A simple and inexpensive method to monitor and minimize exposure from manipulation of cytotoxic drugs.

Pharmacy personnel that manipulate cytotoxic drugs are under continuous exposure risk. Therefore, training and strict adherence to recommended practices should always be promoted. The main objective of this study was to develop and apply a safe, effective and low-cost method for the training and assessment of the safe handling of cytotoxic drugs, using commercially available tonic water. To evaluate the potential of tonic water as a replacement marker for quinine hydrochloride, deliberate spills of 1 mL of four different tonic waters (one coloured and three non-coloured) were analysed under ultraviolet light (300-400 nm). The pigmented sample did not produce fluorescence under ultraviolet (UV) light. The three commercially available tonic waters that exhibited fluorescence were further analysed by UV/Vis spectrophotometry (300-500 nm). Afterwards, a protocol of simulated manipulation of cytotoxic drugs was developed and applied to 12 pharmacy technicians, that prepared 24 intravenous bags according to recommended routine procedures using tonic water. Participants responded to a brief questionnaire to evaluate the adequacy and applicability of the activity. Seven of the participants had spillages during manipulation, the majority of which recorded during manipulation with needles. All participants scored the tonic water manipulation simulation with 4 or 5 points for simplicity, efficiency and feasibility. The obtained results suggest that tonic water can be used to simulate the manipulation of cytotoxic drugs in training and assessment programs. By using this replacement marker for quinine hydrochloride, it is possible to perform a more cost-effective, yet equally effective, assessment.

Sweeping of hydrophobic amines under inhomogeneous electric field and low surfactant concentration in micellar electrokinetic chromatography.

The influence of sample matrix on sample sweeping in MEKC was examined in the presented manuscript. Significant focusing effect was observed for relatively hydrophobic cationic compounds (emetine, strychnine and quinine) using high ionic strength sample matrix (900 mM H3 PO4 /720 mM Tris) which conductivity was about ninefold higher than utilized BGE. Moreover, the results were obtained using BGE composed of comparatively low surfactant concentration (10 mM SDS) and 40 mM H3 PO4 /32 mM Tris buffer solution. About 200 to 300-fold preconcentration of analytes was reached with the presented method. Basing on experimental results and computer simulation using Simul5 software, hypothetical mechanism of observed phenomenon was proposed.

A Microfluidic Platform for the Rapid Determination of Distribution Coefficients by Gravity-Assisted Droplet-Based Liquid-Liquid Extraction.

The determination of pharmacokinetic properties of drugs, such as the distribution coefficient (D) is a crucial measurement in pharmaceutical research. Surprisingly, the conventional (gold standard) technique used for D measurements, the shake-flask method, is antiquated and unsuitable for the testing of valuable and scarce drug candidates. Herein, we present a simple microfluidic platform for the determination of distribution coefficients using droplet-based liquid-liquid extraction. For simplicity, this platform makes use of gravity to enable phase separation for analysis and is 48 times faster and uses 99% less reagents than performing an equivalent measurement using the shake-flask method. Furthermore, the D measurements achieved in our platform are in good agreement with literature values measured using traditional shake-flask techniques. Since D is affected by volume ratios, we use the apparent acid dissociation constant, pK', as a proxy for intersystem comparison. Our platform determines a pK' value of 7.24 ± 0.15, compared to 7.25 ± 0.58 for the shake-flask method in our hands and 7.21 for the shake-flask method in the literature. Devices are fabricated using injection molding, the batchwise fabrication time is <2 min per device (at a cost of $1 U.S. per device), and the interdevice reproducibility is high.

Macromolecular crowding-assisted fabrication of liquid-crystalline imprinted polymers.

A macromolecular crowding-assisted liquid-crystalline molecularly imprinted monolith (LC-MIM) was prepared successfully for the first time. The imprinted stationary phase was synthesized with polymethyl methacrylate (PMMA) or polystyrene (PS) as the crowding agent, 4-cyanophenyl dicyclohexyl propylene (CPCE) as the liquid-crystal monomer, and hydroquinidine as the pseudo-template for the chiral separation of cinchona alkaloids in HPLC. A low level of cross-linker (26%) has been found to be sufficient to achieve molecular recognition on the crowding-assisted LC-MIM due to the physical cross-linking of mesogenic groups in place of chemical cross-linking, and baseline separation of quinidine and quinine could be achieved with good resolution (R(s) = 2.96), selectivity factor (α = 2.16), and column efficiency (N = 2650 plates/m). In contrast, the LC-MIM prepared without crowding agents displayed the smallest diastereoselectivity (α = 1.90), while the crowding-assisted MIM with high level of cross-linker (80%) obtained the greatest selectivity factor (α = 7.65), but the lowest column efficiency (N = 177 plates/m).

