藏药婆婆纳药材薄层鉴别方法学研究

本文建立了藏药婆婆纳药材的薄层鉴别方法。采用单因素试验方法,对影响婆婆纳对照药材的薄层色谱因素进行系统考察,筛选出最佳的薄层色谱条件。该薄层鉴别条件吸附剂:硅胶G薄层板;展开剂:环己烷-丙酮(6∶4);显色剂:10%硫酸乙醇溶液;显色条件:110℃烘箱中加热至斑点显色清晰,日光下检视,所得的薄层色谱,斑点清晰,分离度好。该方法分离度高,重现性好,简便,可用于婆婆纳的质量控制。     

红梅汤薄层色谱鉴别研究

目的建立红梅汤制剂主要药材红梅叶、锡叶藤的鉴别方法。方法采用薄层色谱法对红梅汤制剂中的红梅叶、锡叶藤进行鉴别。结果在确定的条件下,红梅汤中红梅叶、锡叶藤药材的薄层色谱分离效果良好,斑点清晰,重现性好。结论该方法简便可靠,分离度较好,可用于红梅汤的质量控制。     

藏药翼首草薄层鉴别方法研究

本文对藏药翼首草薄层鉴别方法进行系统考察。试验以熊果酸为对照品,采用单因素试验方法,对影响翼首草薄层色谱的因素进行了考察,筛选最佳薄层鉴别方法。结果采用硅胶G板,薄层板置展开缸中饱和10min,以10%硫酸乙醇溶液为显色剂,105℃加热至斑点清晰,置日光和紫外光灯(365 nm)下检视,所得的薄层色谱分离效果较好。研究结果表明本法分离效果好、重复性好,可用于藏药翼首草的质量控制。     

复方新会陈皮含片的鉴别研究

目的建立复方新会陈皮含片的鉴别方法。方法采用薄层色谱与化学鉴别法。结果在选定的条件下,薄层色谱法鉴别陈皮、蛇胆汁、甘草、桔梗的斑点清晰,且阴性对照无干扰;化学方法鉴别氯化钠,反应结果明显。结论建立的鉴别方法结果可靠,且操作简便,可用于复方新会陈皮含片的质量控制。     

不同制剂中相同中草药薄层色谱鉴别探讨

目的探讨不同制剂中相同中药材统一鉴别的可行性。方法采用薄层色谱法,取不同制剂适量,按统一方法制备供试液、层析条件、显色与检视斑点。结果多种含大黄、人参的制剂,薄层色谱斑点清晰明显。结论本实验方法可行,在不同制剂中相同药材鉴别有较统一的方法为好。     

健胃益肠宁与滑肠通便片的鉴别探讨

目的探讨健胃益肠宁与滑肠通便片的鉴别方法。方法采用薄层色谱法和显微鉴别法进行鉴别。结果采用显微鉴别法,滑肠通便片大黄征明显;采用薄层鉴别法,健胃益肠宁检出党参、甘草,滑肠通便片检出决明子,斑点清晰,无干扰。结论两个制剂的鉴别方法简便可行,专属性强,重现性好,可作为该制剂的质量控制方法。     

治糜灵凝胶剂的薄层色谱鉴别研究

目的:将治糜灵栓剂(《中国药典》2010版一部)改为治糜灵凝胶剂,建立治糜灵凝胶剂的薄层色谱鉴别方法,为制定其质量控制标准制定依据。方法:参照《中国药典》2010版一部治糜灵栓剂项下黄柏、苦参、儿茶、冰片的薄层色谱鉴别方法,对处方中的主要药味进行定性鉴别。黄柏鉴别中以甲苯-乙酸乙酯-异丙醇-水(6:3:1.5:1.5)为展开剂;苦参鉴别中以甲苯-乙酸乙酯-甲醇(8:3:0.5)为展开剂;儿茶鉴别中以正丁醇-醋酸-水(3:2:1)为展开剂;冰片鉴别中以环己烷-三氯甲烷-乙酸乙酯(9:1:2)为展开剂。结果:薄层色谱上具有黄柏,苦参,儿茶,冰片的鉴别特征,且阴性对照无干扰。结论:色谱斑点清晰,专属性强;该方法操作简便,稳定性、重现性均很好。可作为质量标准的控制依据。证明治糜灵凝胶剂研究的方法可行。     

