PAD在HPLC分析长春胺中的应用

本文报道用光电二极管阵列检测器(PAD)鉴定小蔓和大蔓长春花中的长春胺。并对经典制备方法得到的长春胺样品进行了纯度判断。利用建立的紫外光谱数据库对长春胺中的分解产物进行光谱检索和鉴定。测定了长春胺的稳定性。     

高效液相色谱-光电二极管阵列检测器联用检测不同产地荆芥中乌索酸和齐墩果酸

建立反相高效液相色谱-光电二极管联用法同时测定了荆芥中乌索酸和齐墩果酸的含量。色谱柱为Kromasil C18柱(250mm×4.6mm,5μm),流动相为甲醇-水(86∶14),流速0.8mL.min-1,光电二极管阵列检测器(PAD),检测波长210nm,柱温25℃。在选定色谱条件下线性范围良好,乌索酸和齐墩果酸的样品加样回收率分别为98.6%和99.2%,RSD为1.1%和1.9%。方法简便,分离效果好,可用于荆芥中乌索酸和齐墩果酸含量的测定。     

高效液相梯度洗脱法测定杏香兔耳风中绿原酸和芦丁

建立了反相高效液相色谱法同时测定不同来源的杏香兔耳风中的绿原酸和芦丁。采用Kromasil C18柱(4.6 mm i.d.×250 mm,5μm),以甲醇-0.5%H3PO4的水溶液为流动相,梯度洗脱程序为0 min(30∶70)-15 min(30∶70)-20min(50∶50)-40 min(50∶50);流速0.8 mL/min;检测波长350 nm;柱温25℃;光电二极管阵列检测器(PAD)。绿原酸线性范围为0.654~5.886μg,r=0.9997,样品的平均加标回收率为98.6%,RSD 1.7%;芦丁线性范围为0.218~1.962μg,r=0.9993,样品的平均加标回收率为97.1%,RSD 2.3%。本法可作为杏香兔耳风中绿原酸和芦丁的定量分析方法。     

Separation and quantification of inulin in selected artichoke (Cynara scolymus L.) cultivars and dandelion (Taraxacum officinale WEB. ex WIGG.) roots by high-performance anion exchange chromatography with pulsed amperometric detection

The profile of fructooligosaccharides and fructopolysaccharides in artichoke heads and dandelion roots was investigated. For this purpose, a suitable method for high-performance anion exchange chromatography with pulsed amperometic detection was developed. The separation of monomers, oligomers and polymers up to a chain length of 79 sugar residues was achieved in one single run. Glucose, fructose, sucrose and individual fructooligosaccharides (kestose, nystose, fructofuranosylnystose) were quantified in six different artichoke cultivars and in dandelion roots. The contents ranged from 12.9 g/kg DM to 71.7 g/kg DM for glucose, from 15.8 g/kg DM to 67.2 g/kg DM for fructose, and from 16.8 g/kg DM to 55.2 g/kg DM for sucrose in the artichoke heads. Kestose was the predominant fructooligosaccharide, followed by nystose and fructofuranosylnystose. In four cultivars fructofuranosylnystose was only detectable in traces and reached its maximum value of 3.6 g/kg DM in the cultivar Le Castel. Furthermore, an average degree of polymerization of 5.3 to 16.7 was calculated for the individual artichoke cultivars, which is noticeably lower than hitherto reported. In contrast, the contents of kestose, nystose and fructofuranosylnystose in dandelion root exceeded that of artichoke, reflecting the short chain characteristic of the inulin, which was confirmed by chromatographic analysis.     

高效阴离子交换色谱-脉冲安培检测法测定乳粉中的低聚果糖

建立了测定乳粉中低聚果糖的高效阴离子交换色谱-脉冲安培检测(HPAEC-PAD)方法。样品经50%(v/v)乙醇水溶液提取,On Guard RP柱除脂,CarboPac PA200阴离子交换色谱柱(250 mm×3 mm)分离。采用水、0.2 mol/L NaOH溶液和0.4 mol/L NaAc溶液三元梯度淋洗,流速为0.4 mL/min,柱温为30 ℃,进样量为25 μL,脉冲安培检测器检测。蔗果三糖、四糖、五糖在0.05~10 mg/L范围内线性关系良好,相关系数(r²)> 0.9993;定量限分别为0.02、0.005和0.02 mg/L。在0.5、1.0、5.0 mg/L添加水平下,蔗果三糖、四糖、五糖的平均回收率为86.0%~114.0%。该方法处理简单、结果准确、灵敏度高,适用于乳粉中低聚果糖的测定。     

Development and validation of a HPAE-PAD method for the quantification of CGP69669A, a sialyl Lewis(x) mimetic, in skin permeation studies

