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ISSN 1420-3049 © 2007 by MDPI www.mdpi.org/molecules Communication
Preparation of Benzalkonium Salts Differing in the Length of a Side Alkyl Chain
Kamil Kuca 1,2,3, Jan Marek 4,5, Petr Stodulka 1, Kamil Musilek 2,5,*, Petra Hanusova 4, Martina Hrabinova 1 and Daniel Jun 1,2
1 2 3
Center of Advanced Studies, Faculty of Military Health Sciences, Hradec Kralove, Czech Republic Department of Toxicology, Faculty of Military Health Sciences, Hradec Kralove, Czech Republic Department of Chemistry, Faculty of Sciences, J.E. Purkinje University, Usti nad Labem, Czech Republic Vakos XT, Prague, Czech Republic Faculty of Pharmacy in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic
* Author to whom correspondence should be addressed; E-mail: firstname.lastname@example.org Received: 3 September 2007; in revised form 3 October 2007 / Accepted: 4 October 2007 / Published: 24 October 2007
Abstract: Benzalkonium salts are widely used as disinfectants, biocides and detergents, among a variety of other applications. The cationic surface-activity of these salts determines their potential to act as a biocide on both target and non-target organisms. In this study, a quick synthesis of a complete set of the benzalkonium salts differing in the length of an alkylating chain (from C2 to C20) is described. Moreover, their 1H-NMR, HPLC-MS, TLC and HPLC analysis were recorded. Keywords: Benzalkonium salt, detergent, surfactant, biocide, HPLC, TLC.
Introduction Quaternary compounds, including the benzalkonium salts, constitute an economically important class of industrial chemicals that are widely distributed among a diverse array of products and users
Thin-Layer Chromatography (TLC) retention factors and High Pressure Liquid Chromatography (HPLC) retention times for of all the salts are also listed in this table. Such mixtures are produced on a large-scale in industry. biocides. The synthesis of long chain derivatives (C8-C20) was described earlier by our group . nasal and parenteral products. and as phase transfer catalysts . However. We have to point out that use of our method (mixture of the pure compounds) might conceivably replace the synthetic “all in one” mixtures now used in industry.Molecules 2007. but rather as mixtures composed of two or three benzalkonium members differing only in the length of the alkyl chains [7-8]. a targeted synthesis of pure individuals could be of interest because of the above-mentioned specificity of each salt against different pathogens. Because of their strong cationic surface activity. the C14-homologue against gram-positive bacteria and C16-homolog against gram-negative bacteria . It is well known that the C12-homolog is most effective against yeast and fungi. Yields achieved for those compounds were also adequate. but also as anti-electrostatics. are shown in Table 1. Preparation of these compounds has been described previously [9-12]. Very important features of benzalkonium salts are their bactericidal and antimicrobial properties.N-dimethylalkyl bromides 3-12 were prepared by the general method shown in Scheme 1. The antimicrobial activity depends on a changing length of the side n-alkyl chain. These compounds are not generally used as single components. Moreover. A new set of the short chain benzalkonium salts was developed and the preparation of the long chain benzalkonium salts was improved. To provide a synthetic route to such compounds. For example. Ajatine® (a C12 analogue used as disinfectant) is generally a mixture of the parent substances (N-benzyl-N. but a general synthesis applicable to the whole series of benzalkonium salts is lacking. Scheme 1. quaternary compounds are used primarily as disinfectants. N EtOH R-Br 2 reflux 3 . 12 2342 from an industrial to the household sector. the C2-C6 derivatives were added to complete the whole C2-C20 set. the preparation of the whole set of these compounds (C2-C20) is now shown. Results and Discussion The N-benzyl-N. The yields achieved in this study were superior to those obtained earlier for C8-C20 derivatives . together with the melting points of the prepared salts. Preparation of benzalkonium salts. They are also used as topical antiseptics and disinfectants for medical equipment [3-6].11 N R Br 1 Yields of all reactions. For these reasons they are widely used as preservatives for ophthalmic.N- . and detergents.
