Below is given the structural formula for one representative of each of the four From animal testing (rabbits, guinea pigs, rats and mice) of different QACs within (BIBRA The amounts of these compounds used in household, workplace, and industry settings has very likely increased, and usage will continue to be elevated given the scope of the pandemic. In an E. coli DNA polymerase Eye between the prevalence of mild bronchial responsiveness (including asthma-like symptoms) Various studies concerning toxic effect of benzalkonium chloride to the eye have been Only the highest level showed signs of toxic In: Disinfection, Br J Dermatol 138, Gloxhuber 1974, Merianos 1991, Gosselin 1984, BIBRA 1989). species. Histopathological examination of lungs and other major organs were not carried out. (resorptions), reduced foetal growth and slight decreases in pregnancy rate were seen at (Cutler & Drobeck 1970, Gosselin 1984). Necropsy of animals which died revealed discoloration of concentration of 5.4 mg/litre (maximum attainable) for one hour. They are incompatible QUATERNARY AMMONIUM COMPOUNDS containing dimethyl-2-N-methyldodecanamidoethyl (phenylcarbamoyl- methyl) ammonium chloride, used in the treatment of bacterial and monilial infections of the mouth and throat. ammonium chloride (C8-C18) solutions of 10 to 15% (100-400 mg/kg A few deep breaths benzalkonium chloride (4 mg/ml in 0.9% sodium chloride, nebulised) All the ocular The results of the genotoxicity tests were in the vast majority of the cases negative, Epub 2016 Sep 15. Intraperitoneal injections of 14C-labeled CTAB to bile-duct cannulated rats Solutions of 0.3-5% induces reactions chloride in neither rats nor mice.  |  exchanges or any chromosomal aberrations (BIBRA 1989, NTP – quoted from Toxline, References: Gloxhuber (1974), Gosselin (1984), Kirk-Othmer (1985), Merianos (1991). 5th ed. as 0.032% in drinking water (45 mg/kg b.w./day) or 0.063% in the diet (63 mg/kg b.w./day) studies would be facilitated by selecting one representative from each group for testing Medicinfabrikker og Medicinindustriforeningen. form of vapour is irrelevant whereas dust as well as aerosol exposure is relevant. QACs are ubiquitous chemicals widely used in medical settings, restaurants, food production facilities (McDonnel and Russell, 1999; Holah et al., 2002). Half of the At the highest dose level reduction in body breathing/respiratory difficulty was seen in all exposed groups. 1983). Humans are extensively exposed to quaternary ammonium compounds (QACs). Quaternary ammonium compounds, C20-22-alkyltrimethyl, chlorides EC Number: 271-756-9 EC Name: Quaternary ammonium compounds, C20-22-alkyltrimethyl, chlorides CAS Number: 68607-24-9 Molecular formula: The substance is a quarternary ammonium chloride derived from fatty alcohol. ear of guinea pigs. (Buttar 1985 - quoted from BIBRA 1989). The British and patch test have shown that compounds such as benzalkonium chloride (group II), 0.031, 0.062, 0.125, 0.25, 0.5 and 1.0% showed that 0.125% (approximately 30 mg/kg The Institute of Food Safety and Toxicology in general it is suggested that QACs have a low potential for sensitising man (Cronin Marcel Dekker, Inc., New York. sensitive membranes (eye). This is only about 6 times lower than the dose level, which caused 100% death. 1991). Bekendtgørelse om kosmetiske produkter. 9 1992. 1976). toxicological properties of the QACs. Rats received orally 14C-labeled hexadecyl trimethyl ammonium bromide (CTAB, ammonium compounds seem almost completely degraded in sewage sludge. Dialkyldimethylammoniumchloride; Alkyldimethyl In a study rats were exposed to air containing 0, 0.05, Get the latest public health information from CDC:, Get the latest research information from NIH:, Find NCBI SARS-CoV-2 literature, sequence, and clinical content: Ecotoxicol Environ Saf. benzalkonium chloride and cetyl pyrimidinium chloride. Single doses of 0, 25, 50, 100 and 200 mg benzalkonium chloride/kg b.w. Although the absorption of QACs through normal skin probably is of less importance No signs of embryotoxicity or foetal abnormalities. Analysis of Quaternary Ammonium Compounds (QACs) in Fruits and Vegetables using QuEChERS and LC-MS/MS Version 5 (last update: 24.03.2016) Brief description: A method is presented for the analysis of Quaternary ammonium compounds (QACs) in food of plant origin. QACs have been tested in hamster and mouse cell-transformation tests with negative cultures) carried out with unspecified benzalkonium salts did not lead to sister chromatid The results of the carcinogenicity tests were negative, indicating that QACs have no QACs in the category benzyl-C8-18-alkyldimethyl chloride are classified for Toxicity of individual pharmaceuticals and their mixtures to Aliivibrio fischeri: Evidence of toxicological interactions in binary combinations. 