Simultaneous determination of albiflorin and paeoniflorin in rat urine by solid-phase extraction and high-performance liquid chromatography following oral administration of Si-Wu decoction

A high performance liquid chromatographic method (HPLC), together with solid phase extraction (SPE), was developed for simultaneous determination of albiflorin and paeoniflorin in rat urine after oral administration of Si-Wu decoction. The samples were pretreated with solid phase extraction using Extract-Cleantrade mark cartridges. Analysis of the extract was performed on a reversed-phase C18 column and a mobile phase made up of acetonitrile and 0.03% formic acid (17:83, v/v). UV detection was set at 230 nm. The assay was linear over the range 2.625-52.50 mg/mL for albiflorin and 3.875-77.50 microg/mL for paeoniflorin. The average percentage recoveries of three spiked urines were 97.01 +/- 3.32 and 102.32 +/- 6.97 for albiflorin and paeoniflorin, respectively. The intra-day precision (RSD) ranged from 0.21 to 1.79% at concentrations of 4.20, 10.50, 26.25 and 39.375 microg/mL of albiflorin and 0.12 to 2.92% at concentrations of 3.875, 10.85, 23.25 and 58.125 microg/mL of paeoniflorin, and inter-day precision (RSD) was from 1.02 to 1.86% for albiflorin and 0.94 to 3.30% for paeoniflorin, at the same four concentrations. This method was applied in order to analyze albiflorin and paeoniflorin in rat urine following oral administration of traditional Chinese medicinal preparation of Si-Wu decoction.     

经典名方当归四逆汤物质基准的关键质量属性研究

该文建立了当归四逆汤物质基准的高效液相色谱(HPLC)特征图谱及多指标含量测定方法,阐明了当归四逆汤物质基准的关键质量属性,完善了其质量控制体系.使用的色谱柱为PomenexLuna?C18(250 mm×4.6 mm,5 μm),流动相为乙腈-0.05％磷酸水溶液,体积流量为1.0 mL·min-1,梯度洗脱,进样量...     

Homotherapy for heteropathy active components and mechanisms of Qiang-Huo-Sheng-Shi decoction for treatment of rheumatoid arthritis and osteoarthritis

Qiang-Huo-Sheng-Shi decoction (QHSSD), a classic traditional Chinese herbal formula, which has been reported to be effective in rheumatoid arthritis (RA) and osteoarthritis (OA). However, the concurrent targeting mechanism of how the aforementioned formula is valid in the two distinct diseases OA and RA, which represents the homotherapy-for-heteropathy principle in traditional Chinese medicine (TCM), have not yet been clarified. In the present study, network pharmacology was adopted to analyze the potential molecular mechanism, and therapeutic effective components of QHSSD on both OA and RA. A total of 153 active ingredients in QHSSD were identified, 142 of which associated with 59 potential targets for the two diseases were identified. By constructing the protein-protein interaction network and the compound-target-disease network, 72 compounds and 10 proteins were obtained as the hub targets of QHSSD against OA and RA. The hub genes of ESR1, PTGS2, PPARG, IL1B, TNF, MMP2, IL6, CYP3A4, MAPK8, and ALB were mainly involved in osteoclast differentiation, the NF-κB and TNF signaling pathways. Moreover, molecular docking results showed that the screened active compounds had a high affinity for the hub genes. This study provides new insight into the molecular mechanisms behind how QHSSD presents homotherapy-for-heteropathy therapeutic efficacy in both OA and RA. For the first time, a two-disease model was linked with a TCM formula using network pharmacology to identify the key active components and understand the common mechanisms of its multi-pathway regulation. This study will inspire more innovative and important studies on the modern research of TCM formulas.     