0 122.p. There are several articles which deal with HPLC analysis of benzalkonium derivatives [14-16].25 Conclusions A quick and easy method for the preparation of pure benzalkonium salts in adequate yields was described.0 34. TLC and HPLC techniques.592 0.634 0. In this study.76 5. melting points.60 18.55 13. retention factors and retention times of prepared bezalkonium salts. Their purities were analyzed by 1H-NMR.84 7. where potential adverse effects of the parent substances would no longer be a factor.92 11. Experimental General All chemicals used in this study were obtained from Sigma-Aldrich (Czech Republic).512 0. Detection: UV254.0 81. Compound 3 4 5 6 7 8 9 10 11 12 Yield (%) 94 44 54 61 61 80 71 58 71 64 m. C12 alkylating chain and solvent) and a reaction product (the appropriate monoquaternary salt).651 0. however.5-121. Such a method could be of interest in the laboratories not equipped by special expensive equipment (HPLC). a TLC method was also developed to distinguish each benzalkonium salt.260 0.0-123.0 TLC Rf 0. Yields.0-40. Such a method could be applicable not only for the benzalkonium salts but also for other long chain quaternary compounds.5 50.612 0.15 21.Molecules 2007.0-56. . (°C) 120. mobile phase n-BuOH/CH3COOH/H2O (5:1:2). None of these.0 43.0-152. TLC (Kieselgel Merck.5-47. HPLC-MS (HP1100 HPLC system . our previously developed HPLC method was modified for a determination of each benzalkonium salt with an aim of distinguishing them .0-53. Our pure products (without parent substances) could be used in more sophisticated applications.0-83. Purity of all products was tested by determination of their melting points (Boetius block) and were uncorrected.Agilent Technologies (Waldbronn.5 53.558 0.664 HPLC Rt 4.083 0. Dragendorff´s reagent).0-88. 12 2343 dimethylamine.5 37. HPLC-MS.0-37.43 15. For quick laboratory analysis of prepared benzalkonium salts.5 150.420 0.0 85.13 8. Table 1. describe HPLC analysis of the whole benzalkonium set.47 9.
Ph).2 [M+] (calc. (CH3)2N+).93 (s. data were collected in positive ion mode with an ESI probe voltage of 4000 V) and NMR analysis (Varian Gemini 300. (CH2)3). 3. 18H.47 Hz. PhCH2N+). CH3). 4.26 (bs. 1. J = 6. (CH2)3).24 (t. Solvent was evaporated and the crude N-benzyl-N.3 [M+] (calc.Molecules 2007. 3. 2H. 3H.55 (bs. ESI-MS: m/z 220. 1. 3H. ESI-MS: m/z 276. ESI-MS: m/z 248. washed with ether and allowed to dry at r. 2H. for [C21H38N]+ 304. 3. CH3). 6H. J = 6.24 (t. 1. CH3). 1H-NMR: δ 0. N-benzyl-N. J=6.96 (s.N-dimethylhexyl-1-ammonium bromide (5). 6H. J = 6. 1. quadrupole mass spectrometer MSD1456 VL. CH2N+).35 (t. 12 2344 Germany). CH2CH2N+). ESI-MS: m/z 164. (CH3)2N+). 2H.15Hz.47 Hz.58 (s. 2H. 2H. J=6. PhCH2N+). The corresponding yields and melting points are summarized in Table 1.N-dimethylethyl-1-ammonium bromide (3). 7.N-dimethylbenzylamine (1. for [C13H22N]+ 192. 1H-NMR: δ 0. 5H. J = 6. J=7.23 (bs.56 (bs.56 (s. 2. 4. N-benzyl-N. (CH3)2N+). 5H. J = 6.55). Ph). CH2CH2N+). for [C19H34N]+ 276. 7. J= 6. 7.88 (t. PhCH2N+).60 Hz. 2H.33Hz.2 [M+] (calc.4 mmol) in dry ethanol (25 ml) was mixed with an equimolar amount of the appropriate 1-bromo-alkane (2. 2H. (CH3)2N+). 2H. N-benzyl-N. 1.t.92 (s. 3H.32Hz. 2H. 2H. CH2N+).33Hz.60 Hz. 3H.78 (m. 10H. PhCH2N+).22 (bs.24 (t. 5H.N-dimethyloctyl-1-ammonium bromide (6).55 (s.54 (s.2 [M+] (calc.N-dimethyldodecyl-1-ammonium bromide (8). 4. (CH2)7).86 (t.83 (t. N-benzyl-N.52 (s. 6H. Spectral data (1H-NMR and MS spectra) are listed below: N-benzyl-N.53 (bs. CH2CH2N+). 3H.33 Hz.38). J = 6. 3. 1. 5H. 3H. 1H-NMR: δ 0. 6H.79 (m. for [C11H18N]+ 164. 2H. 2.28 (m. CH2N+). 2. 1H-NMR: δ 0. 7. 3.24 (t. 2H. CH3CH2N+). CH3). Synthesis Briefly N.83 (t. 6H. for [C17H30N]+ 248. 7.32).77 (m.N-dimethylbutyl-1-ammonium bromide (4). Ph). 1H-NMR: δ 1. 5H. Ph). 6H.90 (t.56 (bs. 1. N-benzyl-N.35 (t. CH2CH2N+). 2H.47 Hz.33 Hz.78 (m.94 (s. 1H-NMR: δ 0. DMSO-d6). 14H. 2.52 (m. CH3). CH2N+).2 [M+] (calc. 2H.53 (bs. 7.59 (s.27).3 [M+] (calc. for [C15H26N]+ 220. 2. 2H.22 (t.77 (m. 2H. 7. (CH2)5). Ph). 5H. J=6. 3. (CH2)9). 2.95 (s. ESI-MS: m/z 304. CH2CH2N+). 300 MHz. 6H. CH2N+). 4.50). 4. 4. ESI-MS: m/z 192. 2H.95 (s. (CH3)2N+). CH2N+). (CH3)2N+). 2H.N-dimethyldecyl-1-ammonium bromide (7). Ph).N-dimethylalkyl-1-ammonium bromides (3-12) were recrystalized from acetone. 1.4 mmol) and the mixture was refluxed for 28 hours. 1. 1. 10. .29 (bs. PhCH2N+). PhCH2N+).44).
