Volume 12,Issue 1,2020 Table of Contents

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  • 1  Inheriting Essence, Keeping Integrity and Innovation
    Chang-xiao Liu De-an Guo
    2020, 12(1):1-2. DOI: https://doi.org/10.1016/j.chmed.2020.01.001
    [Abstract](397) [HTML](0) [PDF 0.00 Byte](23)
    Abstract:
    In China, traditional Chinese medicine (TCM) is a complicated scientific system formulated by the health philosophy and practice in China’s thousands of years of history. It is a treasure of the Chinese civilization and embodies the great wisdom of the Chinese people and nation. Since the founding of the People’s Republic of China in 1949, remarkable achievements have been made in the field of TCM, which has made important contributions to the improvement of people’s health. Today, during the process of advancing the construction of healthy China, Chinese herbal medicine (CHM) must abide by “The Thought of Socialism with Chinese Characteristics for a New Era”, and give full play to the advantages of TCM in an attempt to make a historic contribution to realize the healthy China dream. In addition, one should vigorously push forward with the scientific and technological innovation of Chinese medicine and new drug research & development and make full use of the TCM unique advantage in disease prevention, treatment and rehabilitation, and finally realize high quality development of TCMs amid the inheritance and innovation of TCMs. President Xi Jin-ping called for the importance of "following the development rules of TCM, inheriting the essence and keeping the integrity and innovation", which is the significant and guiding thought to give play to the unique advantages of TCM in the disease prevention and treatment, modernization and industrialization of CHM products, complement each other between traditional and west medicine, promote the coordinated and high quality development of Chinese medicine cause and industry. It is a pivotal strategic guiding thought to build a healthier China system, and to realize the great rejuvenation of the Chinese nation. Following the law of development of Chinese medicine, understanding the thought of "Inheriting essence, keeping integrity and innovation", insisting on cultural confidence, deepening the reform and innovation are the core messages passed by "Central Committee of Chinese Communist Party and State Council Opinion on Preservation and Innovative Development of TCM” (Opinion) with 20 articles containing 125 items released in October 2019. Under the guidance of the “Opinion”, actions should be urgently taken on implementing the ground work in accordance with development road of the characteristics of TCM. It is an imperative historical mission and responsibility to follow President Xi’s philosophy of "Inheriting essence, keeping integrity and innovation" and develop TCM with the historical view and development vision. After thousands of years of development, TCM embodies the profound theory, great wisdom, cognitive mode, development law and exuberant vitality of the Chinese people and nation. In today’s great rejuvenation of the Chinese nation, "carrying forward,inheriting and innovating" has always been the key to promote TCM sticking to its own development. During the development and transformation of TCM industry, we gradually lost ourselves due to the existing problems in TCM research & development and industry supervision, theory, methods and standards for TCM evaluation,as well as lack of culture confidence. The newly released historic "Opinion” by Central Committee of CPC and States Council of China has clearly stated that the spirit of following the development law of TCM should run throughout. That is to say, only by following the development law of TCM could we understand how to inherit the “essence”, keep the “right” and create the “new”. As the document pointed out, it is essential to give play to the unique role of TCM in maintaining and promoting people’s health. We should follow the development rules of TCM, establish and improve the modern management system reflecting the characteristics of TCM, establish the safety and efficacy evaluation methods and technical standards of TCM products with the characteristics of TCM. However, if it deviates from its own law and "right" is not observed, it will be impossible to address the characteristic advantages of TCM. If it does not create "new", it will inevitably deviate from the goal with great clinical demand. Of course, to follow its own development rules does not mean self-isolation. Openness and inclusiveness also need to be followed. It is an important part of the development to accelerate the construction of TCM theory and the establishment of a multidisciplinary scientific research platform in the coordinated