyzed, and some suggestions were put forward. It is hoped that the reference standard can be provided for the limited standard of arsenic in C. sinensis.

Determination of five effective components in medicinal material of Tripterygium by UPLC-ESI-MS/MS / 中国中药杂志

This paper is aimed to develop a method for the determination of five effective components in medicinal material of Tripterygium using ultra performance liquid chromalography coupled with electrospray ionization tandem mass spectrometry(UPLC-ESI-MS/MS), which then was used to study their contents in raw materials from different areas and different sources.The separation was performed on a Waters ACQUITY UPLC-CSH-C18S column(2.1 mm×100 mm,1.7 μm), usingacetonitrile-0.2% ammonium form ateaqueous solutionas mobile phase. The target components were detected in multiple-reaction monitoring(MRM) mode by mass spectrometry with electrospray ionization (ESI) source operated in positive ionization mode. The quantitative results showed that good linearity was achieved in their respective linear ranges and fine determination coefficient (r > 0.997 8),and the overall recoveries ranged from 96.72%-103.2% with the RSD ranging from 1.0%-2.4%.The method is sensitive and accurate, and suitable for the effective components quantification in medicinal material of Tripterygium; contents of five effective components from different sources vary significantly, so the quality and safety of medicinal material of Tripterygium needs to be improved. It is very important to control the quality with multi-index for clinic safety.

Advance in studying early bolting of Umbelliferae medicinal plant / 中国中药杂志

The early bolting has become one of the key issues affecting the Umbelliferae herbs production. The early bolting does not only reduce herbs production but also affect the active ingredients of plants. The concept of early bolting, physiological theory, induce factors of the quality of medicine and, control technology about the early bolting of Umbelliferae are integrative reviewed in the paper. Its occurrence mechanism include: genetic factors, environmental and ecological factors, endogenous hormones, sowing, fertilizer and other production management measures. For the above occurring mechanism, the proposed prevention measures are as follows: preferably germplasm, sowing, rational fertilization, exogenous hormones, shading, reed head cut off, cut fall the leaves, reasonable planting density and other agronomic measures can reduce its bolting rate. And the early bolting problem focused for future research are mentioned, including: development of authentic ingredients cultivation technical specifications, screened early bolting hormone inhibitor, in-depth study of the mechanism of early bolting, reveal its early bolting gene and key ecological factors. The research of vernalization gene, gene regulation of flowering early bolting will solve the problem of quality and production reducing fundamentally caused by the early bolting.

Different metabolites of leaves between Tripterygium wilfordii and Tripterygium hypoglaucum based on UPLC-Q-TOF-MS / 中国中药杂志

To analysis the differences between Tripterygium wilfordii and T. hypoglaucum, specimens of their leaves were collected from five production regions and analyzed by ultra performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS). The data were analyzed by multivariate statistical method, such as hierarchical cluster analysis (HCA) principal component analysis (PCA) and orthogonal signal correction partial least square discrimination (OPLS-DA). Potential markers with VIP values above 5.0 and corresponding r values above 0.85, were selected and further tested by combining mann-Whitney nonparametric. Those with P < 0.001 and AUC = 1 were confirmed as metabolite markers to discriminate them from each other. Results revealed that the two species were obviously different in their leaf metabolites. Based on their mass spectra, 23 potential metabolite markers were identified to distinguish T. wilfordii from T. hypoglaucum.

Comparison of agronomic traits of Panax notoginseng between traditional cultivated fields and new cultivated fields / 中国中药杂志