妇科洗剂的质量控制及HPLC指纹图谱研究

本研究旨在建立妇科洗剂的HPLC指纹图谱分析及其质量控制方法。薄层色谱鉴别法对妇科洗剂中苦参、黄柏、蛇床子进行定性鉴别。应用高效液相色谱-二极管阵列检测技术(HPLC-DAD)进行检测,采用Wondasil C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.1%磷酸水溶液(50∶50)(每100 m L加十二烷基磺酸钠0.2g)为流动相,检测波长265 nm,柱温30℃,测定盐酸小檗碱的含量;以乙腈-水(0.3%磷酸和0.3%二乙胺)为流动相梯度洗脱,检测波长284 nm,柱温30℃,建立妇科洗剂的HPLC指纹图谱。结果表明薄层色谱中斑点清晰,阴性无干扰。盐酸小檗碱在132.40~662.00 ng范围内与峰面积有良好的线性关系(r=0.9999),平均加样回收率为100.97%。10批妇科洗剂的指纹图谱分离效果好,确定15个共有峰,相似度均0.9。研究表明,上述实验方法快速、准确,操作简单,专属性强,建立的指纹图谱可为妇科洗剂的质量评价提供依据,能较好地控制该制剂的质量。     

二十味沉香丸质量控制标准优化

本研究通过建立藏药二十味沉香丸的新型质量控制标准体系,为生产企业质量控制与临床安全使用提供参考。用单因素实验优化没食子酸提取工艺,以高效液相色谱法对其进行定量检测并以薄层色谱法对制剂没食子酸成分进行定性分析。结果显示,薄层色谱所得斑点清晰,分离度好;提取工艺和HPLC色谱条件优化后,在0.1～0.5 mg/mL浓度区间内,没食子酸浓度与HPLC峰面积值呈现y=5 979 576x+27 982的稳定线性关系,r~2值为0.999;方法学实验误差在允许范围内,没食子酸成分在24 h内无明显变化。此方法作为二十味沉香丸质量控制标准,专属性强,定量准确。     

和中消滞合剂中总槲皮素的定性定量研究

目的建立和中消滞合剂中总槲皮素的定性定量方法。方法采用薄层色谱法定性,高效液相色谱法(HPLC)定量。色谱柱为Inertsil ODS-XP柱(4.6 mm×150 mm,5μm),流动相为甲醇-0.4%磷酸溶液(36∶64),检测波长360 nm,流速为1 ml·min~(-1),柱温为30℃。结果薄层色谱斑点清晰;HPLC的槲皮素进样浓度在0.039～0.413μg·ml~(-1)范围内,与峰面积值呈良好的线性关系,回归方程Y=46589X-4756(r=0.9999);平均加样回收率为98.10%,RSD=0.96%(n=6)。结论建立和中消滞合剂中总槲皮素的定性定量测定方法可行,可用于该制剂的质量控制。     

银耳多糖单糖组成分析的三种色谱方法比较

为选择一种准确快捷的方法测定银耳多糖的单糖组成,对薄层色谱法(TLC)、气相色谱法(GC)、高效液相色谱法(HPLC)三种色谱方法进行比较。结果表明,前两种方法的测定结果均不理想,而HPLC法,操作简便,灵敏度高,分离效果好,信息完整。测定结果为由葡萄糖、甘露糖、葡萄糖醛酸、木糖、岩藻糖组成,其摩尔比为0.24∶1.00∶0.06∶0.29∶0.25。HPLC法对酸性杂多糖组成糖分析是一种比较理想的选择。     

