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Dean Guo - One of the best experts on this subject based on the ideXlab platform.
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quantitative analysis of fourteen Bufadienolides in venenum bufonis crude drug and its chinese patent medicines by ultra high performance liquid chromatography coupled with tandem mass spectrometry
Journal of Ethnopharmacology, 2020Co-Authors: Wenlong Wei, Yanzhi Zhang, Yang Zhou, Min Lei, Jianqing Zhang, Dean GuoAbstract:Abstract Ethnopharmacological relevance Venenum Bufonis, a product of the secretions of Bufo gargarizans Cantor or B. melanostictus Schneider, possessed an array of pharmacological activities, such as cardiotonic, anti-tumor, antinociceptive, anti-inflammatory, anesthetic and antimicrobial activities. However, there were few efficient methods for quality evaluation of Venenum Bufonis medicinal materials and its related Chinese patent medicines. Aim of the study To establish an effective method for quality assessment of crude drugs and Chinese proprietary medicines of Venenum Bufonis, and explore the relationship of primary compounds - target - pathway - disease through a series of network databases. Materials and methods An ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-QqQ-MS/MS) method was developed and validated to simultaneously determine 14 Bufadienolides for quantitative analysis of 71 batches of crude drugs and 20 kinds of Chinese patent medicines of Venenum Bufonis. Multiple reaction monitoring with good specificity and accuracy was applied to monitor the 14 Bufadienolides in positive mode. Results The methodology was validated with good specificity, precision, stability, repeatability and recovery. The low limits of quantification were in the range of 0.1–2.7 ng/mL. The relative standard deviation values for intra- and inter-day precisions ranged from 0.98% to 6.3% and from 2.39% to 6.76%, respectively. The recovery was varied from 87.78% to 110.57% for crude drugs and 88.32%–100.96% for Chinese proprietary medicine (Shexiang Baoxin Pill). The contents of 14 analytes in 71 batches of crude drugs and 20 sorts of Chinese proprietary medicines were procured, the results showed that the contents of crude drugs collected from the market exhibited great variations. Furthermore, 13 batches of crude drugs were identified as counterfeit with no Bufadienolides detected. In addition, the total contents of Bufadienolides in the same drug showed great difference among products from various manufacturers or brands. Subsequently, 9 Bufadienolides with the higher contents were applied to screen the anti-tumor effect by network pharmacology, and 8 pathways which had prior correlation with Bufadienolides were disclosed. Conclusion This method could be used for quality assessment of crude drugs and Chinese patent medicines of Venenum Bufonis, and the data could be served as the fundamental basis for drug research and development of Venenum Bufonis.
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simultaneous determination of cytotoxic Bufadienolides in the chinese medicine chansu by high performance liquid chromatography coupled with photodiode array and mass spectrometry detections
Journal of Chromatography B, 2006Co-Authors: Hui Guo, Hongzhu Guo, Jian Han, Dean GuoAbstract:Abstract ChanSu (toad venom) is a traditional Chinese medicine for the treatment of serious liver and gastric cancers. The major cytotoxic compounds in ChanSu are Bufadienolides. In this paper, a strategy combining qualitative LC/MS analysis and quantitative HPLC determination of major Bufadienolides was used for global quality control of ChanSu crude drug. Majority of the Bufadienolides in methanol extract of ChanSu were unambiguously characterized by high-performance liquid chromatography coupled with atmospheric pressure chemical ionization tandem mass spectrometry (HPLC/APCI-MS/MS), and by comparing with pure compounds. In addition, eight major Bufadienolides were simultaneously determined in one single HPLC run within 30 min with photodiode array detection (DAD). All compounds showed good linearity in a wide concentration range, and their limits of detection (LOD) were around 1 ng. Thus, >95% of the Bufadienolides in ChanSu could be characterized, and >90% of them were quantitated. The established method is rapid, simple and sensitive, and could be used for the routine analysis of ChanSu crude drug and its preparations. This research sets a good example for the comprehensive quality control of traditional medicine.
