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Phospholipids, such as phosphatidic acid, suppress bitter taste without affecting other taste qualities. In the present study, we detected and quantified this suppression effect with an electronic tongue whose transducer is composed of several kinds of lipid/polymer membranes with different characteristics. We measured a phospholipid cocktail and various kinds of taste substances with five basic taste qualities. The responses to quinine hydrochloride and L-tryptophan, which have a bitter taste, were reduced as the phospholipid concentration was increased, and the responses to the other taste substances were not affected by the phospholipids, as with the human sensation test. Furthermore, the change of bitter intensity caused by phospholipid was quantified by principal component analysis and the tau scale, which expresses the relationship between taste intensity and taste substance concentration. The results are compared with those of the human sensory test and discussed., (Copyright 2001 Wiley-Liss, Inc. and the American Pharmaceutical Association)

Potentiometric response characteristics were evaluated for quinine selective sensors based on a lipophilic ion-exchanger potassium tetrakis[3,5-bis(trifluoromethylphenyl)]borate (PTFB) immobilized together with plasticizing solvents in polyvinyl chloride membranes. The use of dioctyl phthalate (DOP), 2-nitrophenyl phenyl ether (NPPE), and bis(2-ethylhexyl)adipate (BEHA) plasticizers produced good quality quinine sensors that were sensitive and fast responding, and exhibited near Nernstian responses when used as batch-sensors. These membranes were further tested in a wall-jet flow-through potentiometric flow injection analysis (FIA) detector. Quinine sensors containing BEHA were the most suitable membrane, with no noticeable differences in sensitivity even after 5 h of continuous exposure to solutions. Interference by foreign species such as alkali, alkaline earth metal ions, sugars, and sodium benzoate was minimal in either the batch-mode (log selectivity coefficients <-2.60) or as FIA detector. When the FIA method was used, recoveries averaged 99.70% from mineral water samples spiked with 16.22 and 162.20 ppm quinine. Results for determination of quinine in other drinks such as carbonated (64.88 and 71.37 ppm) and bitter lemon drinks (55.15 ppm) agreed with those of the manufacturer's claimed values of <68, <81, and 51 ppm, respectively. With a sample throughput of 50 samples/h under optimized flow conditions, the method can be routinely used to monitor quinine in such samples.

Background & Objectives: Though a number of barrier methods and potent spermicides are available, most of these have nonoxynol-9 (N-9) as the active ingredient which is observed to cause inflammation and genital ulceration on repeated use. The present study was undertaken to develop a safe spermicide with conjoint microbicidal properties., Methods: A polyherbal pessary was formulated with purified ingredients from neem (Azadirachta indica) leaves, Sapindus mukerossi (pericarp of fruit) and Mentha citrata oil. Spermicidal action on human sperm was tested by Sander-Cramer slide test in vitro and by post coital tests in vivo. Contraceptive action was tested in rabbits., Results: The combination of the three herbal ingredients resulted in the potentiation of the spermicidal action by 8-folds. The post coital tests confirmed the spermicidal properties of the Praneem polyherbal pessary (PPP) in women with high cervical mucous score around mid estrus. It also prevented in most women the migration of sperm into the cervical mucous. In 15 rabbits studied pregnancy was prevented by the intravaginal administration of PPP, whereas 13 of the 15 animals in the control group became pregnant., Interpretation & Conclusion: The Praneem polyherbal pessary has potent spermicidal action on human sperm in vitro and in vivo. When applied in the vagina before mating, it prevented rabbits from becoming pregnant.

Thermal treatment of aqueous solutions of xylose and primary amino acids led to rapid development of a bitter taste of the reaction mixture. To characterize the key compound causing this bitter taste, a novel bioassay, which is based on the determination of the taste threshold of reaction products in serial dilutions of HPLC fractions, was developed to select the most intense taste compounds in the complex mixture of Maillard reaction products. By application of this so-called taste dilution analysis (TDA) 21 fractions were obtained, among which 1 fraction was evaluated with by far the highest taste impact. Carefully planned LC-MS as well as 1D and 2D NMR experiments were, therefore, focused on the compound contributing the most to the intense bitter taste of the Maillard mixture and led to its unequivocal identification as the previously unknown 3-(2-furyl)-8-[(2-furyl)methyl]-4-hydroxymethyl-1-oxo-1H,4H-quinolizinium-7-olate. This novel compound, which we name quinizolate, exhibited an intense bitter taste at an extraordinarily low detection threshold of 0.00025 mmol/kg of water. As this novel taste compound was found to have 2000- and 28-fold lower threshold concentrations than the standard bitter compounds caffeine and quinine hydrochloride, respectively, quinizolate might be one of the most intense bitter compounds reported so far.

