Series of Benzothiazole Derivatives Experimental

Synthesis, Antimicrobial, Antiquorum-sensing and Cytotoxic Activities of New Series of Benzothiazole Derivatives Moustafa T. Gabr

a

a,b

a,

a

, Nadia S. El-Gohary , Eman R. El-Bendary , Mohamed M. Ela b c,d Kerdawy , Nanting Ni , Mona I. Shaaban

Department of Medicinal Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt b

c

Department of Chemistry, Georgia State University, Atlanta, Georgia 30303, USA d Department of Microbiology, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt Department

of Microbiology, College of Pharmacy, Taibah University, Almadinah Almunawwarah 344, Saudi Arabia 

Corresponding author: Tel.: +2 010 00326839, Fax: +2 050 2247496 [email protected]

Experimental Chemistry All melting points (°C) were recorded on Fisher-Johns melting point apparatus and are uncorrected. The infrared -1

spectra were recorded using Nicolet Magna-IR Fourier-Transform 560 Spectrometer (v in cm ) at the Department of 1

Chemistry, Georgia State University, USA. Nuclear magnetic resonance ( H and

13

C NMR) spectra were obtained on

Bruker Avance 400 MHz spectrometer using CDCl3 and DMSO-d6 as solvents at the Department of Chemistry, Georgia State University, USA. The chemical shifts are expressed in δ ppm using tetramethylsilane (TMS) as internal reference. Mass spectra were recorded on nano LC-Q-TOF micro (Waters Micromass) spectrometer in positive or negative ion mode as necessary at the Department of Chemistry, Georgia State University, USA. Reaction times were monitored using TLC plates, Silica gel 60 F254 precoated (E. Merck, Germany) and the spots were visualized by UV (366 nm). Chloroform : methanol (9:1) and hexane : ethyl acetate (3:1) were used as elution solvents. 2-Amino-6fluorobenzothiazole (1) was obtained from Sigma-Aldrich Co., USA. Compound 2 was prepared adopting the reported procedure [1]. Compounds 3a-e were prepared following the literature method [2].

6-Fluoro-2-hydrazinobenzothiazole (2). Hydrochloric acid (10 mL) was added dropwise to hydrazine hydrate 99% (10 g, 0.2 mol) at 5-10 °C, followed by addition of a solution of 2-amino-6-fluorobenzothiazole (1) (3.364 g, 0.02 mol) in ethylene glycol (40 mL). The mixture was heated at reflux temperature for 5 h. On cooling, the precipitated solid was collected by filtration, washed with water, dried and crystallized from ethanol. Yield 89%, m.p. 194-196 °C [1]. Ethyl 3-oxo-2-(2-(substituted phenyl)hydrazono)butanoates 3a-e. General Procedure A. A solution of sodium nitrite (1.035 g, 0.015 mol) in water (15 mL) was added to a stirred ice-cooled solution of the appropriate aniline (0.01 mol) in 5N hydrochloric acid (30 mL). After 10 min, the reaction mixture was rendered alkaline by addition of a saturated

-1-

solution of sodium acetate. The obtained diazonium solution was then added slowly to a stirred cold slurry of ethyl acetoacetate (1.3 g, 0.01 mol) in ethanol (50 mL) and sodium acetate (1.25 g, 0.015 mol) in water (50 mL). The reaction mixture was stirred at room temperature for 1 h and the solid obtained was collected by filtration, washed with water, dried and crystallized from methanol. Ethyl 2-(2-(2-chloro-4-nitrophenyl)hydrazono)-3-oxobutanoate (3a) -1 1 Yield 71%, m.p. 107-108 °C [3]. IR spectrum (KBr, v, cm ): 3107 (NH), 1732 (COOC2H5), 1693 (C=O). H NMR spectrum (400 MHz, DMSO-d6, δ ppm): 1.30 (t, 3H, CH2CH3), 2.45 (s, 3H, CH3), 4.25-4.40 (q, 2H, CH2CH3), 7.808.50 (m, 4H, Ar-H, NH). Ethyl 2-(2-(4-nitrophenyl)hydrazono)-3-oxobutanoate (3b) -1 1 Yield 85%, m.p. 129-130 °C [4]. IR spectrum (KBr, v, cm ): 3157 (NH), 1714 (COOC2H5), 1693 (C=O). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 1.25 (t, 3H, CH2CH3), 2.35 (s, 3H, CH3), 4.25-4.35 (q, 2H, CH2CH3), 7.65 (d, 2H, Ar-H), 8.25 (d, 2H, Ar-H), 11.70 (s, 1H, NH).

