基于白色念珠菌体外抑菌效果的复方苦参洗剂抑菌作用评价

观察复方苦参洗剂中君、臣、佐各药组联合抑菌性质以及协同增效作用。采用加热回流水提法提取各药组获得相应提取液,采用二倍稀释法测得相应各药组单用及联合对白色念珠菌的最低抑菌浓度(ρMIC)，采用联合抑菌指数（IFIC）判定两药组联合抑菌作用性质。结果显示，8味中药水提液的ρMIC范围为7.50 ~30.00 mg/mL,君、臣、佐各药组的 ρMIC范围为4.01 ~21.00 mg/mL，复方组ρMIC为2.22 mg/mL，黄柏+苦参、黄柏+白矾、花椒+白矾的IFIC≤0.5，黄柏+花椒、苦参+花椒、苦参+白矾3组0.5＜IFIC≤1；君药组+臣药组IFIC=0.375 0，君药组+佐药组IFIC=0.750 0，臣药组+佐药组IFIC=1.000 0。复方组及君药组的联合抑菌效果明显强于单味中药的抑菌效果，黄柏、苦参、花椒、白矾4味君药对白色念珠菌抑菌活性较强，复方苦参洗剂抑菌效果的增强与各药组抑菌作用的协同和相加有关。     

Simultaneous Determination of Seven Active Compounds in Radix Salviae Miltiorrhizae by Temperature-Controlled Ultrasound-Assisted Extraction and HPLC

A simple and effective high-performance liquid chromatographic (HPLC) method has been developed for simultaneous quantification of three phenolic acids (3,4-dihydroxyphenyllactic acid (Chinese name danshensu), protocatechuic aldehyde, and salvianolic acid B) and four diterpenes (dihydrotanshinone I, cryptotanshinone, tanshinone I, and tanshinone II_A) in radix salviae miltiorrhizae. Chromatography was performed on a 250 mm × 4.6 mm i.d., 5-μm particle size, C₁₈ column. The mobile phase was a linear gradient prepared from 0.1% (v/v) aqueous formic acid and acetonitrile at a flow-rate of 1.0 mL min⁻¹. All the target components were well separated with high resolution and without interference. Good linearity (R ² > 0.999) was observed over the concentration ranges investigated, and intra-day and inter-day precision were high. Temperature-controlled ultrasound-assisted extraction was used to prevent hydrolysis of thermally unstable components during the sample-extraction procedure, and the extraction conditions were carefully optimized. Recovery of the seven components was from 98.45 to 100.63% and relative standard deviations were always <1.5%. The validated method was successfully used for simultaneous quantification of the three phenolic acids and the four diterpenes in radix salviae miltiorrhizae of different geographic origins.     

最佳抑菌效果的三种药材配比筛选

分别用不同酒精浓度的土茯苓、苦参和白鲜皮提取液对金黄色葡萄球菌和白色念球菌进行抑茵试验,筛选最佳抑茵效果的酒精浓度,再以不同配比的土茯苓、苦参和白鲜皮混合物为原料提取抑茵活性物,筛选最佳原料配比。结果表明,对所试两种茵较合适的酒精浓度是95%。复方抑茵效果明显好于单方。B组(苦参:土茯苓:白鲜皮=1:2.5:1)配方对白色念球菌抑茵效果最好,E组(苦参:土茯苓:白鲜皮=8:2.5:1)配方对金黄色葡萄球菌抑茵效果最好。     

A rapid method for simultaneous determination of 14 phenolic compounds in Radix Puerariae using microwave-assisted extraction and ultra high performance liquid chromatography coupled with diode array detection and time-of-flight mass spectrometry

A microwave-assisted extraction (MAE) and ultra high performance liquid chromatography coupled with diode array detection and time-of-flight mass spectrometry (UHPLC-DAD-TOF-MS) method was developed for simultaneous determination of 14 phenolic compounds in the root of Pueraria lobata (Wild.) Ohwi and Pueraria thomsonii Benth. Operational conditions of MAE were optimized by central composite design (CCD). The optimized result was 65% ethanol as extraction solvent, 17 mL of extraction volume, 100 °C of extraction temperature and 2 min of hold time. A Zorbax SB C₁₈ (50 mm × 4.6 mm I.D., 1.8 μm) and gradient elution were used during the analysis. The chromatographic peaks of 14 investigated compounds in samples were successfully identified by comparing their retention time, UV spectra and TOF mass data with the reference substances. All calibration curves showed good linearity (r > 0.9997) within the test ranges. The intra-day and inter-day variations were less than 1.77% and 2.88%, respectively. The developed method was successfully applied to determine the investigated compounds in 10 samples of Radix Puerariae Lobatae and Radix Puerariae Thomsonii, respectively. The result indicated that MAE and UHPLC-DAD-TOF-MS system might provide a rapid method for the quality control of Radix Puerariae.     

偏最小二乘回归分析法在黄芩抑菌谱-效关系及数学模型研究中的应用

以黄芩药材指纹图谱及其对金黄色葡萄球菌抑菌率为基础数据，采用偏最小二乘回归分析法（PLS）进行数据的谱-效相关性分析，并建立数学模型。结果表明，黄芩中有5个成分对抑菌率的影响较大，建立模型的药效预测值偏差在10%以内，能够较好地预测药效。该方法及所建立的数学模型能成为预测黄芩抑制金黄色葡萄球菌的药效作用的一种有效的手段。     

Preparative enrichment and separation of astragalosides from Radix Astragali extracts using macroporous resins