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analysis of Bufadienolides in the chinese drug chansu by high performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry
Rapid Communications in Mass Spectrometry, 2005Co-Authors: Dean GuoAbstract:The qualitative analysis of Bufadienolides in the Chinese drug ChanSu was performed using high-performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry (APCI-MS/MS). Bufadienolides are the major bioactive constituents of ChanSu, which is used to treat heart failure and cancer in traditional Chinese medicine. The APCI-MS fragmentation behavior of Bufadienolides was studied. For Bufadienolides with only hydroxyl substituents, the fragmentation was characterized by successive eliminations of H2O and CO molecules, and the profile of MS/MS product ions was correlated with the number of hydroxyl groups. If a C-16 acetoxyl group was present, the fragmentation of [M+H]+ ions was triggered by initial loss of 60 Da (HOAc). The elimination of CO was significant for Bufadienolides with a 19-formyl group, and the 19-hydroxyl group could be characterized by the loss of 30 Da (HCHO). These fragmentation rules were applied to the identification of Bufadienolides in a methanolic extract of ChanSu, which was separated on a C18 column with gradient elution. A total of 35 Bufadienolides were identified, including four new constituents. The method established here facilitated the convenient and rapid quality control of ChanSu crude drug and its pharmaceutical preparations. Copyright © 2005 John Wiley & Sons, Ltd.
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analysis of Bufadienolides in the chinese drug chansu by high performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry
Rapid Communications in Mass Spectrometry, 2005Co-Authors: Dean GuoAbstract:The qualitative analysis of Bufadienolides in the Chinese drug ChanSu was performed using high-performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry (APCI-MS/MS). Bufadienolides are the major bioactive constituents of ChanSu, which is used to treat heart failure and cancer in traditional Chinese medicine. The APCI-MS fragmentation behavior of Bufadienolides was studied. For Bufadienolides with only hydroxyl substituents, the fragmentation was characterized by successive eliminations of H(2)O and CO molecules, and the profile of MS/MS product ions was correlated with the number of hydroxyl groups. If a C-16 acetoxyl group was present, the fragmentation of [M+H](+) ions was triggered by initial loss of 60 Da (HOAc). The elimination of CO was significant for Bufadienolides with a 19-formyl group, and the 19-hydroxyl group could be characterized by the loss of 30 Da (HCHO). These fragmentation rules were applied to the identification of Bufadienolides in a methanolic extract of ChanSu, which was separated on a C(18) column with gradient elution. A total of 35 Bufadienolides were identified, including four new constituents. The method established here facilitated the convenient and rapid quality control of ChanSu crude drug and its pharmaceutical preparations.
Xinmiao Liang - One of the best experts on this subject based on the ideXlab platform.
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efficient purification of active Bufadienolides by a class separation method based on hydrophilic solid phase extraction and reversed phase high performance liquid chromatography
Journal of Pharmaceutical and Biomedical Analysis, 2014Co-Authors: Aijin Shen, Chaoran Wang, Jingyu Yan, Weijie Zhao, Xinmiao LiangAbstract:Abstract Traditional Chinese medicines (TCMs) have played a significant role in the process of discovering natural bioactive compounds, especially in anticancer therapeutics. However, the components of TCMs are complex mixtures with wide variation in polarity and content, which leads to inefficiency in the process of active compound discovery from TCMs. In this paper, the popular strategy of utilizing “pre-fractionated natural product libraries” has been improved by a new class separation approach to accelerate the process. As an example, the skin of Bufo bufo gargarizans Cantor, a well-known TCM, mainly contains two distinct Bufadienolide classes: amino acid-conjugated Bufadienolides (AACBs) and free form Bufadienolides (AAUBs). We utilized hydrophilic interaction liquid chromatography solid-phase extraction (HILIC-SPE) to resolve the two types of Bufadienolides, which co-eluted on C18 columns. By this strategy, twelve Bufadienolides of the two types were purified via prep-HPLC from one active fraction, and eight of them were identified by 1 H NMR and 13 C NMR. These results indicated that the class separation method not only overcame the limited orthogonality in a 2D-RPLC × RPLC system but also accelerated the process of active compound discovery.
