The assay of benzalkonium chloride in pilocarpine, hypromellose and polyvinyl alcohol ophthalmic drops by second-order derivative ultraviolet spectrophotometry.
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Benzalkonium Compounds↗
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Mesenteric neurons in the adult rat are responsive to ileal treatment with benzalkonium chloride.
The application of a 2 mM solution of the cationic surfactant benzalkonium chloride (BAC) to an ileal segment produced a selective and extensive myenteric denervation. The aim of the present study was to investigate whether such a selective unbalance of the enteric nervous system in the adult rat elicited any plastic response within the mesenteric nervous structures contacting it. Acetylcholinesterase (AChE) staining, beta-nicotinamide adenine dinucleotide (NADH) and beta-nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry and glyoxylic acid-induced fluorescence were performed on whole-mount preparations of myenteric plexus and mesenteric nerves. In both control and BAC-treated animals nervous elements were detected along the mesenteric nerves. Although rather similar in position, shape and size, these neurons displayed striking differences with regard to their arrangement and density per nerve. In the controls, few small aggregates of neurons could be detected; more commonly, isolated nerve cell bodies were scattered along the nerve trunks. In the BAC-treated animals, numerous spherical or spindle-shaped clusters of neurons were located along the nerves; the mean neuronal density per nerve displayed a two-five-fold increase over the control values. The observed changes within the mesenteric nerves might be involved in an attempt at reinnervation of the BAC-treated intestinal segments from extra-enteric sources.
Acetylcholinesterase↗
Chemical denervation of the myenteric plexus of the muscular stomach of turkeys.
1. The thin caudoventral muscle (TCM) of the muscular stomach of domestic turkeys was surgically exposed and painted with solutions of saline or 0.1, 0.25, 0.5, and 1.0% benzalkonium chloride (BC), a cationic surfactant shown to irreversibly damage neurons but not muscle tissue in mammals. 2. Following fluoroscopic observations of gastric motility for 2 weeks, turkeys were euthanized, the entire muscular stomach was excised and weighed, and serial frozen sections of the TCM were taken for evaluation of the number and size of neurons in the myenteric plexus. 3. Treatment with 0.5 and 1.0% BC resulted in loss of motility in the TCM, significant hypertrophy (P less than 0.001) of the CTM, a 70% decrease in number and 60% decrease in size of myenteric neurons, and a 4-fold increase in thickness of the serosa, compared with saline-treated controls.
Animals↗
Genotoxic effects of selected biocides on RTG-2 fish cells by means of a modified Fast Micromethod Assay.
A sensitive in vitro assay for detecting DNA damage in RTG-2 cells culture is described. This assay employs a dye, PicoGreen double stranded DNA (dsDNA) quantitation reagent, which becomes intensely fluorescent upon binding nucleic acids. The assay includes a simple and rapid 50-min sample lysis in the presence of EDTA, SDS, and high urea concentration at pH 10, followed by time-dependent DNA denaturation at pH 11.6 after NaOH addition. The time course and the extent of DNA denaturation are followed in a microplate fluorescence reader at room temperature for less than 1h. Comparative studies between suspension and fixed RTG-2 cells indicated that it is possible to apply this methodology in both cases with good results. Neutral red assay was used for to determine the cellular viability when RTG-2 cultures were exposed to tetrakis(hydroxymethyl) phosphonium chloride (THPC) and benzalkonium chloride (BC), as biocides used in the disinfection of cooling towers. The results obtained by neutral red assay indicate IC(50(48)) values of 0.017 (0.011-0.028) and 2.71 (1.91-3.86) mg/L for tetrakis(hydroxymethyl) phosphonium chloride and benzalkonium chloride, respectively. DNA damage has been evaluated for both disinfectants in RTG-2 culture, by exposure to 1/10-, 1/25-, 1/50-, and 1/100-IC(50(48)) value, and the results obtained indicate a strain scission factor (SSF) of 0.126+/-0.014, 0.181+/-0.014, 0.217+/-0.013, and 0.245+/-0.013 in cell suspensions, and 0.077+/-0.019, 0.107+/-0.014, 0.151+/-0.014, and 0.202+/-0.015 in attached cells for tetrakis(hydroxymethyl) phosphonium chloride; while the SSF values for benzalkonium chloride are 0.023+/-0.009, 0.033+/-0.017, 0.068+/-0.012, and 0.088+/-0.015 in cell suspensions, and 0.033+/-0.010, 0.044+/-0.011, 0.080+/-0.009, and 0.093+/-0.010 in attached cells. Thus, the assay proposed in this study has made it possible to show DNA damage in RTG-2 cells when exposed to 0.2(1/100 IC(50(48))) and 300(1/10 IC(50(48))) Hg/L of tetrakis(hydroxymethyl) phosphonium chloride and benzalkonium chloride, respectively. The results obtained indicate that the Fast Micromethod Assay, applied on RTG-2 cell line cultures, is a fast and sensitive method for the early DNA damage detection in the aquatic environment.
