Publications by authors named "Baiping Ma"

29 Publications

  • Page 1 of 1

Lycium barbarum mitigates radiation injury via regulation of the immune function, gut microbiota, and related metabolites.

Biomed Pharmacother 2021 Jul 3;139:111654. Epub 2021 May 3.

Tianjin University of Traditional Chinese Medicine, Tianjin, China; Beijing Institute of Radiation Medicine, Beijing, China. Electronic address:

Previous studies have suggested that Lycium barbarum (L. barbarum) has a radioprotective function, although more in-depth investigation is still required. We investigated the radioprotective efficacy of extract of the fruits of L. barbarum (LBE) and its radioprotective mechanisms. Mice were exposed to 8.5 Gy, 5.5 Gy, or 6.0 Gy total body irradiation (TBI), and the survival rate, lymphocyte percentage, amount of cytokines, and viability of the irradiated cells, as well as the gut microbiome and fecal metabolomics were studied. LBE enhanced the survival of the mice exposed to 8.5 Gy γ-ray TBI or 5.5 Gy X-ray TBI. After 6.0 Gy γ-ray TBI, LBE exhibited good immunomodulatory properties, mainly characterized by the accelerated recovery of lymphocyte percentages, and the enhanced expression of immune-related cytokines. LBE reconstituted the gut microbiota of irradiated mice, increased the relative abundance of potentially beneficial genera (e.g., Turicibacter, Akkermansia), and decreased the relative abundance of potentially harmful bacterial genera (e.g., Rikenellaceae_RC9_gut_group). Beneficial regulatory effects of LBE on the host metabolites were also noted, and the major upregulated metabolites induced by LBE, such as Tetrahydrofolic acid and N-ornithyl-L-taurine, were positively correlated with the immune factor interleukin (IL)-6. In vitro, LBE also increased the vitality of rat small intestinal epithelial cells (IEC-6) after 4.0 Gy γ-ray irradiation and promoted the growth of Akkermansia muciniphila. These results confirmed a radioprotective function of LBE and indicated that the radioprotective mechanism may be due to immunomodulation and the synergistically modulating effect on the gut microbiota and related metabolites.
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http://dx.doi.org/10.1016/j.biopha.2021.111654DOI Listing
July 2021

Chemical profiling and multicomponents quantitative analysis of Panzerina lanata by ultra-fast liquid chromatography with tandem mass spectrometry.

J Sep Sci 2021 May 3. Epub 2021 May 3.

School of Pharmacy, Key Laboratory of Ethnomedicine of Ministry of Education, Minzu University of China, Beijing, P. R. China.

Panzerina lanata is a Chinese medicine with the bioactivity of detumescence and detoxification. In this study, novel qualitative and quantitative methods were established by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry and ultra-high-performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry, respectively. As a result, 20 compounds were identified or tentatively characterized including flavonoids, organic acids, alkaloids, and lignans, five of which were identified for the first time based on the reference standards. The quantitative approach exhibited good linearity (R  > 0.995), precision (RSDs < 4.97%), stability (RSDs < 4.77%), and recovery (96.04-104.14%). Afterward, this method was implemented to determine 11 flavonoids in four batches of P. lanata. Among them, seven compounds were quantified for the first time. Narcissin was abundant in each batch of P. lanata (average of 10.890-14.230 mg/g) with the highest quantities. The results provide valuable information for quality evaluation.
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http://dx.doi.org/10.1002/jssc.202000944DOI Listing
May 2021

Screening and Selection of a New Medium for Diosgenin Production via Microbial Biocatalysis of sp.

Pharmaceuticals (Basel) 2021 Apr 21;14(5). Epub 2021 Apr 21.

Division for Medicinal Microorganisms Related Strains, CAMS Collection Center of Pathogenic Microorganisms, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.

Steroidal saponins are widely used as starting precursors and medical intermediates for the semi-/total-synthesis of hundreds of steroidal drugs. One such steroidal saponin is diosgenin, which has attracted significant attention due to the huge market demand in the pharmaceutical industry. Due to water waste and severe environmental pollution, the traditional diosgenin production process based on direct acid hydrolysis is no longer used. In this study, to develop a submerged fermentation (SmF) medium for clean diosgenin production via efficient microbial biocatalysis, the Box-Behnken design (BBD) in combination with the Plackett-Burman design (PBD) was used to determine the medium compositions for strains. Three components (wheat bran, phosphate, and Tween-80) were determined as significant factors by the PBD. Using the BBD, the three significant factors were further optimized, and the optimum values were determined for maximal diosgenin production. With 21.16 g/L of wheat bran, 9.60 g/L of phosphate, and 1.97 g/L of Tween-80, the diosgenin yield was 2.28%, i.e., 3.17 mg/L/h. The experimental values agreed with the predicted values, representing a significant increase in diosgenin production compared to its production using the basic SmF medium. For the first time, we reported the development of a new medium for strains to produce diosgenin via microbial biocatalysis of the root of C. H. Wright (DZW). A simple-composition, low-cost, and high-efficiency medium was developed for the first time for the SmF of strains. The medium is considered useful for large-scale SmF and may be applicable to other fungi. This study lays a solid foundation for diosgenin production in an acid-free and wastewater-free way. It may also provide fundamental support for producing other value-added products via microbial biocatalysis of low-value materials by endophytic fungi.
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http://dx.doi.org/10.3390/ph14050390DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8143133PMC
April 2021

Development of a New Bioprocess for Clean Diosgenin Production through Submerged Fermentation of an Endophytic Fungus.

ACS Omega 2021 Apr 31;6(14):9537-9548. Epub 2021 Mar 31.

Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, 2 Nanwei Road, Beijing 100050, P. R. China.