13

C NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 14.4, 25.5, 62.0,

116.2, 125.7, 135.3, 142.5, 148.5, 162.5, 194.3. Ethyl 2-(2-(2,4-dichlorophenyl)hydrazono)-3-oxobutanoate (3c) -1 1 Yield 74%, m.p. 116-117 °C [5]. IR spectrum (KBr, v, cm ): 3122 (NH), 1724 (COOC2H5), 1690 (C=O). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 1.30 (t, 3H, CH2CH3), 2.40 (s, 3H, CH3), 4.28-4.35 (q, 2H, CH2CH3), 7.407.80 (m, 3H, Ar-H), 12.50 (s, 1H, NH). Ethyl 2-(2-(2,5-dichlorophenyl)hydrazono)-3-oxobutanoate (3d) -1 1 Yield 65%, m.p. 214-216 °C [2]. IR spectrum (KBr, v, cm ): 3148 (NH), 1732 (COOC2H5), 1687 (C=O). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 1.25 (t, 3H, CH2CH3), 2.40 (s, 3H, CH3), 4.22-4.35 (q, 2H, CH2CH3), 7.107.70 (m, 3H, Ar-H), 12.40 (s, 1H, NH). Ethyl 2-(2-(2-methyl-5-nitrophenyl)hydrazono)-3-oxobutanoate (3e) -1 1 Yield 81%, m.p. 201-203 °C [6]. IR spectrum (KBr, v, cm ): 3106 (NH), 1710 (COOC2H5), 1655 (C=O). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 1.25 (t, 3H, CH2CH3), 2.45 (s, 3H, CH3), 2.55 (s, 3H, CH3), 4.20-4.35 (q, 2H, CH2CH3), 7.45-7.90 (m, 3H, Ar-H), 12.10 (s, 1H, NH).

13

C NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 14.3, 17.1,

26.8, 61.3, 108.8, 118.5, 128.5, 130.9, 132.8, 140.9, 147.4, 162.7, 193.7. 1-(6-Fluorobenzothiazol-2-yl)-3-methyl-4-(2-(substituted phenyl)hydrazono)pyrazol-5(4H)-ones 4a-e. General Procedure B. A solution of 6-fluoro-2-hydrazinobenzothiazole (2) (0.55 g, 0.003 mol) in glacial acetic acid (10 mL) was added to a solution of the appropriate ethyl 3-oxo-2-(2-(substituted phenyl)hydrazono)butanoate 3a-e (0.003 mol) in glacial acetic acid (10 mL). The mixture was heated at reflux temperature for 10-16 h, then cooled and allowed to stand overnight. The precipitated solid was collected by filtration, washed with water, dried and crystallized from ethanol to give compounds 4a-e.

-2-

4-(2-(2-Chloro-4-nitrophenyl)hydrazono)-1-(6-fluorobenzothiazol-2-yl)-3-methylpyrazol-5(4H)-one (4a) -1

1

Yield 81%, m.p. 199-201 °C. IR spectrum (KBr, v, cm ): 1688 (C=O). H NMR spectrum: (400 MHz, DMSO-d6, δ -

ppm): 2.05 (s, 3H, CH3), 7.40-8.55 (m, 6H, Ar-H), 13.10 (s, 1H, NH). HRMS: m/z (ESI) calcd for C17H9ClFN6O3S , -

[M-H] : 431.0208; found: 431.0280. 1-(6-Fluorobenzothiazol-2-yl)-3-methyl-4-(2-(4-nitrophenyl)hydrazono)pyrazol-5(4H)-one (4b) -1