development. The "integrity" of TCM innovation lies in the practical significance of strengthening the confidence of TCM culture and theory, keeping an open and inclusive mind and focusing on the innovation goal. For TCM research & development and industry sustainable development, ranging from industry development,medicinal herbs cultivation and breeding, research and development of new products, the recourse of the quality management system, to the medical and health need, to the product safety and effectiveness, must conform to the trend of the new era, social demand and social contribution to the economic development of practical significance. The development of TCM has always been the root of TCM discipline innovation, but also the soul of TCM innovation. Therefore, looking back on the unforgettable 2019 and ushering in the 2020s, we should "inherit essence and keep integrity and innovation", carry forward the innovative development, and blaze a trail with the characteristics of TCM development for a healthier China. Cultivating TCM acclaimed doctors, famous hospitals and classic prescriptions, forming well-known factories, celebrated medicines and branded shops in CHM industry,will be the inevitable trend for the development of TCM in the world. Our international academic journal, Chinese Herbal Medicines (CHM), with the support of colleagues home and abroad, has gone through its full 10 years of history and has made an international impact on published quality articles. With the ushering of 2020s, we would like to convey our sincere gratitude to all who contributed CHM. TCMs from different nationalities are the targeted sources of the research and development, and CHM is a great platform for scientists to promote academic discoveries and carry forward innovative research results.
    2  Chinese herbal medicine resources: Where we stand
    Ran-ran Gao a c Ya-ting Hu b Yang Dan a Li-jun Hao a c Xia Liu b Jing-yuan Song a c d ?
    2020, 12(1):3-13. DOI: https://doi.org/10.1016/j.chmed.2019.08.004
    [Abstract](208) [HTML](0) [PDF 0.00 Byte](19)
    Abstract:
    In recent years, the development of Chinese herbal medicine (CHM) has been challenged by shortages of CHM resources and drug safety concerns related to end products. There have been significant efforts by Chinese scholars to tackle these challenges, which are revealed by analyzing the research trend of CHM resources via surveying Chinese Traditional and Herbal Drugs (Zhong Cao Yao), a representative jour- nal in CHM. Our study focused on 781 articles in CHM resources from 2013 to 2018 and included four subject areas: germplasm resources, quality analysis and evaluation, cultivation, and bioengineering of CHM. Discussion and prospective for future investigations were also presented, including: construct the core germplasm of medicinal plants and expand germplasms; combine molecular research with field ex- periments and promote the deeper study of cultivation of CHM plants; improve the quality evaluation method of CHM and strengthen the identification of Chinese patented medicines; promote the sustain- able development of CHM resources by utilizing bioengineering and synthetic biology. This study helps international scholars understand the status quo of CHM research and provides theoretical support for the healthy, modern, and international development of CHM, and it will facilitate the sustainable devel- opment of the traditional Chinese medicine industry
    3  New insights into roles of basophils in initiating T helper type 2 immunity
    Tian-xiang Ma a Yuan Gao b ?
    2020, 12(1):14-18. DOI: https://doi.org/10.1016/j.chmed.2019.09.006
    [Abstract](203) [HTML](0) [PDF 0.00 Byte](21)
    Abstract:
    Allergic diseases, mainly mediated by T helper type 2 (Th2) immunity, have become a worldwide public health problem. Traditional Chinese medicine (TCM) has long been used in treating and preventing allergic symptoms. As the new target of anti-allergy TCM, basophils, after approximately 140 years since their discovery, are just now gaining respect as important contributors in the pathogenesis underlying allergic inflammation and disease. In addition to their role as effector cells, basophils can release early IL-4, migrate from circulatory system into draining lymph nodes, present antigen to naive CD4 + T cells,and promote the differentiation of Th2 cells. Herein, we briefly summarized the recent research advances of the essential contributions of basophils in the initiation of Th2 immune responses.
    4  Integrated chemical and transcriptomic analyses unveils synthetic characteristics of different medicinal root parts of Angelica sinensis
    Ran Xu a b Jiang Xu b Yong-chang Li c Yun-tao Dai b Shao-peng Zhang a Guang Wang a Zhi-guo Liu a Lin-lin Dong b ? Shi-lin Chen b ?