Diosgenin is used widely to synthesize steroidal hormone drugs in the pharmaceutical industry. The conventional diosgenin production process, direct acid hydrolysis of the root of C. H. Wright (DZW), causes large amounts of wastewater and severe environmental pollution. To develop a clean and effective method, the endophytic fungus sp. CPCC 400226 was screened for the first time for the microbial biotransformation of DZW in submerged fermentation (SmF). Statistical design and response surface methodology (RSM) were implemented to develop the diosgenin production process using the strains. The environmental variables that significantly affected diosgenin yield were determined by the two-level Plackett-Burman design (PBD) with nine factors. PBD indicates that the fermentation period, culture temperature, and antifoam reagent addition are the most influential variables. These three variables were further optimized using the response surface design (RSD). A quadratic model was then built by the central composite design (CCD) to study the impact of interaction and quadratic effect on diosgenin yield. The values of the coefficient of determination for the PBD and CCD models were all over 0.95. -values for both models were 0.0024 and <0.001, with -values of ∼414 and ∼2215, respectively. The predicted results showed that a maximum diosgenin yield of 2.22% could be obtained with a fermentation period of 11.89 days, a culture temperature of 30.17 °C, and an antifoam reagent addition of 0.20%. The experimental value was 2.24%, which was in great agreement with predicted value. As a result, over 80% of the steroidal saponins in DZW were converted into diosgenin, presenting a ∼3-fold increase in diosgenin yield. For the first time, we report the SmF of a strain used to produce diosgenin through the microbial biotransformation of DZW. A practical diosgenin production process was established for the first time for strains. This bioprocess is acid-free and wastewater-free, providing a promising environmentally friendly alternative to diosgenin production in industrial applications. The information provided in the current study may be applicable to produce diosgenin in SmF by other endophytic fungi and lays a solid foundation for endophytic fungi to produce natural products.
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http://dx.doi.org/10.1021/acsomega.1c00010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8047649PMC
April 2021

Comprehensive Investigation on Ginsenosides in Different Parts of a Garden-Cultivated Ginseng Root and Rhizome.

Molecules 2021 Mar 18;26(6). Epub 2021 Mar 18.

Beijing Institute of Radiation Medicine, No. 27 Taiping Road, Beijing 100850, China.

Background: Ginseng is widely used as herb or food. Different parts of ginseng have diverse usages. However, the comprehensive analysis on the ginsenosides in different parts of ginseng root is scarce.

Methods: An ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) combined with UNIFI informatics platform and ultra-high-performance liquid chromatography-charged aerosol detection (UHPLC-CAD) were employed to evaluate the different parts of cultivated ginseng root.

Results: 105 ginsenosides including 16 new compounds were identified or tentatively characterized. 22 potential chemical markers were identified, 20, 17, and 19 for main root (MR) and fibrous root (FR), main root (MR) and branch root (BR), and main root (MR) and rhizome (RH), respectively. The relative contents of Re, Rb, 20(R)-Rh, Rd, and Rf were highest in FR. The relative content of Rg was highest in RH. The total relative content of pharmacopoeia indicators Rg, Re, and Rb was highest in FR.

Conclusion: The differences among these parts were the compositions and relative contents of ginsenosides. Under our research conditions, the peak area ratio of Rg and Re could distinguish the MR and FR samples. Fibrous roots showed rich ingredients and high ginsenosides contents which should be further utilized.
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http://dx.doi.org/10.3390/molecules26061696DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8003075PMC
March 2021

Comparative analysis of chemical components in different parts of Epimedium Herb.

J Pharm Biomed Anal 2021 May 17;198:113984. Epub 2021 Feb 17.

Beijing Institute of Radiation Medicine, Beijing 100850, China. Electronic address:

Epimedium herb is a well-known traditional Chinese medicine (TCM) that is used for treating kidney-yang deficiency, impotence and rheumatism, and flavonoids are the main active ingredients. The leaves and rhizomes of Epimedium herb are two separate kinds of medicinal materials with different functional indications and clinical applications. This study aimed to comprehensively analyze the chemical components of different parts of the herb from three Epimedium species (Epimedium sagittatum, E. pubescens and E. myrianthum) by using ultra high-performance liquid chromatography coupled with photo-diode array and quadrupole time-of-flight mass spectrometry (UHPLC-PDA-Q-TOF/MS) and multivariate statistical analysis to clarify the differences. Firstly, the workflow of UHPLC-Q-TOF/MS combined with UNIFI informatics was developed for characterizing the chemical compounds in different parts of Epimedium herb. Based on the exact mass information, the fragmentation characteristics and the retention times of compounds, all chromatographic peaks (74 chemical components) were identified. Secondly, 21 potential chemical markers for differentiating different parts of Epimedium herb were selected through PCA and PLS-DA analysis. The characteristic components in the leaves included flavonoids with Anhydroicaritin (type A, C-4' linked methoxy) as the backbone, and the characteristic components in the stems and rhizomes were Magnoline and flavonoids with Demethylanhydroicaritin (type B, C-4' linked hydroxyl) as the backbone. Thirdly, the UHPLC-PDA combined with heatmap visualization was employed to clarify the distribution of chemical components with high content in different parts of Epimedium herb. The results showed clear differences in the contents of chemical components in leaves, stems and rhizomes. The levels of flavonoids with Anhydroicaritin backbone were high in the leaves, and levels of flavonoids with Demethylanhydroicaritin backbone were high in the rhizomes. The levels of Magnoline in stems and rhizomes were higher than that in leaves. The contents of most of the compounds in stems remained low. The leaves and the other two parts (stems and rhizomes) can be distinguished by qualitative and semi-quantitative analysis of Magnoline and Epimedoside A (type B backbone). These results indicated that the different plant parts of Epimedium herb can be quickly and accurately distinguished by this method, establishing a foundation for the application of Epimedium herb.
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http://dx.doi.org/10.1016/j.jpba.2021.113984DOI Listing
May 2021

Anemarrhena asphodeloides modulates gut microbiota and restores pancreatic function in diabetic rats.

Biomed Pharmacother 2021 Jan 29;133:110954. Epub 2020 Nov 29.