中药四物汤提取物调控鼠骨髓间充质干细胞增殖活性的化学成分分析

用石油醚、乙酸乙酯、氯仿和二次蒸馏水分别提取四物汤药剂,得到不同极性部位的溶剂提取物。促进骨髓间充质干细胞(rMSCs)增殖活性用四甲基偶氮唑盐(MTT)法,结果表明,四物汤乙酸乙酯提取部位(A-2)具有促进MSCs增殖的活性。用硅胶柱层析的方式对A-2作梯度洗脱得到20个组分,记为F-1~F-20。经MTT法和流式细胞技术评价,选择具有促进MSCs增殖的活性的F-4、F-7、F-10和F-11组分。采用HPLC、红外光谱和质谱检测发现,组分F-4为藁本内酯;用HPLC-MSn发现,F-11中含量达84.47%的成分为6,7-二羟基-烯丙基苯酞。HPLC结果显示,F-7和F-10有较大含量的脂肪酸酯类化合物。选用4个酯类标准品,先用MTT法和流式细胞技术对其评价;结果显示浓度为90mg/L时,十六酸甲酯(S-1)和十八酸乙酯(S-4)的增殖指数(PI)均高于对照组,具有促进MSCs体外增殖作用;而十六酸乙酯(S-2)和十八酸甲酯(S-3)增殖指数(PI)均低于对照组,具有抑制MSCs体外增殖作用。对F-7鉴定中检出含有S-1(3.04%),S-2(3.92%),S-4(9.61%),在F-10中检出S-1(14.46%),S-2(17.47%),S-3(3.19%),S-4(1.03%)。生物活性显示F-7活性强于F-10。初步推测在酯类混合物中,对MSCs起促进作用主要为S-1和S-4。结果表明,四物汤乙酸乙酯提取部位具有促进MSCs增殖作用的成分是藁本内酯、十六酸甲酯和十八酸乙酯。     

高效液相色谱-质谱法鉴定中药复方小续命汤有效成分组中醇溶性成分

在对中药复方小续命汤进行了活性研究之后,建立了适宜的高效液相色谱-电喷雾电离-串联质谱法(HPLC-ESI-MS/MS)对小续命汤抗脑缺血有效成分组中16个主要的醇溶性成分进行鉴定。这些成分分别属于生物碱类、黄酮类和苷类。样品采用MS和MS/MS全扫描两种模式进行分析。结果显示:在电喷雾离子化过程中生物碱类成分易于质子化而成为正离子,黄酮与苷类成分则易于失去质子成为负离子。因而运用了两种流动相系统以得到这些化合物的适宜离子进行检测。对于中药复方小续命汤中有效成分进行鉴定,能为更好地阐明中药复方的有效成分及作用机制提供依据。     

Comparative pharmacokinetics of three monoester‐diterpenoid alkaloids after oral administration of Acontium carmichaeli extract and its compatibility with other herbal medicines in Sini Decoction to rats

Sini decoction (SND) is an important traditional Chinese multiherbal formula, which is widely used to treat cardiovascular disease. Acontium carmichaeli (AC) is a leading herb in SND, whose main components are monoester‐diterpenoid alkaloids (MDAs). The aim of this study is to compare the pharmacokinetics of three MDAs in rat plasma after oral administration of AC extract and its compatibility with other herbal medicines in SND. A sensitive, accurate and specific LC‐MS/MS method was developed to determine the contents of three MDAs in rat plasma. Male Sprague–Dawley rats were randomly assigned to four groups: AC, AC + ZO, AC + GU and SND groups. There were significant differences in the pharmacokinetic parameters (C_max, T_max, t_1/2, AUC_(0–24), MRT and CL). Compared with the AC group, C_max, AUC_(0–24) and CL of three MDAs increased and t_1/2 decreased in AC + ZO, AC + GU and SND groups. Little changed in the AC + GU group in comparison with AC + ZO group, which indicated that other ingredients in ZO may promote the absorption rate and accelerate excretion rate of MDAs. The results could be helpful for revealing the compatibility mechanism of Chinese multiherbal medicine and providing clinical medication guidance on AC and its compatibility with other herbal medicines in SND. Copyright © 2014 John Wiley & Sons, Ltd.     

Rapid identification of the quality decoction pieces by partial least squares ‐based pattern recognition: grade classification of the decoction pieces of Saposhnikovia divaricata

Herbal medicines are commonly used in many countries after they undergo processing. Quality decoction pieces are a guarantee of the efficacy and safety of the herbal medical products. Here, a strategy based on chemical analysis combined with chemometric techniques was proposed for the classification and prediction of the different grades of the decoction pieces. Considering the necessity for a shared and simple method for the grade classification for the public, in this paper, the characterization of the chemical constituents was determined by utilizing high‐performance liquid chromatography (HPLC)/diode array detection. HPLC was first established for the characterization of the chemical constituents of the different grade decoction pieces. Furthermore, a simultaneous quantification of several of the marker compounds in these decoction pieces was obtained. Finally, a partial least squares‐based pattern recognition method was utilized to obtain a predictive model for the grade classification of the decoction pieces. Saposhnikovia divaricata (Turcz.) Schischk was used as a case study. The partial least squares ‐based pattern recognition for the grade classification of the decoction pieces of S. divaricata demonstrated good sensitivity, specificity and prediction performance, which may efficiently validate the identification results of appearance assessment. The proposed strategy is expected to provide a new insight for the grade classification and quality control of the decoction pieces. Copyright © 2016 John Wiley & Sons, Ltd.     