    2020, 12(1):19-28. DOI: https://doi.org/10.1016/j.chmed.2019.07.003
    [Abstract](702) [HTML](0) [PDF 0.00 Byte](17)
    Abstract:
    Objective: Why are different medicinal parts including heads, bodies and tails of Angelicae Sinensis Radix (ASR) distinct in pharmaceutical activities? Here we explored their discrepancy in chemical constituents and transcriptome. Methods: ASR were separated into three medicinal parts: heads (rootstocks with petiole traces of ASR), bodies (taproots of ASR) and tails (lateral roots of ASR), and chemical and transcriptomic analyses were conducted simultaneously. Results: High performance liquid chromatography (HPLC) fingerprint results showed that five widely used active ingredients (ferulic acid, senkyunolide H, senkyunolide A, n-butylphathlide, and ligustilide) were distributed unevenly in the three ASR medicinal parts. Partial least squares-discriminant analysis (PLS-DA) demonstrated that the heads can be differentiated from the two other root parts due to different amounts of the main components. However, the content of ferulic acid (a main quality marker) was significantly higher in tails than in the heads and bodies. The transcriptome analysis found that 25,062, 10,148 and 29,504 unigenes were specifically expressed in the heads, bodies and tails, respectively. WGCNA analysis identified 17 co-expression modules, which were constructed from the 19,198 genes in the nine sam- ples of ASR. Additionally, we identified 28 unigenes involved in two phenylpropanoid biosynthesis (PB) pathways about ferulic acid metabolism pathways, of which 17 unigenes (60.7%) in the PB pathway were highly expressed in the tails. The expression levels of PAL, C3H, and CQT transcripts were significantly higher in the tails than in other root parts. RT-qPCR analysis confirmed that PAL, C3H, and CQT genes were predominantly expressed in the tail parts, especially PAL, whose expression was more than doubled as compared with that in other root parts. Conclusion: Chemical and transcriptomic analyses revealed the distribution contents and pivotal tran- scripts of the ferulic acid biosynthesis-related pathways. The spatial gene expression pattern partially explained the discrepancy of integral medicinal activities of three medicinal root parts.
    5  Critical pharmaceutical process identification considering chemical composition, biological activity, and batch-to-batch consistency: A case study of notoginseng total saponins
    Jing-jing Pan a Si-yuan He a Jing-yuan Shao a Ning Li b Yun-qi Gong b Xing-chu Gong a ?
    2020, 12(1):29-35. DOI: https://doi.org/10.1016/j.chmed.2019.11.002
    [Abstract](485) [HTML](0) [PDF 0.00 Byte](19)
    Abstract:
    Objective: Critical pharmaceutical process identification (CPPI) is an important step in the implementa- tion of quality by design concept to traditional Chinese medicines (TCMs). Risk assessment methods are usually used in CPPI. However, risk evaluation is usually subjective. The purpose of this work is to present a more objective CPPI method. Methods: A CPPI method considering chemical composition, biological activity, and batch-to-batch con- sistency was presented in this work. The manufacturing process of notoginseng total saponins (NTS) was investigated as an example. The changes of chemical composition, biological activity, and chemical com- position consistency after main processes were measured and compared. A significant change of them indicated a critical process. Results: After extraction process and chromatography process, saponin purity and chemical composition similarity remarkably increased, and saponin content variations decreased. Thrombin inhibitory activity was remarkably decreased after chromatography process. Because of the large influences on NTS quality, extraction process and chromatography process were identified to be critical processes of NTS. Conclusion: Based on a comprehensive and objective examination of the role of each process, critical pharmaceutical processes can be identified. A similar method can also be applied to other TCM processes.