<p><b>OBJECTIVE</b>In order to provide scientific basis for introduction breeding and production regionalization of Panax notoginseng, the environment of producing area, agronomic traits and medicinal material output were investigated.</p><p><b>METHOD</b>Using field survey sampling at harvest time, agronomic traits indicators of leaf segment, stem segment and roots segment etc. of commodity P. notoginseng, longitude and latitude, elevation, soil type, landform of producing area were measured and observed.</p><p><b>RESULT</b>The P. notogiseng cultivation was expanding from traditional area like Wenshang to new areas as Honghe, Kunming, Qujing at large scale. Comparing with traditional cultivated fields, the elevation of new fields, which are red soil of moderate or low mountain slopes and gentle hills between 1 800-2 130 m, increases markedly. The agronomic traits of new cultivated fields such as plant height, stem diameter, the ground and underground biomass were better than those of traditional cultivated fields in varying degree. Furthermore, the root weight, taproot weight, taproot length and other agronomic index augmented more than 20%. Comparison among different cultivated fields showed P. notoginseng of Honghe Shiping Niujie, Kunming Shilin Guishan, Honghe Jiangshui Guanting etc. had better agronomic traits, the plant were thick and tall, the taproot and the top of reed were large, the number of root was more. Inversely, P. notoginseng of Kunming Guandu Xiaoshao, Guangxi Jingxi Lutong, Wenshang Yanshan Jiangna were thin and small. Cluster analysis showed that cultivated fields of P. notoginseng which was across clustered by traditional and new cultivated fields can be divided into three groups, including a high-yielding region insist of three new cultivated fields Honghe Shiping Niujie, Kunming Shilin Guishan, Honghe Jiangshui Guanting and one traditional cultivated field Wenshan Yanshan Jiangna. Correlation analysis indicated that the size and weight of taproot and top of reed were significantly positive correlated with plant height, stem diameter, leaf size, leaf weight and stem weight. Regression analysis pointed out that stem diameter, leaf width, leaf length/width, leaf weight were the main factors affecting the dry weight of taproot, and the main factors influencing the dry weight of top of reed were plant height, petiole, leaf length, leaf width and other agronomic traits.</p><p><b>CONCLUSION</b>From agronomic traits, P. notoginseng in new cultivated fields were more robust than that in traditional fields, but besides the length of taproot, the difference of rest agronomic traits didn't reach the significant level. It is suggested that, in the west and the north of traditional area, the red soil land of moderate or low mountain slopes and gentle hills where the elevation is between 1 800-2 130 m is suitable for cultivating P. notoginseng. Honghe Shiping Niujie, Honghe Jianshui Guanting, Kuming Shilin Gongshan can be used as vital development area for commodity P. notoginseng.</p>

Protection and bidirectional effect of rhubarb anthraquinone and tannins for rats' liver / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To compare the bidirectional effect of rhubarb total anthraquinone (TA) and total tannins (TT) on rats' liver.</p><p><b>METHODS</b>One hundred rats were randomly divided into 10 groups, i.e., the blank group, the model group, the blank + high dose TA group, the blank +low dose TA group, the blank + high dose TT group, the blank + low dose TT group, the model + high dose TA group, the model + low dose TA group, the model +high dose TT group, and the model + low dose TT group, 10 in each group. The carbon tetrachloride (CCI4) was used to prepare the acute liver injury rat model. TA and TT of rhubarb (at 5.40 g crude drugs/kg and 14.69 g crude drugs/kg) were intragastrically administrated to rats in all groups except the blank group and the model group, once daily for 6 successive days.The general state of rats, biochemical indices such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), laminin (LN), hyaluronic acid (HA), transforming growth factor beta1 (TGF-beta1), as well pathological results of rat liver tissues. Finally the protection laws of TA and TT for rats' liver were analyzed using factor analysis.</p><p><b>RESULTS</b>Compared with the blank control group, all biochemical indices increased in the blank group (P < 0.05, P < 0.01). HA also increased in the blank + high dose TA group; AST, ALT, and HA also increased in the blank +high dose TT group (P < 0.05). Compared with the model group, AST, ALT, ALP, HA, and TGF-beta1 significantly decreased in the model + low dose TA group, the model + high dose TA group, the model + low dose TT group (P < 0.05, P < 0.01). Serum AST, ALT, and ALP also decreased in the model + high dose TT group (P < 0.05, P < 0.01). Pathological results showed that mild swollen liver cells in the model + high dose TA group. Fatty degeneration and fragmental necrosis around the central veins occurred in the blank + high dose TA group. The pathological injury was inproved in the model +low dose TA group. Two common factors, liver fibrosis and liver cell injury, were extracted by using factor analysis. TA showed stronger improvement of the two common factors than TT.</p><p><b>CONCLUSIONS</b>Rhubarb TA and TT showed protective and harmful effects on rats' liver. At an equivalent dosage, TA had better liver protection than TT. High dose TT played a role in liver injury to some extent.</p>

Determination and analysis of heavy metals content in Panax notoginseng of different origination / 中国中药杂志

Six heavy metals, including As, Cu, Hg, Cd, Pb and Cr in Panax notoginseng were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) combined with wet digestion method. The samples of P. notoginseng were collected in 12 different regions, including Yunnan and Guangxi Province. Green Standards of Foreign Trading Medicinal Plants & Preparations was used as the standard to evaluate the pollution status of As, Cu, Hg, Cd, Pb and Cr in P. notoginseng. The results showed that content of As and Cd exceeded the limit of the standard and the percentage was 32.4% and 29.7%, respectively, while Cu, Hg and Pb were all bellow the limit. The SPSS 16.0 software was used to analyze the data. The occurrence of contained heavy metals has been discussed.