Library identifications for Lemnaceae

Transcription

1 Library identifications for Lemnaceae

2 NP ESI+ NP present in, enriched in (15.47min)

3 NP MS^E CH 3 Moupinamide H NH

4 NP NP ESI- s NP NP NP present in, enriched in (8.98min), its isomer at 8.80min is present too, higher intensity than identified compound, only as a impurity in ; NP is false positive

5 NP MS^E Interfering compound H

6 NP ESI- NP isomers NP NP present in, enriched in (28.30min), its isomer at 27.97min is present too, higher intensity than identified compound, only as an impurity in, third isomer detected at 28.15min.

7 NP MS H Gingerglycolipid A H H H H 3 C H

8 NP MS^E Compound identification confirmed by MS fragmentation pattern

9 NP MS^E - isomers isomer 28.15min isomer NP Compounds at 27.97min and 28.15min show identical mass fragmentation Isomers of NP ; signal at 27.97min is also present as impurity in

10 NP ESI- NP isomers NP NP present in, enriched in (29.55min), its isomer at 29.23min is present too, higher intensity than identified compound, only as an impurity in, two more isomers detected at 29.45min & 29.73min.

11 NP MS^E [M-H+FA]- Co-eluting masses H 3 C Gingerglycolipid derivative H

12 NP isomers Isomer 29.73min [M-H+FA]- [M-H]- Isomer 29.44min Isomer 29.25min

13 NP ESI- NP present in, enriched in (12.08min)

14 NP MS Enrichment of intensity of m/z in compared to

15 NP MS^E H H H H 3 C H CH 3

16 NP ESI- NP present in, enriched in (12.88 min)

17 NP MS^E Apigenin-7--glycoside H H

18 NP ESI- NP present in, enriched in (12.27min)

19 NP MS H Kampferol-3--glucoronoside H H

20 NP MS^E

21 NP ESI- NP present in, enriched in (5.03 min)

22 NP MS Catechin H

23 NP ESI- NP NP NP present in, enriched in (8.78 min); double peak in and, relative intensity and enrichment corresponds to ratio in versus (probably two isomers), other possible hit NP is a false hit

24 NP MS H Apigenin-4,6-C-glycoside H H H

25 NP ramp & isomers 8.69min 8.69min 8.78min 8.78min Signals at 8.69 and 8.78 min in and show identical fragments

26 NP ESI- NP present in, nice co-elution, enriched in (16.72min)

27 NP MS Luteolin H

28 NP ESI- NP present in, enriched in (10.53min), but not perfectly coeluted, peak shape differs slightly MSMS confirmation

29 NP MS NP m/z clearly enriched

30 NP MS^E H H Quercetin-3-glyciside H

31 NP ESI- NP present in (11.34min), good co-elution and little increase in -injection

32 NP MS Interfering mass H 3, 5 glucopyranosylphloretin H H H

33 NP MS^E Compound identification confirmed by MS fragmentation pattern

34 NP ESI- NP present in (13.02min), good co-elution and increase in -injection

35 NP MS CH 3 Rhoifolin (Apigenin-7--diglycoside) H H H H

36 NP MS^E Compound identification confirmed by MS fragmentation pattern

37 NP ESI- Isomers NP present in (10.25min), good co-elution and increase in -injection

38 NP MS Rutin H H H H CH 3

39 NP MS^E Compound identification confirmed by MS fragmentation pattern

40 NP ESI- NP present in (35.04min), good co-elution and increase in -injection

41 NP MS Rutamarin H 3 C H 3 C H 3 C H 3 C CH 3 CH 2

42 NP MS^E Compound identification confirmed by MS fragmentation pattern

43 NP ESI- NP present in (13.02min), good co-elution and increase in -injection

44 NP MS Apigenin diglycoside H CH 3 H

45 NP MS^E Compound identification confirmed by MS fragmentation pattern

46 NP ESI- 2nd Isomer NP present in (8.63min), good co-elution and increase in -injection, second isomer present as impurity in

47 NP MS Luteolin-6-C-glycoside H H H

48 NP MS - isomers 8.99min 8.63min 8.99min 8.63min Signals at 8.63 and 8.99 min in and show identical m/z Isomer of library

49 NP MS^E Compound identification confirmed by MS fragmentation pattern

50 NP ESI- NP present in (10.3min), good co-elution and increase in -injection

51 NP MS Apigenin-6-C-glycoside H H H H

52 NP MS^E Compound identification confirmed by MS fragmentation pattern

53 NP ESI- NP present in (8.63min), good co-elution and increase in -injection, but identical signal as for NP , compounds are regioisomers and perfectly coelute, it is not possible to distinguish between them!

