葛根HPLC指纹图谱及模式识别研究

采用高效液相色谱法,建立9种不同产地的葛根样品的指纹图谱,并利用化学计量学中的主成分分析和聚类分析方法对所得到的葛根HPLC指纹图谱进行模式识别的研究,实现了9种不同产地葛根样品的分类.本研究获得的指纹图谱较全面的反映不同产地的葛根的特征差异,可为葛根的质量控制及综合评价奠定基础.     

UPLC-ELSD法测定黄芪中的黄芪甲苷含量

采用Waters Acquity UPLC BEH C18色谱柱（2.1mm×100mm,1.7μm）,流动相为乙腈-水（30∶70）,流速为0.2mL·min^-1,柱温30℃,用UPLC-ELSD法测定黄芪中黄芪甲苷的含量。黄芪甲苷在0.10-0.50mg·mL^-1范围内有良好的线性关系（r=0.9990）,平均回收率（n=6）为98.9%。该方法快速、准确、重复性好,为黄芪的质量控制提供了一种快速准确的检测方法。     

Determination of 12 potential nephrotoxicity biomarkers in rat serum and urine by liquid chromatography with mass spectrometry and its application to renal failure induced by Semen Strychni

In previous nephrotoxicity metabonomic studies, several potential biomarkers were found and evaluated. To investigate the relationship between the nephrotoxicity biomarkers and the therapeutic role of Radix Glycyrrhizae extract on Semen Strychni‐induced renal failure, 12 typical biomarkers are selected and a simple LC–MS method has been developed and validated. Citric acid, guanidinosuccinic acid, taurine, guanidinoacetic acid, uric acid, creatinine, hippuric acid, xanthurenic acid, kynurenic acid, 3‐indoxyl sulfate, indole‐3‐acetic acid, and phenaceturic acid were separated by a Phenomenex Luna C₁₈ column and a methanol/water (5 mM ammonium acetate) gradient program with a runtime of 20 min. The prepared calibration curves showed good linearity with regression coefficients all above 0.9913. The absolute recoveries of analytes from serum and urine were all more than 70.4%. With the developed method, analytes were successfully determined in serum and urine samples within 52 days. Results showed that guanidinosuccinic acid, guanidinoacetic acid, 3‐indoxyl sulfate, and indole‐3‐acetic acid (only in urine) were more sensitive than the conventional renal function markers in evaluating the therapeutic role of Radix Glycyrrhizae extract on Semen Strychni‐induced renal failure. The method could be further used in predicting and monitoring renal failure cause by other reasons in the following researches.     

黄连与甘草化学成分的相互作用研究之一——混浊汤剂中沉淀部分的成分研究

以黄连与甘草为配伍组成的复方是一种沉淀性配伍, 它属于仲景古方系列之一。其汤剂呈混浊状, 分别对离心分离后的溶液与沉淀物的成分进行研究。运用高效液相色谱(HPLC)、傅里叶变换红外光谱(FTIR)和飞行时间质谱(TOF-MS)的方法, 对单煎黄连、单煎甘草和混煎黄连与甘草的溶液和沉淀物的结构组成进行研究。文章着重报道沉淀物成分的研究, 同时还将混煎沉淀物和溶液的谱学行为做了比较。结果发现, 在混煎黄连与甘草的沉淀物中, 黄连的有些化学成分被保留下来, 有些成分却被抑制了;而甘草的化学成分基本都消失了;同时在混煎黄连与甘草的沉淀物中还产生了在单煎黄连与单煎甘草中所没有的一些新物质。此结果说明黄连与甘草在混煎后其沉淀物的结构组成发生了变化, 这是由于在此复方中发生了甘草与黄连化学成分的相互作用所致。根据飞行时间质谱的数据, 推测黄连与甘草发生的化学反应可能有加成反应、分解反应和缔合反应等类型。它表明甘草在此复方中起到了极为重要的作用。通过混煎沉淀物与溶液的研究结果比较还表明, 在沉淀物中可能含有比溶液中更多的化学活性成分,它为古方的药用和疗效提供了一定科学依据。     

Rapid Screening of Lipase Inhibitors from Ophiopogonis Radix Using High-Performance Thin Layer Chromatography by Two Step Gradient Elution Combined with Bioautographic Method

In this study, a high-performance thin layer chromatography (HPTLC) method by two step gradient elution with two mobile phases was developed for the simultaneous analysis of seven constituents in Ophiopogonis Radix. The chromatography was performed on silica gel 60 F₂₅₄ plate with dichloromethane-methanol-ethyl acetate-water (70:25:12:3, v/v/v/v) and dichloromethane-methanol (300:1, v/v) as the mobile phase for two step gradient elution. Then, the HPTLC profiles were observed after derivatization with 10% sulfuric acid in ethanol solution. The obtained HPTLC images were further analyzed by chemometric approaches and the samples could be clustered based on regions and/or growth years, which were two important factors affecting the constituents in Ophiopogonis Radix. Furthermore, five compounds including ophiopogonin D, ophiopojaponin C, ophiopogonin D’, ophiopogonin C’ and methylophiopogonanone B were screened as potential lipase inhibitors from Ophiopogonis Radix by the HPTLC-bioautographic method. The binding modes and interactions between the five compounds and lipase were further explored by molecular docking analysis. The developed HPTLC method could be used for quality control of Ophiopogonis Radix and screening of the potential lipase inhibitors.     