The enrichment and separation of astragalosides I–IV (AGs I–IV) were studied on eight macroporous resins in the present study. SA‐3 resin offered the best adsorption and desorption capacities for AGs I–IV than other resins. The models of adsorption kinetics were investigated in order to elucidate the mechanism of adsorption. The pseudo‐second‐order model was the better choice than the pseudo‐first‐order model to describe the adsorption behavior of AGs I–IV onto SA‐3 resin. The equilibrium experimental data were well fitted to Langmuir and Freundlich isotherms. SA‐3 resin adsorption chromatography tests were carried out to optimize the separation process of AGs I–IV from Radix Astragali extracts. With the optimum parameters for adsorption and desorption, the contents of AGs I–IV were 8.78‐, 11.60‐, 10.52‐ and 11.28‐fold increased with the recovery yields being 65.88, 90.92, 84.25 and 94.17%, respectively. The preparative enrichment and separation of AGs I–IV from Radix Astragali extracts can be easily and effectively achieved by SA‐3 resin adsorption chromatography. The developed methodology can also be referenced for the separation of other active constituents from herbal materials and manufacture of Radix Astragali products.     

泰山四叶参对小鼠肺癌的作用及机制初探

研究泰山四叶参对乌拉坦诱导的小鼠肺癌的作用及其可能的作用机理.用乌拉坦诱导小鼠建立肺癌模型,随机分组在不同时间饲喂泰山四叶参粗多糖,观察小鼠生存状况,记录其体重变化、内脏(胸腺、脾、肝脏、肺、心脏)指数和存活时间,检测巨噬细胞吞噬功能和血清、肝脏、肺中SOD活性.结果发现泰山四叶参能改善乌拉坦所致肺癌小鼠的生存质量,提高了小鼠的胸腺指数和脾脏指数,增强肝脏中SOD活性,显著延长了其存活时间.因此,泰山四叶参对乌拉坦诱导的小鼠肺癌有一定的预防和抑制作用,主要机制可能与其免疫调节、抗氧化作用有关.     

Simultaneous determination of ten flavonoids of crude and wine‐processed Radix Scutellariae aqueous extracts in rat plasma by UPLC‐ESI‐MS/MS and its application to a comparative pharmacokinetic study

Radix Scutellariae (RS) is a herbal medicine with various pharmacological activities to treat inflammation, respiratory and gastrointestinal infections, etc. In this study, a rapid, sensitive and selective UPLC‐ESI‐MS/MS method was developed for simultaneous determination of 10 flavonoids – scutellarin, scutellarein, chrysin, wogonin, baicalein, apigenin, wogonoside, oroxylin A‐7‐O‐glucuronide, oroxylin A and baicalin – from RS aqueous extracts in rat plasma with propyl paraben as internal standard (IS). Chromatographic separation was achieved on a C₁₈ column using gradient elution with the mobile phase consisting of methanol and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min. The detection was performed in multiple reaction monitoring mode using electrospray ionization in negative mode. The validated method showed good linearity over a wide concentration range (r >0.9935). The intra‐ and interday assay variabilities were <9.5% and <12.4% for all analytes, respectively. The extraction recovery ranged from 71.2 to 89.7% for each analyte and IS. This method was successfully applied to pharmacokinetic comparision after oral administration of crude and wine‐processed RS aqueous extracts. There were significant differences in some pharmacokinetic parameters of most analytes between crude and wine‐processed RS. This suggested that wine‐processing exerted effects absorption of most flavonoids. Copyright © 2014 John Wiley & Sons, Ltd.     

A practical strategy for the characterization of coumarins in Radix Glehniae by liquid chromatography coupled with triple quadrupole-linear ion trap mass spectrometry

The aim of the present study was to develop a practical method for the characterization of coumarins in Radix Glehniae by liquid chromatography–mass spectrometry (LC–MS). First, 10 coumarin standards (including two pairs of isomers) were studied, and mass spectrometry fragmentation patterns and elution time rules for the coumarins were found. Then, an extract of Radix Glehniae was analyzed by the combination of two scan modes, i.e., multiple ion monitoring-information-dependent acquisition-enhanced product ion mode (MIM-IDA-EPI) and precursor scan information-dependent acquisition-enhanced product ion mode (PREC-IDA-EPI) on a hybrid triple quadrupole-linear ion trap mass spectrometer. A total of 41 coumarins were identified on the basis of their mass spectrometry fragmentation patterns. This is the first time that these two scan modes have been combined to characterize chemical constituents in traditional Chinese medicine. This new method allowed the identification of coumarins in Radix Glehniae in trace amounts. The methodology proposed in this study could be valuable for the structural characterization of coumarins from complex natural and synthetic sources.     

Identification and determination of the saikosaponins in Radix bupleuri by accelerated solvent extraction combined with rapid‐resolution LC‐MS

A method based on accelerated solvent extraction combined with rapid‐resolution LC–MS for efficient extraction, rapid separation, online identification and accurate determination of the saikosaponins (SSs) in Radix bupleuri (RB) was developed. The RB samples were extracted by accelerated solvent extraction using 70% aqueous ethanol v/v as solvent, at a temperature of 120°C and pressure of 100 bar, with 10 min of static extraction time and three extraction cycles. Rapid‐resolution LC separation was performed by using a C₁₈ column at gradient elution of water (containing 0.5% formic acid) and acetonitrile, and the major constituents were well separated within 20 min. A TOF‐MS and an IT‐MS were used for online identification of the major constituents, and 27 SSs were identified or tentatively identified. Five major bioactive SSs (SSa, SSc, SSd, 6″‐O‐acetyl‐SSa and 6″‐O‐acetyl‐SSd) with obvious peak areas and good resolution were chosen as benchmark substances, and a triple quadrupole MS operating in multiple‐reaction monitoring mode was used for their quantitative analysis. A total of 16 RB samples from different regions of China were analyzed. The results indicated that the method was rapid, efficient, accurate and suitable for use in the quality control of RB.