Mouth rinsing with a bitter solution without ingestion does not improve sprint cycling performance.

PURPOSE: Recently, we have shown that combining mouth rinsing with the ingestion of a 2 mM quinine solution immediately before a 30-s cycling sprint significantly improves performance. However, the strong bitterness of such a solution produces an unpleasant taste and evokes nausea at higher concentrations. Given the possibility that mouth rinsing with quinine without ingesting it may not produce nausea, a mouth rinse only protocol may be a more practical approach to administer quinine for improving exercise performance. The purpose of the present study was to determine whether mouth rinsing with quinine without ingesting it improves 30-s sprint cycling performance. METHODS: Twelve competitive male cyclists performed a 30-s maximal cycling sprint immediately after rinsing their mouth for 10 s with either a 10 mM bitter quinine solution (QUI), plain water (WAT), a 7.1 % w/v sweet glucose solution (GLU), or no solution at all (control; CON). Sprint performance was assessed, and heart rate, ratings of perceived exertion and blood variables were measured pre- and post-exercise. RESULTS: Mean power output during the 30-s sprint (QUI 888 ± 38; CON 873 ± 39; WAT 885 ± 37; GLU 873 ± 42 W; p = 0.431) as well as peak power (QUI 1230 ± 61; CON 1,208 ± 65; WAT 1,220 ± 70; GLU 1,202 ± 59 W; p = 0.690) were similar between the four conditions. There were no significant differences in any other performance measures, heart rate, subjective ratings or blood variables between conditions. CONCLUSIONS: Mouth rinsing with a bitter tasting quinine solution without ingestion does not improve 30-s sprint cycling performance.

A case of fixed drug eruption secondary to quinine in tonic water presenting to a sexual health clinic.

Fixed drug eruption (FDE) is a cutaneous drug reaction which occurs repeatedly at a given mucocutaneous site after exposure to the causative agent. Lesions typically occur on extremities, oral mucosa and genital skin. Quinine is a common food additive and is recognised as a rare cause of FDE. We report a case of FDE with oral and genital lesions presenting to a sexual health clinic due to quinine contained in tonic water.

Effect of downscaling on the linearity range of a calibration curve in spectrofluorimetry.

Interest in the microfluidic environment, owing to its unique physical properties, is increasing in much innovative chemical, biological, or medicinal research. The possibility of exploiting and using new phenomena makes the microscale a powerful tool to improve currently used macroscopic methods and approaches. Previously, we reported that an increase in the surface area to volume ratio of a measuring cell could provide a wider linear range for fluorescein (Kwapiszewski et al., Anal. Bioanal. Chem. 403:151-155, 2012). Here, we present a broader study in this field to confirm the assumptions we presented before. We studied fluorophores with a large and a small Stokes shift using a standard cuvette and fabricated microfluidic detection cells having different surface area to volume ratios. We analyzed the effect of different configurations of the detection cell on the measured fluorescence signal. We also took into consideration the effect of concentration on the emission spectrum, and the effect of the surface area to volume ratio on the limit of linearity of the response of the selected fluorophores. We observed that downscaling, leading to an increase in the probability of collisions between molecules and cell walls with no energy transfer, results in an increase in the limit of linearity of the calibration curve of fluorophores. The results obtained suggest that microfluidic systems can be an alternative to the currently used approaches for widening the linearity of a calibration curve. Therefore, microsystems can be useful for studies of optically dense samples and samples that should not be diluted.

Quinine sensitivity influences the acceptance of sea-buckthorn and grapefruit juices in 9- to 11-year-old children.

The acceptance of novel foods by children is related to a number of factors, and differences in taste sensitivity may form some specific challenges. High sensitivity might be a barrier to the acceptance of sour/bitter products by children. This study investigated the effect of sensitivity to bitter, sour, sweet, and salty tastes on the acceptance of Nordic juices in 9- to 11-year-old children. A total of 328 children were subjected to two taste sensitivity tests for quinine, citric acid, sucrose, and NaCl. Their acceptance of six juices (carrot, rosehip, sea-buckthorn, lingonberry, grapefruit, and aronia) was measured. Bitter sensitivity was found to be significantly correlated to the intake of the sweet sea-buckthorn and lingonberry juices; the most bitter-sensitive children exhibited the highest intake of these juices. The opposite relationship was found for bitter sensitivity and the intake of the bitter grapefruit juice. Sour, sweet, and salt sensitivities did not affect the intake of any of the juices. Liking scores were not affected by sensitivity. In conclusion, bitter sensitivity appears to influence food intake in children to a greater extent than sour, sweet, or salt sensitivity. Bitter-sensitive children exhibited a reduced intake of grapefruit juice and a higher intake of sucrose-sweetened juices. Thus, bitter sensitivity might be a challenge in the acceptance of certain bitter foods.