Beijing Institute of Radiation Medicine, Beijing, China. Electronic address:

Anemarrhena asphodeloides is an herb widely used to treat symptoms associated with diabetes in traditional Chinese medicine. However, its key components and metabolites have low bioavailability and poor host absorption. To clarify the anti-diabetic mechanism of A. asphodeloides extract (AAE), we examined the anti-diabetic effects of AAE in rats with diabetes induced by a high-fat diet and streptozotocin. Faeces levels of the main components and metabolites of AAE were significantly higher than levels in plasma, which indicated that gut microbiota might play important roles in its anti-diabetic effect. Microbiological studies showed that unabsorbed components increased the diversity of the gut microbiota, enriched potentially beneficial bacteria, and suppressed potentially harmful bacteria. In vitro studies showed that AAE promoted the proliferation of Blautia coccoides, a bacterium with positive implication for diabetes, in a dose-dependent manner. AAE also promoted pancreatic cell regeneration and restored the function of pancreatic islet cells via peroxiredoxin 4 overexpression. Overall, these results suggest that AAE alleviates diabetes via modulating gut microbiota and protein expression.
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http://dx.doi.org/10.1016/j.biopha.2020.110954DOI Listing
January 2021

Development of a New High-Cell Density Fermentation Strategy for Enhanced Production of a Fungus β-Glucosidase in .

Front Microbiol 2020 21;11:1988. Epub 2020 Aug 21.

Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.

Traditional diosgenin manufacturing process has led to serious environmental contamination and wastewater. Clean processes are needed that can alternate the diosgenin production. The β-glucosidase FBG1, cloned from sp. CPCC 400709, can biotransform trillin and produce diosgenin. In this study, production of recombinant FBG1 was implemented to investigate various conventional methanol induction strategies, mainly including DO-stat (constant induction DO), μ-stat (constant exponential feeding rate) and m-stat (constant methanol concentration). The new co-stat strategy combining μ-stat and m-stat strategies was then developed for enhanced FBG1 production during fed-batch high-cell density fermentation on methanol. The fermentation process was characterized with respect to cell growth, methanol consumption, FBG1 production and methanol metabolism. It was found that large amounts of formaldehyde were released by the enhanced dissimilation pathway when the co-stat strategy was implemented, and therefore the energy generation was enhanced because of improved methanol metabolism. Using co-stat feeding, the highest volumetric activity reached ∼89 × 10 U/L, with the maximum specific activity of ∼90 × 10 U/g. After 108 h induction, the highest volumetric production reached ∼403 mg/L, which was ∼91, 154, and 183 mg/L higher than the maximal production obtained at m-stat, μ-stat, and DO-stat strategies, respectively. FBG1 is the first produced recombinant enzyme for diosgenin production through the biotransformation of trillin. Moreover, this newly developed co-stat induction strategy represents the highest expression of FBG1 in , and the strategy can be used to produce FBG1 from similar strains harboring gene, which lays solid foundation for clean and sustainable production of diosgenin. The current work provides unique information on cell growth, substrate metabolism and protein biosynthesis for enhanced β-glucosidase production using a strain under controlled fermentation conditions. This information may be applicable for expression of similar proteins from strains.
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http://dx.doi.org/10.3389/fmicb.2020.01988DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7472535PMC
August 2020

Furostanol Saponins from Promote the Expansion of Human Cord Blood Hematopoietic Stem and Progenitor Cells.

J Nat Prod 2020 09 1;83(9):2567-2577. Epub 2020 Sep 1.

Beijing Institute of Radiation Medicine, Beijing 100850, People's Republic of China.

is a medicinal plant known to biosynthesize steroidal saponins. A phytochemical investigation of the rhizomes of led to the isolation of nine new furostanol saponins (-) and 11 known analogues (-). Five of these new compounds were shown to have hydroxy groups at the C-5 and C-6 positions, while two possess a rare aglycone containing carbonyl groups at the C-16 and C-22 positions as well as a Δ double bond, and the others have conjugated double bonds in the E-ring or have different sugar chains at the C-3 position. All the isolates were tested for their effect on the expansion of human cord blood (CB) CD34 hematopoietic stem and progenitor cells. It was found that CB CD34 cells treated with compounds , , , , , , and showed increased numbers of rigorously phenotype-defined hematopoietic stem cells. Notably, compounds , , , and demonstrated an enhanced ability to increase the percentages and numbers of CB CD34CD38 cells and multipotential progenitors. The present study is the first to report that furostanol saponins from rhizomes can promote hematopoietic stem/progenitor cell (HSPC) expansion.
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http://dx.doi.org/10.1021/acs.jnatprod.9b01268DOI Listing
September 2020

Nasal timosaponin BII dually sensitive in situ hydrogels for the prevention of Alzheimer's disease induced by lipopolysaccharides.

Int J Pharm 2020 Mar 8;578:119115. Epub 2020 Feb 8.

Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, Beijing 100850, China; Anhui Medical University, Hefei 230032, China. Electronic address:

Alzheimer's disease (AD) is a common and severe brain disease with a high mortality among the elders, but no highly efficient medications are currently available. For example, timosaponin BII, an efficient anti-AD agent, has low oral bioavailability. Here, timosaponin BII was formulated in a temperature/ion-sensitive in situ hydrogel (ISG) that was well transformed into gels in the nasal environment. Timosaponin BII protected the PC12 cells injured by lipopolysaccharides (LPS) by decreasing TNF-α and IL-1β and stabilizing F-actin. Timosaponin BII ISGs were intranasally administered to the mice every day for 38 days. On Day 36, LPS was injected to the mice to establish an AD model. Morris water maze experiments showed that the number of the animals that were able to cross the platform returned to normal and the total distance over which the animals moved in the open field also increased, which demonstrated that the spatial memory and spontaneous behavior were improved after treatment compared to the model. Moreover, an AD improver, inducible nitric oxide synthase (iNOS) in the brain, was reduced after treatment. High brain targeting effect of timosaponin BII ISGs was confirmed by in vivo fluorescence imaging. The nasal timosaponin BII dually sensitive ISGs can serve as a promising medication for local prevention of AD.
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http://dx.doi.org/10.1016/j.ijpharm.2020.119115DOI Listing
March 2020

Study on steroidal saponins in crude and stir-fried Fructus Tribuli by ultra-high-performance liquid chromatography-mass spectrometry coupled with multivariate statistical analysis.

