World Congress on Biosensors 2014

World Congress on Biosensors 2014
Biosensors 2014

Tuesday, 18 December 2012

Just Published: Journal of Pharmaceutical and Biomedical Analysis


A new issue of this journal has just been published. To see abstracts of the papers it contains (with links through to the full papers) click here:
Journal of Pharmaceutical and Biomedical Analysis
http://rss.sciencedirect.com/publication/science/5266
Selected papers from the latest issue:

Application of an efficient strategy for discovery and purification of bioactive compounds from Chinese herbal medicines, a case study on the Puerariae thomsonii Flos

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

In this study, an efficient strategy based on bioassay-guided fractionation, high-performance liquid chromatography/electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC-ESI-Q/TOF-MS) and high-speed counter-current chromatography (HSCCC) was established to screen and purify bioactive compounds from Chinese herbal medicines (CHMs). This screening system was efficient and successfully applied to reveal anti-prostate cancer candidates from Puerariae thomsonii Flos. As a result, an active fraction with strong in vitro anti-prostate cancer activity was obtained, and the main compounds in the fraction were purified by HSCCC, giving 82mg of tectoridin, 36mg of tectorigenin-7-O-[β-d-xylopyranosyl-(1→6)-β-d-glucopyranoside and 64mg of tectorigenin. Among them, tectorigenin, possessing the highest anti-prostate cancer activity with IC50 value of 0.08μM, has priority to be lead compound. The results of this work demonstrated that the developed method was efficient and could be employed for the rapid screening, identification and purification of active components from CHMs.

Graphical abstract

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Highlights

► An efficient strategy based on bioassay-guided fractionation, HPLC-ESI-Q/TOF-MS and HSCCC was established to screen and purify bioactive compounds from CHMs. ► The screening system succeeded in discovering anti-prostate cancer candidates from Puerariae thomsonii Flos. ► Tectoridin, tectorigenin-7-O-[β-d-xylopyranosyl-(1→6)-β-d-glucopyranoside and tectorigenin were prepared from Puerariae thomsonii Flos by HSCCC for the first time. ► Tectorigenin possessed potent inhibition effect on growth of RM-1 prostate cancer cells with IC50 value of 0.08μM.

Assay at low ppm level of dimethyl sulfate in starting materials for API synthesis using derivatization in ionic liquid media and LC–MS

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

Dimethyl sulfate (DMS) is frequently used in pharmaceutical manufacturing processes as an alkylating agent. Trace levels of DMS in drug substances should be carefully monitored since the compound can become an impurity which is genotoxic in nature. Derivatization of DMS with dibenzazepine leads to formation of the N-methyl derivative, which can be retained on a reversed phase column and subsequently separated from other potential impurities. Such derivatization occurs relatively slowly. However, it can be substantially speed up if ionic liquids are used as reaction media. In this paper we report the use of 1-butyl-1-methylpyrrolidinium bis(trifluoromethylsulfonyl)imide (IL1) and 1-butyl-4-methylpyridinium tetrafluoroborate (IL2) as reaction media for the derivatization of DMS with dibenzazepine. It was determined that the stoichiometry between the substrate and DMS may be 1:1 or 2:1, in relation with the nature of the reaction media. An (+)ESI-MS/MS approach was used for quantitation of the derivatized product. Alternatively, DMS derivatization may be carried out with pyridine in acetonitrile (ACN). The N-methylpyridinium derivative was separated by hydrophilic interaction liquid chromatography (HILIC) and detected through (+)ESI-MS (in the SIM mode). In both cases, a limit of quantitation (LOQ) of 0.05μg/ml DMS was achievable, with a linearity range up to 10μg/ml. Both analytical alternatives were applied to assay DMS in 4-(2-methoxyethyl)phenol, which is used as a starting material in the synthesis of metoprolol.

Graphical abstract

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Highlights

► LC/MS–MS analysis at sub-ppm level of dimethyl sulfate in starting materials for APIs is proposed. ► Derivatization of DMS with dibenzazepine or pyridine was achieved. ► Derivatization kinetics was studied in acetonitrile and ionic liquids (pseudo-first-order). ► Derivatization with dibenzazepine in ionic liquids is much faster than in acetonitrile. ► RPLC (for N-methyl-dibenzazepine) and HILIC (for N-methyl-pyridine) separation modes were used.