1

Yield 82%, m.p. 218-219 °C. IR spectrum (KBr, v, cm ): 1672 (C=O). H NMR spectrum: (400 MHz, DMSO-d6, δ -

ppm): 2.30 (s, 3H, CH3), 7.20-8.35 (m, 7H, Ar-H), 13.05 (s, 1H, NH). HRMS: m/z (ESI) calcd for C17H10FN6O3S , [M-

H] : 397.0597; found: 397.0663. 4-(2-(2,4-Dichlorophenyl)hydrazono)-1-(6-fluorobenzothiazol-2-yl)-3-methylpyrazol-5(4H)-one (4c) -1 1 Yield 72%, m.p. 186-188 °C. IR spectrum (KBr, v, cm ): 1687 (C=O). H NMR spectrum: (400 MHz, CDCl3, δ ppm): -

-

2.50 (s, 3H, CH3), 7.10-8.10 (m, 6H, Ar-H), 13.40 (s, 1H, NH). HRMS: m/z (ESI) calcd for C17H9Cl2FN5OS , [M-H] : 419.9967; found: 420.0038. 4-(2-(2,5-Dichlorophenyl)hydrazono)-1-(6-fluorobenzothiazol-2-yl)-3-methylpyrazol-5(4H)-one (4d) -1 1 Yield 71%, m.p. 207-208 °C. IR spectrum (KBr, v, cm ): 1673 (C=O). H NMR spectrum: (400 MHz, CDCl3, δ ppm): -

-

2.45 (s, 3H, CH3), 7.20-8.10 (m, 6H, Ar-H), 13.45 (s, 1H, NH). HRMS: m/z (ESI) calcd for C17H9Cl2FN5OS , [M-H] : 419.9967; found: 420.0038. 1-(6-Fluorobenzothiazol-2-yl)-3-methyl-4-(2-(2-methyl-5-nitrophenyl)hydrazono)pyrazol-5(4H)-one (4e) -1 1 Yield 82%, m.p. 194-196 °C. IR spectrum (KBr, v, cm ): 1689 (C=O). H NMR spectrum: (400 MHz, CDCl3, δ ppm): 2.35 (s, 3H, CH3), 2.55 (s, 3H, CH3), 7.15-8.10 (m, 6H, Ar-H), 13.20 (s, 1H, NH).

13

C NMR spectrum: (400 MHz,

CDCl3, δ ppm): 12.1, 17.1, 107.5, 107.8, 109.8, 114.9, 120.3, 123.7, 129.1, 132.0, 132.1, 139.6, 146.7, 148.2, 151.9, -

-

157.3, 158.7, 165.7. HRMS: m/z (ESI) calcd for C18H12FN6O3S , [M-H] : 411.0754; found: 411.0685. 6-Fluoro-2-(2-(5-(un)substituted-2-oxoindolin-3-ylidene)hydrazino)benzothiazoles 5a-c. General Procedure C. A mixture of compound 2 (0.183 g, 0.001 mol), the appropriate isatin (0.001 mol) and glacial acetic acid (0.1 mL) in ethanol (20 mL) was heated at reflux temperature for 4-6 h. The precipitated yellow to orange solid was collected by filtration, washed with ethanol, dried and crystallized from methanol to yield compounds 5a-c. 6-Fluoro-2-(2-(2-oxoindolin-3-ylidene)hydrazino)benzothiazole (5a) -1

1

Yield 62%, m.p. 202-204 °C. IR spectrum (KBr, v, cm ): 3145, 3123 (2NH), 1691 (C=O). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 6.90-7.95 (m, 7H, Ar-H), 11.25 (s, 1H, NH), 13.40 (s, 1H, NH). HRMS: m/z (ESI) calcd for +

+

C15H10FN4OS , [M+H] : 313.0557; found: 313.0555. 2-(2-(5-Bromo-2-oxoindolin-3-ylidene)hydrazino)-6-fluorobenzothiazole (5b) -1

1

Yield 66%, m.p. 195-197 °C. IR spectrum (KBr, v, cm ): 3172, 3134 (2NH), 1686 (C=O). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 6.95 (d, 1H, Ar-H), 7.28 (t, 1H, Ar-H), 7.52 (d, 1H, Ar-H), 7.66-7.78 (m, 2H, Ar-H), 7.91 (d, 1H, Ar-H), 11.40 (s, 1H, NH), 13.40 (s, 1H, NH).