J Sep Sci 2020 Apr 29;43(7):1208-1223. Epub 2020 Jan 29.

Beijing Institute of Radiation Medicine, Beijing, P. R. China.

Fructus Tribuli is a traditional Chinese medicine used clinically for many years. Crude Fructus Tribuli and stir-fried Fructus Tribuli are recorded in the Pharmacopoeia of the People's Republic of China. However, the differences between steroidal saponins in crude Fructus Tribuli and stir-fried Fructus Tribuli have not been compared. In this study, ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry along with multivariate statistical analysis was developed to discriminate the chemical profiles and identify the steroidal saponins of crude Fructus Tribuli and stir-fried Fructus Tribuli. Additionally, an ultra-high-performance liquid chromatography triple-quadrupole mass spectrometer was used for the simultaneous quantification of nine major steroidal saponins to analyze the variations between crude Fructus Tribuli and stir-fried Fructus Tribuli. Finally, a total of 30 steroidal saponins whose structures or contents changed significantly after processing were found and identified. The mechanism of structural transformations deduced indicated that during the stir-frying of Fructus Tribuli, C-22 hydroxy furostanol saponins were converted to the corresponding furostanol saponins containing C-20-C-22 double bonds by dehydroxylation and deglycosylation reactions that occurred in the spirostanol saponins causing the generation of steroidal sapogenins. This study was successfully applied to the global analysis of crude Fructus Tribuli and stir-fried Fructus Tribuli. The results of this research will be beneficial to explore the processing mechanism of Fructus Tribuli.
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http://dx.doi.org/10.1002/jssc.201900961DOI Listing
April 2020

Liupao tea extract alleviates diabetes mellitus and modulates gut microbiota in rats induced by streptozotocin and high-fat, high-sugar diet.

Biomed Pharmacother 2019 Oct 31;118:109262. Epub 2019 Jul 31.

Beijing Institute of Radiation Medicine, No.27 Taiping Road, Haidian District, Beijing, 100850, China. Electronic address:

Liupao tea, a popular dark tea, exhibited anti-diabetic activity in Chinese tea culture. However, the bioavailability of the main active components is low. Therefore, the relationship between the anti-diabetic activity and the modulation of gut microbiota was investigated in this study for the first time. Liupao tea extract (LTE) could significantly alleviate hyperglycemia, insulin-resistance, and plasma lipid profiles. The hypoglycemic effect of high-dose LTE was comparable to that of metformin. Importantly, LTE exhibited beneficial effect on modulating the diabetic-induced disorder of gut microbiota, such as increasing Bacteroidetes/Firmicutes ratio and up-regulating putative short-chain fatty acid (SCFA)-producing bacteria, including Prevotella and Bacteroides. Moreover, 29 key phylotypes were significantly correlated with at least one diabetic index. In conclusion, LTE could be developed into an anti-diabetic functional beverage and its beneficial effects is likely to be associated with the prebiotic effects for increasing the level of SCFAs and the relative abundance of SCFA-producing bacteria.
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http://dx.doi.org/10.1016/j.biopha.2019.109262DOI Listing
October 2019

The qualitative and quantitative analyses of Gelsemium elegans.

J Pharm Biomed Anal 2019 Aug 7;172:329-338. Epub 2019 May 7.

Beijing Institute of Radiation Medicine, Beijing 100850, China; School of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. Electronic address:

Gelsemium elegans is a traditional Chinese medicine that has been used to treat eczema, bruises, rheumatoid arthritis and skin ulcers for many years, and alkaloids are its major active and toxic constituents. This study aimed to comprehensively assess the quality of G. elegans samples including different plant parts and origins using ultra high-performance liquid chromatography coupled with photo-diode array and quadrupole time-of-flight mass spectrometry (UHPLC-PDA-QTOF/MS) and high-performance liquid chromatography coupled with UV detector (HPLC-UV). Firstly, the UHPLC-PDA-QTOF/MS approach was developed for the characterization of alkaloids in G. elegans and understanding the differences between multiple groups of samples. Based on the exact mass information, the fragmentation characteristics and the retention time of compounds, 38 alkaloids were identified or tentatively identified. 24 potential chemical markers for differentiating different plant parts of G. elegans were selected through PCA and OPLS/PLS-DA analysis. Secondly, a heatmap visualization was employed for clarifying the distribution of 24 selected alkaloids with high response in the UV. The roots, stems and leaves from Yunnan Province possess relatively consistent alkaloids composition, respectively. Most compounds in the root have a higher content than stems and leaves. Thirdly, a HPLC-UV approach was developed for quantitative analysis of three major alkaloids (gelsemine, koumine and gelsenicine) of G. elegans, and the results showed remarkable variation in the contents of these constituents. While, the contents of three alkaloids fluctuate relatively less in the stem. These results indicated that integrated chemical profiling and quantitative analysis of alkaloids in G. elegans from different plant parts and origins could be assessed by this method, which would establish the foundation for the application of G. elegans.
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http://dx.doi.org/10.1016/j.jpba.2019.05.015DOI Listing
August 2019

An optimized LC-MS/MS method for determination of HYNIC-3PRGD, a new promising imaging agent for tumor targeting, in rat plasma and its application.

J Chromatogr B Analyt Technol Biomed Life Sci 2018 Sep 21;1095:142-148. Epub 2018 Jul 21.

Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, Beijing 100850, China.

