Simultaneous Determination of Pioglitazone and Glimepiride by High-Performance Liquid Chromatography

A rapid and accurate HPLC method has been developed for simultaneous determination of pioglitazone and glimepiride. Chromatographic separation of the two pharmaceuticals was performed on a Cosmosil C₁₈ column (150 mm × 4.6 mm, 5 m) with a 45:35:20 (v/v) mixture of 0.01 m triammonium citrate (pH adjusted to 6.95 with orthophosphoric acid), acetonitrile, and methanol as mobile phase, at a flow rate of 1.0 mL min^–1, and detection at 228 nm. Separation was complete in less than 10 min. The method was validated for linearity, accuracy, precision, limit of quantitation, and robustness [1, 2]. Linearity, accuracy, and precision were found to be acceptable over the ranges 2.50–30.00 g mL^–1 for pioglitazone and 0.10–10.00 g mL^–1 for glimepiride.     

表面活性剂增敏2.5次微分极谱测定香豆素

拟定了表面活性剂增敏、2 .5次微分极谱测定香豆素的方法。在含 6.0× 10 -6 mol·L-13 氯 2 羟丙基 三甲基氯化铵的 0 .1mol·L-1KH2 PO4 Na2 HPO4 (pH 6.8)缓冲液中 ,香豆素还原波 [峰电位Ep=- 1.60V(vs .SCE) ]的 2 .5次微分极谱峰峰电流e″p 与其浓度在 4 .0× 10 -7～ 6.0× 10 -5mol·L-1范围内呈良好线性关系 (r =0 .9998,n =10 ) ,检出限为 1.2× 10 -7mol·L-1。 13次平行测量 8.0× 10 -6 mol·L-1香豆素的峰电流 ,RSD为 1.5 %。该方法可用于中药白芷中香豆素的直接测定     

Analysis of Zoledronic Acid and Its Related Substances by Ion-Pair RP-LC

A simple high-performance liquid chromatographic method – ion-pair reversed- phase high performance liquid chromatography (RPIC) has been developed and employed for the analysis of zoledronic acid and its related substances in bulk material and commercial dosage forms. The mobile phase was a mixture of methanol (20%) and 5 mmol L^–1 phosphate buffer (80%) containing 6 mmol L^–1 tetrabutylammonium bromide, adjusted to pH 7.0 with sodium hydroxide. C₈ column was used as the stationary phase. The chromatographic conditions were optimized. The active ingredient – zoledronic acid was successfully separated from its related substances, including remained imidazol-1-yiacetic acid in the synthesis of zoledronic acid and other possible impurities of oxidation and decomposition. The excipients did not interfere with the determination of zoledronic acid in commercial dosage formulations. The method was rapid, simple, accurate and reproducible. It was not only successfully employed for the assay of zoledronic acid in bulk material and pharmaceutical dosage forms but also for the determination of its related substances.     

Application of the pKa rule to synthesize salts of bezafibrate

ABSTRACT

Co-crystallization frequently employs forces such as hydrogen bonds, halogen bonds, and π-π stacking to assemble molecules in a multi-component crystal. In an effort to increase the strength of the intermolecular interaction between the anti-cholesterol drug bezafibrate (BEZA), a wastewater contaminant, and hydrogen-bond-acceptor molecules, we modified the pK_a values of the acceptors. Here, we describe the first series of salts incorporating BEZA and achieve a variety of supramolecular architectures including discrete assemblies, 1D chains, tapes, and 2D sheets. We discuss exceptions to the pK_a rule, and demonstrate that the presence of hydrogen-bond-donor atoms on the acceptor molecule supports salt formation.     

A novel technique to specifically analyze the secretome of cells and tissues

The secretome of cells and tissues may reflect a broad variety of pathological conditions and thus represents a rich source of biomarkers. The identity of secreted proteins, usually isolated from cell supernatants or body fluids, is hardly accessible by direct proteome analysis, because these proteins are often masked by high amounts of proteins actually not secreted by the investigated cells. Here, we present a novel method for the specific detection of proteins secreted by human tissue specimen as well as cultured cells and chose liver as a model. The method is based on the metabolic labelling of proteins synthesized during a limited incubation period. Then, the cell supernatant is filtered, precipitated, and subjected to two-dimensional gel electrophoresis. Whereas fluorography detected a large number of proteins derived from residual plasma and dead cells, the autoradiographs selectively displayed genuinely secreted proteins. We demonstrate the feasibility of this approach by means of the secretomes of the hepatocellular carcinoma-derived cell line HepG2 and human liver slices. The selective identification of cell- and tissue-specific protein secretion profiles may help to identify novel sets of biomarkers for wide clinical applications.     