Assessment of field fluorometers.

Two field fluorometers, devoted either to natural organic matter (NOM) or to tryptophan-like fluorescing substances, were tested for the characterization of a large set of water samples (n = 263) impacted to various degrees by untreated or poorly treated urban sewage. Both fluorometers yielded consistent results when testing discrete samples. A nonlinear correlation (coefficient of determination = 0.98) was found between the tryptophan concentration given by the tryptophan field fluorometer and the fluorescence intensity given by a bench-top fluorometer (excitation = 285 nm, emission = 335 nm), corresponding to tryptophan-like fluorescing substances. A linear correlation with a mediocre coefficient of determination (0.63) was found between the NOM concentration given by the NOM field fluorometer and the fluorescence intensity given by the bench-top fluorometer (excitation = 355 nm, emission = 405 nm). This could be related to the diversity of NOM present, as illustrated by the different shapes of synchronous fluorescence spectra collected for the same samples.

Solid-state electrochemical assay of heme-binding molecules for screening of drugs with antimalarial potential.

The interaction between heme and ligands is the basis for a variety of tests aimed at the discovery of antiplasmodial molecules. Two electrochemical methods for the screening of molecules with potential antimalarial activity through heme-binding mechanism are described. The first method is applicable to lipophilic environment, by using solution phase electrochemistry in DMSO solutions of Fe(III)-heme plus the tested compounds at carbon electrodes. This method provides well-defined voltammetric signals, characteristic of the heme-ligand (L) interaction. The second method involves aqueous media at biological pH and the use of voltammetry of immobilized particles, by means of microparticulate films of the tested compounds immersed into Fe(III)-heme solutions with no need of prior incubation. These methodologies are applied to the testing of heme-binding activity in macromolecular level systems like hemoglobin, or much more complex mixtures like total blood, erythrocytes, or hemolyzed samples.

On-line coupled capillary isotachophoresis-capillary zone electrophoresis in hydrodynamically closed separation system hyphenated with laser induced fluorescence detection.

An analytical method, based on a column coupling capillary ITP and CZE in a hydrodynamically closed separation mode hyphenated with the detection in the modular arrangement, was developed in this work. Analytical possibilities of this approach are demonstrated on the direct and ultrasensitive quantitative determination of quinine (QUI) in diluted real multicomponent ionic matrices (beverages, urine). The detection cell interface, with the rectangular arrangement of the optical channels inside, connected the separation capillary with the LIF detector via optical fibers in the on-column detection arrangement. ITP enabled the direct large volume (30 µL) injections of the diluted real matrices with an on-line sample pretreatment (preseparation, preconcentration) so that no external sample preparation (except for the dilution) was necessary for the separation of the analyte in the multicomponent ionic matrices. Due to the ITP sample preconcentration and intrinsic sensitivity of the LIF detection, very low concentration LOD (as low as 77 pg/mL), were reached at the same time. This was ca. two orders lower than the corresponding LOD achieved by the same 2D separation system with UV absorbance detection. Compared to the single column CE-LIF methods applied for this model analyte and matrix, this method was found to be superior in terms of concentration LOD, with acceptable selectivity and benefits of the on-line sample preparation. A food control and bioanalytical application clearly illustrates great practical possibilities and routine use of the proposed modular ITP-CZE-LIF technique.

Chemical contamination during the preparation of cytotoxics: validation protocol for operators in hospital pharmacies.

BACKGROUND AND OBJECTIVES: The chemical contamination during the preparation of cytotoxics remains a serious problem in hospital pharmacies and the operators could contribute to this risk during their manipulations. A validation protocol was developed using a non-toxic, highly detectable tracer, quinine dihydrochloride. METHOD: Further, a method for a high recovery extraction and quantification of this marker, and a protocol covering the critical operations of cytotoxic preparation, was developed and validated. Various devices were used to fill the syringes and perfusion bags. All the filled containers and used materials were collected at the end of the protocol and the tracer was extracted in water. The contaminated water was analyzed by fluorimetry. The number of spots on the working pads was counted under ultraviolet light. During a total of 28 sessions, the procedure was applied by 20 different operators. RESULTS: The mean cumulated quantities of contamination were 6.2 µL (0.6-23.8) and >10 spots (0-20), which was considered as high. No correlation was observed between the contamination rate and the operator's experience. CONCLUSION: This validation protocol facilitates controlling the operators' working 'cleanliness' and helps to improve the initial and continuing training. This simple test presents an effective answer for the important issue of the chemical safety of operators.