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Purification of Bufadienolides from the skin of Bufo bufo gargarizans Cantor with positively charged C18 column
Journal of pharmaceutical and biomedical analysis, 2014Co-Authors: Chaoran Wang, Aijin Shen, Weijie Zhao, Xinmiao LiangAbstract:As a kind of promising anticancer compounds, the preparation of Bufadienolides is a hot study spot. However, due to the complexity of biological sample, the purification of Bufadienolides from a crude sample (toad skin) is a tough work. In this paper, we reported a new way based on positively charged C18 material (XCharge C18) to quickly separate and purify Bufadienolides from toad skin. By this method, the different ionic feature of the amino acid conjugated Bufadienolides (AACBs) and the free form Bufadienolides (AAUBs) was firstly utilized to obtain distinct separation selectivity on the XCharge C18 column. Additionally, the peak tailing problem of AACBs on conventional C18 was resolved and better resolutions were achieved on the XCharge C18, thus, two kinds of Bufadienolides on one column were successfully purified respectively. Taking F13 as an example, the method was validated by liquid chromatography-mass spectrometry (LC-MS), and then 4 AACBs as well as 4 AAUBs were simultaneously purified by preparative XCharge C18. In addition, the application of this method in other fractions was also validated. The results suggested that the developed method is a practical and promising tool for efficient separation and purification of Bufadienolides from toad skin.
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systematic screening and characterization of novel Bufadienolides from toad skin using ultra performance liquid chromatography electrospray ionization quadrupole time of flight mass spectrometry
Rapid Communications in Mass Spectrometry, 2010Co-Authors: Yuansheng Xiao, Xingya Xue, Xinmiao LiangAbstract:during the discovery process of novel compounds, it is of significant importance to differentiate novel from known compounds in crude extracts before starting the time-consuming process of purification. Bufadienolides are the main active components of the skin of the toad bufo bufo gargarizans cantor (toad skin), an important traditional chinese medicine. the fragmentation behavior and mass spectra profiles of Bufadienolides standards were investigated using ultra-performance liquid chromatography/electrospray ionization quadrupole time-of-flight mass spectrometry (uplc/esi-q-tofms). several fragmentation rules were summarized and applied to characterize novel and known Bufadienolides in toad skin. characteristic substituent groups could be identified by both diagnostic ions and their relative abundance. Bufadienolide stereoisomers could be differentiated from positional isomers by comparing fragment abundance profiles. this was used to characterize new stereoisomers; for known Bufadienolides. a total of 39 Bufadienolides were screened out using a systematic method developed in our laboratory. in addition to 19 known Bufadienolides, 20 putative novel compounds, including 8 stereoisomers, were characterized. uplc/q-tofms was demonstrated to be a powerful tool for the characterization of low-abundance Bufadienolides in complex samples. this study provides guidelines for the targeted isolation of novel Bufadienolides from natural products. copyright (c) 2010 john wiley & sons, ltd.
Ren-wang Jiang - One of the best experts on this subject based on the ideXlab platform.
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An Efficient Strategy Based on Liquid–Liquid Extraction with Three-Phase Solvent System and High Speed Counter-Current Chromatography for Rapid Enrichment and Separation of Epimers of Minor Bufadienolide from Toad Meat
2018Co-Authors: Denglang Zou, Xuelin Zhu, Fan Zhang, Ren-wang JiangAbstract:This study presents an efficient strategy based on liquid–liquid extraction with three-phase solvent system and high speed counter-current chromatography for rapid enrichment and separation of epimers of minor Bufadienolide from toad meat. The reflux extraction conditions were optimized by response surface methodology first, and a novel three-phase solvent system composed of n-hexane/methyl acetate/acetonitrile/water (3:6:5:5, v/v) was developed for liquid–liquid extraction of the crude extract. This integrative extraction process could enrich minor Bufadienolide from complex matrix efficiently and minimize the loss of minor targets induced by repeated extraction with different kinds of organic solvents occurring in the classical liquid two-phase extraction. As a result, four epimers of minor Bufadienolide were greatly enriched in the middle phase and total content of these epimers of minor Bufadienolide was increased from 3.25% to 46.23%. Then, the enriched four epimers were separated by HSCCC with a two-phase solvent system composed of chloroform/methanol/water (4:2:2, v/v) successfully. Furthermore, we tested Na+,K+-ATPase (NKA) inhibitory effect of the four epimers. 3β-Isomers of Bufadienolide showed stronger (>8-fold) inhibitory activity than 3α-isomers. The characterization of minor Bufadienolide in toad meat and their significant difference of inhibitory effect on NKA would promote the further quantitative analysis and safety evaluation of toad meat as a food source
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isolation and identification of l d lactate conjugated Bufadienolides from toad eggs revealing lactate racemization in amphibians
Organic and Biomolecular Chemistry, 2017Co-Authors: Shiwen Zhou, Ren-wang Jiang, Lei Wang, Qingfei Zheng, Xiuyong Huang, Yong Wang, Sifan Luo, Hai-yan TianAbstract:Three pairs of Bufadienolide l/d-lactate epimers (1-6) were isolated from the eggs of the toad Bufo bufo gargarizans. The structures were elucidated by using spectroscopic methods, X-ray diffraction analysis and a modified Mosher's method. Compounds 1-6 represent the first occurrence of lactate-conjugated Bufadienolides in nature, and illustrate the existence of an enzyme-controlled epimerization from l- to d-lactate in amphibians. The biosynthetic pathways, in which two key enzymes might be involved (i.e., lactate racemase and acyltransferase), were proposed. In addition, the biological assays revealed that compounds 1-4 are potent cytotoxic agents against human gastric cancer cells BGC-823 and human lung cancer cells A549 with IC50 values in a range of 8.0 to 80.0 nM.