a UV1000 detector (Spectra-Physics Analytical.47 Hz.0. 7.6 mm I. ESI-MS: m/z 388. This mixture was prepared as 0. 3H.83 (t. 2.54 (s. for [C27H50N]+ 416. 7. for [C23H42N]+ 332. 3.2 mm layer of silica gel 60 F254 (Merck. 2. N-benzyl-N.5 mM solutions) were applied 1 cm from the bottom edge of the plate.71).4 [M+] (calc. 2H.61).4 [M+] (calc.22 (t.1 M sodium acetate solution. for [C27H50N]+ 388.78). 7. The HPLC system consisted of a P200 gradient pump (Spectra-Physics Analytical.25 (t. Fremont. 5H. Germany). 1H-NMR: δ 0. Acknowledgements Support of the Grant FVZ0000501 (Ministry of Defence. 2H. Ph). Ph).78 (m. PhCH2N+).78 (m. Germany) and developed with a methanol/chloroform/acetic acid (25:5:0. 2.54 (s. 3. CH2CH2N+). For analyses a 250 × 4. . CH2N+). Cotati.0. 6H. 1H-NMR: δ 0.52 (m. 3. Fremont. Czech Republic) is gratefully acknowledged. 2H. It was delivered isocratically at a flow-rate of 1 mL/min.84 (t.60 Hz. N-benzyl-N. Plates was streaked with a CAMAG Linomat IV automatic applicator (Camag. Development was stopped when the mobile phase front reached 1 cm from the top of the plate.4 [M+] (calc. 4.71Hz.93 (s. 4. CH2CH2N+).22 (bs.22 (bs. 6H. PhCH2N+). CH2CH2N+).74 (m. a 7125 injection valve – 10 μΛ loop (Rheodyne. (CH2)15).5 v/v)) mobile phase in a twin trough chamber with a stainless steel lid (Camag. Samples (5 μL from 2.23 (t.52 (s. The mobile phase contained 45% acetonitrile and 55 % water. 7. J = 6. Berlin. 2H.53 (bs. 2H.95 (s. (CH3)2N+). 5H. Darmstadt. (CH2)13).75 (m. Finally the pH was adjusted with acetic acid to 5. Compounds were dissolved in acetonitrile/water (45:55 v/v) as 0.Molecules 2007. (CH2)11).52 (s. J=7. Czech Republic). N-benzyl-N. 2H. 5H. USA).25Hz. 12 2345 N-benzyl-N. 6H.94 (s. Ph).CH2N+).26 Hz.64). (CH3)2N+). 26H.D. 2H. 3H.N-dimethyltetradecyl-1-ammonium bromide (9). (CH3)2N+). ESI-MS: m/z 416.24 (t. for [C25H46N]+ 360. (CH3)2N+).39 Hz. 5H. 1. Prague. 4. J=8. CH3). PhCH2N+). Subsequently. J = 6. ESI-MS: m/z 332. The absorbance was measured at 263 nm . 1. 2H. CH2N+). 1.N-dimethyloctadecyl-1-ammonium bromide (11). CH2CH2N+). J = 6. Ph).3 [M+] (calc. J = 6. 2H. USA) and a CSW Chromatography Station 1. ESI-MS: m/z 360.83 (t. Waters Spherisorb Cyano (5 μm) column was used (Supelco Inc. 6H.N-dimethyleicosyl-1-ammonium bromide (12). J = 8. 34H. pH 5. Germany).N-dimethylhexadecyl-1-ammonium bromide (10) 1H-NMR: δ 0. 2H. USA). 3H. 3. 1. 2H. TLC was performed on 140 mm × 140 mm plates coated with a 0. 1H-NMR: δ 0. 22H.24 (bs. 4. CH3).52 (m.5 software (Data Apex. (CH2)17). 2. Berlin.94 (s. PhCH2N+). 3H.52 (bs. 1.60 Hz. 1. 30H. CH3). CH3). J=8.84 (t. 1. 2H.1 M acetate buffer.). 1. TLC and HPLC separations of all prepared compounds were developed (Table 1).24 (bs.47 Hz. CH2N+).
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