Simultaneous separation of atovaquone, proguanil and its metabolites on a mixed mode high-performance liquid chromatographic column

An isocratic high-performance liquid chromatographic (HPLC) method for simultaneous separation of the components in the antimalarial combination drug Malarone^® with UV detection is described. An HPLC system using a mixed mode column composed of 50% C₁₈ phase and 50% strong cation-exchanger has been optimised for the simultaneous separation of atovaquone, proguanil and its two main metabolites. The mobile phase was optimised for factors such as pH, counter ion concentration and acetonitrile. Elimination of interferences from other antimalarial drugs was achieved by adding sodium perchlorate to the mobile phase. With a mobile phase of acetonitrile-phosphate buffer (60:40, v/v) pH 6.8, 50.7 mmol l⁻¹ K⁺ and 10 mmol l⁻¹ Na·ClO₄, separation was achieved within a run time shorter than 17 min.     

Mortality patterns in trout (Salmo trutta and S. gairdneri) exposed to an anionic detergent in relation to concentration and mechanisms of toxic action

SUMMARY. Histological and electron microscopical study of the gills of fish poisoned by the anionic detergent sodium lauryl sulphate indicates that the nature of the toxic action changes at a concentration of about 120 mg I⁻¹. Toxicity tests were conducted to determine whether this change could be detected in the test data, using brown trout ( Salmo trutta ) and rainbow trout ( S. gairdneri ).
Toxicity curves showed no indication of the change in toxic action, but at concentrations of 120 and 130 mg I⁻¹ of sodium lauryl sulphate 'split' probit lines occurred, and the lines for 150 mg I⁻¹ consistently differed in slope from other lines.     

A histological study of the response of the interrenal cells of the goldfish (Carassius auratus) to treatment with sodium lauryl sulphate

Treatment of goldfish ( Carassius auratus ) with sub-lethal concentrations of an anionic detergent sodium lauryl sulphate for 1, 2 or 4 weeks produced histological changes in the interrenal (steroidogenic) cells of the head kidney which are indicative of a cellular activation. Significant increases in both the nuclear diameter of these cells and the number of nucleoli contained in the nuclei occurred after 1 week's treatment, although these effects were more pronounced after 2 and 4 weeks. There were little associated alterations in chromaffin cell activity. Preliminary experiments with another pollutant, zinc sulphate, suggest that it exerts a similar action to sodium lauryl sulphate on the interrenal cells. It is suggested the increases in corticosteroid production which occur after exposure of fish to sub-lethal concentrations of pollutants may be important in the development of a generalized disease syndrome and to the long-term success of fish populations.     

Sulphate ion influx and efflux inhydrodictyon reticulatum

Sulphate ion is accumulated in the cells of the algaHydrodidyon reticulatum to a concentration approximately 30 mmol 1^-1. The rate of SO₄ ^2- uptake is increased markedly in the light, however, the effect is observable only after the cells have been kept in the light for about one hour. Uptake of sulphate is strongly inhibited by phosphorylation uncouplers and also by DIDS. The two-phase sulphate uptake kinetics, reported earlier was confirmed in experiments in which competition with phosphate anion was tested. Phosphate competes with sulphate only in the range of higher substrate concentrations (from 0.2 mmol 1^-1) and does not affect the system which works at low substrate concentrations. Efflux of SO₄ ^2- is very slow; the reason of the flux hindrance is not yet known.     

Attachment of fibroblasts to a polyanionic surface promoted by adenosine and prostaglandin E1

Dextran sulphate, which inhibits the growth of normal fibroblasts, marginally reduces their rate of initial attachment to polystyrene dishes. Transformed cells grow readily in dextran sulphate but their attachment is markedly inhibited. Adenosine and prostaglandin E₁ promote the attachment of transformed cells in the presence and absence of dextran sulphate. On dishes pretreated with serum and dextran sulphate, concentrations of both adenosine and PGE₁ as low as 10⁻⁸ M are active. Other purine derivatives and prostaglandins are inactive. The effect is discussed in relation to the properties common to these two substances in other cell systems.     

Identification and quantification of dehydroepiandrosterone sulphate in saliva

3β-Hydroxy-5-androsten-17-one (dehydroepiandrosterone) sulphate has been separated from an extract of human saliva by ion exchange gel chromatography and identified by high resolution gas chromatography-high resolution mass spectrometry of the $\text{tert}$-butyldimethylsilyl derivative of the neutral steroid obtained by enzymic hydrolysis. Quantitative analyses, employing 7,7?²H-dehydroepiandrosterone sulphate as the internal standard, have indicated concentrations in the saliva of young adult subjects to be generally in the range 0.9?5.7 nmol/l, though concentrations as high as 1O.7 nmol/l have been observed.     