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structures and inhibitory activity against breast cancer cells of new Bufadienolides from the eggs of toad bufo bufo gargarizans
RSC Advances, 2016Co-Authors: Pengwei Zhang, Hai-yan Tian, Dong-mei Zhang, Lei Wang, Shiwen Zhou, Qiulin Nie, Ren-wang JiangAbstract:Two new 19-norBufadienolides (1 and 2), one new 14,15-epoxy Bufadienolide (3), and eight rare Bufadienolide–fatty acid conjugates (4–11), together with four known ones (12–15) were isolated from the eggs of toad Bufo bufo gargarizans. Their chemical structures were elucidated by extensive spectroscopic methods in combination with X-ray diffraction analyses. Furthermore, we tested the inhibitory effect of these compounds against breast cancer cell lines MCF-7 and MDA-MB-231. Most of them showed strong cytotoxicity with IC50 values less than 0.1 μM. The further mechanistic study showed that they could induce apoptosis and cycle arrest in the G2/M phase in MCF-7 cells.
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C23 Steroids from the Venom of Bufo bufo gargarizans
2016Co-Authors: Hai-yan Tian, Jun-shan Liu, Dong-mei Zhang, Lei Wang, Shi-lin Luo, Ying Wang, Xiao-qi Zhang, Ren-wang JiangAbstract:Five new C23 steroids (1–5) together with five known Bufadienolides (6–10) were isolated from the venom of Bufo bufo gargarizans (ChanSu in Chinese). The structures of the new steroids were elucidated by extensive spectroscopic methods in combination with X-ray diffraction analysis. Among these C23 steroids, only compound 3 showed cytotoxicities against HepG2 and A549 cancer cells, with respective IC50 values of 26.8 ± 8.3 and 45.6 ± 2.5 μM. In contrast, the Bufadienolides (7–10) displayed potent inhibitory activities against these cancer cells, with respective IC50 values in the ranges 0.5–5.5 and 0.6–6.5 μM, but relatively less cytotoxicity on normal mouse spleen cells. In addition, the Na+/K+-ATPase inhibitory activities of 2, 5, and 7 revealed that the lactone moiety of a Bufadienolide was important for the inhibitory activity
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Bufadienolides with cytotoxic activity from the skins of Bufo bufo gargarizans.
Fitoterapia, 2015Co-Authors: Hai-yan Tian, Yu-he Lei, Dong-mei Zhang, Lei Wang, Ren-wang JiangAbstract:Abstract Twelve new Bufadienolides (1–12), along with fourteen known analogues (13–26) were isolated from the skins of Bufo bufo gargarizans Cantor. Their chemical structures were elucidated on the basis of NMR, HRESIMS and X-ray diffraction analysis. Compound 1 was an unusual Bufadienolide with 3,19-epoxy moiety and A/B trans ring junction. Compounds 2–4 were rare Bufadienolides possessing 10-H or 10-carboxyl units. All the isolated compounds were tested for their cytotoxic effects on HepG2, A549 and HeLa cells. Six new compounds (2, 3, 5, 6, 10 and 12) displayed significant anti-proliferative activities with IC50 values ranging from 0.049 to 1.856 μM. Arenobufagin (24) exhibited the most potent cytotoxic activity with IC50 value 0.011 μM. In addition, the present data provided more insight into the structure–activity relationships of Bufadienolides.