黄芪中微量元素的形态分析

按照传统煎煮法对中药黄芪中铜、锌、铁、镁、钙和铬6种元素进行提取;用微孔滤膜分离提取液中的可溶态与悬浮态;利用大孔吸附树脂柱对可溶态中的有机态与无机态进行分离;采用火焰原子吸收光谱法对各种形态中的6种元素进行测定。结果显示:黄芪中6种元素的总提取率在44.0%~74.4%,浸留比在79·4%~293.8%,悬浮态颗粒吸附率在10%左右,可溶态中铬的有机态与无机态的比例为114.6%,铜、锌、铁、镁、钙的有机态与无机态的比例在3.7%~43.5%。该法对各元素的加标回收率在96.8%~103.1%;相对标准偏差小于1.5%。     

溶剂浮选法分离富集葛根中大豆甙元的研究

采用溶剂浮选法分离富集葛根中的大豆甙元。考察了浮选溶剂、氮气流速、试液pH、浮选时间及电解质（KC1）等因素对浮选效率的影响,优选出最佳浮选条件;对最佳条件下的浮选效果进行了评价,并与溶剂萃取法进行了对照,前者明显优于后者。     

ICP-AES法测定丹参中的镉和铅

建立了利用电感耦合等离子体原子发射光谱(ICP-AES)法测定丹参中重金属元素Cd和Pb的方法,比较了干灰化法和湿化法两种样品处理方法对分析结果的影响,并测定了山东一些地区丹参中的Cd和Pb含量。结果表明：采用电感耦合等离子体原子发射光谱法检测丹参中的Cd和Pb,最低检出限(DL)分别为1.92和1.07 ng·mL-1;相对标准偏差(RSD)分别为3.14%和1.83%;回收率分别为103.05%和96.24%。利用湿法消解样品,精密度好,回收率高,优于干灰化法。干灰化法适合测定丹参中Pb,但不适合测定Cd,其回收率仅为0.1%。用湿法消解样品,测定不同产地丹参的Cd和Pb含量,结果表明：所测地区丹参的Cd, Pb含量大大低于《中华人民共和国药典》(2005)规定的中药材重金属含量标准,符合GAP生产的要求。     

Rapid Sample Pretreatment,Identification and Determination of Active Constitutents in Water‐soluble Radix isatidis Extract via MAE,ESI‐MS and RODWs‐HPLC

In this study, we successfully studied water‐soluble extract from Radix isatidis. Optimized conditions of MAE were listed, the sample can be extracted completely in 10 minutes under microwave power of 400W and solid/liquid ratio of 1:80. Active compounds in water‐soluble extract from R. isatidis were identified with HPLC‐DAD/ESI‐MS, these compounds followed by cytidine, uridine, guanosine, (R,S)‐goitrin and adenosine. RODWs–HPLC as a new sensitive chromatography were also first proposed and investigated, we favoringly used this method for simultaneous determination of these active constitutents in water‐soluble R. isatidis extract. Chromatographic separation was performed on a Diamonsil C18 column (5 μm, 150 mm × 4.6 mm) with a mobile phase gradient consisting of methanol and water at a flow‐rate of 1.0 mL/min, detection wavelengths 240, 250, 260 and 270 nm, the retention times of the tested five compounds were about 4.2, 5.8, 11.1, 14.2 and 20.8 min respectively, the limits of detection were 15, 12, 20, 5.8 and 24 ng/mL for cytidine, uridine, guanosine, (R,S)‐goitrin and adenosine respectively, their linear ranges were between 0.045 and 350 μg/mL with correlation coefficient (R) of 0.9998‐0.9999. The relative standard deviations (RSDs) of intra‐day and inter‐day assays were 0.30‐2.36% and 0.86‐2.54% respectively. Extraction recoveries were 94.25‐106.21%. This novel analytical method was shown to be simple, low‐cost, sensitive and reliable for multiple components in complex or undeveloped materials via MAE, ESI‐MS and RODWs‐HPLC.     

液质联用技术研究人参与干姜或赤芍配伍前后人参皂苷及其抗氧化活性的变化

采用高效液相色谱与电喷雾质谱联用技术(HPLC-ESI-MS),对不同质量比的人参与干姜或赤芍配伍过程中人参皂苷的变化进行了研究,发现随加入的干姜量增加,共煎液中的人参皂苷含量依次降低;少量的赤芍可以使各皂苷的溶出量增加;同时测定了人参单煎液、人参与干姜、赤芍共煎液中正丁醇提取物和水提物的抗氧化活性。 以抗坏血酸(500 μmol/L)作对照,人参与干姜、赤芍配伍溶液的抗氧化活性比人参单煎液要好,同时人参与干姜、赤芍共煎液中正丁醇提取物的FRAP(铁离子还原/抗氧化能力测定)值分别为1562.29和2969.78 μmol/L,高于人参与2种药单煎液（1260.27和2502.07 μmol/L）之和。