Effects of zinc and cholesterol/choleate on serum lipoproteins and the liver in rats

The effects of short-term treatment with orally-administered zinc sulphate and/or a mixture of cholesterol/choleate on serum lipoprotein and hepatic enzyme levels were studied. Administration of graded doses of zinc sulphate (20 or 40 mg/kg, as zinc ion) for 5 days, dose-dependently increased serum and hepatic zinc levels but depressed the serum high-density lipoprotein-cholesterol (HDL-C) concentration and liver cytochrome P-450 activity. However, it did not affect hepatic concentrations of malondialdehyde and free beta-glucuronidase. Cholesterol/choleate treatment for 5 days markedly damaged the liver, as reflected by elevations of hepatic concentrations of malondialdehyde (both in the mitochondrial and microsomal fractions) and of free beta-glucuronidase; total cholesterol and low-density lipoprotein-cholesterol in the blood were increased, whereas HDL-C was decreased significantly. Concomitant administration of zinc sulphate with cholesterol/choleate further lowered HDL-C levels, but reversed the high hepatic concentrations of both malondialdehyde and free beta-glucuronidase. The present study indicates that both zinc ions and cholesterol can decrease circulatory HDL-C levels and that zinc protects against cholesterol-induced hepatic damage by reducing lysosomal enzyme release and preventing lipid peroxidation in the liver.     

Bile and the absorption of strontium and iron

1. Strontium absorption was studied in vivo with loops of ileum in rachitic chicks and found to be increased by vitamin D(3), bile salts and sodium lauryl sulphate. 2. Bile salts and sodium lauryl sulphate rendered strontium soluble in butanol-benzene (1:1, v/v). 3. Bile was not concerned in the absorption of iron in rats from its water-soluble form, ferrous sulphate. 4. Ligation of the bile ducts in rats caused a decrease in the absorption of iron presented as its sparingly soluble phosphate. 5. The effect of bile on cation absorption is discussed.     

Toxic action of several lethal concentrations of an anionic detergent on the gills of the brown trout (Salmo trutta L.)

The pathological effects often lethal concentrations of the anionic detergent, sodium lauryl sulphate, on the gills of Salmo trutta L. have been studied by light and electron microscopy. At concentrations to 120 mg/1 (medium survival times >1 h), epithelial cell death is associated with lysosome formation. Acute inflammation of the gill tissue, extensive detachment of the epithelium and, except at the lowest concentrations, collapse of the pillar cell system occur. At concentrations above 120 mg/1 (medium survival times <1 h) very rapid lysis of cells results in the complete disruption of cellular and tissue structure. Changes in the gross structure of the gills are explainable in terms of the rate and nature of toxic action at the cellular level. Review of the biomedical literature suggests the observed effects of sodium lauryl sulphate on gill cells correspond to the two mechanisms by which detergents cause death in isolated cells. These are autolysis, i.e. lysis by the action of the cell's own enzymes, induced by an initial lesion in the cell membrane whose precise nature is not known; and rapid lysis by the direct action of the detergent on the cell constituents.     

Increased susceptibility of injured spores of Bacillus subtilis to cationic and other stressing agents

Sublethally heated spores of Bacillus subtilis NCTC 8236 were susceptible to posttreatment concentrations in agar of polymyxin B sulphate, sodium hydroxide, cetylpyridinium chloride and sodium lauryl sulphate that did not prevent colony formation to untreated spores. The non-ionic surfactants polysorbates 20 and 80 were not inhibitory when used at high concentrations against both heated and unheated spores. The method has been developed for detecting sublethal injury in biocide-exposed spores, since iodine-treated spores became highly susceptible to polymyxin contained in recovery agar.     

Relapses after Withdrawal of Proguanil Treatment in Tropical Splenomegaly Syndrome

After the remission of symptoms and reduction in spleen size while on proguanil therapy four patients with the tropical splenomegaly syndrome defaulted from treatment. The withdrawal of proguanil caused a recrudescence of original symptoms, splenomegaly, and a return of the initially raised serum IgM. Complete return to normal values was again effected with proguanil therapy.The role of the spleen in the tropical splenomegaly syndrome in the production of the raised serum IgM is discussed. These patients should be educated as to the nature of their disease and the importance of continued medical treatment.     