The peak shape and retention of some basic probes together with a neutral reference compound were investigated as a function of temperature and flow-rate using a reversed-phase HPLC column at both pH 3.0 and pH 7.0. The retention of bases often showed an anomalous increase with temperature; retention mechanisms are complex as shown by studies of the effect of buffer cation concentration on retention. Considerable improvements in column efficiency for bases may result from operation at elevated temperature. Improvements did not seem attributable either to incidental changes in the retention factor, or (in this particular study where low sample masses were utilised) to the influence of sample load. The optimum flow-rate for highest efficiency is generally lower for basic compounds than neutrals, and due to the steepness of the Van Deemter curves obtained, high flow-rates appear to be particularly detrimental in the chromatography of basic compounds.

The stability of quinine was determined in various diluents and in polyvinyl chloride (PVC) containers. The release of diethyhexyl phthalate (DEHP) from PVC bags into intravenous infusions of quinine was also measured. We used an injection of two doses of quinine; quiniforme at 500 mg and quinimax at 400 mg in either 250- or 500-ml PVC infusion bags containing 5% dextrose, to give initial nominal concentrations of 2 or 1 mg ml(-1) quiniforme and 1.6 or 0.8 mg ml(-1) quinimax, the mean concentrations commonly used in clinical practice. Samples were assayed by stability-indicating high-performance liquid chromatography (HPLC) and the clarity was determined visually. Experiments were conducted to determine whether the stability and compatibility of quinine would be compromised, and whether DEHP would be leached from PVC bags and PVC administration sets during storage and simulated infusion. There was no substantial loss of quiniforme and quinimax over 1- or 2-h simulated infusion irrespective of the diluent, and storage during 8 h at 22 degrees C, 48 or 72 h at 4 degrees C and 96 h at 45 degrees C. Leaching of DEHP was also detected during simulated infusion delivery using PVC bags and PVC administration sets. The quantity was less than 2 microg ml(-1). During storage at 4 degrees C and room temperature the leaching of DEHP was low, but when the temperature was 45 degrees C the quantity was high, 21 microg ml(-1). To minimise patient exposure to DEHP, quinine solutions with all drugs should be infused immediately or stored for a maximum of 48 h at 4 degrees C.

A high-performance liquid chromatographic (HPLC) method with ultraviolet detection for the determination of quinine in rat biological fluids is described. Due to its selectivity and sensitivity, the proposed method can be used in the case of such rat biological fluids as cerebrospinal fluid (CSF) and perilymph for which the accessible volumes are limited to 100 microl and 10 microl, respectively. Consequently, the assay method has been applied to the measurements of quinine concentration in rat plasma, CSF and perilymph samples.

Aims: Our previous studies using in vitro hepatic microsomal preparations suggested that the hepatic metabolism of quinine to form the major metabolite 3-hydroxyquinine is most likely catalysed by human P450 3A (CYP3A). The present study was carried out to investigate the kinetics and to identify and further characterise the human liver CYP isoforms involved in the metabolism of quinine., Methods: In vitro human microsomal techniques were employed., Results: The mean apparent Km value for 3-hydroxyquinine formation was 83 +/- 19 (s.d.) microM, ranging from 57 microM to 123 microM in microsomes from ten human livers. There was a 6.7-fold variation in Vmax values (mean 547 +/- 416 pmol min-1 mg-1). Quinine 3-hydroxylation was inhibited by the specific CYP3A inhibitors, troleandomycin, midazolam and erythromycin. Inhibitors selective for CYP1A1/2, CYP2D6, CYP2E1, CYP2C9/10 or CYP2C19 had little or no effect on quinine 3-hydroxylation. Using microsomes from a panel of livers, significant correlations were found only between 3-hydroxyquinine activity and other CYP3A activities (caffeine 8-oxidation, omeprazole sulphoxidation, midazolam 1'-hydroxylation and midazolam 4-hydroxylation) and immunoreactive CYP3A content. There were no statistically significant correlation with activities selective for CYP1A2, CYP2C9 and CYP2E1. Competitive inhibition of quinine 3-hydroxylation was observed with a substrate known to be specifically metabolized by human CYP3A, i.e. midazolam, with an apparent Ki value of 11.0 microM., Conclusions: The present results strongly indicate that the conversion of quinine to 3-hydroxyquinine is the major metabolic pathway in human liver in vitro and that the reaction is catalysed by CYP3A isoforms.