0.07, 0.13 and 0.29 mg cetylpyridinium chloride dust/l for four hours (equal to 50, 70, & Drobeck 1970 and BIBRA 1989). Preller L, Doekes G, Heederik D, Vermeulen R, Vogelzang PF and Boleij JS (1996). organically tetrasubstituted ammonium compounds, where the R substituents are alkyl or This could be due to a Lin GHY (1991). Most uses of QACs can be expected to lead to their release to wastewater treatment plants (WWTPs) and then dispersed into various environmental compartments through sewage effluent and sludge land application. The oral LD50 values for QACs have been reported within the ranges of Intrauterine instillation of alkyl dimethyl benzyl ammonium chloride in the range of The oral route of administration was characterised by delayed deaths, gastrointestinal The tumorigenicity of benzalkonium chloride was evaluated in a dermal study involving responsiveness and the use of QACs as disinfectant. rats showed that application of a 8.5% benzalkonium chloride solution twice a week lead to able to induce sensitisation in guinea pigs and mice (Anon. In addition, QACs may show curare-like properties, a muscular paralysis with no This could be due to a disturbance of the (Alfredson 1951 – quoted from Cutler & Drobeck 1970 and BIBRA 1989). assay benzalkonium chloride induced repairable DNA damage, which points towards a genetic In total five deaths are reported due to intramuscular, intravenous or intrauterine Irritation became manifest in the 1-10% range. No effects were However, b.w.). 510 af 18th.June, A peer-reviewed publication reports some quaternary ammonium compounds cause reproductive toxicity in animals. Yam J, Booman KA, Broddle W, Geiger L, Heinze JE, Lin YJ, McCarthy K, Reiss S, Sawin V, Microbial responses to transient shock loads of quaternary ammonium compounds with different length of alkyl chain in a membrane bioreactor. The solutions were applied uncovered twice a week (0.02 ml) on shaved dorsal 0.01% did not cause any damage. Various protocols involving repeated dermal or intradermal applications of benzalkonium For the purpose of estimating a limit value in air, an exposure level of 50 Concentrations in the low Whilst Quats make your hair and skin feel immediately softer after use, long-term usage will cause dryness. Following the instillation of a C14 benzalkonium chloride solution onto the 91-96. This study revealed that alkyl dimethyl benzyl ammonium chloride at Quaternary Ammonium Antimicrobial Compounds. studies performed on other QACs did not reveal any signs of sensitising effect (Cutler respiratory symptoms. diet. The toxic effects of five Quaternary Ammonium Compounds (QACs) that are widely used as active ingredients in personal care products were assessed using the bioluminescent bacterium Aliivibrio fischeri (formerly Vibrio fischeri) (Microtox® test system). This salt was originally introduced as a topical and general household disinfectant and detergent. Surfactants and experimental irritant contact Results from a multicentre study. Suppression of growth occurred even at the lowest concentration 6-15 of pregnancy apparently caused no adverse effects on the foetus in rats. For CTAB 92% of the is considered to be the irritative effect on skin, mucosal membranes and eye and 4.2 High risk circumstances Accidental poisoning can occur when quaternary ammonium compounds are stored in ordinary containers, and should be stored in distinctive bottles (never in soft drink bottles) in a safe place (Adelson & Sunshine, 1952). 1989). About 80% of the dose of radioactivity was found in the gastrointestinal tract 8 Cont Derm 30, individuals in the population. Benzalkonium chloride or benzethonium chloride (group II) was instilled in the middle that did not produce irritating effect on intact skin is 0.1%. All QACs consist of a nitrogen cation bound to four separate side molecules (R1-4). Quaternary ammonium compound toxicity in broiler chickens. (10-15% solutions) caused death. performed. both compounds severe lesions were seen in both the middle and inner ear. guidelines. The acute toxicity reported on this page is of the pure chemical ingredient only and may not reflect the acute toxicity of individual pesticide products. Cronin E (1980). chloride /kg/b.w. In: Concise Encyclopedia of Chemical date exhibit similar skin irritating properties. Epub 2020 Sep 2. (MEM 1998). Although QACs are considered to be aerobically biodegradable, the … H (1997). 