Analysis of Variance↗
On the possibility of molecular recognition of taste substances studied by Gábor analysis of oscillations.
A liquid membrane oscillator containing nitromethane as membrane material has been investigated. The influence of substances responsible for taste belonging to four classes (sweetness, saltiness, sourness and bitterness) on oscillation patterns of liquid membrane oscillators with cationic surfactant benzyldimethyltetradecylammonium chloride (BDMTAC) was examined. A new approach based on Gábor transformation is proposed for obtaining the power spectra of the observed oscillating signals and for establishing "fingerprints" of the investigated substances. It was shown that two-dimensional form of these power spectra might be used efficiently for this purpose.
Benzalkonium Compounds↗
Removal of arsenic from groundwater by micellar-enhanced ultrafiltration (MEUF).
The removal characteristics of arsenate using micellar-enhanced ultrafiltration (MEUF) were investigated. Among four different cationic surfactants used, hexadecylpyridinium chloride (CPC) showed the highest removal efficiency of arsenic (96%), and the removal efficiency with hexadecyltrimethylammonium bromide (CTAB) was 94%. But the removal efficiency with benzalkonium chloride (BC) was the lowest (57%) due to higher critical micelle concentration (CMC) of BC than those of other surfactants. Over 80% of arsenic was removed with octadecylamine acetate (ODA). On the effect of solution pH on the arsenic removal, since the valance of arsenate decreases from trivalent to monovalent as pH decreases, the removal was reduced at lower pH. The presence of 0.45mM of nitrate and 0.01mM of phosphate reduced the removal efficiency by 5-8%. This decrease was because of the competition between the arsenate, nitrate and phosphate for the binding sites of the surfactant micelle. Similar decrease in the removal of arsenate was observed with CPC, CTAB and ODA in the presence of these anions. In cross-flow filtration, the removal efficiency of arsenic was similar to that in the dead-end system. However, the decline in flux was less than that in dead-end filtration. In order to lower the concentration of the surfactant in the effluent, the effluent was treated with powdered activated carbon (PAC) before discharging to the environment. Over 98% surfactant was removed with 1gl(-1) of PAC. In conclusions, the MEUF is considered as a feasible process using CPC or CTAB to remove the arsenate from groundwater compared with the other solid based adsorbent processes.
Arsenic↗
Determination of quaternary ammonium biocides by liquid chromatography-mass spectrometry.