A flow-through optosensor for cinchona alkaloids with C18 silica gel as a substrate is proposed. The sensor is developed in conjunction with a flow-injection analysis system and is based on the retention of the cinchona alkaloids on a C18 column and the enhancement of their fluorescence. The analytical performance characteristics of the proposed sensor for the detection and quantification of these alkaloids were as follows: the detection limits of quinine, cinchonine, quinidine and cinchonidine were 2.3, 31.6, 2.3 and 31.6 ng ml-1, respectively, with relative standard deviations of 0.9% for quinine and quinidine (20 ng ml-1, n = 7) and 1.1% for cinchonine and cinchonidine (4.0 micrograms ml-1, n =7), respectively. Most of the common species did not interfere. The recommended method has been successfully tested for determination of quinine in pharmaceutical preparations and soft drinks.

A rapid method for the determination of quinine in bitter drinks by micellar electrokinetic capillary chromatography (MECC) with ultraviolet (UV) detection is described. The beverage is simply diluted with deionized water, filtered, and analyzed using a 75 cm x 75 micrometer uncoated fused-silica capillary column with a buffer consisting of 15% methanol and 85% of a mixture of 0.05 M cetyltrimethylammonium bromide (CTAB), 0.01 M sodium tetraborate, and 0.01 M potassium dihydrogen orthophosphate, pH 8.6, with an operating voltage of -25 kV. The levels of quinine determined by MECC were in good agreement with those determined by HPLC. The CVs for area calculation (2.1%, n = 7) and migration time variation (1.3%, n = 20) for multiple injections of a sample solution by MECC were satisfactory.

The analysis of quinine in whole blood, plasma, urine, and samples dried on filter paper is described. Extraction was made with toluene followed by back-extraction into phosphate buffer. A reversed-phase liquid chromatography system with fluorescence detection was used. The within-day coefficient of variation of the method was 4-10% at the lower limit of determination (2 nM in plasma and 50 nM in whole blood, dried samples, and urine) and 2-4% at 10 microM. The quinine concentration was found to be lower in whole blood than in plasma (mean ratio, plasma-whole blood, 1.17). The concentration in capillary blood was lower than that in venous blood (mean ratio, capillary blood-venous blood, 0.93).

A new simple, selective and reproducible high-performance liquid chromatographic method for the determination of quinine in plasma, saliva and urine is described. The ion-pair method was carried out on a reversed-phase C18 column, using perchlorate ion as the counter ion and ultraviolet detection at 254 nm. Quinine was well resolved from its major metabolite, 3-hydroxyquinine, and the internal standard, primaquine. The limit of detection was 10 ng/ml and the recovery was greater than 90% from the three biological fluids.

A high-performance liquid chromatographic method with fluorescence detection is described for the simultaneous measurement of quinine, chloroquine and mono- and bidesethylchloroquine in human plasma, erythrocytes and urine. After a liquid-solid extraction on a Bond Elut C8 cartridge, the compounds are separated on an Inertsil silica column by gradient elution; the mobile phase is a mixture of acetonitrile and methanol-25% ammonia solution (92.7:7.5, v/v). The eluent was monitored with a fluorescence detector (excitation wavelength 325 nm and emission wavelength 375 nm). The limit of detection was ca. 5 ng/ml for chloroquine and ca. 23 ng/ml for quinine. No chromatographic interferences could be detected from endogenous compounds or other antimalarial drugs. The method is accurate with inter- and intra-assay coefficients of variation lower than 7%. Hydroxychloroquine is used as an internal standard because of its structural similarity to chloroquine. The procedure requires 30 min and can be used for therapeutic drug monitoring.