Qualitative and quantitative analysis of the chemical constituents in Mahuang‐Fuzi‐Xixin decoction based on high performance liquid chromatography combined with time‐of‐flight mass spectrometry and triple quadrupole mass spectrometers

High‐performance liquid chromatography coupled with time‐of‐flight mass spectrometry (HPLC‐TOF/MS) and high‐performance liquid chromatography–triple quadrupole mass spectrometry (HPLC‐QQQ/MS/MS) were utilized to clarify the chemical constituents of Mahuang‐Fuzi‐Xixin Decoction. There are 52 compounds, including alkaloids, amino acids and organic acids were identified or tentatively characterized by their characteristic high resolution mass data by HPLC‐QQQ/MS/MS. In the subsequent quantitative analysis, 10 constituents, including methyl ephedrine, aconine, songrine, fuziline, neoline, talatisamine, chasmanine, benzoylmesaconine, benzoylaconine and benzoylhypaconine were simultaneously determined by HPLC‐QQQ/MS/MS with multiple reaction monitoring mode. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.9992). The relative standard deviations (RSD) of inter‐ and intra‐day precisions were <3%. This method was also validated by repeatability, stability and recovery with RSD <3% respectively. A highly sensitive and efficient method was established for chemical constituents studying, including identification and quantification of Mahuang‐Fuzi‐Xixin decoction.     

Determination and pharmacokinetics of geniposidic acid in rat plasma after oral administration of Gardenia jasminoides fruit crude extract and Zhi‐zi‐chi decoction

A simple and efficient liquid chromatography–mass spectrometry method was developed and validated for the determination of geniposidic acid in rat plasma. After the addition of internal standard salidroside and acidification (0.1% formic acid, pH = 3.2), plasma samples were carried out by protein precipitation with acetonitrile and separated on a Kromasil C₁₈ column (150 × 4.6 mm, 5 µm) within a run time of 9.0 min. Analysis was performed in selected ion monitoring mode with a positive electrospray ionization interface. No endogenous interference was observed at retention times of the analytes because of the high specificity of selected ion monitoring mode. The linear range was 0.02–4.0 µg/mL and the lower limit of quantification was 0.02 µg/mL. The mean extraction recoveries of geniposidic acid and internal standard from rat plasma were all >88.0% and the matrix effects were within acceptance criteria (90–110%). The validated method was successfully applied to the pharmacokinetic study of geniposidic acid in rat plasma after oral administration of G. jasminoides fruit crude extract and Zhi‐zi‐chi decoction, respectively. Copyright © 2013 John Wiley & Sons, Ltd.     

葛根芩连汤联合西药治疗小儿毛细支气管炎的疗效研究

目的研究葛根芩连汤联合西药治疗小儿毛细支气管炎的疗效。方法将2012年3月—2014年5月期间住院治疗的120例毛细支气管炎患儿纳入研究,根据治疗方式不同随机分为两组,观察组患者接受葛根芩连汤联合西药治疗,对照组患者接受常规西药治疗,比较两组患者的临床症状和体征缓解时间、血氧饱和度以及免疫功能。结果 (1)症状和体征缓解时间:观察组患儿的咳嗽、喘憋缓解时间以及两肺啰音消失时间分别为(3.35±0.42)、(1.75±0.23)、(3.58±0.45)d,均短于对照组的(4.95±0.62)、(2.84±0.40)、(5.91±0.71)d;(2)血氧饱和度:治疗后1、2、3 d时,观察组患儿的血氧饱和度分别为(95.33±12.24)%、(97.58±11.94)%、(99.13±12.85)%,高于对照组的(91.98±10.85)%、(93.54±12.38)%、(95.16±14.23)%;(3)免疫功能:观察组患儿的血清Ig G(6.25±0.82)、Ig M(0.82±0.10)、Ig A(0.42±0.05)g/L,高于对照组的(4.34±0.65)、(0.58±0.06)、(0.29±0.03)g/L。结论葛根芩连汤联合西药治疗有助于促进临床症状和体征缓解、提高血氧饱和度、增强免疫功能,是治疗小儿毛细支气管炎的理想方法。