Enhancers of the non-peroxidative metal porphyrin chemiluminescence reaction

Metal porphyrins catalyse luminol chemiluminescence at pH 13 without added peroxide. The effects of 22 different surface active compounds on this reaction were studied using six metal porphyrins and one metal porphyrin conjugate. The most active catalyst was Mn-meso-tetra(4-sulphonatophenyl)porphine. Tween-20 enhanced the activity of this catalyst best at a Tween-20 to luminol ratio of 74:1. However, lauryl sulphate enhanced best at an optimum lauryl sulphate to luminol ratio of over 1000:1 and both detergents enhanced the reaction when present below their critical micelle concentrations. Negatively charged aliphatic compounds such as fatty acids enhanced the reaction but positive-charged aliphatic compounds inhibited it. Small differences in enhancer structure resulted in differing enhancement. For example, linoleic acid enhanced Mn-meso-tetraphenyl porphine more than 10-fold, yet linolenic acid inhibited this catalyst. Conjugation of a metal porphyrin to antibody did not influence its enhancement by detergents. The results indicate that the enhancement mechanism does not require formation of pure detergent micelles but that direct association between enhancer and catalyst may be important.     

Isolation of a pregnancy-associated globulin (PAG) from the rat and preparation of a specific antiserum (anti-PAG)

Isolation of a glycoprotein from serum of pregnant rats has been accomplished by ammonium sulphate precipitation, gel filtration, ion exchange and affinity chromatography. Low concentrations of the protein were detectable in the serum of some of the male rats tested, while somewhat higher concentrations were detected also in the serum of non-pregnant female rats. No relationship could be established between this protein and the well known pregnancy-specific proteins or the alpha 2-acute phase-macroglobulin of the rat, whereas evidence was obtained for a strong cross-reaction with a serum protein of pregnant mice.     

Chemical and Isotopic Switching within the Subglacial Environment of a High Arctic Glacier

Natural environmental isotopes of nitrate, sulphate and inorganic carbon are discussed in conjunction with major ion chemistry of subglacial runoff from a High Arctic glacier, Midre Lovénbreen, Svalbard. The chemical composition of meltwaters is observed to switch in accordance with subglacial hydrological evolution and redox status. Changing rapidly from reducing to oxidizing conditions, subglacial waters also depict that ¹⁵N/¹⁴N values show microbial denitrification is an active component of nutrient cycling beneath the glacier. ¹⁸O/¹⁶O ratios of sulphate are used to elucidate mechanisms of biological and abiological sulphide oxidation. Concentrations of bicarbonate appear to be governed largely by the degree of rock: water contact encountered in the subglacial system, rather than the switch in redox status, although the potential for microbiological activity to influence ambient bicarbonate concentrations is recognised. Glaciers are therefore highlighted as cryospheric ecosystems supporting microbial life which directly impacts upon the release of solute through biogeochemically mediated processes.     

Effects of sulphate and pH on the plant-availability of phosphate adsorbed on goethite

The adsorption of phosphate on metal (hydr)oxides may be influenced by the pH and by the adsorption of other ions. In this study, the influence of sulphate and pH on phosphate adsorption on goethite and the availability to plants of adsorbed phosphate was examined. Maize plants were grown on suspensions of goethite with adsorbed phosphate, containing the same total amount of phosphate and either 0.11 mM or 2.01 mM sulphate at pH 3.7, 4.6 or 5.5. The uptake of phosphorus by the plants increased with the larger sulphate concentration and decreasing pH. Mean P uptake in the treatment with 2.01 mM sulphate and pH 3.7 was 55 µmol plant^-1, whereas in the treatment with 0.11 mM sulphate and pH 5.5 it was 2 µmol plant^-1. Batch adsorption experiments using³² P and speciation modelling of ion adsorption showed that in the presence of sulphate, the phosphate concentration in solution strongly increased with decreasing pH, due to competitive adsorption between sulphate and phosphate on goethite. Modelled phosphate concentrations in solution in the uptake experiment were all below 0.6 µM and correlated well with the observed P uptake. This correlation indicates that the strong influence of the sulphate concentration and pH on the plant-availability of adsorbed phosphate results from the competition between sulphate and phosphate for adsorption on goethite.     