    6  Mangiferin exerts neuroprotective activity against lead-induced toxicity and oxidative stress via Nrf2 pathway
    Hao-wen Li a c d Tai-jin Lan b d Chen-xia Yun b Ke-di Yang c Zheng-cai Du d Xue-fei Luo e Er-wei Hao d Jia-gang Deng d
    2020, 12(1):36-46. DOI: https://doi.org/10.1016/j.chmed.2019.12.002
    [Abstract](675) [HTML](0) [PDF 0.00 Byte](15)
    Abstract:
    Objective: The study was designed to assess the beneficial role of mangiferin (MGN) in lead (Pb)-induced neurological damages in the activation of Nrf2-governed enzymes, genes and proteins. Methods: A total of 96 weaned Wistar rats (48 males and 48 females, 26- to 27-day-old), weighing 50?80 g were used. The experiment was performed in six groups: normal group (control, n = 16), model group (chronic Pb exposed, n = 16), Dimercaptosuccinic acid (DMSA)-treated group (positive control, Pb + DMSA, n = 16), three MGN-treated groups with different doses (Pb + MGN, n = 48). Normal group freely had access to purified water. DMSA-treated group was given DMSA, which was clinically used as the standard treatment for moderate Pb poisoning, at 50 mg/kg (2 mL suspension with purified water) by intragastric gavage (ig) 4 continual days a week for 4 weeks, MGN-treated groups were given MGN at 50, 100, or 200 mg/kg (2 mL suspension with purified water) by ig daily for 4 weeks. At the end of the treatment, all rats were sacrificed and the brain samples were collected. The haematoxylin and eosin (H&E) staining was used for observation of histopathology. Commercial kit, real-time quantitative poly- merase chain reaction (RT-qPCR), Western-blot and immunohistochemistry (IHC) detection were used to detect the mRNA and protein expression. Results: Eight weeks exposure to Pb-containing water resulted in pathological alterations, anti-oxidative system disorder in the brain, all of which were blocked by MGN in a Nrf2-dependent manner. Nrf2 down- stream enzymes such as HO-1, NQO1, γ -GCS were activated. Nrf2, GCLC, GCLM, HO-1 mRNA and total Nrf2, Nuclear Nrf2, γ -GCS, HO-1 protein expression were affected too. Conclusion: MGN ameliorated morphological damage in the hippocampus. Its neuroprotective effects were achieved by the activation of the Nrf2 downstream genes. The data from this in vitro study indi- cates that MGN targeting Nrf2 activation is a feasible approach to reduce adverse health effects associated with Pb exposure. Thus, MGN could be an effective candidate agent for the Pb-induced oxidative stress and neurotoxicity in the human body.
    7  Phytochemical and anti-neuropathic investigations of Crocus sativus via alleviating inflammation, oxidative stress and pancreatic beta-cells regeneration
    Karim Raafat Maha Aboul-Ela Abdalla El-Lakany
    2020, 12(1):47-55. DOI: https://doi.org/10.1016/j.chmed.2019.07.004
    [Abstract](333) [HTML](0) [PDF 0.00 Byte](20)
    Abstract:
    Objective: The aim of this study is to investigate the phytochemical and the long-term anti-neuropathic potentials of Crocus sativus cultivated in the University botanical garden, and explore its most bioactive compounds and their underlying mechanisms of action. Methods: Phytochemical analysis and bio-guided isolation-procedures including RP-HPLC and 1 H and 13 C NMR utilizing biological models of diabetes, inflammation, and diabetic-neuropathy were used. Cultivated saffron (S-RCED) and Spanish-saffron stigma (S-SP) alone or in combination with Camellia sinus (CS) were investigated. Results: The RP-HPLC analyses showed the presence of picrocrocin, crocin I, crocin II, crocin I’, crocin II’, and safranal (SAF) in both S-SP and S-RCED extracts with higher-concentrations. It had been shown that SAF was the most bioactive-compound in Crocus sativus. Both S-SP and S-RCED possessed significant (P < 0.05) anti-diabetic activities in acute (6 h), subchronic (8 d) and chronic (8 weeks) models. S-RCED had been proven with more hypoglycemic potentials when compared to S-SP and SAF. S-SP, S-RCED, and SAF produced significant anti-inflammatory and anti-nociceptive activities against carrageenan-induced inflammatory, hyperalgesic and tactile diabetic-neuropathy models, respectively. S-SP, S-RCED, and SAF elevated serum catalase, reduced glutathione, and insulin serum levels, ameliorated lipid peroxidation and HbA1c levels, and histopathologically regenerated the pancreatic beta-cells. Combinations with CS showed more significant efficacy than the single component. Conclusion: The oxidative stress reduction, insulin secretagogue, and pancreatic beta-cells regeneration potentials might be responsible for the mechanism underlying the anti-diabetic, anti-inflammatory and anti-diabetic neuropathy activities. Thus, the cultivated Crocus sativus might be clinically useful for pro- tecting against many serious-disorders.