Quantitative analysis of major dibenzocyclooctane lignans in Schisandrae fructus by online TLC-DART-MS

Introduction – Direct analysis in real time (DART) ion source is a powerful ionising technique for the quick and easy detection of various organic molecules without any sample preparation steps, but the lack of quantitation capacity limits its extensive use in the field of phytochemical analysis. Objective – To improvise a new system which utilize DART‐MS as a hyphenated detector for quantitation. Methodology – A total extract of Schisandra chinensis fruit was analyzed on a TLC plate and three major lignan compounds were quantitated by three different methods of UV densitometry, TLC‐DART‐MS and HPLC‐UV to compare the efficiency of each method. To introduce the TLC plate into the DART ion source at a constant velocity, a syringe pump was employed. The DART‐MS total ion current chromatogram was recorded for the entire TLC plate. The concentration of each lignan compound was calculated from the calibration curve established with standard compound. Results – Gomisin A, gomisin N and schisandrin were well separated on a silica‐coated TLC plate and the specific ion current chromatograms were successfully acquired from the TLC‐DART‐MS system. The TLC‐DART‐MS system for the quantitation of natural products showed better linearity and specificity than TLC densitometry, and consumed less time and solvent than conventional HPLC method. Conclusion – A hyphenated system for the quantitation of phytochemicals from crude herbal drugs was successfully established. This system was shown to have a powerful analytical capacity for the prompt and efficient quantitation of natural products from crude drugs. Copyright © 2010 John Wiley & Sons, Ltd.     

Synthesis of a Precursor Dipeptide of Thymopentin in Organic Solvents by an Enzymatic Method

Abstract

The protease‐catalyzed, kinetically controlled synthesis of a precursor dipeptide of thymopentin(TP‐5), Z‐Arg‐Lys‐NH₂ in organic solvents was studied. Z‐Arg‐OMe was used as the acyl donor and Lys‐NH₂ was used as the nucleophile. An industrial alkaline protease alcalase and trypsin were used to catalyze the synthesis of the target dipeptide in water‐organic cosolvent systems. The conditions of the synthesis reaction were optimized by examining the effects of several factors, including organic solvents, water content, temperature, pH, and reaction time on the yield of Z‐Arg‐Lys‐NH₂. The optimum conditions using alcalase as the catalyst are pH 10.0, 35°C, in acetonitrile/DMF/Na₂CO₃‐NaHCO₃ buffer system (80∶10∶10, V/V), 6 h, with the dipeptide yield of 71.1%. Compared with alcalase, the optimum conditions for trypsin are pH 8.0, 35°C, in ethanol/Tris‐HCl buffer system (80∶20, V/V), 4 h, with the dipeptide yield of 76.1%.     

川楝子中香草酸含量测定研究

本实验采用薄层色谱法对川楝子中香草酸、异香草酸进行定性鉴别;采用高效液相色谱法对川楝子中香草酸进行含量测定。色谱柱:Hypersil BDS C18(250 mm×4.60 mm,5μm),柱温:30℃,流动相:甲醇∶水∶冰乙酸=25∶75∶0.5,流速:1.0 mL.min-1,检测波长:260 nm。最终,定性鉴别斑点清晰。结果表明香草酸含量在1.022～16.352μg范围内,进样量与峰面积呈良好线性关系,相关系数r=0.9998;香草酸的平均回收率为102.45%,RSD=1.22%(n=6)。     

鲟鱼鱼肠抗氧化肽的提取及抗氧化性

探讨酶法制备具有抗氧化活性的鲟鱼鱼肠抗氧化肽的方法,并进行体外抗氧化活性的测定。结果表明,比较4种蛋白酶酶解产物的抗氧化能力,确定胃蛋白酶为制备鲟鱼鱼肠抗氧化肽的最佳水解用酶;通过单因素试验和正交实验分析得出最适酶解工艺是:胃蛋白酶加酶量3 200 U/g,酶解时间1.5 h,料液比1∶20,温度35℃。其体外抗氧化能力随肽质量浓度增大而增大,在浓度为1.5 mg/mL时,鲟鱼鱼肠抗氧化肽清除DPPH·能力达到Vc的83.64%,对·OH清除率为78.06%,还原力大小约为Vc溶液的1/3。胃蛋白酶酶解鲟鱼鱼肠制备的抗氧化肽具有较好的抗氧化活性,其作为一种潜在的商业抗氧化剂具有良好的应用前景。     

