Metformin Inhibits the Urea Cycle and Reduces Putrescine Generation in Colorectal Cancer Cell Lines

The urea cycle (UC) removes the excess nitrogen and ammonia generated by nitrogen-containing compound composites or protein breakdown in the human body. Research has shown that changes in UC enzymes are not only related to tumorigenesis and tumor development but also associated with poor survival in hepatocellular, breast, and colorectal cancers (CRC), etc. Cytoplasmic ornithine, the intermediate product of the urea cycle, is a specific substrate for ornithine decarboxylase (ODC, also known as ODC1) for the production of putrescine and is required for tumor growth. Polyamines (spermidine, spermine, and their precursor putrescine) play central roles in more than half of the steps of colorectal tumorigenesis. Given the close connection between polyamines and cancer, the regulation of polyamine metabolic pathways has attracted attention regarding the mechanisms of action of chemical drugs used to prevent CRC, as the drug most widely used for treating type 2 diabetes (T2D), metformin (Met) exhibits antitumor activity against a variety of cancer cells, with a vaguely defined mechanism. In addition, the influence of metformin on the UC and putrescine generation in colorectal cancer has remained unclear. In our study, we investigated the effect of metformin on the UC and putrescine generation of CRC in vivo and in vitro and elucidated the underlying mechanisms. In nude mice bearing HCT116 tumor xenografts, the administration of metformin inhibited tumor growth without affecting body weight. In addition, metformin treatment increased the expression of monophosphate (AMP)-activated protein kinase (AMPK) and p53 in both HCT116 xenografts and colorectal cancer cell lines and decreased the expression of the urea cycle enzymes, including carbamoyl phosphate synthase 1 (CPS1), arginase 1 (ARG1), ornithine trans-carbamylase (OTC), and ODC. The putrescine levels in both HCT116 xenografts and HCT116 cells decreased after metformin treatment. These results demonstrate that metformin inhibited CRC cell proliferation via activating AMPK/p53 and that there was an association between metformin, urea cycle inhibition and a reduction in putrescine generation.     

气泡富集-高效液相色谱法测定地表水样中微量土霉素

建立了气泡富集-高效液相色谱（HPLC）测定地表水样中微量土霉素的方法。采用新型样品前处理方法--气泡富集法,对水溶液中的微量土霉素进行富集,考察了气泡富集条件对富集效果的影响。研究发现,在优化的气泡富集和色谱条件下,土霉素的富集倍数可达37.06,土霉素含量测定的RSD为4.8%（n=11）,LOD为0.038 mg/L。将该方法用于地表水样中土霉素的测定,平均加标回收率为101.9%。可见,气泡富集法对土霉素的富集效果好,能与色谱结合用于地表水样中土霉素的快速、灵敏、准确检测。同时,在进行样品前处理时无需任何有机溶剂,而且装置简单、成本低廉、易操作。可见气泡富集法是一种非常有研究和推广价值的绿色样品前处理方法,有望用于复杂样品中其他微量甚至痕量物质的分析。     

Development and Validation of an HPLC Method for Simultaneous Determination of Capsaicinoids and Camphor in Over-the-Counter Medication for Topical Use