The development of a method to quantify encapsulated and free prednisolone phosphate in liposomal formulations

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

This paper presents the development of a new method for the simple and reliable quantification of the free drug amount in liposomal preparations of prednisolone phosphate (PP). In this method the free drug is distinguished from the encapsulated drug by means of hydrolysis of the free PP into prednisolone (P) by alkaline phosphatase (AP). During method development reaction progress curves were recorded to determine the required AP concentration and the corresponding incubation time to achieve hydrolysis of all free PP. Reaction progress curves also showed that small changes in the amount of weighted AP and the incubation periods used do not cause a change in outcome. Further, several organic solvents were tested as precipitation solvent and the use of tetrahydrofuran (THF) yielded clean chromatograms, rapid AP deactivation and complete liposome rupture avoiding under- and overestimations of the encapsulated and free drug concentrations. Method accuracy was evaluated during a cross-validation involving dialysis. Intra- and interday precision were evaluated by determining the standard deviation (SD) and relative standard deviation (RSD) after applying the new method on one day (n =4) and on different days (n =3). The accuracy of the developed method is comparable to the accuracy determined by dialysis, while clearly the method using AP is more precise. In conclusion, comprehensive method development yielded an accurate and precise method, which can replace traditional methods like dialysis and solid phase extraction (SPE). With little effort the method can be upgraded and become part of the liposome certification prior to human use. The overall principle behind the method offers possibilities for many drug carrier systems.

Highlights

► A method for the quantification of the free drug amount in liposomal preparations of prednisolone phosphate was developed. ► The free drug is distinguished from the encapsulated drug by means of enzymatic hydrolysis. ► In this way difficulties accompanying traditional separation techniques were circumvented. ► Thismethod is reliable, accessible, inexpensive,fast and suitable for small sample volumes and a large number of samples. ► The principle behind this method offers also possibilities for the free drug determination of many drug carrier systems.

A separation strategy combining three HPLC modes and polysaccharide-based chiral stationary phases

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

Nine polysaccharide-based chiral stationary phases (CSPs) were used to define a separation strategy that combines normal-phase (NP), reversed-phase (RP) and polar organic solvent chromatography (POSC) modes. After limiting ourselves to two CSPs per mode, in total, five CSPs, Chiralpak AD (NP), Chiralcel OD (RP and POSC), Lux Cellulose-1 (NP), Lux Cellulose-2 (POSC) and Lux Cellulose-3 (RP), showed the broadest enantioselectivity and most complementarity. Six sequences of the three modes were considered to decide which sequence is the most successful for screening a set of 56 pharmaceutical compounds. Starting the strategy with the NP mode, followed by RP and finally POSC was found preferable from both the number of cumulative separations and of baseline separations. Two approaches were considered for strategy fine tuning using an additional set of eight racemic mixtures. In both approaches, seven of the eight compounds were baseline resolved, on one of the examined columns at either screening or optimization conditions of a mode. One approach was finally preferred because of its lower workload.

Highlights

► 56 compounds and nine CSPs were used to update strategies in three common HPLC modes. ► An HPLC chiral method development strategy that combines NPLC, RPLC and POSC modes was defined. ► Success rate: all compounds were separated with the updated strategy in less experiments. ► The updated strategy was fine tuned.

A microflow chemiluminescence sensor for indirect determination of dibutyl phthalate by hydrolyzing based on biological recognition materials

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

A microflow chemiluminescence (CL) sensor for determination of dibutyl phthalate (DBP) based on magnetic molecularly imprinted polymer (MMIP) as recognition element was fabricated. Briefly, a hydrophilic molecularly imprinted polymer layer was produced at the surface of Fe3O4@SiO2 magnetic nanoparticles (MNPs) via combination of molecular imprinting and reversible stimuli responsive hydrogel. In this protocol, the initial step involved co-precipitation of Fe2+ and Fe3+ in an ammonia solution. Silica was then coated on the Fe3O4 nanoparticles using a sol–gel method to obtain silica shell magnetic nanoparticles. The MMIP was synthesized using methacrylic acid (MAA) as functional monomer and ethylene glycol dimethacrylate (EGDMA) as cross-linker and 2, 2-azobisisobutyronitrile (AIBN) as initiator in chloroform. Then the synthesized MMIP and magnetic non-molecular imprinted polymers (MNIP) were employed as recognition by packing into lab-made straight shape tubes, connected in CL analyzer for establishing the novel sensor with a single channel syringe pump. And a mixer for hydrolyzing of DBP was followed. Based on this experiment principle, DBP was determined indirectly. And the MMIP showed satisfactory recognition capacity to DBP, resulting to the wide linear range of 3.84×10−8 to 2.08×10−5 M and the low detection limit of 2.09×10−9 M (3σ) for DBP. The relative standard deviation (RSD) for DBP (3.20×10−6 M) was 1.40% (n =11). Besides improving sensitivity and selectivity, the sensor was reusable. The proposed DBP–MMIP–CL sensor has been successfully applied to determine DBP in drink samples.