HYNIC-3PRGD is used to prepare a new Tc-radiolabeled tracer. HYNIC-3PRGD, which has a high binding affinity for the integrin αβ due to its special structure, has become a promising tumor imaging agent for diagnosis and monitor of the clinical response to therapeutic effects of anti-tumor agents. Here, we developed and validated a method for determination of HYNIC-3PRGD concentration in rat plasma using ultra-high performance liquid chromatography-tandem mass spectrometry system. Following sample extraction by methanol precipitation, satisfactory separation through chromatography was achieved on an hydrophilic reverse-phase C column AQ (2.1 mm × 100 mm, 2.7 μm) at a flow rate of 0.2 mL·min with an gradient elution using mobile phase consisting of ultrapure water and acetonitrile fortified with 0.1% formic acid respectively. The calibration curve was developed over a linear range of 3.125-100 ng·mL with the lower limit of quantification of 3.125 ng·mL. The HYNIC-3PRGD and its internal standard c(RGDfK)(RK5) were detected and quantified with the multiple reaction monitoring (MRM) mode on a triple-quadrupole tandem mass spectrometer. This method was successfully validated and applied for pharmacokinetic evaluation of HYNIC-3PRGD during pre-clinical experiments.
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http://dx.doi.org/10.1016/j.jchromb.2018.07.026DOI Listing
September 2018

Isolation of endophytic fungi from Dioscorea zingiberensis C. H. Wright and application for diosgenin production by solid-state fermentation.

Appl Microbiol Biotechnol 2018 Jul 3;102(13):5519-5532. Epub 2018 May 3.

Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100050, China.

In this study, endophytic fungi were isolated from Dioscorea zingiberensis C.H. Wright (DZW), and a novel clean process to prepare diosgenin from DZW was developed. A total of 123 strains of endophytic fungi were isolated from different plant tissues of DZW. Among them, the strain Fusarium sp. (CPCC 400709) showed the best activity of hydrolyzing steroidal saponins in DZW into diosgenin. Thus, this strain was used to prepare diosgenin from DZW by solid-state fermentation. The fermentation parameters were optimized using response surface methodology, and a high yield of diosgenin (2.16%) was obtained at 14.5% ammonium sulfate, an inoculum size of 12.3%, and 22 days of fermentation. Furthermore, the highest diosgenin yield (2.79%) was obtained by co-fermentation with Fusarium sp. (CPCC 400709) and Curvularia lunata (CPCC 400737), which was 98.9% of that obtained by β-glucosidase pretreated acid hydrolysis (2.82%). This process is acid-free and wastewater-free, and shows promise as an effective and clean way to prepare diosgenin for use in industrial applications from DZW.
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http://dx.doi.org/10.1007/s00253-018-9030-5DOI Listing
July 2018

Qualitative and quantitative analysis of furofuran lignans, iridoid glycosides, and phenolic acids in Radix Dipsaci by UHPLC-Q-TOF/MS and UHPLC-PDA.

J Pharm Biomed Anal 2018 May 3;154:40-47. Epub 2018 Mar 3.

Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China. Electronic address:

Radix Dipsaci (RD), the dried root of Dipsacus asper, is used in traditional Chinese medicine as a remedy for bone fractures, traumatic hematoma, threatened abortion, and uterine bleeding. A novel ultra high-performance liquid chromatography coupled with quadrupole-time-of-flight tanderm mass spectrometry (UHPLC-Q-TOF/MS) approach was performed to rapidly characterize the chemical constituents of RD. Consequently, 21 compounds, including 12 iridoid glycosides (IGs), 4 furofuran lignans (FLs), and 5 phenolic acids (PAs) were discovered and identified from RD. Among these compounds, 3 IGs were previously unreported. Furthermore, a rapid and reliable UHPLC-DAD-based method was developed. The linearity (R > 0.999), repeatability (RSDs < 3.0%), intra-day and inter-day precision (RSDs < 1.6%), recovery (98.9%-102.5%), limits of detection (0.2-2.75 ng), and limits of quantification (0.75-8.5 ng) of the method was validated. The method was successfully applied for the simultaneous determination of 11 compounds in 20 batches of RD collected from various geographical regions in China. Different RD samples exhibited significantly varied contents of 11 analytes, among which IGs and PAs are abundant compounds that could be used as suitable quality markers for RD. The present study provided a useful approach for comprehensively evaluating the chemical composition and quality of RD.
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http://dx.doi.org/10.1016/j.jpba.2018.03.002DOI Listing
May 2018

The reason leading to the increase of icariin in Herba Epimedii by heating process.

J Pharm Biomed Anal 2018 Feb 22;149:525-531. Epub 2017 Nov 22.

Beijing Institute of Radiation Medicine, Beijing 100850, China. Electronic address:

Icariin (ICA), a major flavonoid responsible for the pharmacological actions of Herba Epimedii, was found to be increased significantly after the heating process. The increase of ICA was considered to be due to the transformation of other flavonoids, in particular, epimedin (EPI) A-C. This novel study demonstrated that the increase of ICA in Epimedium. koreanum was induced mainly through the conversion of 3'''-carbonyl-2''-β-l-quinovosyl icariin (CQICA) by deglycosylation during the heating process. Furthermore, the comparative analysis of the changes of CQICA and ICA in four raw Epimedium species (EPs) designated in the Chinese Pharmacopoeia revealed that the increase of ICA during the heating process was associated with the presence of CQICA in EPs. The present study provided scientific evidence for the conversion of CQICA into ICA, correcting the understanding of the transformation of EPI A-C into ICA. The results provided a reasonable scientific basis for the traditional use of processed Herba Epimedii, and might be helpful for the quality assessment of EPs.
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http://dx.doi.org/10.1016/j.jpba.2017.11.050DOI Listing
February 2018

Acute toxicity, 28-day repeated-dose toxicity and toxicokinetic study of timosaponin BII in rats.

Regul Toxicol Pharmacol 2017 Nov 22;90:244-257. Epub 2017 Sep 22.