A reverse-phase high-performance liquid chromatography method was developed to determine and quantify capsaicin (trans-8-methyl-N-vanillyl-6- nonenamid), dihydrocapsaicin (8-methyl-N-vanillylnonanamide), and camphor (trimethylbicyclo[2.2.1]heptan-2-one). It is applicable in analyses of over-the-counter (OTC) medications for topical use and raw materials such as chili pepper oleoresin. Chromatographic separation was carried out on a C18 column using an isocratic flow of the mobile phase containing acetonitrile and ultrapure water in a ratio of 2:3, with pH adjusted to 3.2 using glacial acetic acid, and a flow rate of 1.5 mL/min. The concentration of the eluting compounds was monitored by a diode-array detector at a wavelength of 281 nm. The method was evaluated for several validation parameters, including selectivity, accuracy (confidence intervals < 0.05%), repeatability, and intermediate precision. The limit of detection (LOD) was determined to be 0.070 µg/mL for capsaicin, 0.211 µg/mL for dihydrocapsaicin, and 0.060 µg/mL for camphor. The limit of quantification (LOQ) was determined to be 0.212 µg/mL for capsaicin, 0.640 µg/mL for dihydrocapsaicin, and 0.320 µg/mL for camphor. Linearity was set in the range of 2.5–200 µg/mL for capsaicin and dihydrocapsaicin and 25–2000 µg/mL for camphor. The suggested analytical method can be used for quality control of formulated pharmaceutical products containing capsaicinoids, camphor, and propolis.     

水中痕量土霉素的亲水溶剂浮选-分光光度测定

本文利用自制的浮选装置,选择四氢呋喃和无水乙醇(4+1)混合溶液作溶剂,氯化钠作分相剂,将Fe(Ⅲ)与土霉素(OTC)形成的疏水性缔合物浮选至有机相,然后采用分光光度法测定,方法线性线性范围为6.6×10~(-7)～3.4×10~(-5) mol/L;检出限为3.63×10~(-7) mol/L;平均回收率为99.95%;表观摩尔吸光系数ε=2.028×10~5 L·5moL~(-1)·5cm~(-1).该方法设备简单、测定快速、灵敏度高、分离物质易于提取和不产生二次污染,可用于环境水样中痕量土霉素的快速测定.     

双水相气浮浮选光度法分离/测定废水中痕量土霉素

利用自制的浮选装置,选择四氢呋喃作溶剂,氯化钠作分相剂,NaOH溶液调节酸度,将Zn(II)与OTC形成的疏水性缔合物浮选至有机相,浮选完毕后经分光光度法分析,方法线性回归方程为A=2.038×105c(mol/L)+0.005,相关系数r=0.9996,线性范围：1.1×10-7~9.7×10-5 mol/L;检出限7.36×10-8 mol/L;回收率99.8~100.4%;表观摩尔吸光系数ε=2.038×105 L/(mol•cm),适用于废水中痕量土霉素的分离/富集及分析测定。     

Development and validation of a generic RP‐HPLC PDA method for the simultaneous separation and quantification of active ingredients in cold and cough medicines

A novel generic reverse phase high performance liquid chromatography (RP‐HPLC) method is developed and validated for simultaneous determination of seven pharmaceutically active ingredients, namely, acetaminophen, dextromethorphan, doxylamine, phenylephrine, guaifenesin, caffeine and aspirin. All seven ingredients were quantified in soft gel, syrup and tablet formulations of the over‐the‐counter US‐marketed products, as per the guidelines of the International Conference on Harmonization. The separation was achieved in a 16 min run time on an Agilent Zorbax Phenyl column using a gradient method with two mobile phases. Mobile phase A was 0.15% trifluoro acetic acid in purified water and while mobile phase B was a mixture of acetonitrile and methanol (750:250 v/v) with 0.02% trifluoro acetic acid. The flow rate was 1.0 mL min^?1 and injection volume was 10 μL. Detection was performed at 280 nm using a photodiode array detector. As part of the method validation, specificity, linearity, precision and recovery parameters were verified. The concentration and area relationships were linear (R² > 0.999), over the concentration ranges 20–120 μg mL^?1 for acetaminophen, 75–450 μg mL^?1 for dextromethorphan, 31.25–187.5 μg mL^?1 for doxylamine, 25–150 μg mL^?1 for phenylephrine, 25–150 μg mL^?1 for aspirin, 6.5–39 μg mL^?1 for caffeine and 12–72 μg mL^?1 for guaifenesin. The relative standard deviations for precision and intermediate precision were <1.5%. The proposed RP‐HPLC generic method is applicable for routine analysis of cold and cough over‐the‐counter products.     