An improved method for the extraction of prostaglandins

Present methods of extracting prostagalndins (PGs) give poor recoveries from synovial fluid, probably because the PGs bind to protein and are lost in the precipitation stage of extraction. Addition of the nonpolar detergent sodium lauryl sulphate prior to extraction improves recovery of PGs. It is suggested that sodium lauryl sulphate competes with PGs for the binding sites.     

Sulphate reduction in the hypolimnion and sediments of Lake Kinneret, Israel

1. Lake Kinneret is a warm (13–30°C) monomictic lake. Between January and June a heavy annual bloom of the dinoflagellate Peridinium gatunense dominates phytoplankton biomass (250 g m^?2). At the beginning of the summer, degradation and decomposition of the Peridinium biomass occurs, serving as a trigger for intense sulphate reduction in the hypolimnion and sediments. 2. The rates of sulphate reduction in the sediments varied seasonally from 12 to 1700 nmol SO₄.^?2 reduced cm^?3 day^?1 in December and July, respectively. The availability of organic matter and sulphate is high in June after the crash of the Peridinium bloom and the beginning of stratification and is lowest in December before overturn. 3. Sulphate concentrations in the hypolimnion range between 0.52 mM and 0.20 mM during mixing (January-April) and before overturn (December), respectively. The depletion in sulphate in the hypolimnion is stoichiometrically correlated to the increase in sulphide. The lake is not depleted of sulphate at any time, so the sulphate reduction process in Lake Kinneret is not limited by sulphate concentrations except in the sediments just before overturn.     

Nutrient fluxes in a snow-dominated, semi-arid forest: Spatial and temporal patterns

We tested five hypotheses regarding the potential effects of precipitation change on spatial and temporal patterns of water flux, ion flux, and ion concentration in a semiarid, snowmelt-dominated forest in Little Valley, Nevada. Variations in data collected from 1995 to 1999 were used to examine the potential effects of snowpack amount and duration on ion concentrations and fluxes. Soil solution NO₃ ^–, NH₄ ⁺, and ortho-phosphate concentrations and fluxes were uniformly low, and the variations in concentration bore no relationship to snowmelt water flux inputs of these ions. Weathering and cation exchange largely controlled the concentrations and fluxes of base cations from soils in these systems; however, soil solution base cation concentrations were affected by cation concentrations during snowmelt episodes. Soil solution Cl^– and SO₄ ^2– concentrations closely followed the patterns in snowmelt water, suggesting minimal buffering of either ion by soils. In contrast to other studies, the highest concentration and the majority of ion flux from the snowpack in Little Valley occurred in the later phases of snowmelt. Possible reasons for this include sublimation of the snowpack and dry deposition of organic matter during the later stages of snowmelt. Our comparison of interannual and spatial patterns revealed that variation in ion concentration rather than water flux is the most important driver of variation in ion flux. Thus, it is not safe to assume that changes in total precipitation amount will cause concomitant changes in ion inputs to this system.     

Phase separation in an aqueous quaternary system

(1) We have measured the incompatible phase separation that occurs in a polyethylene glycol-sodium dextran sulphate-sodium chloride-water system and have determined a critical point. (2) We have measured the activity coefficients of sodium chloride in critical-point concentrations of polyethylene glycol and sodium dextran sulphate respectively, and the osmotic coefficient of sodium dextran sulphate at the critical-point concentration. (3) With use of the relevant thermodynamic equations for a quaternary ionic system, we have determined the interaction coefficients between polyethylene glycol and dextran sulphate and between polyethylene glycol and sodium chloride. The former could be due mainly to volume exclusion, but the latter is too large to be explained on that basis.     

Intermittent preventive therapy for malaria: arguments in favour of artesunate and sulphamethoxypyrazine - pyrimethamine combination

The case of a 32-year-old Caucasian female with multi-drug resistant HIV-1 subtype B infection treated with a salvage regimen including maraviroc, raltegravir, etravirine and unboosted saquinavir who started atovaquone/proguanil prophylaxis, is reported. The potential interactions between atovaquone/proguanil and these anti-retroviral drugs are investigated. Pharmacokinetic analyses documented a marked increase in etravirine and saquinavir plasma concentrations (+55% and +274%, respectively), but not in raltegravir and maraviroc plasma concentrations. The evidence that atovaquone/proguanil significantly interacts with etravirine and saquinavir, but not with raltegravir and maraviroc, suggests that the mechanism of interaction is related to cytochrome P450.