In this study we have developed a method for the direct determination of benzalkonium chloride (BAC) homologues and didecyldimethylammonium chloride (DDMAC), generally used as biocides. The chromatographic separation was performed using a Luna C18 column and gradient elution. A 50 mM formic acid-ammonium formate buffer at pH 3.5 was used as aqueous phase to allow ion-pair formation with the quaternary ammonium biocides. The detection was carried out using an ion trap mass analyser and electrospray ionisation (ESI) source. Parameters such as the magnitude and duration of the resonant excitation voltage and the magnitude of the trapping RF voltage for full scan tandem mass spectrometry (MS-MS) experiments were studied to establish the optimal experimental conditions. Instrumental quality parameters of both liquid chromatography coupled to mass spectrometry (LC-MS) and LC-MS-MS methods were studied and good run-to-run and day-to-day precision values (relative standard deviations, RSDs lower than 11%) and LODs down to 0.1 microg L(-1) (LC-MS-MS) were obtained. Finally, the applicability of the LC-MS-MS method was demonstrated by analysis of a spiked water sample and some commercial products containing BAC.
Benzalkonium Compounds↗
Adsorption of mycotoxins by organozeolites.
Adsorption of zearalenone (ZEN), ochratoxin A (OCHRA) and aflatoxin B1 (AFB1) on natural zeolite, clinoptilolite, modified with different amounts of octadecyldimethylbenzyl ammonium (ODMBA) ions was investigated. Results showed that adsorption of hydrophobic ionizable ZEN on unmodified zeolite tuff was very low and that adsorption on organozeolites increased with increasing hydrophobicity of the zeolitic surface. The adsorption was independent of the form of ZEN in solution and the solution pH, indicating that hydrophobic interactions with ODMBA are responsible for ZEN adsorption. Adsorption of low polar ionizable OCHRA on organozeolites also increased with increasing hydrophobicity of the zeolitic surface, however, OCHRA showed moderate adsorption on unmodified zeolitic tuff at pH 3. OCHRA adsorption on unmodified zeolite as well as on lower surface coverage of organozeolite was dependent on the form of OCHRA in solution; there was a decrease of adsorption at high pH, where OCHRA is in the anionic form. It indicated that at acidic pH, low surface coverage allows some combination of hydrophobic interaction with ODMBA and interactions with the surface of the zeolite. At higher surface coverage, the OCHRA adsorption was higher and practically independent of pH, indicating that the hydrophobic interactions of OCHRA with ODMBA are responsible for its adsorption. Nonionizable low polar AFB1 had a high affinity for the unmodified zeolitic tuff and the adsorption of AFB1 was greatly reduced for organozeolites, indicating that AFB1 does not have high tendency for hydrophobic interactions with ODMBA. pH dependence of AFB1 adsorption, while AFB1 has the same form at all pHs, demonstrated that the surface modification of the zeolite depends on pH and that these modifications have influence on its adsorption. The calculated dipole moments of neutral mycotoxin molecules: AFB1-9.5D, OCHRA-6.9D and ZEN-2.2D are in qualitative agreement with adsorption experimental data.
Adsorption↗
Innocuity of tetradecyl-dimethyl-benzyl ammonium fluoride on the DNA of human spermatozoa.
Tetradecyl-dimethyl-benzyl-ammonium fluoride (TDBAF) is a powerful new spermicide. In cases of failure of local contraception, there is a theoretical risk that a spermatozoon exposed to a sublethal concentration of spermicide might suffer DNA strand breaks, with transmission of damaged DNA to the oocyte. The present study aims to ascertain the innocuity of TDBAF in this regard. Spermatozoa from 32 fertile men were exposed to a sublethal concentration of TDBAF. The degree of DNA fragmentation was measured by modified terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin end labeling (TUNEL) assay, pulsed field gel electrophoresis of purified sperm DNA and radioenzymatic labeling. With the TUNEL assay, a statistically significant increase in sperm labeling was observed after TDBAF treatment compared to the controls. However, pulsed field gel electrophoresis and enzymatic labeling of purified DNA showed no evidence of augmented DNA fragmentation in TDBAF-treated sperm. Under electron microscopy, TDBAF caused sperm chromatin decondensation. In the TUNEL assay, this phenomenon would allow easier access of terminal transferase to its DNA substrate, explaining the apparent increase in DNA fragmentation in the presence of TDBAF. In conclusion, the use of TDBAF as a spermicide does not place progeny at heightened risk of abnormalities should pregnancy occur. In regard to the TUNEL assay, we suggest that pretreatment of spermatozoa with TDBAF promotes a more accurate measurement of sperm DNA fragmentation.