Dielectric Barrier Discharge Ionization in Characterization of Organic Compounds Separated on Thin-Layer Chromatography Plates

A new method for on-spot detection and characterization of organic compounds resolved on thin layer chromatography (TLC) plates has been proposed. This method combines TLC with dielectric barrier discharge ionization (DBDI), which produces stable low-temperature plasma. At first, the compounds were separated on TLC plates and then their mass spectra were directly obtained with no additional sample preparation. To obtain good quality spectra the center of a particular TLC spot was heated from the bottom to increase volatility of the compound. MS/MS analyses were also performed to additionally characterize all analytes. The detection limit of proposed method was estimated to be 100 ng/spot of compound.     

Isolation and identification of nine sulfated glycosphingolipids containing two unique sulfated gangliosides from the African green monkey kidney cells, Verots S3, and their possible metabolic pathways

Verots S3 cells derived from the African green monkey kidney were revealed to contain nine types of sulfoglycolipids by incorporating [35S]sulfate. These sulfated glycolipids were separated by DEAE-Sephadex column chromatography and preparative thin-layer chromatography (TLC). The major sulfoglycolipids were characterized using TLC, gas-liquid chromatography (GLC), mass spectrometry, solvolysis, TLC immunostaining, and nuclear magnetic resonance spectra as follows: V1, SM4s (GalCer I3-sulfate); V2, SM3 (LacCer II3-sulfate); V3, SM2a (Gg3Cer II3-sulfate); V4, globopentaosyl ceramide sulfate (Gb5Cer V3-sulfate); V5, (Gg4Cer II3-sulfate, IV3-NeuAc); V6, SB1a (Gg4Cer II3, IV3-bis-sulfate); and V8, (Gg4Cer II3-NeuAc, IV3-sulfate). Both V5 and V8 were sulfated gangliosides comprising both N-acetyl neuraminic acid and sulfate, and this was the first report on V8. A minor component V7 was identified as SM1a (Gg4Cer II3-sulfate) based on its behavior in TLC, GLC, and liquid secondary ion mass spectroscopy. It was postulated that this substance was a precursor of V6 (SB1a) and V5 (Gg4Cer II3-sulfate, IV3-NeuAc), and to date, its presence has not been demonstrated in nature. Another minor component V9 was identified as glucosyl ceramide sulfate based on its migration in TLC and GLC. This renal cell line was shown to be an excellent model for studying the metabolism and function of sulfoglycolipids.     

固相萃取-反相高效液相色谱法分析粗裂地钱中的黄酮类化合物

建立了粗裂地钱中四种黄酮类化合物的固相萃取高效液相色谱分析方法。样品经95%乙醇溶液提取后,用C18固相萃取小柱预处理,采用甲醇∶乙腈∶0.5%乙酸(150∶100∶150,V/V)为流动相,C18色谱柱(150 mm×4.6 mm,5μm)进行分离与测定。低、中、高浓度下方法的回收率为89.70%～97.95%,RSD小于6%。结果表明,该法简便快速,节省溶剂,分析结果准确可靠,重复性好,可用于粗裂地钱中四种黄酮类化合物的含量分析。     

叶黄素循环组分的分离鉴定及紫黄质的大量制备

以薄层层析法(TLC)分离鉴定植物组织中叶黄素循环组分,建立了大量制备紫黄质的方法。此法所需时间短,分离效果好,检出灵敏度高,既适于少量样品的分离,也适于大样品的提纯制备,因此适合用于紫黄质的提取及大量制备。