13

C NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 109.4, 113.6, 114.6, 114.8, 119.9, 122.7, 129.2, 129.4, 133.3, 140.8, 142.0, 153.3, 157.4, 165.4, 166.1. HRMS: m/z (ESI) calcd for -

-

C15H7BrFN4OS , [M-H] : 388.9508; found: 388.9507.

-3-

6-Fluoro-2-(2-(5-methyl-2-oxoindolin-3-ylidene)hydrazino)benzothiazole (5c) -1

1

Yield 63%, m.p. 211-213 °C. IR spectrum (KBr, v, cm ): 3187, 3125 (2NH), 1687 (C=O). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 2.30 (s, 3H, CH3), 6.85-7.95 (m, 6H, Ar-H), 11.15 (s, 1H, NH), 13.35 (s, 1H, NH).

13

C NMR

spectrum: (400 MHz, DMSO-d6, δ ppm): 21.0, 109.4, 111.4, 114.8, 120.9, 122.1, 124.6, 128.7, 131.9, 132.1, 134.6, -

-

140.0, 141.5, 159.6, 163.5, 166.7. HRMS: m/z (ESI) calcd for C16H10FN4OS , [M-H] : 325.0654; found: 325.0563. 2-(2-Arylidenehydrazino)-6-fluorobenzothiazoles 6a-r. General Procedure D. A mixture of compound 2 (0.0549 g, 0.0003 mol), the appropriate aromatic aldehyde (0.00033 mol) and glacial acetic acid (0.1 mL) in ethanol (5 mL) was heated under microwave (20 W) at 80 °C for 10 min. On cooling, the precipitated solid was collected by filtration, washed with water, dried and crystallized from the appropriate solvent to give the desired compounds 6a-r. 6-Fluoro-2-[2-((furan-2-yl)methylene)hydrazino]benzothiazole (6a) -1

1

Crystallized from ethanol : water (2:1). Yield 72%, m.p. 183-184 °C. IR spectrum (KBr, v, cm ): 3170 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 7.10-7.95 (m, 6H, Ar-H), 8.10 (s, 1H, CH=N), 12.30 (s, 1H, NH). HRMS: m/z -

-

(ESI) calcd for C12H7FN3OS , [M-H] : 260.0294; found: 260.0293. 6-Fluoro-2-[2-((5-nitrofuran-2-yl)methylene)hydrazino]benzothiazole (6b) 1

Crystallized from ethanol : water (2:1). Yield 68%, m.p. 201-202 °C. H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): -

-

6.60-8.10 (m, 6H, Ar-H, CH=N), 12.90 (s, 1H, NH). HRMS: m/z (ESI) calcd for C12H6FN4O3S , [M-H] : 305.0145; found: 305.0144. 6-Fluoro-2-[2-((1H-pyrrol-2-yl)methylene)hydrazino]benzothiazole (6c) -1

1

Crystallized from chloroform. Yield 79%, m.p. 165-167 °C. IR spectrum (KBr, v, cm ): 3171, 3101 (2NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 6.10-7.75 (m, 6H, Ar-H), 8.00 (s, 1H, CH=N), 11.30 (s, 1H, NH), 11.70 (s, 1H, -

-

NH). HRMS: m/z (ESI) calcd for C12H8FN4S , [M-H] : 259.0450; found: 259.0446. 2-(2-(2-Chloro-5-nitrobenzylidene)hydrazino)-6-fluorobenzothiazole (6d) -1

1

Crystallized from ethyl acetate : ethanol (3:1). Yield 70%, m.p. 205-206 °C. IR spectrum (KBr, v, cm ): 3117 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 7.00-8.10 (m, 6H, Ar-H), 8.60 (s, 1H, CH=N), 12.90 (s, 1H, NH). -

-

HRMS: m/z (ESI) calcd for C14H7ClFN4O2S , [M-H] : 349.0049; found: 348.9976. 6-Fluoro-2-(2-(3-methoxybenzylidene)hydrazino)benzothiazole (6e) -1