    8  Quantification of Chinese yam processing methods based on pyrolysis characteristics and its relation to Maillard reaction
    Xiang-long Meng a b Bo Wang a Chen-zi Lv a Cong Hu c Mei-jing He a Shuo-sheng Zhang a
    2020, 12(1):56-66. DOI: https://doi.org/10.1016/j.chmed.2019.12.001
    [Abstract](839) [HTML](0) [PDF 0.00 Byte](20)
    Abstract:
    Objective: Chinese yam (Shanyao in Chinese, SY) as one of the representatives for Chinese medicines can be used as both of medicine and food with rich nutritional and medicinal value. Most of Chinese herbal medicines need to be processed prior to be used in clinical practice. SY was divided into Maoshanyao (Hairy Shanyao, MSY) and Guangshanyao (Smooth Shanyao, GSY) based on different processing meth- ods at the place of origin, and it also could be processed as stir-fried SY and bran stir-fried SY to meet the different clinical use. Moreover, during the processing of Chinese herbal medicines, more compli- cated Maillard reaction occurs compared to food processing. Therefore, the objective of this research is to quantify the firepower of SY processing, and combined this with the relevant parameters of Maillard reaction. Methods: The MSY and GSY produced in Shanxi and Henan Provinces were chosen as the research ob- jects. By using thermal analysis technology, we first established the correlation between pyrolysis and processing of SY and its mixtures. We also quantified the firepower of Shaoyao processing, and com- bined this with the relevant parameters of Maillard reaction (pH value, amino acid, and 5-HMF) and the changes in medicinal ingredients (allantoin). Results: The SY was mainly fried with moderate-fire (190 °C?200 °C), and the starting temperatures of different SY–ingredient mixtures were (176.3 ± 5.33) °C for (honey) bran, and (205.9 ± 8.05) °C for rice. The upper limits of processing temperature were (289.9 ± 6.47) °C for (honey) bran and (298.9 ± 1.15) °C for rice. The cooking time was (10.80 ± 1.76) min for soil stir-fry, (10.31 ± 1.06) min for bran stir-fry, and (8.43 ± 0.68) min for rice stir-fry. Moreover, the pH values and the content of 5-HMF were increased (P < 0.001), while the content of glycine was decreased significantly (P < 0.001) after processing. Conclusion: The results verified and quantified the firepower of traditional processing of SY, and also provided scientific reference for other studies related to SY processing.