Adult↗
Thymosin beta4 inhibits benzalkonium chloride-mediated apoptosis in corneal and conjunctival epithelial cells in vitro.
Thymosin beta-4 (Tbeta(4)) is known to promote ocular wound healing, to decrease ocular inflammation, and to have anti-apoptotic effects on corneal epithelium. In this study, the effect of Tbeta(4) on the survival of human ocular surface epithelial cells exposed to benzalkonium chloride (BAK) was measured. Human conjunctival epithelial cells (HC0597) or human corneal epithelial cells (HCET) were treated with 0%, 0.001%, 0.01%, or 0.1% BAK for 15 min. After 3 or 24h of recovery in culture medium containing 1 microg/ml Tbeta(4), a dosage that has been demonstrated effective in several published studies, DNA synthesis was measured using a colorimetric BrdU incorporation assay. Both conjunctival and corneal epithelial DNA synthesis was inhibited by BAK in a dose-dependent manner. Tbeta(4) did not protect the epithelial cells from BAK-induced inhibition of proliferation. To assess the ability of Tbeta(4) to prevent apoptosis, epithelial cells were treated with 0.01% BAK+Tbeta(4) and cell death was measured using a colorimetric assay. BAK-induced apoptosis increased throughout the duration of the assay, which was carried out to 5 days in culture. Treatment of HC0597 cells with Tbeta(4) significantly inhibited the apoptosis shown to be initiated by BAK. Treatment of non-transformed human corneal epithelial cells (HCEC) with Tbeta(4) also significantly inhibited the apoptosis shown to be initiated by BAK at later times in culture. Ocular solutions containing BAK as a preservative are typically used for extended periods of time. This study suggests that Tbeta(4) may be able to overcome the apoptotic side effect of BAK, and may be a useful additive to solutions containing this preservative.
Apoptosis↗
Antimicrobial effects of sanitizers against planktonic and sessile Listeria monocytogenes cells according to the growth phase.
This study was designed to investigate the individual or combined effects of sanitizers on survival of planktonic or sessile Listeria monocytogenes cells at different phase of growth. The sanitizers tested included: (i) acetic acid (pH 5.0), (ii) NaOH (pH 12.0), (iii) 10% Na2SO4, (iv) 10% Na2SO4 and acetic acid (pH 5.0), (v) 10% Na2SO4 and NaOH (pH 12.0), (vi) a quaternary ammonium (20 ppm) and (vii) glyceryl monolaurate (75 ppm). Results revealed a great efficacy of alkaline treatments on both sessile and planktonic cells with a slightly higher resistance of 6 h biofilms. Quaternary ammonium appeared very effective in killing more than 98% of cells, but a resistance of 7 days biofilm was observed. Other sanitizers did not succeed in inhibiting totally the pathogen but acted in a similar way on both sessile and planktonic cells. Renewing the medium or not do not seem to be the major cause of a resistance emergence.
Acetic Acid↗
Peroxinectin gene transcription of the giant freshwater prawn Macrobrachium rosenbergii under intrinsic, immunostimulant, and chemotherapeutant influences.
Peroxinectin (PE) gene expressions were determined using real-time PCR in the giant freshwater prawn Macrobrachium rosenbergii based on moulting; prawns were fed diets containing different concentrations of sodium alginate, and were exposed to different concentrations of copper sulphate, benzalkonium chloride (BKC), and trichlorfon. Results showed that PE mRNA expression of prawns was the highest in stage A, significantly decreased in stage B, and reached the lowest level in stages D0/D1. The PE transcript was significantly higher in prawns fed the 1.0 gkg(-1) sodium alginate-containing diet than those fed the 2.0 gkg(-1) sodium alginate-containing diet and those fed the control diet. PE transcripts significantly decreased in prawns exposed to 0.1-0.4 mgL(-1) copper sulphate after 96 h, 0.3-1.0 mgL(-1) BKC after 96 h, and 0.2-0.4 mgL(-1) trichlorfon after 48 h. It was concluded that the status of PE gene expression was seriously affected by the moult cycle, immunostimulant, and chemotherapeutants.