Hai-yan Tian - One of the best experts on this subject based on the ideXlab platform.
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isolation and identification of l d lactate conjugated Bufadienolides from toad eggs revealing lactate racemization in amphibians
Organic and Biomolecular Chemistry, 2017Co-Authors: Shiwen Zhou, Ren-wang Jiang, Lei Wang, Qingfei Zheng, Xiuyong Huang, Yong Wang, Sifan Luo, Hai-yan TianAbstract:Three pairs of Bufadienolide l/d-lactate epimers (1-6) were isolated from the eggs of the toad Bufo bufo gargarizans. The structures were elucidated by using spectroscopic methods, X-ray diffraction analysis and a modified Mosher's method. Compounds 1-6 represent the first occurrence of lactate-conjugated Bufadienolides in nature, and illustrate the existence of an enzyme-controlled epimerization from l- to d-lactate in amphibians. The biosynthetic pathways, in which two key enzymes might be involved (i.e., lactate racemase and acyltransferase), were proposed. In addition, the biological assays revealed that compounds 1-4 are potent cytotoxic agents against human gastric cancer cells BGC-823 and human lung cancer cells A549 with IC50 values in a range of 8.0 to 80.0 nM.
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structures and inhibitory activity against breast cancer cells of new Bufadienolides from the eggs of toad bufo bufo gargarizans
RSC Advances, 2016Co-Authors: Pengwei Zhang, Hai-yan Tian, Dong-mei Zhang, Lei Wang, Shiwen Zhou, Qiulin Nie, Ren-wang JiangAbstract:Two new 19-norBufadienolides (1 and 2), one new 14,15-epoxy Bufadienolide (3), and eight rare Bufadienolide–fatty acid conjugates (4–11), together with four known ones (12–15) were isolated from the eggs of toad Bufo bufo gargarizans. Their chemical structures were elucidated by extensive spectroscopic methods in combination with X-ray diffraction analyses. Furthermore, we tested the inhibitory effect of these compounds against breast cancer cell lines MCF-7 and MDA-MB-231. Most of them showed strong cytotoxicity with IC50 values less than 0.1 μM. The further mechanistic study showed that they could induce apoptosis and cycle arrest in the G2/M phase in MCF-7 cells.
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C23 Steroids from the Venom of Bufo bufo gargarizans
2016Co-Authors: Hai-yan Tian, Jun-shan Liu, Dong-mei Zhang, Lei Wang, Shi-lin Luo, Ying Wang, Xiao-qi Zhang, Ren-wang JiangAbstract:Five new C23 steroids (1–5) together with five known Bufadienolides (6–10) were isolated from the venom of Bufo bufo gargarizans (ChanSu in Chinese). The structures of the new steroids were elucidated by extensive spectroscopic methods in combination with X-ray diffraction analysis. Among these C23 steroids, only compound 3 showed cytotoxicities against HepG2 and A549 cancer cells, with respective IC50 values of 26.8 ± 8.3 and 45.6 ± 2.5 μM. In contrast, the Bufadienolides (7–10) displayed potent inhibitory activities against these cancer cells, with respective IC50 values in the ranges 0.5–5.5 and 0.6–6.5 μM, but relatively less cytotoxicity on normal mouse spleen cells. In addition, the Na+/K+-ATPase inhibitory activities of 2, 5, and 7 revealed that the lactone moiety of a Bufadienolide was important for the inhibitory activity
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Bufadienolides with cytotoxic activity from the skins of Bufo bufo gargarizans.