54 NP MS 8-C-galactosylluteolin H

55 NP MS^E Compound identification confirmed by MS fragmentation pattern

56 NP ESI- 2nd Isomer NP present in (10.65min), good co-elution and increase in -injection, further isomer identified at 9.72min

57 NP MS H H H H CH 3 H 3 C H

58 NP MS^E Compound identification confirmed by MS fragmentation pattern

59 NP MS^E - isomer Compound at 9.74min shows identical fragmentation pattern as NP and the corresponding impurity in Isomer of NP

60 NP ESI- NP present in, as in injection multiple signals are detected, note: axes are not linked due to the very low intensity of compound in

61 NP MS tri-hydroxy-octadecanoic acid C H 3

62 NP ESI- NP present in, increased in, co-elution is not perfect due to UPLC system, see shift of compound at 21.87min, note: axes are not linked due to the very low intensity of compound in

63 NP MS tri-hydroxy-octadecanoic acid C H 3 H

64 NP MS^E Compound identification confirmed by MS fragmentation pattern, note: axes are not linked due to the very low intensity of compound in

65 NP ESI- NP present in (35.29min), good co-elution and increase of intensity in -injection

66 NP MS CH 3 H 3 C

67 NP MS^E Compound identification confirmed by MS fragmentation pattern

68 NP ESI- NP present in (36.68min), good co-elution and increase in injection compared to, but higher in injection (maybe due to ion suppression in matrix)

69 NP MS Co-eluting mass Sesquiterpenoyl-coumarin CH 3 CH 2 H H 3 C CH 3

70 NP MS^E Compound identification confirmed by MS fragmentation pattern

71 NP ESI- injection shows a systematic shift to earlier retention times compared to injection, difference is min!

72 NP (ESI-) NP present in (9.67min), good co-elution and increase in injection (considering the systematic retention time shift)

73 NP MS H Quercetin--glycoside H

74 NP MS^E Compound identification confirmed by MS fragmentation pattern

75 NP ESI- NP present in (36.68min), good co-elution, systematic shift of retention time ( & -0.05min) and increase in -injection, note: axes are not linked due to the very low intensity of compound in

76 NP MS Co-eluting mass Formic acid adduct n-hexanoyl-diglycoside C H 3 H

77 NP MS^E Compound identification confirmed by MS fragmentation pattern

78 MS-MS validation of library hits Several library hits were further verified via MS-MS experiments since identification of the compounds was not clear due to absence of MS ramp spectrum because of low intensity or co-eluting compounds that interfered with the molecules fragmentation patterns. All MS-MS spectra were acquired using a UPLC-QTF mass spectrometer (QuanTF) and obtained spectra were compared to the previously recorded MS^E spectra of the authentic s, collision energy was identical to MS^E. Experiments were performed on different instruments, hence retention time differs, peaks were identified by comparison of retention times of a 15 compound mix (mix15)

79 NP ESI- NP present in (7.42min), good co-elution and increase in -injection

80 NP MS H Quercetin--glycoside H

81 NP MS^E

82 NP MS-MS spectrum of m/z Compound identification confirmed by MS fragmentation pattern

83 NP ESI- NP present in (10.65min), good co-elution and increase in injection, note: axes are not linked due to the very low intensity of compound in

84 NP MS Co-eluting mass H Pyracanthoside H H H

85 NP MS^E

86 NP MS-MS spectrum of m/z Compound identification confirmed by MS fragmentation pattern

87 NP ESI- NP present in (9.96min), good co-elution and increase in injection

88 NP MS H H Apigenin-6-C-diglycoside H

89 NP MS^E

90 NP MS-MS spectrum of m/z Compound identification confirmed by MS fragmentation pattern

91 NP ESI- NP present in (9.93min), good co-elution and increase in injection, note: axes are not linked due to the very low intensity of compound in

92 NP MS H

93 NP MS^E

94 NP MS-MS spectrum of m/z Compound identification confirmed by MS fragmentation pattern