Quinine localizes to a non-acidic compartment within the food vacuole of the malaria parasite Plasmodium falciparum.

BACKGROUND: The naturally fluorescent compound quinine has long been used to treat malaria infections. Although some evidence suggests that quinine acts in the parasite food vacuole, the mechanism of action of quinine has not yet been resolved. The Plasmodium falciparum multidrug resistance (pfmdr1) gene encodes a food vacuolar membrane transporter and has been linked with parasite resistance to quinine. The effect of multiple pfmdr1 copies on the subcellular localization of quinine was explored. METHODS: Fluorescence microscopy was used to evaluate the subcellular localization of quinine in parasites containing different pfmdr1 copy numbers to determine if copy number of the gene affects drug localization. The acidotropic dye LysoTracker Red was used to label the parasite food vacuole. Time-lapse images were taken to determine quinine localization over time following quinine exposure. RESULTS: Regardless of pfmdr1 copy number, quinine overlapped with haemozoin but did not colocalize with LysoTracker Red, which labeled the acidic parasite food vacuole. CONCLUSIONS: Quinine localizes to a non-acidic compartment within the food vacuole possibly haemozoin. Pfmdr1 copy number does not affect quinine subcellular localization.

Polymer-modified glassy carbon electrode for the electrochemical detection of quinine in human urine and pharmaceutical formulations.

Glassy carbon electrode was modified by electropolymerization of 4-amino-3-hydroxynaphthalene sulfonic acid. Cyclic voltammetric study of quinine showed higher current response at the modified electrode compared to the bare and activated glassy carbon electrodes in pH 7.0 phosphate buffer solution. Under optimized conditions, a calibration curve was obtained by square wave voltammetry at the modified electrode. The linear relationship between the peak current and the concentration of quinine in the range of 1.0 × 10(-7) to 1.0 × 10(-5) M was I (pa) (in microamperes) = 6.26C (in micromolars) + 0.2997 (R (2) = 0.999). The detection limit calculated (S/N = 3) was 1.42 × 10(-8) M, which is much lower than similar reports. The method was successfully applied for the determination of quinine in spiked human urine, and pharmaceutical formulations and recovery values >90 % were obtained.

Determination of quinine in beverages by online coupling capillary isotachophoresis to capillary zone electrophoresis with UV spectrophotometric detection.

The present study illustrates the possibilities of capillary isotachophoresis (CITP) online coupled with capillary zone electrophoresis (CZE) and hyphenated with fiber-based spectrophotometric diode array detection (DAD) for the direct, highly reliable, and ultrasensitive determination of quinine (QUI) in real multicomponent ionic matrices (beverages). Here, the CITP provided an effective online sample pretreatment (preseparation and preconcentration) prior to the CZE separation. Due to the CITP sample preconcentration, a simple UV-visible absorbance spectrophotometric detection was sufficient for obtaining very low concentration limits of detection (~2.3 ng/mL). Enhanced separation selectivity due to the combination of different separation mechanisms (CITP vs. CZE) enabled to obtain a pure analyte zone, suitable for its detection and quantitation in the directly injected real samples. The spectrophotometric DAD, unlike single wavelength UV detection, enabled to characterize the purity (i.e. spectral homogeneity) of the analyte zone and preliminary data indicate structurally related compounds via characteristic spectra recorded in the interval of 200-600 nm. The proposed CITP-CZE-DAD method was characterized by favorable performance parameters (sensitivity, linearity, precision, recovery, accuracy, robustness, and selectivity) and successfully applied to the control of QUI and potential QUI impurities in commercial beverages. This method is proposed as a routine automatized method for the highly reliable quality food control.

Rapid and green analytical method for the determination of quinoline alkaloids from Cinchona succirubra based on Microwave-Integrated Extraction and Leaching (MIEL) prior to high performance liquid chromatography.