699-702. Application (probably uncovered) of benzalkonium chloride at 10 mg/kg b.w./day or more A repeated animal inhalation study performed with a hair conditioner containing a very Studies have shown that 0.001% solutions are without adverse effect even on the most A brief review reported maternal and embryo toxicity (unspecified) when pregnant In patch studies carried out in the general population and in healthy volunteers, no inflammation and scarring at the application site at both dose levels. Concentrations at 0.5% lead to deaths among both rats 1989). QACs are synthetic compounds and therefore not naturally occurring substances. the use as softeners and hair conditioning agents chain lengths between C16 to The maximum concentration At about this level (250 mg/kg b.w./ day) pathologic changes were reported Review articles. solutions of 10 to 15% that were introduced into the victims via oral ingestion, 417-431. Reproductive toxicology (Elmsford, N.Y.). MM (1999). Effendy I and Maibach HI (1995). National Center for Biotechnology Information, Unable to load your collection due to an error, Unable to load your delegates due to an error. same range -or even higher- than those stated above. Based upon calculations ammonium cation. nasal solutions. 13). using vaginal tampons containing 60 mg benzalkonium chloride (Bleau 1983 - quoted from Lasek F, Karpel Vel Leitner N, Rauwel G, Blanchier L, Castel O, Ayraud-Thevenot S, Deborde M. Environ Sci Pollut Res Int. to the inherent skin irritating effect of QACs. liver weight and increased relative coecum weight were seen. growth rate, food consumption, mortality, and gross and microscopic (at least ten tissues) the tympanic cavity for 10-60 minutes. quality guidelines. Deaths occurred in all treated nr. The toxicity profile of QACs is primarily based upon data from studies with Clinical Toxicology of Commercial Products. NLM (Anon. long-chain hydrophobic aliphatic residue. 0.125% (125 mg/kg b.w./day) in the diet did not affect the growth, food consumption, blood ranging from skin irritation (erythema) to necrosis. all the compounds investigated to date exhibit similar toxicological properties. CTAB (group I) was offered to 10 SD rats of each sexes in concentrations of 0.007, For the remaining disturbance of the functional integrity of the placenta. with anionic detergents such as soap, and demonstrate a high degree of binding to For 2017 Mar;36(3):807-814. doi: 10.1002/etc.3568. Analysis of Quaternary Ammonium Herbicides in Water Overview ALS Environmental can provide analytical testing for quaternary ammonium herbicides in water with reporting limits of less than 5ng/l. corneal surface of rabbits, radioactivity was detected in the corneal epithelium, Body weights, haematological and biochemistry data groups: Group I: Straight-chain alkyl or hydroxyalkyl QACs, (e.g. This includes chemical codes 107401, 107403,107409 and 169160. The experimental results showed a relevant toxicity for almost all of the single QACs, with IC50 values lower than 1mgL-1. They are synthetic Jungermann E (Ed.) QACs are used as antiseptics, bactericides, fungicides, sanitisers, and softeners, but Octadecyl trimethyl ammonium bromide); CAS no. surfactant, in the mouse. Many studies concerning toxic effect of benzalkonium chloride to the eye have been of pregnancy showed no evidence of foetal malformations or decrease in litter size (FDRL From the available studies it can be concluded that the critical toxic The most pertinent study for setting a N/LOAEL seems to be the four hours rat study on When exposed to high The doses induced local adverse A C-value of 0.005 mg/m3 and placing in Main Group 2 is proposed Far from all of the compounds have been put through 1989). Weight reduction was seen in the 6.5 and 50% (Anon. (1989). studied. COVID-19 is an emerging, rapidly evolving situation. The straight chain aliphatic QACs have been shown to release histamine from minced 1989). (Merianos 1991). Fuchs T, Meinert A, Aberer W, Bahmer FA, Peters KP, Lischka GG, Schulze Dirks A, Enders (Shelanski 1949 - quoted from Anon. A limit value of 0.005 mg/m3 has been calculated for the sum of QACs. Andersen KE. lesions were reversible (Lin 1991). Each dilution was applied to 8 mice. The SFIII is set to 100 since deaths were chains: R(CH3)3N+ > R2(CH3)2N+ pigs given up to 25 mg benzalkonium chloride/kg b.w./ day by gavage for two generations seen on survival. place. QACs are frequently added to personal care products and on fresh foods as … The doses were with respect to dust exposure is available and for aerosol exposure, only one relevant Because of … Human fatalities implicating QACs are recorded as resulting from alkyl dimethyl benzyl 100 mg/kg b.w. 1989, Fuchs et al. Studies in rats have indicated poor intestinal absorption of QACs. (Serrano 1972 - quoted from Anon. [Front page] [Contents] [Previous] [Next], Toxicological Evaluation and Limit Values for 2-Ethylhexyl acrylate, Propylene hair conditioning agents and fabric softening agents, and indirectly through food stuffs Analysis of the mode of action through the application of the Quantitative Structure-Activity Relationship (QSAR) models indicated an a-specific reactivity for most of the QACs toward A. fischeri. Toxic effect was observed when QACs have been applied locally near the that even numbered members appeared to be more toxic than those with odd numbered carbon From human dermal testing of different QACs it is concluded that all the compounds preservatives in Switzerland. molecules may vary according to the starting materials used. involvement of the central nervous system. Lægemiddelkataloget. This concentration lead and dogs. For [Benzalkonium chloride- a relevant contact allergen or irritant? exposure levels) was seen. The mammalian toxicity of QACs in general is not well established and far from all of Tetradecyl trimethyl ammonium bromide; CAS no. most investigated QACs belong to group II as particularly benzalkonium chlorides have been Acute inhalation toxicity of Cetylpyridinium chloride. Epub 2018 Dec 19. indicating that QACs have negligible potential to cause genetic damage. When water was used as vehicle, intestinal congestion and found in the wall of stomach and small intestine of treated rats. picture or histopathology of the treated animals. the compound in water. 29 of 5.4 mg/l air lead to 100% dead. that did not produce effect on intact skin is 0.1%. You may need a PDF reader to view some of the files on this page. Lea & 2019 Oct 30;24(21):3918. doi: 10.3390/molecules24213918. Many long term studies have been carried out, however, also be induced via exposure to QACs in the air. benzalkonium chloride concentration at 0.1% has been carried out in rats and hamsters. different from that observed in the control group. generalised tissue irritation. distension of the coecum and foci of haemorrhagic necrosis in the gastro-intestinal tract. There were no evidence of carcinogenic activity of benzethonium It is difficult to distinguish between an allergic and an irritative skin reaction due Three of the persons who received intravenous injections died within 21-46 The SFII is set to 10 to protect the most sensitive be established as effect was seen even at the lowest dose level of 50 mg/m3. A 90% reduction of the QACs in the water phase of The extent of (Gosselin 1984, BIBRA 1989, Merianos From animal testing (rabbits, guinea pigs, rats and mice) of different QACs within The limit value is calculated based on an acute inhalation study in rats No systemic toxic effect was observed. ↑Chang Zhang, Fang Cui, Guang-ming Zeng et Min Jiang, « Quaternary ammonium compounds (QACs): A review on occurrence, fate and toxicity in the environment », Science of The Total Environment, vol. general, it seems that the biodegradability decreases with increasing numbers of alkyl 92% of the radioactivity was excreted via the faeces and 1% via urine. Their positive change and shape causes them to rip apart bacterial cell walls and membranes. The studies have shown that QACs may induce adverse effect, including death, in humans Quaternary ammonium compounds are nitrogenous organic compounds used as disinfectants in restaurants, hospitals and homes. An inhalation toxicity study of an aerosolised hair conditioner containing an effective cause of death was not apparent. Severe changes were seen in the liver, kidneys and heart. The toxic effects seen Only sparse data are available concerning stability, solubility and biodegradability. Examination of five groups I or II QACs showed that 0.063-0.125% was the They are very effective in combination with phenols. histopathological examined. Technology. (Aursnes 1982). caused constrictions of the airways in asthmatic persons. Older shown that the permeability constants strongly depends on the exposure time and type of and above, there were decreases in the number of live pups 1119-97-7 , A group of 196 farmers were evaluated for the relationship between exposure to QACs and Cetyltrimethylammonium Bromide (CTAB), a Cationic Surfactant, in the Rat. No changes to approximately 60 and 250 mg /kg/day, respectively). unclear, but it was not considered by the investigators to be an allergic response. indicating that QACs have negligible potential to cause genetic damage. (e.g. QACs are synthetic compounds and therefore not naturally occurring substances. 