The use of high-performance liquid chromatography (HPLC) interfaced with thermospray (TSP) mass spectrometry is described for the separation and identification of various alkaloids from Cinchona ledgeriana extracts. The use of water-acetonitrile-acetic acid (71:25:4) with 0.01 M ammonium acetate (pH 3.0) as the mobile phase gave good HPLC separation and good TSP sensitivity. The specificity obtained by single-ion monitoring allowed the analysis of commercially important alkaloids such as quinine and quinidine in plant material, transformed roots and in cells from tissue culture, with relatively simple extraction and work-up procedures. TSP gave protonated species with few fragment ions but collision-induced dissociation offers the promise of increased analytically specificity from the fragment ion data. This work has important implications for the biotechnological production of pharmaceuticals normally obtained from plant sources.

The isocratic reversed phase high performance liquid chromatographic method proposed for quinidine metabolic studies facilitates particularly the separation of 10(R) and (S) isomers of quinidine 10,11-dihydrodiols. The finding of each of these forms following a new synthetic pathway allows us to identify and quantify them in biological fluids. These two isomers have especially been observed in rat bile and hepatocyte secretions. The metabolic inducing effect of phenobarbital on the oxidative metabolism of quinidine is verified in rat isolated hepatocytes. Simultaneous secretion of the two dihydrodiols is also verified in human urine by a gas chromatography/mass spectrometry procedure.

Quinine is determined quantitatively with a liquid-chromatographic method in various solutions using silica as a stationary and methanol + 0.5% ammonia as a mobile phase. The approximate concentrations of quinine were found to be between 50.0 to 72.0 mg/1 in the soft-drinks investigated. Beers, which were also investigated do not content quinine. The method presented is also useful for determining quinine in aqueous solutions of various origin.

Three thin-layer chromatographic systems have been selected on the basis of their discriminating power on which to standardise for the identification of basic drugs. They are systems of silica gel sprayed with 0.1 N NaOH, dried and run using one of the following solvents: cyclohexane-toluene-diethylamine (75:15:10), chloroform-methanol (90:10) or acetone. They can be used in combination since their correlation coefficients are low. Four reference compounds should be used, equally spaced across the plate. The inter-laboratory variation of measurement of RF values has been ascertained and the use of a graphical RF correction method reduced the mean deviation of measurement to 0.02. The measurement of RF values was found to be less reproducible in the middle of the chromatogram than in regions of very low or very high RF values.

A liquid chromatography method for the determination quinidine in serum and urine has been developed. This method uses isocratic conditions, ambient temperature, a conventional fixed wavelength, 254-nm detector, and is free of potential interference from quinidine metabolites. Sample pretreatment involves extraction of quinidine along with quinine, an internal standard, into an organic phase and reextraction into an aqueous acidic phase. In this manner, interference due to commonly used drugs are eliminated. The method is capable of accurately measuring quinidine to levels as low as 0.5 mg/L.

A liquid chromatographic (LC) method is described for the determination of quinine, hydroquinine, sodium saccharin, and sodium benzoate in soft drinks. The method involves simple sample preparation, direct injection onto an octadecylsilane column, and elution with a methanol-acetonitrile-water-acetic acid (20 + 10 + 70 + 1) mobile phase. Eluted constituents are measured spectrophotometrically at 254 nm. The relationship between peak height and concentration was linear between 20 and 120 micrograms/mL for quinine. A relative standard deviation of 0.82% was obtained for commercial samples spiked with quinine, and the average recovery was 100.3%. The proposed procedure is accurate and rapid and can also detect hydroquinine (a natural contaminant of quinine), sodium saccharin, and sodium benzoate. Linear responses ranged from 0.45 to 20 micrograms/mL for hydroquinine, from 54.8 to 219 micrograms/mL for sodium saccharin, and from 10.1 to 145.1 micrograms/mL for sodium benzoate. The reproducibility of the LC method was evaluated with standard solutions of hydroquinine, sodium saccharin, and sodium benzoate, which produced relative standard deviations of 0.42, 0.46, and 1.13%, respectively. The average recoveries for sodium saccharin and sodium benzoate from spiked samples were 99.4 and 100.2%, respectively.

Quinine can easily be determined in soft drinks by means of reversed-phase ion-pair chromatography. Sample preparation is not necessary. Quinine is detected by fluorescence and UV spectrophotometry by continuous monitoring of the column effluent. A rectilinear response is obtained in the concentration range of 20--100 mg/l. The complete assay procedure takes about 8 min.