    9  Effects of total iridoid glycosides of Picrorhiza scrophulariiflora against non-alcoholic steatohepatitis rats induced by high-fat and high-sugar diet through regulation of lipid metabolism
    Xu Xu a Wei-ting Wang b Zhuan-you Zhao b Wen-gong Xi b Bing Yu b Chun-hua Hao b Xin Li a Wen-bin Hou c Li-da Tang b
    2020, 12(1):67-72. DOI: https://doi.org/10.1016/j.chmed.2019.12.005
    [Abstract](296) [HTML](0) [PDF 0.00 Byte](22)
    Abstract:
    Objective: To investigate the therapeutic effect of total iridoid glycosides of Picrorhiza scrophulariiflora (TIGP) on non-alcoholic steatohepatitis (NASH). Methods: SD rats were fed with high-fat and high-sugar diet for 8 weeks to establish NASH. TIGP were given orally at doses of 20, 40 and 80 mg/kg/d for 4 weeks. Triglycerides assay (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), aspar- tate aminotransferase (AST), alanine aminotransferase (ALT), fasting plasma glucose (FPG), fasting insulin (FINS), tumor necrosis factor- α (TNF- α ), interleukin-6 (IL-6), chemokine-1 (MCP-1), leptin (LEP) in serum were tested. TG, TC, superoxide dismutase (SOD), malondialdehyde (MDA), and free fatty acid (FFA) in liver tissue were determined by colorimetric methods. Steatosis of hepatocytes and inflammation was performed by pathological examination. Results: The results showed that TIGP significantly decreased TC, TG and FFA in liver tissue, increased SOD activity, decreased MDA content, decreased serum levels of TG, TC, HDL-C/LDL-C, ALT, AST, GLU, HOMA-IR, TNF- α and LEP, and in addition, improved steatosis of liver cells compared to NASH. Conclusion: TIGP had anti-fatty liver effect against NASH rats induced by high-fat and high-sugar diet. Its mechanism was related to the regulation of lipid metabolism and reduction of insulin resistance, through inhibition of oxidative stress and inflammation.
    10  Attenuation of inflammation in streptozotocin-induced diabetic rabbits by Matricaria chamomilla oil: A focus on targeting NF- κ B and NLRP3 signaling pathways
    Saeid Saghahazrati a Seyed Abdulmajid Ayatollahi a b c Farzad Kobarfard a d Bagher Minaii Zang e
    2020, 12(1):73-78. DOI: https://doi.org/10.1016/j.chmed.2019.12.003
    [Abstract](381) [HTML](0) [PDF 0.00 Byte](13)
    Abstract:
    Objectives: The present study was conducted to investigate the protective effects of chamomile oil from Matricaria chamomilla against type 1 diabetes mellitus (T1DM) and its potential mechanisms. Methods: T1DM was established in male New Zealand white rabbits via a single intraperitoneal infusion of streptozotocin (STZ) (80 mg/kg body weight ?1 , dissolved in 0.2 mL of normal saline). Different doses of chamomile oil (25, 50 and 100 mg/kg) were orally administrated to STZ induced diabetic rabbits for 21 consecutive days. The expression of pro-inflammatory cytokines was determined using ELISA assay. The expression of NF- κ B and NLRP3 was measured using Western blot assay. Results: Compared with normal rabbits, STZ-induced diabetic rabbits exhibited significant increased levels of blood glucose and decreased levels of serum insulin that were reversed using middle and high tested dose of chamomile oil. Likewise, STZ-induced diabetic rabbits showed a significant increased expression of NF- κ B and NLRP3 proteins in the pancreas tissue that was reversed by high tested dose of chamomile oil. Conclusion: Collectively, our findings demonstrated that chamomile oil possesses anti-hyperglycemic, and anti-inflammatory activities in STZ-induced diabetic rabbits by targeting inflammatory cytokines and NF- κ B and NLRP3 signaling pathways.