Graphical abstract

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Highlights

► Fe3O4@SiO2 was used as support for easier to control. ► Microscale Syringe Pump was used for saving reagent. ► A mixer for hydrolyzing was used for chemiluminescence.

Dispersive liquid–liquid microextraction combined with ultra-high performance liquid chromatography for the simultaneous determination of 25 sulfonamide and quinolone antibiotics in water samples

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

In this work, a dispersive liquid–liquid microextraction (DLLME) procedure combined with ultra-high performance liquid chromatography with diode-array detection was developed to determine 25 antibiotics in mineral and run-off waters. Optimum DLLME conditions (5mL of water at pH=7.6, 20% (w/v) NaCl, 685μL of CHCl3 as extractant solvent, and 1250μL of ACN as disperser solvent) allowed the repeatable, accurate and selective determination of 11 sulfonamides (sulfanilamide, sulfacetamide, sulfadiazine, sulfathiazole, sulfadimidin, sulfamethoxypyridazine, sulfadoxine, sulfamethoxazole, sulfisoxazole, sulfadimethoxine and sulfaquinoxaline) and 14 quinolones (pipemidic acid, marbofloxacin, fleroxacin, levofloxacin, pefloxacin, ciprofloxacin, lomefloxacin, danofloxacin, enrofloxacin, sarafloxacin, difloxacin, moxifloxacin, oxolinic acid and flumequine). The method was validated by means of the obtention of calibration curves of the whole method as well as a recovery study at two levels of concentration. The LODs of the method were in the range 0.35–10.5μg/L with recoveries between 78% and 117%.

Graphical abstract

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Highlights

► A DLLME–UHPLC-DAD method was developed to determine antibiotics in water. ► A total of 25 compounds (11 sulfonamides and 14 quinolones) was analyzed. ► Relative recoveries values of the method were between 74% and 117%. ► The procedure is simple, fast and reliable to determine the selected antibiotics.

Proteomics-based identification of plasma biomarkers in oral squamous cell carcinoma

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

Oral squamous cell carcinoma (OSCC) is an aggressive cancer and its occurrence is closely related to betel nut chewing in Taiwan. However, there are few prognostic and diagnostic biomarkers for this disease especially for its association with betel nut chewing. Recent progresses in quantitative proteomics have offered opportunities to discover plasma proteins as biomarkers for tracking the progression and for understanding the molecular mechanisms of OSCC. In present study, plasma samples from OSCC patients with at least 5-year history of betel nut chewing and healthy donors were analyzed by fluorescence 2D-DIGE-based proteomic analysis. Totally, 38 proteins have been firmly identified representing 13 unique gene products. These proteins mainly function in inflammatory responses (such as fibrinogen gamma chain) and transport (Apolipoprotein A–I). Additionally, the current quantitative proteomic approach has identified numerous OSCC biomarkers including fibrinogen (alpha/beta/gamma) chain, haptoglobin, leucine-rich alpha-2-glycoprotein and ribosomal protein S6 kinase alpha-3 (RSK2) which have not been reported and may be associated with the progression and development of the disease. In summary, this study reports a comprehensive patient-based proteomic approach for the identification of potential plasma biomarkers in OSCC. The potential of utilizing these markers for screening and treating OSCC warrants further investigations.

Graphical abstract

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Highlights

► Oral squamous cell carcinoma (OSCC) is an aggressive cancer closely related to betel nut chewing. ► Quantitative proteomics have offered opportunities to discover plasma OSCC protein biomarkers. ► This study identified numerous unreported OSCC markers such as RSK2. ► The utilization of these markers for screening and treating OSCC warrants further investigations.

Pre-column derivatization combined with UHPLC–MS/MS for rapid and sensitive quantification of bakuchiol in rat plasma

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

Psoralea corylifolia L. (Fabaceae) is a traditional Chinese medicine with many beneficial effects in medical therapies. Bakuchiol was the main active ingredient of Psoralea corylifolia L. In this study, a novel method of pre-column derivatization with dansyl chloride followed by analysis of ultra high-performance liquid chromatographic–tandem mass spectrometry (UHPLC–MS/MS) was established and validated for quantification of bakuchiol in rat plasma. The linearity of this approach was confirmed to be within the concentration range of 0.5–1000ng/mL and the lower limit of quantification was at 0.5ng/mL. The total analysis time was 1.5min for each pretreated sample. Also, the precision, accuracy, stability, recovery and matrix effect of this method were proved to meet the requirements for bioanalysis. The intravenous and oral pharmacokinetic profiles of bakuchiol were obtained by utilizing this approach. The oral bioavailability of bakuchiol in rats (3.2%) was identified.