We propose a method to quantify quinine in biological fluids using liquid chromatography. After alkalinisation, samples are extracted with a chloroform-isoamyl alcohol mixture; in the case of urinary samples, a preliminary enzymatic hydrolysis is performed. Extracts are analyzed by adsorption chromatography. A mobile phase composed of methanol-acetonitril-ammonia in used to elute. A post-column reaction with sulfuric acid allows a fluorimetric detection. With this method concentration as low as 0.05 mg/l can be measured. Variation coefficient is about 2.5 p. cent Quinine, its metabolites and dihydroquinine are well separated; thus it is a very useful method for pharmacokinetic and metabolic studies.

Among the analytical techniques available for the determination of toxic products, fluorimetry is, at present, one of the most frequently employed. But in many cases, measurements at low concentrations are interfered with by Rayleigh and Raman scattering. We propose to eliminate, in great part, this interference and hence increase the sensitivity of the method, by using the new synchronous excitation spectrofluorimetry. Application of this technique to the identification and determination of drugs during recent forensic investigations demonstrates its advantages over classical spectrofluorimetry.

The two characteristic erythroquinine and thalleioquin tests for quinine and quinidine were studied to optimize the experimental conditions for quantitative analysis. Both methods were quantitatively sensitive for either quinine or quinidine in a concentration range of 0.1--10 microgram/ml with the erythroquinine method and of 3--50 microgram/ml with the thalleioquin method. A TLC--colorimetric method also is described for the assay of quinine and quinidine in the presence of cinchonine, cinchonidine, and other cinchona alkaloids. The results were compared with those obtained with a spectrophotometric method.

Binary mixtures of weak and strong UV-absorbing alkaloids were analyzed by a charge-transfer spectrophotometric method, utilizing iodine in ethylene dichloride as the acceptor. In the uncomplexed form, the strong absorbing alkaloid (papaverine, quinine, ergotamine, or reserpine) was measured at a wavelength where there was no interference from weak absorbers (at 335, 332, 315, or 300 nm, respectively). The weak absorbing alkaloid (ephedrine, codeine, atropine, or homatropine methylbromide) was determined by computing its contribution to the total charge-transfer band at 295 nm where absorbance was linearly additive for mixtures. The greater increase in the original epsilon-values of the weak absorbers upon complexation with iodine relative to the corresponding increase in the epsilon-values of the strong absorbers led to good recoveries even at the low dose ratios of the weakly absorbing, and often more potent, alkaloids.

The suitability of isotachophoresis for the determination of quinine in different samples was investigated. The operational conditions were 0.01 M potassium-morpholinoethanesulphonic acid (MES) (pH 6.0) with 0.05% Mowiol as the leading electrolyte and ca. 0.005 M creatinine-MES as the terminating electrolyte. The analyses were carried out at 25 microA in a 0.2 mm I.D. PTFE capillary with UV and conductivity detection. Quinine-containing beverages were degassed by sonification and directly injected. The limit of detection was 5 mg/l with a 4 microliter injection volume. The allowed concentrations could be determined with sufficient accuracy. Analgesic preparations were dissolved in a solution of 5 X 10(-3) M MES with sonification. The quinine levels found agreed well with the declared values. The other constituents of the pharmaceuticals did not interfere with the analysis. Urine samples from volunteers were analysed after consumption of tonic. The samples were extracted with dichloromethane-isopropanol (95:5), vortexed, centrifuged, evaporated to dryness, the residue dissolved in 5 X 10(-3) M MES and analysed. At a concentration factor of 33, the limit of detection was ca. 60 micrograms in 48-h urine: 2-15% of the quinine consumed was excreted as the parent compound in the first 48 h after consumption. The combination of the extraction procedure and the operational system makes the method suitable for the determination of a number of other alkaloids in physiological samples.

Antimalarial therapy is directed more and more towards the use of associated substances: Pyrimethamine, sulfadoxime, quinine et chloroquine. The authors propose a method of direct determination of these four molecules mixed in biological fluids. The recommended extraction process followed by a spectrophotometric UV determination is fast, reproductible and sufficiently sensitive to study the evolution of a therapy.

A method is presented for the positive identification of quinine and quinidine. The method requires extraction of the compounds from an alkaline solution into an organic solvent and concentration of the sample by evaporation of the solvent. The sample is chromatographed without derivatization and may be detected by the flame ionization detector (FID) or mass spectrometry (MS). Limits of sensitivity are less than 5 ng per assay for the underivatized compounds. Analysis of urine extracts is discussed.