Alginates↗
Mechanisms of resistance in Salmonella enterica adapted to erythromycin, benzalkonium chloride and triclosan.
The potential for adaptive resistance of S. enterica serovar Enteritidis, Typhimurium and Virchow to increasing sub-lethal concentrations of erythromycin, benzalkonium chloride and triclosan was investigated to identify mechanisms underlying resistance. Permeability changes of the outer membrane, including LPS, cell surface charge, hydrophobicity and the presence of an active efflux in the adapted strain compared with the parent were studied. Examination of the outer membrane and LPS did not reveal any significant changes, although most of the pre-adapted strains were notably less hydrophobic than resistant strains. More than one type of active efflux was identified in all strains investigated, on the basis of restored sensitivity in the presence of the inhibitors reserpine and carbonyl cyanide 3-chlorophenylhydrazone (CCCP). Cell surface hydrophobicity and the presence of active efflux could contribute to the resistance of S. enterica to the antibacterial agents studied here.
Adaptation, Physiological↗
Pluronic gels for nasal delivery of Vitamin B12. Part I: preformulation study.
Thermoreversible nasal gels of Vitamin B(12) using pluronic PF 127 were aimed to improve absorption and patient compliance. In the present research work, effects of Vitamin B(12) and gel additives, viz. PF concentration, osmolarity, polyethylene glycol (PEG 15000) on thermodynamic properties of phase transitions at gelation (T(1)) and gel melting (T(2)) is reported. Aqueous PF 127 gels prepared by cold method containing pluronic (20-24%, w/w), vitamin, sorbitol, PEG, and benzalkonium chloride. T(1) decreases and T(2) increases with vitamin and PF concentration. Gelation range narrows with sorbitol and PEG. Suppression of T(2) is significantly higher than T(1) with both the additives. The linearity was observed only for semilogarithmic plot of PF concentration and 1/T(2) for sorbitol and PEG, which reveals significant interaction of both at gel melting. Enthalpy of both transitions remains unchanged with vitamin indicating no interaction with polymer. Benzalkonium chloride decreased gelation onset temperature. Thermodynamic properties of PF 127 gels are significantly altered with polymer concentration and water-soluble formulation additives.
Administration, Intranasal↗
Rapid-onset intranasal delivery of metoclopramide hydrochloride. Part I. Influence of formulation variables on drug absorption in anesthetized rats.
Intranasal (IN) administration is a promising approach for rapid-onset delivery of medications and to circumvent their first-pass elimination when taken orally. Metoclopramide (MCP) is a potent antiemetic, effective even for preventing emesis induced by cancer chemotherapy. The feasibility of developing an efficacious intranasal formulation of metoclopramide has been undertaken in this study. The nasal absorption of MCP was studied in anesthetized rats over 60min using the in vivo in situ technique. The influence of several formulation variables, vis., pH and the addition of preservative, viscosity and absorption enhancing agents on the nasal MCP absorption was examined. The data obtained showed that MCP was well absorbed nasally where almost 90% of the drug was absorbed after 60min from the rat nasal cavity. The MCP absorption was pH-dependant such that the apparent first-order rate constant of absorption (K(app)) was almost tripled when the pH of the solution was increased from 5 to 8. However, deviation from the classical pH-partition theory was observed pointing to the role of aqueous pore pathway in MCP nasal absorption. The K(app) was significantly increased (P<0.05) by incorporation of 0.01% of the preservative benzalkonium chloride. Conversely, increasing the solution viscosity by the use of hydroxylpropyl methylcellulose adversely affected the rate of absorption. The use of enhancers namely sodium deoxycholate, sodium cholate, chitosan low and high molecular weight, protamine sulphate and poly-l-arginine resulted in significant increase in MCP absorption. The highest promoting effect was observed with the bile salt sodium deoxycholate where about 92% of the drug was absorbed in 25min from the rat nasal cavity and the K(app) showed more than two-fold increase as compared to control (from 0.0452 to 0.1017min(-1)).