Beijing Institute of Radiation Medicine, Beijing 100850, People's Republic of China. Electronic address:

Timosaponin BII (TBII), a major steroidal saponin isolated from Anemarrhena asphodeloides Bge., displays a variety of promising pharmacological activities, such as neuroprotection, enhancement of learning and memory, vascular protection and inhibition of platelet aggregation; therefore, it has been developed as a pharmaceutical for prevention or treatment of dementia. Given the safety concerns surrounding timosaponins and the absence of studies on the safety of TBII, the potential toxicity of TBII was evaluated in toxicity and toxicokinetic studies in rats. In the acute oral toxicity study, loose stools were observed in rats receiving 4000 mg/kg, and the symptoms recovered within 1 day. In the 28-day repeated-dose oral toxicity and toxicokinetic study, rats receiving 540 mg/kg showed loose stools and a slight deceleration of body weight growth in both sexes, and the females also showed a slight decrease in food consumption. Moreover, urinalysis indicated reversible treatment-related toxicity in rats receiving 540 mg/kg. The toxicokinetic study demonstrated a dose-dependent increase in systematic exposure to TBII after 28 successive days of oral treatment with TBII. The accumulation coefficients of TBII were 4.35, 1.70 and 1.81, respectively, in rats that received 60, 180 and 540 mg/kg. The no-observed-adverse-effect level (NOAEL) is proposed to be 180 mg/kg.
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http://dx.doi.org/10.1016/j.yrtph.2017.09.021DOI Listing
November 2017

Rapid Characterization of Constituents in Tribulus terrestris from Different Habitats by UHPLC/Q-TOF MS.

J Am Soc Mass Spectrom 2017 11 1;28(11):2302-2318. Epub 2017 Aug 1.

Beijing Institute of Radiation Medicine, No. 27 Taiping Road, Beijing, 100850, China.

A strategy for rapid identification of the chemical constituents from crude extracts of Tribulus terrestris was proposed using an informatics platform for the UHPLC/Q-TOF MS data analyses. This strategy mainly utilizes neutral losses, characteristic fragments, and in-house library to rapidly identify the structure of the compounds. With this strategy, rapid characterization of the chemical components of T. terrestris from Beijing, China was successfully achieved. A total of 82 steroidal saponins and nine flavonoids were identified or tentatively identified from T. terrestris. Among them, 15 new components were deduced based on retention times and characteristic MS fragmentation patterns. Furthermore, the chemical components of T. terrestris, including the other two samples from Xinjiang Uygur Autonomous region, China, and Rome, Italy, were also identified with this strategy. Altogether, 141 chemical components were identified from these three samples, of which 39 components were identified or tentatively identified as new compounds, including 35 groups of isomers. It demonstrated that this strategy provided an efficient protocol for the rapid identification of chemical constituents in complex samples such as traditional Chinese medicines (TCMs) by UHPLC/Q-TOF MS with informatics platform. Graphical Abstract ᅟ.
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http://dx.doi.org/10.1007/s13361-017-1761-5DOI Listing
November 2017

Separation of furostanol saponins by supercritical fluid chromatography.

J Pharm Biomed Anal 2017 Oct 19;145:71-78. Epub 2017 Jun 19.

Beijing Institute of Radiation Medicine, Beijing 100850, China. Electronic address:

Supercritical fluid chromatography (SFC) has good separation efficiency and is suitable for separating weakly polar compounds. Furostanol saponins, as an important kind of steroidal saponins, generally have two sugar chains, which are polar and hydrophilic. The hydroxyl group at the C-22 position of furostanol saponins is active and easily reacts with lower alcohols under appropriate conditions. The separation of hydrophilic furostanol saponins was tested by SFC in this study. The effects of chromatographic conditions on the separation of the mixed furostanol saponins and their hydroxyl derivatives at the C-22 position were studied. The conditions for SFC, which included different column polarity, modifier, additive, and column temperature, were tested. After optimization, the mixed 10 similar structures of furostanol saponins were separated in 22min on the Diol column at a temperature of 40°C. The mobile phase was CO (mobile phase A) and methanol (containing 0.2% NH∙HO and 3% HO) (mobile phase B). The backpressure was maintained isobarically at 11.03MPa. SFC was found to be effective in separating the furostanol saponins that shared the same aglycone but varied in sugar chains. SFC was sensitive to the number and type of sugars. The resolution of furostanol saponin isomers was not ideal. The extract of Dioscorea zingiberensis C. H. Wright was profiled by SFC-quadrupole time-of-flight mass spectrometry. The main saponins of the extract were well separated. Therefore, SFC could be used for separating hydrophilic furostanol saponins and analyzing traditional Chinese medicines that mainly contained steroidal saponins.
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http://dx.doi.org/10.1016/j.jpba.2017.05.023DOI Listing
October 2017

Timosaponin AIII induces antiplatelet and antithrombotic activity via Gq-mediated signaling by the thromboxane A2 receptor.

Sci Rep 2016 12 9;6:38757. Epub 2016 Dec 9.

Department of Pathophysiology, Beijing Key Laboratory for Radiobiology (BKLRB), Beijing Institute of Radiation Medicine, No. 27 Taiping Road, Beijing 100850, China.

The thromboxane (Tx) A pathway is a major contributor to the amplification of initial platelet activation and is therefore a key drug target. To identify potent small-molecule inhibitors of the thromboxane prostaglandin (TP) receptor, we screened a small steroidal saponin library using U46619-induced rat platelet aggregation assays. Timosaponin AIII (TAIII) was identified as a potent inhibitor of U46619-induced rat platelet aggregation and exhibited superior selectivity for the TP receptor versus other G protein-coupled receptors and a PKC activator. TAIII inhibited U46619-induced rat platelet aggregation independent of increases in cAMP and cGMP and the inhibition of TxA2 production. Both PKC and PLC activators restored TAIII-inhibited platelet aggregation, whereas TAIII did not inhibit platelet aggregation induced by co-activation of the G and G pathways. Furthermore, TAIII did not affect the platelet shape change or ROCK2 phosphorylation evoked by low-dose U46619. In vivo, TAIII prolonged tail bleeding time, reduced the mortality of animals with acute pulmonary thromboembolism and significantly reduced venous thrombus weight. Our study suggests that TAIII, by preferentially targeting Gq-mediated PLC/PKC signaling from the TP receptor, induces stronger in vitro antiplatelet activity and in vivo antithrombotic effects and may be an excellent candidate for the treatment of thrombotic disorders.
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http://dx.doi.org/10.1038/srep38757DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5146924PMC
December 2016

Quantitative and Qualitative Analysis of Flavonoids and Phenolic Acids in Snow Chrysanthemum (Coreopsis tinctoria Nutt.) by HPLC-DAD and UPLC-ESI-QTOF-MS.

