Development of new extraction method based on liquid–liquid–liquid extraction followed by dispersive liquid–liquid microextraction for extraction of three tricyclic antidepressants in plasma samples

In the present study, a new extraction method based on a three–phase system, liquid–liquid–liquid extraction, followed by dispersive liquid–liquid microextraction has been developed and validated for the extraction and preconcentration of three commonly prescribed tricyclic antidepressant drugs – amitriptyline, imipramine, and clomipramine – in human plasma prior to their analysis by gas chromatography–flame ionization detection. The three phases were an aqueous phase (plasma), acetonitrile and n–hexane. The extraction mechanism was based on the different affinities of components of the biological sample (lipids, fatty acids, pharmaceuticals, inorganic ions, etc.) toward each of the phases. This provided high selectivity toward the analytes since most interferences were transferred into n–hexane. In this procedure, a homogeneous solution of the aqueous phase (plasma) and acetonitrile (water–soluble extraction solvent) was broken by adding sodium sulfate (as a phase separating agent) and the analytes were extracted into the fine droplets of the formed acetonitrile. Next, acetonitrile phase was mixed with 1,2–dibromoethane (as a preconcentration solvent at microliter level) and then the microextraction procedure mentioned above was performed for further enrichment of the analytes. Under the optimum extraction conditions, limits of detection and lower limits of quantification for the analytes were obtained in the ranges of 0.001–0.003 and 0.003–0.010 μg mL⁻¹, respectively. The obtained extraction recoveries were in the range of 79–98%. Intra– and inter–day precisions were < 7.5%. The validated method was successfully applied for determination of the selected drugs in human plasma samples obtained from the patients who received them.     

Simultaneous Determination of Caffeine and Theophylline in Human Plasma with a Weak Cation Monolithic SPE-column

A simple reversed‐phase high performance liquid chromatography (RP‐HPLC) method was developed to determine the level of caffeine and theophylline in human plasma samples. The sample clean‐up step involved the on‐line solid‐phase extraction (SPE) of the analytes from plasma samples into a weak cation monolithic column using a column switching system. Separation was performed on a C₁₈ column (5 µm, 150 mm×4.6 mm) with ultraviolet detection at 274 nm. The mobile phase consisted of methanol‐water (32/68, V/V) under isocratic conditions at a flow rate of 0.6 mL·min⁻¹. The measured concentration of caffeine and theophylline showed a good linear relationship over the concentrations range, 0.1–80.0 µg·mL⁻¹. The absolute recoveries ranged from 77.10% to 85.39%, and the inter‐day and intra‐day relative standard deviations (RSD) were all less than 5%. This method avoids a tedious pretreatment and provides an economic, repeatable and effective method for assaying trace drugs in biological samples.     

Dodging matrix effects in liquid chromatography tandem mass spectrometric assays—compilation of key learnings and perspectives

Triple quad liquid chromatography mass spectrometric assays (LC/MS/MS) have revolutionized the analysis of drug(s)/metabolite(s) with exceptional speed, sensitivity and selectivity features. From inception to date, several new and innovative features have been regularly proposed by researchers to further enhance the value in the applicability of this analytical tool. However, owing to such compressed run times and scanty sample preparation procedures, LC/MS/MS assays that are not fully optimized generally have issues of matrix effects, where ionization potential is either suppressed or enhanced due to the presence of other materials (endogenous/exogenous) in the matrix. By definition, even co‐medications, isomeric or isobaric impurities, and drug excipients used in dosing solutions could also potentially contribute to matrix effects. This article captures some of the interesting work carried out by researchers to understand and handle matrix effects. Additionally, it provides perspectives to effectively deal with matrix effects. Copyright © 2008 John Wiley & Sons, Ltd.     

Unstable plasma characteristics in mirror field electron cyclotron resonance microwave ion source

Electron cyclotron plasma reactor are prone to instabilities in specific input power [3–7] region (150–450 watts). In this region power absorption by gas molecules in the cavity is very poor and enhanced input power gets reflected substantially without increasing ion density. There are abrupt changes in plasma characteristics when input power was decreased from maximum to minimum, it was observed that reflected power changed from <2% to ∼50%. Minimum two jumps in reflected power were noticed in this specific power region and these appear to be highly sensitive to three stub tuner position in the waveguide for this particular input power zone. Unstable plasma region of this source is found to be dependent upon the magnetic field strength. Some changes in reflected power are also noticed with pressure, flow and bias and they are random in nature.     

镧在细胞膜上键合形态研究

研究了一种简单快速的富膜细胞组分的提取和分离方法，并运用这种方法研究了大豆幼苗根细胞膜上La结合形态，La末与膜蛋白结合，而是与一非膜蛋白组分呈优势结合。     

Limitation of immunoaffinity column for the removal of abundant proteins from plasma in quantitative plasma proteomics

In plasma proteomics, before a proteome analysis, it is essential to prepare protein samples without high‐abundance proteins, including albumin, via specific preparation techniques, such as immunoaffinity capture. However, our preliminary experiments suggested that functional changes with use alter the ability of the immunoaffinity column. Thus, in this study, to evaluate the changes of the removal ability of abundant proteins from plasma by the immunoaffinity column, plasma proteome analysis was performed for the long‐term test for the reproducibility of the affinity column using the fluorogenic derivatization–liquid chromatography–tandem mass spectrometry method combined with an IgY column. The specific adsorption for albumin decreased with an increase in the number of the column usage before its expiration date. Moreover, it was demonstrated that hydrophobic high molecular weight compounds in plasma adsorbed onto the column materials surface contributed to the functional changes from specific immunoaffinity adsorption into hydrophobic interaction. These results suggested that, in quantitative plasma proteomics studies, it is important to keep in mind the risk of not only the nonselective loss but also the changes in the adsorption ability of the immunoafinity column. Copyright © 2008 John Wiley & Sons, Ltd.