Quinas contains several compounds, such as quinoline alkaloids, principally quinine, quinidine, cinchonine and cichonidine. Identified from barks of Cinchona, quinine is still commonly used to treat human malaria. Microwave-Integrated Extraction and Leaching (MIEL) is proposed for the extraction of quinoline alkaloids from bark of Cinchona succirubra. The process is performed in four steps, which ensures complete, rapid and accurate extraction of the samples. Optimal conditions for extraction were obtained using a response surface methodology reached from a central composite design. The MIEL extraction has been compared with a conventional technique soxhlet extraction. The extracts of quinoline alkaloids from C. succirubra obtained by these two different methods were compared by HPLC. The extracts obtained by MIEL in 32 min were quantitatively (yield) and qualitatively (quinine, quinidine, cinchonine, cinchonidine) similar to those obtained by conventional Soxhlet extraction in 3 hours. MIEL is a green technology that serves as a good alternative for the extraction of Cinchona alkaloids.

[Therapeutic drug monitoring of quinine]. / Suivi thérapeutique pharmacologique de la quinine.

Quinine is an antimalarial agent whose main mechanism of action on Plasmodium is to inhibit the transformation of toxic haem to polymeric non-toxic haemozoin. After oral and intramuscular administration, quinine is well absorbed, with peak plasma concentration reached in 1 to 3 hours. The pharmacokinetic of quinine differs depending on the severity of the disease: the volume of distribution and the clearance decrease proportionally to the infection, while the half-life increases. Plasma concentrations are approximately 50% higher in patients in the acute phase than in convalescence. Quinine is metabolized primarily by CYP3A4, implying changing the dosage when combined with inhibitors or inducers of CYP. The efficacy of quinine has been proved for residual concentrations above 5 mg/L (15 µmol/L) throughout the duration of treatment. Some side effects are concentration-dependent and a concentration of 20 mg/L (60 µmol/L) is considered as the threshold for toxicity. The 2007 consensus conference of the French Language Infectious Diseases Society calls for daily monitoring of plasma concentrations during the first 3 days of treatment targeting a trough concentration between 10 and 12 mg/L (30-36 µmol/L). For this compound, the level of evidence of the interest of therapeutic drug monitoring has been evaluated and the latter is recommended.

An interlaboratory investigation on the use of high-performance thin layer chromatography to perform assays of lamivudine-zidovudine, metronidazole, nevirapine, and quinine composite samples.

Two laboratories extensively investigated the use of HPTLC to perform assays on lamivudine-zidovudine, metronidazole, nevirapine, and quinine composite samples. To minimize the effects of differences in analysts' technique, the laboratories conducted the study with automatic sample application devices in conjunction with variable-wavelength scanning densitometers to evaluate the plates. The HPTLC procedures used relatively innocuous, inexpensive, and readily available chromatography solvents used in the Kenyon or the Global Pharma Health Fund Minilabs TLC methods. The use of automatic sample applications in conjunction with variable- wavelength scanning densitometry demonstrated an average repeatability or within-laboratory RSD of 1.90%, with 73% less than 2% and 97% at 2.60% or less, and an average reproducibility or among-laboratory RSD of 2.74%.

Portable and benchtop Raman spectrometers coupled to cluster analysis to identify quinine sulfate polymorphs in solid dosage forms and antimalarial drug quantification in solution by AuNPs-SERS with MCR-ALS.

This paper proposes for the first time: (a) a qualitative analytical method based on portable and benchtop backscattering Raman spectrometers coupled to hierarchical cluster analysis (HCA) and multivariate curve resolution - alternating least-squares (MCR-ALS) to identify two polymorphs of antimalarial quinine sulfate in commercial pharmaceutical tablets in their intact forms and (b) a quantitative analytical method based on gold nanoparticles (AuNPs) as active substrates for surface-enhanced Raman scattering (SERS) in combination with MCR-ALS to quantify quinine sulfate in commercial pharmaceutical tablets in solution. The pure concentration and spectral profiles recovered by MCR-ALS proved that both formulations present different polymorphs. These results were also confirmed by two clusters observed in the HCA model, according to their similarities within and among the samples that provided useful information about the homogeneity of different pharmaceutical manufacturing processes. AuNPs-SERS coupled to MCR-ALS was able to quantify quinine sulfate in the calibration range from 150.00 to 200.00 ng mL-1 even with the strong overlapping spectral profile of the background SERS signal, proving that it is a powerful ultrahigh sensitivity analytical method. This reduced linearity was validated throughout a large calibration range from 25.00 to 175.00 µg mL-1 used in a reference analytical method based on high performance liquid chromatography with a diode array detector (HPLC-DAD) coupled to MCR-ALS for analytical validation purposes, even in the presence of a coeluted compound. The analytical methods developed herein are fast, because second-order chromatographic data and first-order SERS spectroscopic data were obtained in less than 6 and 2 min, respectively. Concentrations of quinine sulfate were estimated with low root mean square error of prediction (RMSEP) values and a low relative error of prediction (REP%) in the range 1.8-4.5%.