Molecules 2016 Sep 30;21(10). Epub 2016 Sep 30.

Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China.

A simple, accurate and reliable high performance liquid chromatography coupled with photodiode array detection (HPLC-DAD) method was developed and then successfully applied for simultaneous quantitative analysis of eight compounds, including chlorogenic acid (), (/)-flavanomarein (), butin-7--β-d-glucopyranoside (), isookanin (), taxifolin (), 5,7,3',5'-tetrahydroxyflavanone-7--β-d-glucopyranoside (), marein () and okanin (), in 23 batches of snow chrysanthemum of different seed provenance and from various habitats. The results showed total contents of the eight compounds in the samples with seed provenance from Keliyang (Xinjiang, China), are higher than in samples from the other five provenances by 52.47%, 15.53%, 19.78%, 21.17% and 5.06%, respectively, which demonstrated that provenance has a great influence on the constituents in snow chrysanthemum. Meanwhile, an ultra performance liquid chromatography coupled with electrospray ionization and quadrupole time-of-flight-mass spectrometry (UPLC-ESI-QTOF-MS) was also employed to rapidly separate and identify flavonoids and phenolic acids in snow chrysanthemum from Keliyang. As a result, a total of 30 constituents, including 26 flavonoids and four phenolic acids, were identified or tentatively identified based on the exact mass information, the fragmentation characteristics, and retention times of eight reference standards. This work may provide an efficient approach to comprehensively evaluate the quality of snow chrysanthemum.
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http://dx.doi.org/10.3390/molecules21101307DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6272912PMC
September 2016

Galactosylation of steroidal saponins by β-galactosidase from Lactobacillus bulgaricus L3.

Glycoconj J 2016 Feb 7;33(1):53-62. Epub 2015 Nov 7.

Beijing Institute of Radiation Medicine, 27# Tai-ping Road, Haidian District, Beijing, 100850, China.

The galactosylation of furostanosides and spirostanosides were investigated by using β-galactosidase from Lactobacillus bulgaricus L3 as a catalyst and lactose as a sugar donor. Four novel galactosylated furostanoside products (compounds 1-4) from compound F, compound G, compound I, and compound H were obtained and their structures were identified by HR-ESI-MS, 1D and 2D NMR spectra. The result showed that galactosyl moiety was found to be added to the 6-OH of the 26-O-glucosyl in these four furostanoside substrates.
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http://dx.doi.org/10.1007/s10719-015-9632-4DOI Listing
February 2016

Analytical and semipreparative separation of 25 (R/S)-spirostanol saponin diastereomers using supercritical fluid chromatography.

J Sep Sci 2013 Oct 23;36(19):3270-6. Epub 2013 Aug 23.

Department of Biotechnology, Beijing Institute of Radiation Medicine, Beijing, P. R. China.

Spirostanol saponins are a class of steroidal saponins with many pharmacological activities. The structural complexity of the spirostanol saponins presents a daunting challenge in separating their 25 R/S diastereomers. Using two CHIRALPAK IC columns coupled in series, six 25 (R/S)-spirostanol saponin diastereomers from the Trigonella foenum-graecum L. seed were successfully separated using supercritical fluid chromatography (SFC) for the first time. In addition, three 25 (R/S)-spirostanol saponin diastereomers were isolated into their respective individual isomers. The structures of the isolated isomers were unambiguously confirmed by NMR analysis. The SFC method development strategy and its associated underlying principles presented in this paper are generally applicable. SFC is a viable addition to the natural product research toolbox, especially for stereoselective analysis and purification.
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http://dx.doi.org/10.1002/jssc.201300482DOI Listing
October 2013

Pennogenin tetraglycoside induces rat myometrial contraction and MLC20 phosphorylation via PLC-IP(3) and RhoA/Rho kinase signaling pathways.

PLoS One 2012 12;7(12):e51536. Epub 2012 Dec 12.

Department of Pathophysiology, Beijing Institute of Radiation Medicine, Beijing, China.

Background: Total steroidal saponins extracted from the rhizome of Paris polyphylla Sm. var. yunnanensis (TSSPs) have been widely used in China for the treatment of abnormal uterine bleeding. We previously studied the main active constituents of TSSPs and their structure-activity relationships with respect to rat myometrial contractions. Tg (pennogenin tetraglycoside) was identified as one of the active ingredients in TSSPs able to induce rat myometrial contractions. However, the mechanisms underlying the pharmacological actions on uterine activity have not been described clearly.

Methods: Here Tg was screened for effects on contractile activity in isolated uterine strips from estrogen-primed rats and on MLC20 phosphorylation and related signaling pathways in cultured rat myometrial cells as determined by Western blot. Intracellular calcium ([Ca(2+)](i)) was monitored under a confocal microscope using Fluo-4 AM-loaded myometrial cells.

Results: Tg dose-dependently stimulated rat myometrial contractions as well as MLC20 phosphorylation in vitro, which could be completely suppressed by an inhibitor of myosin light chain kinase (MLCK). Use of Ca(2+) channel blockers and kinase inhibitors demonstrated that Tg-induced myometrial contractions are mediated by activation of the phospholipase C (PLC)-inositol triphosphate (IP3) signaling pathway, resulting in increased MLC20 phosphorylation. Furthermore, Y27632, a specific inhibitor of Rho kinase (ROK), notably suppressed Tg-stimulated myometrial contractions and decreased MLC20 phosphorylation.

