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Analysis of E. toletana extracts demonstrating presence of 3-oxo-C6-HSL

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Analysis of E. toletana extracts demonstrating presence of 3-oxo-C6-HSL

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  1. Figure SM1. LC MS/MS analysis ethyl acetate extracted AHLs from P. agglomerans, P. savastanoi and E. toletana. The top rows of every section refer to the reverse-phase chromatographic profile. The last rows are the MS/MS fragmentation patterns. A and B, E. toletana DAPP-PG 736 wild type extract; C and D, P. agglomerans DAPP-PG 734 wild type extract; E, P. savastanoi DAPP-PG 722 wild type; F, G, H and I are the AHL standards. AHLs were identified by retention time analysis and by comparison of transition with the AHL synthetic standards. Comparisons were also made with synthetic AHL standards as published by (Gould et al., 2006).

  2. A Analysis of E. toletana extracts demonstrating presence of 3-oxo-C6-HSL

  3. B Analysis of E. toletana extracts demonstrating presence of 3-oxo-C8-HSL

  4. C Analysis of P. agglomerans extracts demonstrating presence of C4-HSL

  5. D Analysis of P. agglomerans extracts demonstrating presence of C6-HSL

  6. E Analysis of P. savastanoi extracts demonstrating presence of 3-oxo-C6-HSL 113.1

  7. F Intens. 11022402.D: EIC 102 +MS2(172.1) 5 x10 0.5 4 x10 11022402.D: EIC 71 +MS2(172.1) 1 5 11022402.D: EIC 102 +MS2(200.0) x10 2 4 x10 11022402.D: EIC 99 +MS2(200.0) 4 2 5 x10 11022402.D: EIC 102 +MS2(214.0) 1.0 0.5 4 x10 11022402.D: EIC 113 +MS2(214.0) 1 5 x10 11022402.D: EIC 102 +MS2(242.0) 4 2 5 x10 11022402.D: EIC 141 +MS2(242.0) 1 5 x10 11022402.D: EIC 102 +All MS 1 0 +MS2(172.1), 2.0-2.6min #(1364-1394) -3 0 3 6 9 12 15 Time [min] 5 x10 1.0 102.1 standard 250 ng HSL_C4 154.0 71.3 144.0 75 100 125 150 175 200 225 250 m/z

  8. G Intens. 11022402.D: EIC 102 +MS2(172.1) 5 x10 0.5 4 x10 11022402.D: EIC 71 +MS2(172.1) 1 5 x10 11022402.D: EIC 102 +MS2(200.0) 2 4 x10 11022402.D: EIC 99 +MS2(200.0) 4 2 5 x10 11022402.D: EIC 102 +MS2(214.0) 1.0 0.5 4 x10 11022402.D: EIC 113 +MS2(214.0) 1 5 x10 11022402.D: EIC 102 +MS2(242.0) 4 2 5 11022402.D: EIC 141 +MS2(242.0) x10 1 5 x10 11022402.D: EIC 102 +All MS 1 0 -3 0 3 6 9 12 15 Time [min] 5 x10 +MS2(200.0), 12.8-13.1min #(3127-3147) 102.1 1.5 standard 250 ng HSL_C6 1.0 182.0 0.5 99.1 74.3 172.0 71.3 270.9 0.0 75 100 125 150 175 200 225 250 m/z

  9. H Intens. 11022402.D: EIC 102 +MS2(172.1) 5 x10 0.5 4 x10 11022402.D: EIC 71 +MS2(172.1) 1 5 x10 11022402.D: EIC 102 +MS2(200.0) 2 4 x10 11022402.D: EIC 99 +MS2(200.0) 4 2 5 x10 11022402.D: EIC 102 +MS2(214.0) 1.0 0.5 4 x10 11022402.D: EIC 113 +MS2(214.0) 1 5 x10 11022402.D: EIC 102 +MS2(242.0) 4 2 5 11022402.D: EIC 141 +MS2(242.0) x10 1 5 x10 11022402.D: EIC 102 +All MS 1 0 -3 0 3 6 9 12 15 Time [min] 5 x10 +MS2(214.0), 8.1-8.8min #(2400-2436) 1.2 102.1 1.0 0.8 standard 250 ng 3-oxo-HSL_C6 0.6 0.4 0.2 113.1 74.3 186.0 0.0 75 100 125 150 175 200 225 250 m/z

  10. I Intens. 11022402.D: EIC 102 +MS2(172.1) 5 x10 0.5 4 x10 11022402.D: EIC 71 +MS2(172.1) 1 5 x10 11022402.D: EIC 102 +MS2(200.0) 2 4 x10 11022402.D: EIC 99 +MS2(200.0) 4 2 5 x10 11022402.D: EIC 102 +MS2(214.0) 1.0 0.5 4 x10 11022402.D: EIC 113 +MS2(214.0) 1 5 x10 11022402.D: EIC 102 +MS2(242.0) 4 2 5 x10 11022402.D: EIC 141 +MS2(242.0) 1 5 x10 11022402.D: EIC 102 +All MS 1 0 -3 0 3 6 9 12 15 Time [min] 5 x10 +MS2(242.0), 14.5-14.9min #(3419-3443) 102.1 3 standard 250 ng 3-oxo-HSL_C8 2 141.1 1 74.3 184.0 214.0 224.0 84.2 98.2 0 75 100 125 150 175 200 225 250 m/z

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