A simple, rapid, precise and specific isocratic HPAE‐PAD method for quantification of CGP69669A was developed and validated. CGP69669A is a glycomimetic of sialyl Lewis^x and an antagonist of E‐selectin with potential application in the treatment of inflammatory skin disease. Quantification was performed using a Dionex CarboPac^TM PA‐200 anion‐exchange column (3 × 250 mm) with 100 mm NaOH solution as mobile phase, a flow rate of 0.50 mL/min and an injection volume of 10 μL. A quadruple potential waveform was used to detect the carbohydrate (+0.1 V from 0.00 to 0.40 s, ?2.0 V from 0.41 to 0.42 s, +0.6 V at 0.43 s and ?0.1 V from 0.44 to 0.50 s with current integrated between 0.20 and 0.40 s for detection) and rafinose was employed as an internal standard. The optimized conditions enabled rapid elution of CGP69669A (at 3.0 min) without interference from solvent peaks or substances present in the skin. The method showed good intra‐ and inter‐day precision and accuracy and the response was linear from 1.0 to 25 µg/mL. This is the first validated direct method for the quantification of CGP69669A. It will now be employed in studies investigating the topical and transdermal delivery of CGP69669A in vitro and in vivo and it should also be of use for other applications of this molecule. Copyright © 2011 John Wiley & Sons, Ltd.     

Sensitive high-performance anion-exchange chromatographic determination of paeoniflorin and albiflorin by pulsed amperometric detection after solid-phase extraction

We developed a method to simultaneously determine paeoniflorin and albiflorin levels using high-performance anion-exchange liquid chromatography with pulsed amperometric detection (HPAEC-PAD). The main principle of our method includes solid-phase extraction step using Amberlite XAD-2 sorbent to remove sugars and to selectively determine glycosides by PAD. Under these conditions, the linear dynamic range was 0.01–100 μg/mL, and the albiflorin and paeoniflorin detection limits (S/N = 3) were 5 and 10 pg, respectively. The intra- and inter-day precisions (RSDs) were <5.07%, and the average recoveries from Paeoniae Radix and Si-ni-san ranged from 97.12 to 101.15%. Our method showed high selectivity, high sensitivity, and good repeatability for analyzing albiflorin and paeoniflorin in oriental medicinal preparation.     

BEPCⅡ直线加速器相控系统PAD的研制

相位和幅度测量是BEPCⅡ直线加速器相控系统中最重要的部分之一. 基于模拟和 数字I/Q解调器, 构建了两种PAD, 并分别在实验室和速调管长廊进行了测试. 基于LMS方法的校准算法用于补偿模拟I/Q解调器的各种不匹配误差. 实验 证明, 两种PAD都能正常稳定地运行, 可以满足相控系统对相位和幅度测量的要求.     

高效阴离子交换色谱-脉冲安培检测法分析啤酒和麦汁中的糖

建立了高效阴离子交换色谱-脉冲安培检测(HPAEC-PAD)同时测定单糖、二糖和多种低聚糖的方法。采用水、0.25 mmol/L NaOH溶液和1 mol/L NaAc溶液三元梯度淋洗,在CarboPac PA-100色谱柱上,11种糖在40 min内达到良好分离;采用积分脉冲安培检测方法,无需对样品进行复杂的前处理或衍生反应便可直接检测。11种糖的检出限(S/N=3)在13～88 pg范围内。将该方法用于啤酒和麦汁样品中单糖、二糖及低聚糖的分析取得了很好的结果,样品中的加标回收率为81%～107%。     

Discovery of new inhibitor for the protein arginine deiminase type 4 (PAD4) by rational design of α-enolase-derived peptides

Rheumatoid arthritis (RA) is an inflammatory autoimmune disease affecting about 0.24 % of the world population. Protein arginine deiminase type 4 (PAD4) is believed to be responsible for the occurrence of RA by catalyzing citrullination of proteins. The citrullinated proteins act as autoantigens by stimulating an immune response. Citrullinated α-enolase has been identified as one of the autoantigens for RA. Hence, α-enolase serves as a suitable template for design of potential peptide inhibitors against PAD4. The binding affinity of α-enolase-derived peptides and PAD4 was virtually determined using PatchDock and HADDOCK docking programs. Synthesis of the designed peptides was performed using a solid phase peptide synthesis method. The inhibitory potential of each peptide was determined experimentally by PAD4 inhibition assay and IC₅₀ measurement. PAD4 assay data show that the N-P2 peptide is the most favourable substrate among all peptides. Further modification of N-P2 by changing the Arg residue to canavanine [P2 (Cav)] rendered it an inhibitor against PAD4 by reducing the PAD4 activity to 35 % with IC₅₀ 1.39 mM. We conclude that P2 (Cav) is a potential inhibitor against PAD4 and can serve as a starting point for the development of even more potent inhibitors.