Conclusions: These data provide evidence that rat myometrial contractility induced by Tg results from enhanced MLC20 phosphorylation, while both PLC-IP3 and RhoA/ROK signaling pathways mediate the process. These mechanisms may be responsible for the therapeutic effects of TSSPs on abnormal uterine bleeding.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0051536PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3520837PMC
June 2013

Rapid isolation of new furostanol saponins from fenugreek seeds based on ultra-performance liquid chromatography coupled with a hybrid quadrupole time-of-flight tandem mass spectrometry.

J Sep Sci 2012 Jun;35(12):1538-50

Beijing Institute of Radiation Medicine, Beijing, P R China.

An ultra-performance liquid chromatography coupled with a hybrid quadrupole time-of-flight tandem mass spectrometry method was established to rapidly identify and guide the isolation of target saponins from fenugreek seeds. Based on the online screening performance, totally forty-six furostanol saponins were detected and elucidated. Among them, twenty compounds were predicted to be new. To rapidly obtain new furostanol saponins from these seeds, a further phytochemical study was carried out under the guidance of the ultra-performance liquid chromatography coupled with a hybrid quadrupole time-of-flight tandem mass spectrometry. Finally, six new furostanol saponins, named as trigoneosides XIV (1), XV (2), XVI (3), XVIIa (4), XVIIb (5), and XIV (6), together with one known furostanol saponin, parvifloside (7), were rapidly obtained, and their definitive structures were determined by NMR and chemical evidence.
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http://dx.doi.org/10.1002/jssc.201200020DOI Listing
June 2012

Pennogenin tetraglycoside stimulates secretion-dependent activation of rat platelets: evidence for critical roles of adenosine diphosphate receptor signal pathways.

Thromb Res 2012 May 7;129(5):e209-16. Epub 2012 Mar 7.

Department of Pathophysiology, Beijing Institute of Radiation Medicine, Beijing, China.

Background: Total steroidal saponins extracted from the rhizome of Paris polyphylla Sm. var. yunnanensis (TSSPs) have been demonstrated to promote hemostasis in vivo and induce platelet aggregation in vitro. Pennogenin tetraglycoside (Tg) has been identified as one of the active ingredients in TSSPs and can induce rat platelet aggregation.

Objective: To investigate the functional role of Tg in platelets and the signaling pathway mechanisms which mediate Tg-induced platelet aggregation.

Methods And Results: Using scanning electron microscopy, the turbidimetric method and flow cytometry, we demonstrated that Tg induces shape change and concentration-dependently induces aggregation, dense granule secretion and a-granule secretion in rat platelets. The activation characteristics were comprehensively confirmed using transmission electron microscopy. Apyrase and antagonists of the platelet adenosine diphosphate (ADP) receptors, P2Y1 and P2Y12, completely inhibited Tg-induced platelet aggregation, which was not sensitive to indomethacin or SQ29548 inhibition. Furthermore, ADP receptor antagonists inhibited Tg-induced a-granule secretion, and blockade of the P2Y1 receptor prevented Tg-induced platelet shape changes. Tg-induced dense granule secretion was not affected by ADP receptor antagonists or various various pharmacological inhibitors of the intracellular effectors involved in dense granule secretion signaling pathways.

Conclusion: We identified that Tg directly induces platelet activation and demonstrated that Tg-induced platelet activation depends on dense granule secretion of ADP, which in turn activates the P2Y1 and P2Y12 receptor signaling pathways.
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http://dx.doi.org/10.1016/j.thromres.2012.02.001DOI Listing
May 2012

Rapid profiling and identification of triterpenoid saponins in crude extracts from Albizia julibrissin Durazz. by ultra high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.

J Pharm Biomed Anal 2011 Jul 8;55(5):996-1009. Epub 2011 Apr 8.

Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, No. 88 Yuquan Road, Tianjin 300193, PR China. hanlifeng

Ultra high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC/ESI-Q-TOF-MS/MS) was applied to separate and identify triterpenoid saponins in crude extract from the stem bark of Albizia julibrissin Durazz. The molecular weights were determined by comparing quasi-molecular ions [M+NH(4)](+) in positive mode and [M-H](-) and [M-2H](2-) ions in negative mode. The MS/MS spectra of the [M-H](-) ions for saponins provided a wealth of structural information related to aglycone skeletons, sugar types and linked sequence. On the basis of the fragmentation behavior of known saponins isolated before, saponins from this plant were identified, even though references were not available. As a result, a total of twenty-eight saponins in the crude extract were identified, which all had a common basic skeleton of the triterpene oleanolic acid and eight of them were new compounds.
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http://dx.doi.org/10.1016/j.jpba.2011.04.002DOI Listing
July 2011

[Role of Radix Platycodi in Tianwang buxinwan decoction analyzed by fingerprints].

Zhongguo Zhong Yao Za Zhi 2009 Jan;34(2):156-60

Pharmacy of Traditional Chinese Medicine, PLA General Hospital, Beijing 100853, China.

Objective: To establish the fingerprints of the active fractions from Tian-wang-bu-xin-wan decoction by HPLC, and to explain the role of Radix Platycodi in the decoction prescription.

Method: The experimental conditions of the HPLC method were established as follows: Hanbon Lichrospher C18 column (4.6 mm x 250 mm, 5 microm), mobile phase were methanol and 0.012 5 mol L(-1) ammonium acetate/acetic acid buffer, eluted with a linear gradient, flow rate was 1.0 mL min(-1), the photodiode array detector (PDA) and evaporative light scattering detector (ELSD) were connected in series.

Result: In the same conditions and used same method, the extract amount of whole prescription was higher than that of excepting Radix Platycodi. The method provided two kinds of fingerprints with satisfied separation, which were the HPLC-PDA (Max Plot) fingerprint and the HPLC-ELSD fingerprint. And, they had good correlation and complementaritiy.

Conclusion: The Radix Platycodi can enhance the decoction yield of the prescription; and increase the dissolution and the contents of some ingredients in the decoction.
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January 2009