Rapid Electrochemical Detection of Propranolol and Metoprolol in Pharmaceutical Preparations Using Stabilized Lipid Films

This work reports a technique for the rapid electrochemical detection of propranolol and metoprolol in pharmaceutical preparations using stabilized lipid films. Microporous filters composed of glass fibers (nominal pore sizes 0.7 and 1.0 μm) were used as supports for the formation and stabilization of these devices. The lipid film is formed on the filter by polymerization prior to its use. This stabilized after storage in air. Lipid films composed of phosphatidylcholine were used for the detection of propranolol and metoprolol in pharmaceutical preparations. The stabilized lipid membranes provided artificial ion gating events in the form of transient signals within about 60 and 34 s after exposure of the membranes to propranolol and metoprolol, respectively. The magnitude of the transient current signal was related to the concentration of propranolol and metoprolol in bulk solution in the micromolar range. The mechanism of signal generation was investigated by differential scanning calorimetric studies. These studies revealed that the adsorption of the drug is through the hydrophobic aryl terminal of the compound, whereas the hydrophilic groups were directed towards the electrolyte solution. This adsorption caused a rapid alteration of the electrochemical double layer of the lipid film (i.e., capacitance changes) that resulted in the transient ion current signal. The present technique was used for the rapid detection of propranolol and metoprolol in pharmaceutical preparations and can function for repetitive uses after storage in air. Future research is targeted to the determination of these chemicals in human biofluids such as urine of athletes.     

双嘧达莫的荧光光谱分析法

以荧光光谱法研究了双嘧达莫在溴化十六烷基三甲基铵(CTMAB)、β_环糊精 (β_CD)、十二烷基硫酸钠 (SDS)等介质体系中的荧光性质 ,发现CTMAB、SDS和 β_CD对双嘧达莫均有不同程度的荧光增敏作用 ,提出了在CTMAB、SDS和 β_CD水溶液中测定双嘧达莫的荧光光谱分析法 ;该法灵敏度高 ,检出限低(3.20×10 -9mol/L) ,在6.40×10 -8～3.20×10 -6mol/L范围内荧光强度与双嘧达莫的浓度呈良好线性关系     

Oxidative cyclization of 2-pyrrolidinyl-acetamide and 2-pyrrolidinyl-propionamide local anaesthetics

2 - Pyrrolidinyl - acetamide and -propionamide local anaesthetics (1a–d) on oxidation with mercuric acetate, potassium hexacyanoferrate(III) and potassium permanganate underwent oxidative cyclization to give new bicyclic compounds, hexahydro - 1H - pyrrolo[1,2 - a]imidazolin - 2 - ones (2a–d). The propionamides (1c–d) yielded mixtures of the two possible diastereoisomers of 2c and 2d. These were separated; in solution and above their melting points they epimerized via ring opening and reclosure between the 7a-carbon and 1-nitrogen atoms.     

Site-specific detection of S-nitrosylated PKB alpha/Akt1 from rat soleus muscle using CapLC-Q-TOF(micro) mass spectrometry

Protein Kinase Balpha(PKBalpha, or Akt1) is believed to play a crucial role in programmed cell death, cancer progression and the insulin-signaling cascade. The protein is activated by phosphorylation at multiple sites and subsequently phosphorylates and activates eNOS. Free cysteine residues of the protein may capture reactive, endogenously produced nitric oxide (NO) as S-nitrosothiols. Site-specific detection of S-nitrosylated cysteine residues, usually at low stoichiometry, has been a major challenge in proteomic research largely due to the lack of mass marker for S-nitrosothiols that are very labile under physiologic conditions. In this report we describe a sensitive and specific MS method for detection of S-nitrosothiols in PKB alpha/Akt1 in rat soleus muscle. PKB alpha/Akt1 was isolated by immunoprecipitation and 2D-gel electrophoresis, subjected to in-gel tryptic digestion, and cysteinyl nitrosothiols were reacted with iodoacetic acids [2-C(12)/C(13) = 50/50] under ascorbate reduction conditions. This resulted in the production of relatively stable carboxymethylcysteine (CMC) immonium ions (m/z 134.019 and m/z 135.019) within a narrow argon collision energy (CE = 30 +/- 5 V) in the high MS noise region. In addition, free and disulfide-linked cysteine residues were converted to carboxyamidomethylcysteines (CAM). Tryptic S-nitrosylated parent ion was detected with a mass accuracy of 50 mDa for the two CMC immonium ions at the triggered elution time during capillary liquid chromatography (LC) separation. A peptide containing Cys(296) was discriminated from four co-eluting tryptic peptides under lock mass conditions (m/z 785.8426). S-nitrosothiol in the tryptic peptide, ITDFGLBKEGIK (B: CAM, [M + 2H](2+) = 690.86, Found: 690.83), is believed to be present at a very low level, since the threshold for the CMC immonium trigger ions was set at 3 counts/s in the MS survey. The high levels of NO that are produced under stress conditions may result in increased S-nitrosylation of Cys(296) which blocks disulfide bond formation between Cys(296) and Cys(310) and suppresses the biological effects of PKB alpha/Akt1. With the procedures developed here, this process can be studied under physiological and pathological conditions.     

药物分析

本文对国内药物分析在１９９３－１９９５年间的主要进展作了评述。内容包括下述方法在药物分析中的应用：分光光度法、高效液相色谱法、气相色谱法、薄层色谱法、电化学分析法和其它方法等。共引用文献７６１篇。