Fitoterapia, 2015Co-Authors: Hai-yan Tian, Yu-he Lei, Dong-mei Zhang, Lei Wang, Ren-wang JiangAbstract:Abstract Twelve new Bufadienolides (1–12), along with fourteen known analogues (13–26) were isolated from the skins of Bufo bufo gargarizans Cantor. Their chemical structures were elucidated on the basis of NMR, HRESIMS and X-ray diffraction analysis. Compound 1 was an unusual Bufadienolide with 3,19-epoxy moiety and A/B trans ring junction. Compounds 2–4 were rare Bufadienolides possessing 10-H or 10-carboxyl units. All the isolated compounds were tested for their cytotoxic effects on HepG2, A549 and HeLa cells. Six new compounds (2, 3, 5, 6, 10 and 12) displayed significant anti-proliferative activities with IC50 values ranging from 0.049 to 1.856 μM. Arenobufagin (24) exhibited the most potent cytotoxic activity with IC50 value 0.011 μM. In addition, the present data provided more insight into the structure–activity relationships of Bufadienolides.
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Two new Bufadienolides from the rhizomes of Helleborus thibetanus with inhibitory activities against prostate cancer cells
Natural product research, 2014Co-Authors: Wei Cheng, Hai-yan Tian, Ya-fang Tan, Xiang-wen Gong, Ke-li Chen, Ren-wang JiangAbstract:Two new Bufadienolide glycosides (1 and 2) with an A/B trans ring fusion together with nine known compounds (3–11) were isolated from the rhizomes of Helleborus thibetanus. The structures of new compounds were elucidated by extensive spectroscopic analyses in combination with single-crystal X-ray diffraction. The Bufadienolides 1 and 3–6 exhibited potent cytotoxic activities against the prostate cancer cells.
Aijin Shen - One of the best experts on this subject based on the ideXlab platform.
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efficient purification of active Bufadienolides by a class separation method based on hydrophilic solid phase extraction and reversed phase high performance liquid chromatography
Journal of Pharmaceutical and Biomedical Analysis, 2014Co-Authors: Aijin Shen, Chaoran Wang, Jingyu Yan, Weijie Zhao, Xinmiao LiangAbstract:Abstract Traditional Chinese medicines (TCMs) have played a significant role in the process of discovering natural bioactive compounds, especially in anticancer therapeutics. However, the components of TCMs are complex mixtures with wide variation in polarity and content, which leads to inefficiency in the process of active compound discovery from TCMs. In this paper, the popular strategy of utilizing “pre-fractionated natural product libraries” has been improved by a new class separation approach to accelerate the process. As an example, the skin of Bufo bufo gargarizans Cantor, a well-known TCM, mainly contains two distinct Bufadienolide classes: amino acid-conjugated Bufadienolides (AACBs) and free form Bufadienolides (AAUBs). We utilized hydrophilic interaction liquid chromatography solid-phase extraction (HILIC-SPE) to resolve the two types of Bufadienolides, which co-eluted on C18 columns. By this strategy, twelve Bufadienolides of the two types were purified via prep-HPLC from one active fraction, and eight of them were identified by 1 H NMR and 13 C NMR. These results indicated that the class separation method not only overcame the limited orthogonality in a 2D-RPLC × RPLC system but also accelerated the process of active compound discovery.
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Purification of Bufadienolides from the skin of Bufo bufo gargarizans Cantor with positively charged C18 column
Journal of pharmaceutical and biomedical analysis, 2014Co-Authors: Chaoran Wang, Aijin Shen, Weijie Zhao, Xinmiao LiangAbstract:As a kind of promising anticancer compounds, the preparation of Bufadienolides is a hot study spot. However, due to the complexity of biological sample, the purification of Bufadienolides from a crude sample (toad skin) is a tough work. In this paper, we reported a new way based on positively charged C18 material (XCharge C18) to quickly separate and purify Bufadienolides from toad skin. By this method, the different ionic feature of the amino acid conjugated Bufadienolides (AACBs) and the free form Bufadienolides (AAUBs) was firstly utilized to obtain distinct separation selectivity on the XCharge C18 column. Additionally, the peak tailing problem of AACBs on conventional C18 was resolved and better resolutions were achieved on the XCharge C18, thus, two kinds of Bufadienolides on one column were successfully purified respectively. Taking F13 as an example, the method was validated by liquid chromatography-mass spectrometry (LC-MS), and then 4 AACBs as well as 4 AAUBs were simultaneously purified by preparative XCharge C18. In addition, the application of this method in other fractions was also validated. The results suggested that the developed method is a practical and promising tool for efficient separation and purification of Bufadienolides from toad skin.