Absorption↗
Kinetic characteristics and toxic effects of benzalkonium chloride following intravascular and oral administration in rats.
Kinetic characteristics and toxic effects of benzalkonium chloride (BZK) following injection via jugular vein (JV), femoral artery (FA) and oral administration (PO) were experimentally investigated using rats. The BZK concentrations in blood and tissues (lung, liver and kidney) were determined by high-performance liquid chromatography with solid phase extraction. Toxic doses of 15 and 250 mg/kg of BZK were used for intravascular (JV and FA) and PO administration, respectively. The fatal effects appeared soon after the dose in JV-rats, while delayed in FA- or PO-rats. The blood BZK concentrations and the elimination half-lives were similar between JV- and FA-rats, while the distribution of BZK in tissues was slightly different. In PO administration, the rats that aspirated BZK into their lungs had some symptoms, while the rats that did not aspirate BZK appeared to be normal. The BZK concentrations in blood and tissues were significantly higher in the aspirated PO-rats. The toxic degree of BZK was correlated with the BZK concentration in orally dosed rats. Lung and kidney had higher BZK concentrations compared to blood or liver, and they could be the target organs of BZK.Keyword: Benzalkonium chloride
Administration, Oral↗
Effect of alcohol addition to the aqueous phase on the thermal effects of micellization of cationic benzyldimethyldodecylammonium bromide and its adsorption onto porous and nonporous silicas.
Titration calorimetry has been used to study the effect of the addition of two primary alcohols, 1-butanol and 1-heptanol, to the aqueous phase on the thermal effects of micellization of benzyldimethyldodecylammonium bromide (BDDAB) as well as its adsorption onto nonporous Spherosil XO15M and onto porous aluminosilicate SiAl32d22 possessing uniformly sized mesopores. A linear decrease of the critical micelle concentration (CMC) of the cationic surfactant with the additive content was inferred from the specific conductivity measurements. All adsorption and calorimetry experiments were carried out at 298 K and at a fixed alcohol content (0.01 mol kg(-1)) either in deionized water or in a 0.01 mol kg(-1) NaBr solution. Dilution calorimetry measurements allowed determination of the cumulative molar enthalpy changes and a new analysis of these data was proposed to calculate easily the enthalpy of micellization per mole of BDDAB, Delta(mic)h, and the CMC value. The alcohol addition was shown to render the micellization phenomenon more exothermic, the effect being larger as the chain length of alcohol increased. These effects were attributed to the location of alcohol molecules between the surfactant units, their hydroxyl groups close to the surfactant head-groups, in competition with the surfactant counterions. The individual isotherms of alcohol and surfactant adsorption onto XO15M and SiAl32d22 were determined. The plots of the pseudo-differential molar enthalpy of displacement, Delta(dpl)h, against the surface coverage by the surfactant cation, Theta(BDDA+), were derived from the titration calorimetry data. The formation of surface-bound aggregates was thought to be a prerequisite for alcohol coadsorption at the solid-solution interface. At least two different types of adsolubilization sites were postulated, one of the sites being the same as in micelles and the other related to the contact area between the hydrophobic surfactant tails and the equilibrium bulk solution. Coadsorption (adsolubilization) of alcohol molecules at such sites was found to increase the exothermic contribution to the enthalpy of displacement per mole of the BDDA+ adsorbed.
Adsorption↗
Pseudophakic preoperative maculopathy.
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Aqueous Humor↗