1

Crystallized from ethyl acetate : ethanol (3:1). Yield 75%, m.p. 182-184 °C. IR spectrum (KBr, v, cm ): 3123 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 3.80 (s, 3H, OCH3), 6.98 (d, 1H, Ar-H), 7.18 (t, 1H, Ar-H), 7.25-7.41 (m, 3H, Ar-H), 7.48 (s, 1H, Ar-H), 7.78 (d, 1H, Ar-H), 8.10 (s, 1H, CH=N), 12.40 (s, 1H, NH). HRMS: m/z (ESI) calcd -

-

for C15H11FN3OS , [M-H] : 300.0685; found: 300.0620. 6-Fluoro-2-(2-(4-methoxybenzylidene)hydrazino)benzothiazole (6f) -1

1

Crystallized from ethanol : water (2:1). Yield 72%, m.p. 209-210 °C. IR spectrum (KBr, v, cm ): 3165 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 3.85 (s, 3H, OCH 3), 7.10-7.95 (m, 7H, Ar-H), 8.15 (s, 1H, CH=N), 12.15 (s, -

-

1H, NH). HRMS: m/z (ESI) calcd for C15H11FN3OS , [M-H] : 300.0685; found: 300.0609. 6-Fluoro-2-(2-(2-nitrobenzylidene)hydrazino)benzothiazole (6g) -1

1

Crystallized from ethyl acetate : ethanol (3:1). Yield 82%, m.p. 221-222 °C. IR spectrum (KBr, v, cm ): 3121 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 7.18 (t, 1H, Ar-H), 7.52 (s, 1H, Ar-H), 7.61-7.70 (m, 1H, Ar-H), 7.73-

-4-

7.90 (m, 2H, Ar-H), 8.01-8.10 (m, 2H, Ar-H), 8.51 (s, 1H, CH=N), 12.80 (s, 1H, NH). HRMS: m/z (ESI) calcd for -

-

C14H8FN4O2S , [M-H] : 315.0430; found: 315.0480. 6-Fluoro-2-(2-(3-hydroxybenzylidene)hydrazino)benzothiazole (6h) -1

1

Crystallized from chloroform. Yield 69%, m.p. 188-190 °C. IR spectrum (KBr, v, cm ): 3316 (OH), 3125 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 6.75-7.85 (m, 7H, Ar-H), 8.10 (s, 1H, CH=N), 9.65 (s, 1H, OH), 12.10 -

-

(s, 1H, NH). HRMS: m/z (ESI) calcd for C14H9FN3OS , [M-H] : 286.0529; found: 286.0444. 2-(2-(2-Bromobenzylidene)hydrazino)-6-fluorobenzothiazole (6i) -1

1

Crystallized from ethanol : water (2:1). Yield 76%, m.p. 194-195 °C. IR spectrum (KBr, v, cm ): 3189 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 7.10-7.90 (m, 7H, Ar-H), 8.40 (s, 1H, CH=N), 12.50 (s, 1H, NH).

13

C NMR


-

156.9, 159.3, 167.5. HRMS: m/z (ESI) calcd for C14H8BrFN3S , [M-H] : 347.9685; found: 347.9611. 6-Fluoro-2-(2-(3,4-dihydroxybenzylidene)hydrazino)benzothiazole (6j) -1

Crystallized from ethanol : water (2:1). Yield 81%, m.p. 180-182 °C. IR spectrum (KBr, v, cm ): 3348 (2OH), 3165 1

(NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 6.77-6.93 (m, 2H, Ar-H), 7.00-7.19 (m, 2H, Ar-H), 7.40-7.73 (m, 2H, Ar-H), 7.95 (s, 1H, CH=N), 9.20 (s, 1H, OH), 9.40 (s, 1H, OH), 12.00 (s, 1H, NH). HRMS: m/z (ESI) calcd for -

-

C14H9FN3O2S , [M-H] : 302.0478; found: 302.0395. 2-(2-(2-Cyanobenzylidene)hydrazino)-6-fluorobenzothiazole (6k) -1

Crystallized from ethanol : water (2:1). Yield 79%, m.p. 205-206 °C. IR spectrum (KBr, v, cm ): 3165 (NH), 2160 1

(C≡N). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 7.10-8.00 (m, 7H, Ar-H), 8.35 (s, 1H, CH=N), 12.70 (s, 1H, NH).