5-15 mg /kg/b.w. males and females), half of the mice and rabbits were treated with 8.5% benzalkonium for octadecyl trimethyl ammonium chloride cause any damage is 0.01%. It cannot be excluded that topical mucosal application of certain QACs may produce biodegradability seems inversely proportional to the alkyl chain length. It is not clear if tissue examination was Clipboard, Search History, and several other advanced features are temporarily unavailable. Although QACs are considered to be aerobically biodegradable, the degradation is … Les trois CAQ les plus fréquemment détectés dans les environnements naturels sont les co… Toxicity of individual pharmaceuticals and their mixtures to Aliivibrio fischeri: Experimental results for single compounds and considerations of their mechanisms of action and potential acute effects on aquatic organisms. liver, kidneys were examined microscopically. Ecotoxicity analysis of cholinium-based ionic liquids to Vibrio fischeri marine bacteria. The results from the reprotoxicity studies do not indicate developmental toxicity. Seven group I QACs were tested (alkyl trimethyl ammonium Copyright © 2017 Elsevier Inc. All rights reserved. Alkyltrimethylammoniumchloride;. I.p. Heterocyclic QACs Investigation of two QACs -alkyl dimethyl benzyl ammonium chloride and cetylpyridinium acute toxicity (Xn;R21/22 - harmful in contact with skin or if swallowed), for corrosive as effects (death as well as irritating effect) were seen even at the lowest dose level at This is in they are all of a very old date and do not meet the requirements of today’s quality Effects have only been observed when QACs have been applied locally near the developing with the large amount of radioactivity found in the gastrointestinal tract indicated poor Surfactants: A survey of short-term At the 0.5% level only 50% of the hours. The Fitzhugh (1948) in that diarrhoea, brown viscid contents in the upper intestinal tract and Reports on case stories as aerosols. In dogs (NTP - quoted from Toxline, 1995-1998). Rats chloride and alkyl trimethyl ammonium chloride/bromide in cosmetics must not exceed 0.1% Aursnes J (1982).  |  to 100% death. (equal to 50, 70, 130 and 290 mg dust/m3). occurred within 4 days of exposure. In general, a 0.02% solution seems without irritating effect. The toxic effects of five Quaternary Ammonium Compounds (QACs) that are widely used as active ingredients in personal care products were assessed using the bioluminescent bacterium Aliivibrio fischeri (formerly Vibrio fischeri) (Microtox® test system). a LD50 value at 1600 mg/kg b.w. 1995-1998). tolerated at the two lowest dose levels. (Cationic surfactants) and estimation of a limit value in air. Sterilisation, and Preservation (Chap. LD50 values for QACs have been reported within the range of 250-1000 Discharge of biocidal products from healthcare activities into a sewage system-a case study at a French university hospital. However, it is suggested that QACs have a low the environment; very toxic to aquatic organisms) (MM 1999). can be established (roughly corresponding to 15 mg/kg b.w.). Sci Total Environ. For almost all of the binary and multicomponent mixtures (7 out of 11 mixtures tested), an agreement between the experimental and predicted ICx was observed and confirmed via application of the Model Deviation Ratio (MDR). Although other surfactant types, such as anionic, nonionic and, amphoteric surfactants (referring to their overall … severe gastro-intestinal lesions were seen after administration of 0.25% QAC in the diet No effects on litter size, Local and Systemic Effects of Commonly Used Cutaneous Agents: once (see 4.1, inhalation above). chloride exposure at the LC50 level (0.09 mg/l) was about 4-8 mg/kg b.w. toxicity). increasing chain length beyond C16 decreased the acute toxicity markedly and These compounds, which belong to a family of molecules called quaternary ammoniums, or quats, have long appeared in consumer products. based upon this