Graphical abstract

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Highlights

A novel method of pre-column derivatization with dansyl chloride followed by analysis of ultra high-performance liquid chromatographic–tandem mass spectrometry (UHPLC–MS/MS) was established and validated for quantification of bakuchiol in rat plasma. The intravenous and oral pharmacokinetic profiles of bakuchiol were obtained by utilizing the established method.
► An UHPLC–MS/MS method with pre-column derivatization was built for bakuchiol. ► The method demonstrated high sensitivity, wide linear range and short runtime. ► The method was used to explore the pharmacokinetic profile of bakuchiol in rats. ► The oral bioavailability of bakuchiol (3.2%) was reported for the first time.

Development of generic immunoassay for the detection of a series of aminoglycosides with 6′-OH group for the treatment of genetic diseases in biological samples

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

Over the last two decades, a growing number of scientific evidences highlighted the potential therapeutic value of several structures of aminoglycoside antibiotics (including gentamicin and G418) for the treatment of various genetic diseases caused by nonsense mutations. These findings resulted in a fast evolvement of synthetic derivatives of aminoglycosides which were shown to be more target specific and less toxic than the clinically used antibiotics. The emerging progress in drug design and development has necessitated the urge to develop a fast, easy and accurate procedure for the determination of these potential therapeutic agents in various biologically derived matrices. Here we describe the preparation of a generic polyclonal antibody that was used for the development of homologous and heterologous immunoassays for the detection of a wide range of natural and synthetic aminoglycoside derivatives, highlighted today as potential therapeutic agents for the treatment of various genetic diseases. A common two-ring scaffold, NB82, present in the majority of compounds exhibiting potent biological activity, was used as a generic immunization hapten for the immunization of two rabbits. By using a series of chemical steps, NB82 was selectively conjugated via the N-1 position through glutaric acid linker to a carrier protein. Sensitivity (I 50) values for the recognition of three representative compounds NB82, NB84 and NB124 were determined to be 10±3ngmL−1, 0.5±0.04μgmL−1 and 1±0.12μgmL−1, respectively. Limits of detection were determined to be 1±0.3ngmL−1 for NB82, 20±7ngmL−1 for NB84 and 15±8ngmL−1 for NB124. The developed assays were further exploited for the in vivo monitoring of the therapeutic compounds in mice serum. Serum experimentations exhibited similar detection limits as observed for the PBS calibration experiments, demonstrating no interference with assays sensitivity, with rather high recovery ratios ranging from 92 to 107% in whole blood samples.

Graphical abstract

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Highlights

► A broad-specific ELISA for the detection of aminoglycosides with 6′-OH was developed. ► These 6′-OH aminoglycosides are best candidates for the treatment of genetic diseases. ► A two-ring scaffold was selectively conjugated to a carrier protein at N-1 position. ► A generic antibody allowed highly sensitive monitoring of therapeutic agents in vivo. ► The assay will aid in further development of therapeutic compounds in clinic.

Structural characterization of minor metabolites and pharmacokinetics of ganoderic acid C2 in rat plasma by HPLC coupled with electrospray ionization tandem mass spectrometry

18 December 2012, 07:00:59
5 March 2013
Publication year: 2013
Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 75

The metabolites and pharmacokinetics of ganoderic acid C2 (GAC2), a bioactive triterpenoid in Ganoderma lucidum in rat plasma were investigated by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC–ESI-MS/MS). Totally, ten minor phase I metabolites of GAC2 were characterized after oral administration of GAC2, on the basis of their mass fragmentation pathways or direct comparison with authentic compounds by high-performance liquid chromatography coupled with diode array detection and electrospray ion trap tandem mass spectrometry (HPLC–DAD–ESI-MS n ), and liquid chromatography coupled with electrospray ionization hybrid ion trap and time-of-flight mass spectrometry (LC–ESI-IT-TOF/MS) methods. Moreover, a rapid and specific method for quantification of GAC2 in rat plasma after oral administration was developed by using a liquid–liquid extraction procedure and HPLC–ESI-MS/MS analysis. It is the first time to report the metabolites and pharmacokinetics of GAC2.

Graphical abstract

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Highlights

Proposed metabolic pathways of ganoderic acid C2 (M0) in rats.
► Ten minor phase I metabolites were characterized. ► A rapid quantification method was developed by LC–MS/MS. ► The metabolites and pharmacokinetic of GAC2 were reported firstly.

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