13

C NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 108.9, 109.1, 109.8, 113.9, 114.2, 117.8, 126.8, 130.2, 131.4, -

-

133.9, 134.3, 137.0, 141.7, 157.1, 168.0. HRMS: m/z (ESI) calcd for C15H8FN4S , [M-H] : 295.0532; found: 295.0455. 2-(2-(4-N,N-Dimethylaminobenzylidene)hydrazino)-6-fluorobenzothiazole (6l) -1

1

Crystallized from ethyl acetate : ethanol (3:1). Yield 68%, m.p. 200-201 °C. IR spectrum (KBr, v, cm ): 3196 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 2.95 (s, 6H, N(CH3)2), 6.71-6.82 (m, 2H, Ar-H), 7.08-7.16 (m, 1H, ArH), 7.33-7.51 (m, 2H, Ar-H), 7.61-7.71 (m, 2H, Ar-H), 8.15 (s, 1H, CH=N), 11.90 (s, 1H, NH). HRMS: m/z (ESI) calcd -

-

for C16H14FN4S , [M-H] : 313.1001; found: 313.0922. 2-(2-(4-N,N-Diethylamino-2-hydroxybenzylidene)hydrazino)-6-fluorobenzothiazole (6m) -1

1

Crystallized from chloroform. Yield 64%, m.p. 184-186 °C. IR spectrum (KBr, v, cm ): 3315 (OH), 3166 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 1.10 (t, 6H, 2CH2CH3), 4.15-4.25 (q, 4H, 2CH2CH3), 7.10-8.25 (m, 7H, -

-

Ar-H, CH=N), 9.95 (s, 1H, OH), 12.15 (s, 1H, NH). HRMS: m/z (ESI) calcd for C18H18FN4OS , [M-H] : 357.1264; found: 357.1164. 6-Fluoro-2-[2-((pyridin-2-yl)methylene)hydrazino]benzothiazole (6n) -1

1

Crystallized from ethyl acetate : ethanol (3:1). Yield 69%, m.p. 221-222 °C. IR spectrum (KBr, v, cm ): 3181 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 7.10-8.20 (m, 7H, Ar-H), 8.60 (s, 1H, CH=N), 12.50 (s, 1H, NH). -

-

HRMS: m/z (ESI) calcd for C13H8FN4S , [M-H] : 271.0454; found: 271.0466.

-5-

2-(2-[(3,5-Dichloropyridin-4-yl)methylene)hydrazino]-6-fluorobenzothiazole (6o) -1

1

Crystallized from ethanol : water (2:1). Yield 74%, m.p. 199-201 °C. IR spectrum (KBr, v, cm ): 3124 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 7.22 (t, 1H, Ar-H), 7.59 (s, 1H, Ar-H), 7.82 (s, 1H, CH=N), 8.40 (d, 1H, ArH), 8.72 (s, 2H, pyridine-H), 13.00 (s, 1H, NH).

13

C NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 109.0, 109.3, 114.0, -

-

114.3, 130.1, 136.2, 141.7, 145.3, 149.1, 157.3, 160.3. HRMS: m/z (ESI) calcd for C13H6Cl2FN4S , [M-H] : 338.9675; found: 338.9676. 6-Fluoro-2-[2-((isoquinolin-5-yl)methylene)hydrazino]benzothiazole (6p) -1

1

Crystallized from chloroform. Yield 72%, m.p. 153-155 °C. IR spectrum (KBr, v, cm ): 3157 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 7.18 (t, 1H, Ar-H), 7.53 (s, 1H, Ar-H), 7.76-7.85 (m, 2H, Ar-H), 8.10 (d, 1H, Ar-H), 8.21 (d, 1H, Ar-H), 8.63-8.80 (m, 3H, Ar-H, CH=N), 9.40 (s, 1H, isoquinoline-C1-H), 12.50 (s, 1H, NH). HRMS: m/z (ESI) -