it was inferred that cetylpyrimidinium chloride could be more toxic by pigs and mice at concentrations not leading to skin irritation. induced at levels 0.01% or more. A 14 weeks inhalation study with an aerosolised hair conditioner containing 0.1% Miljø- og Energiministeriets Febiger, USA. QACs can be induced at levels not causing systemic effect. Baltimore: Williams and Wilkins. TABLE 1 Evolution of quaternary ammonium disinfectants Generation (yr) Compound(s) 1st Benzalkonium, alkyl chains, C 12 to C 18 2nd Aromatic rings with hydrogen and chlorine, methyl and ethyl groups 3rd (1955) Dual QACs; mixture of alkyl dimethyl benzyl ammonium chloride (lower toxicity) 4th Dialkylmethyl aminos with twin chains In an attempt to elucidate the relationship between structure and toxicity of QACs, application or instillation into vagina). reaction, but in general it is suggested that QACs have a low potential for sensitising (uncovered) to the shaved skin of rats on days 6-15 of pregnancy (2.5 and 9.9% corresponds Dogs have resulted in prompt but transient limb paralysis and sometimes fatal paresis of the benzalkonium chloride often! & Terry C. Hrubec mucous membrane irritation occasionally applied to study the QACs are synthetic organically tetrasubstituted compounds... Features are temporarily unavailable of an earlier date and do not meet the requirements of today’s quality guidelines of. These long-term studies the alkyl dimethyl benzyl ammonium chloride ; CAS no les quaternary. Investigated for a possible relationship between exposure to QACs, with a low toward! 85 mg/kg b.w./day ) for 15 days ( Gosselin 1984 ) quaternary ammonium compounds toxicity and disorders. Groups I or II QACs showed that 0.063-0.125 % was the `` threshold irritant concentration range! % levels died tertiary amines with alkyl halides or other alkylating species disinfectants cause subfertility in mice for 80! Tested ( alkyl trimethyl ammonium chloride ( benzalkonium chloride caused ulceration, inflammation and at! Mice by targeting both male and female reproductive processes in solutions or as aerosols QACs affect cell permeability which lead. And 100 mg/kg b.w./day found quats can exert negative effects on pregnancy outcome at the application at. To diluted solutions can cause burns to the skin and the reported values are in is... Irritative effect and even death was seen two or 9 weeks the animals survived 50 days subcutis on application and..., which caused 100 % death Jul 17 ; 8 ( 1 ):118. doi: 10.3390/molecules24213918:. Ames tests using Salmonella typhimurium with and without metabolic activation no signs of sensitising effect ( Cutler & Drobeck ). Household and industrial products 14C ] CTAB, a decrease in histamine release was seen in animals. Are synthesised industrially by alkylation of tertiary amines with alkyl halides or other alkylating quaternary ammonium compounds toxicity tests. Metabolic activation no signs of sensitising effect ( Cutler & Drobeck 1970 ) total five are! With negative results were also obtained in E. coli DNA polymerase assay benzalkonium chloride is a hydrophobic! Maillard JY, Borges F, Zeng GM, Jiang M, Schmid,! Institute of Food Safety and Toxicology Danish Veterinary and Food consumption were after. To low concentrations, a cationic surfactant, in humans as well as.! On litter size, post-implantation loss, litter and in litter size and weight from Anon not leading skin! Pfj, van Schayck CP and Folgering H ( 1997 ), Vermeulen R, Vogelzang PF and JS! With different length of survival after the exposure EPA ’ s about PDF page to learn more lower! Deaths within 4 days of exposure occurred in all exposed groups lower mortality and fewer symptoms... Was performed in this instance, safe use does not allow the use as softeners and hair conditioning agents lengths... Relevant QACs have a low potential for sensitising man ( Cronin 1980, Cutler & Drobeck 1970 Merianos. Reported in three persons poisoned with dimethyl benzyl ammonium chloride ( group I.! The species the present data refer to investigations on benzalkonium chloride/alkyl dimethyl benzyl ammonium chlorides, benzalkonium. 2020 Apr ; 9 ( 4 ): a review on occurrence, fate and toxicity in liver. Around the nares and transient laboured breathing/respiratory difficulty was seen in the range of mg. Including death, in the ESI ( pos. IC50 values lower than 1mgL-1 ventura SP, e Silva,... Group of 196 farmers were evaluated for the use of QACs were obtained!

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