Supercritical fluid chromatography as a tool for enantioselective separation; A review

Supercritical fluid chromatography (SFC) has become popular in the field of enantioselective separations. Many works have been reported during the last years. This review covers the period from 2000 till August 2013. The article is divided into three main chapters. The first one comprises a basic introduction to SFC. The authors provide a brief explanation of general principles and possibilities of this method. The advantages and drawbacks are also listed. Next part deals with chiral separation systems available in SFC, namely with the commonly used chiral stationary phases. Properties and interaction possibilities of the chiral separation systems are described. Recent theoretical papers are emphasized in this chapter. The last part of the paper gives an overview of applications of enantioselective SFC in analytical chemistry, in both analytical and preparative scales. Separation systems and conditions are summed up in tables so that they provide a helpful tool for analysts who search for a particular method of analysis.     

新型反相限进材料及其性能

以溴代硅胶为引发剂,CuBr/2,2-联吡啶为催化体系,在改性硅胶上经二步表面引发原子转移自由基聚合(SIATRP)制备内表面接枝甲基丙烯酸十八烷基酯(C18)、外表面接枝甲基丙烯酸环氧丙酯(GMA)、水解得到表面含大量二醇基的新型反相限进材料。使用傅里叶变换红外光谱(FT-IR)、元素分析和热重分析(TGA)对其表征,采用静态吸附实验研究反相限进材料的吸附性能,其对磺胺二甲氧嘧啶和土霉素的最大吸附量分别为18.02和4.80mg/g。结合固相萃取(SPE)评价其对大分子蛋白质的排阻性能,以牛血清白蛋白(BSA)作为排阻大分子模型,排阻能力达90%。将其用于牛奶中土霉素的分离富集,经高效液相色谱(HPLC)检测,土霉素的平均加标回收率为89.19%,相对标准偏差为3.03%。有望将新型反相限进材料和HPLC或液相色谱-质谱(LC-MS)等分析系统结合应用于生物样品的处理和检测。     

融合点测量信息的光学层析方法

光学计算机层析术的困难在于不能得到足够的信息以完全确定探测目标,理论上,将先验知识融入层析术可以在一定程度上降低病态程度,而实际中场的一组孤立点的测量值是一种更为直接的可得信息。利用ＡＲＴ３算法对于含噪声、甚至部分的投影数据具有很好的鲁棒性,并且能友好地融合来自其他测量系统的信息的特点,提出一种利用点测量信息的自适应方法,避免了对重建过程引入原本不存在的高频信息,以及在迭代过程中对预期的低频分量的     

Rapid colorimetric sensing of tetracycline antibiotics with in situ growth of gold nanoparticles

A colorimetric assay utilizing the formation of gold nanoparticles was developed to detect tetracycline antibiotics in fluidic samples. Tetracycline antibiotics showed the capability of directly reducing aurate salts into atomic gold which form gold nanoparticles spontaneously under proper conditions. The resulted gold nanoparticles showed characteristic plasmon absorbance at 526 nm, which can be visualized by naked eyes or with a spectrophotometer. UV–vis absorbance of the resulted gold nanoparticles is correlated directly with the concentrations of tetracycline antibiotics in the solution, allowing for quantitative colorimetric detection of tetracycline antibiotics. Reaction conditions, such as pH, temperature, reaction time, and ionic strength were optimized. Sensitivity of the colorimetric assay can be enhanced by the addition of gold nanoparticle seeds, a LOD as low as 20 ng mL⁻¹ can be achieved with the help of seed particles. The colorimetric assay showed minimum interference from ethanol, methanol, urea, glucose, and other antibiotics such as sulfonamides, amino glycosides etc. Validity of the method was also evaluated on urine samples spiked with tetracycline antibiotics. The method provides a broad spectrum detection method for rapid and sensitive detection of reductive substances such as tetracycline antibiotics in liquid and biological samples.