    11  Pharmacokinetic studies of multi-bioactive components in rat plasma after oral administration of Xintiantai I extract and effects of guide drug borneol on pharmacokinetics
    Shao-yu Liang a b Yong-chang Zeng a b Qian-qian Jiang a b Jun-hong Wu a b Zheng-zhi Wu a b
    2020, 12(1):79-87. DOI: https://doi.org/10.1016/j.chmed.2019.06.003
    [Abstract](573) [HTML](0) [PDF 0.00 Byte](17)
    Abstract:
    Objective: To investigate the in vivo pharmacokinetic characteristics of 17 bioactive components includ- ing ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, dehydroevodiamine, rutaecarpine, limonin, hyperin, curcumin, demethoxycurcumin and bis- demethoxycurcumin in rat plasma after oral administration of Xintiantai I extract powder (XI) and Xin- tiantai I without guide drug borneol extract powder (XI without borneol), and study the compatibility effects of guide drug borneol on the pharmacokinetics. Methods: A UHPLC-MS/MS method was established and fully validated for the comparative pharmacoki- netics of 17 bioactive components. The pharmacokinetics parameters of 17 bioactive components after oral administration of XI and XI without borneol were calculated by the software of DAS 3.0 and inter- compared. Results: The specificity, linearity, lower limit of quantification (LLOQ), precision, accuracy, extraction re- covery rates, matrix effects, and stability of the UHPLC-MS/MS assay were good within the acceptance criteria from FDA guidelines. Guide drug borneol can significantly increase AUC of G-Rd, palmatine, hy- perin, curcumin, demethoxycurcumin, bisdemethoxycurcumin and C max of 16 bioactive components ex- cept for dehydroevodiamine (P < 0.05), decrease T max of G-Rd, berberine, columbamin, coptisine, limonin and MRT of 17 bioactive components in XI group (P < 0.05). Conclusion: Guide drug borneol enhanced the absorption of G-Rd, palmatine, hyperin, curcumin, demethoxycurcumin and bisdemethoxycurcumin.
    12  A reliable and rapid pharmacokinetic study of pueraria isoflavones using pueraria reference extractive substance in beagle plasma: Application to study of Yufeng Ningxin Tablets
    Qin Zha a b Sha Chen a Jun Zhang a Xiang-yu Zhang b Pei-jun Ruan b Heba M. Amer c An Liu a
    2020, 12(1):88-94. DOI: https://doi.org/10.1016/j.chmed.2019.11.003
    [Abstract](515) [HTML](0) [PDF 0.00 Byte](15)
    Abstract:
    Objective: In order to evaluate the reliability and feasibility of pueraria reference extractive substance (RES) used in biological sample, the pharmacokinetics of 3 ? –hydroxy puerarin (3 ? -HP), puerarin, 3 ? – methoxy puerarin (3 ? -MP), and daidzein-8-C-apiosyl-(1-6)-glucoside (DAG) in beagle plasma following oral administration of Yufeng Ningxin Tablet were quantitated. Methods: A reliable and sensitive high-performance liquid chromatography-tandem triple quadrupole mass spectrometry (HPLC-QQQ-MS/MS) method developed with chromatographic separation was oper- ated on a Merck C 18 column, and acetonitrile-5 mmol/L ammonium was used as mobile phase in gradient elution. The plasma samples were deproteinized by acetone, detected by triple quadrupole mass spec- trometry with an electrospray ionization interface, and quantified using selected ion monitoring mode. The pharmacokinetic parameters were calculated by Winnonlin 4.1. Results: The calibration curves of the reference extractive substance and standard substance methods were linear over the ranges 0.0417–11.3309 μg/mL and 0.0394–10.0000 μg/mL. The intra-day and inter- day precision of the two methods at three concentrations were less than 13.63%, and the average recov- eries of 3 ? -HP, puerarin, 3 ? -MP, and DAG were more than 70.67%. The RSD of the mean plasma concentra- tions of the analytes calculated by the two methods was less than 5%, and cos (?) = =1.000. Among the analytes, puerarin showed the highest blood concentration [(940 ± 185) ng/mL] and the longest retention time [(5 ± 1) h] in the dog’s bodies. Conclusion: Pueraria reference extractive substance can be seen as an alternative to the standard sub- stance to overcome the scarcity of standard substance for the analysis of biological samples.

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