-

calcd for C17H10FN4S , [M-H] : 321.0767; found: 321.0772. 2-[2-((2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)methylene)hydrazino]-6-fluorobenzothiazole (6q) -1

1

Crystallized from ethyl acetate : ethanol (3:1). Yield 81%, m.p. 176-178 °C. IR spectrum (KBr, v, cm ): 3119 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 4.15 (s, 4H, 2CH2), 7.15-8.25 (m, 6H, Ar-H), 8.65 (s, 1H, CH=N), 12.25 -

-

(s, 1H, NH). HRMS: m/z (ESI) calcd for C16H11FN3O2S , [M-H] : 328.0550; found: 328.0553. 6-Fluoro-2-[2-((pyren-2-yl)methylene)hydrazino]benzothiazole (6r) -1

1

Crystallized from ethyl acetate : ethanol (3:1). Yield 72%, m.p. 144-146 °C. IR spectrum (KBr, v, cm ): 3054 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 7.19 (t, 1H, Ar-H), 7.50 (s, 1H, Ar-H), 7.80 (d, 1H, Ar-H), 8.09-8.55 (m, 9H, Ar-H), 8.83 (s, 1H, CH=N), 12.50 (s, 1H, NH).

13

C NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 109.2, 113.8, 123.0, 124.7, 125.4, 125.7, 126.2, 126.5, 127.1, 127.4, 127.9, 128.7, 129.2, 130.7, 131.4, 132.0, 156.9, 167.5. HRMS: -

-

m/z (ESI) calcd for C24H13FN3S , [M-H] : 394.0814; found: 394.0822. 6-Fluoro-2-[2-(1-(substituted phenyl)ethylidene)hydraizno]benzothiazoles 7a-d. General Procedure E. A mixture of compound 2 (0.183 g, 0.001 mol), the appropriate acetophenone (0.001 mol) and glacial acetic acid (0.1 mL) in ethanol (20 mL) was heated at reflux temperature for 6-8 h. On cooling, the precipitated solid was collected by filtration, washed with ethanol, dried and crystallized from ethyl acetate : ethanol (3:1) to furnish compounds 7a-d. 2-[2-(1-(5-Bromo-2-hydroxyphenyl)ethylidene)hydrazino]-6-fluorobenzothiazole (7a) -1

1

Yield 63%, m.p. 187-189 °C. IR spectrum (KBr, v, cm ): 3323 (OH), 3108 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 2.35 (s, 3H, CH3), 6.95-7.90 (m, 6H, Ar-H), 12.10 (s, 1H, OH), 12.85 (s, 1H, NH).

13

C NMR


-

156.8, 157.7, 159.1, 166.5. HRMS: m/z (ESI) calcd for C15H10BrFN3OS , [M-H] : 377.9717; found: 377.9720. 6-Fluoro-2-[2-(1-(3-hydroxy-2-methoxyphenyl)ethylidene)hydrazino]benzothiazole (7b) -1

1

Yield 73%, m.p. 204-206 °C. IR spectrum (KBr, v, cm ): 3289 (OH), 3170 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 1.90 (s, 3H, CH3), 3.90 (s, 3H, OCH3), 6.80-7.70 (m, 6H, Ar-H), 10.00 (s, 1H, OH), 11.90 (s, 1H, NH).

13

C NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 21.4, 55.9, 108.3, 113.8, 117.9, 118.0, 119.9, 123.8, 129.3, -

-

129.7, 146.6, 147.9, 152.1, 156.8, 168.5, 172.6. HRMS: m/z (ESI) calcd for C16H13FN3O2S , [M-H] : 330.0722; found: 330.0722.

-6-

6-Fluoro-2-[2-(1-(2-hydroxy-5-methylphenyl)ethylidene)hydrazino]benzothiazole (7c) -1

1

Yield 61%, m.p. 194-196 °C. IR spectrum (KBr, v, cm ): 3317 (OH), 3123 (NH). H NMR spectrum: (400 MHz, DMSOd6, δ ppm): 2.15 (s, 3H, CH3), 2.35 (s, 3H, CH3), 6.85-7.80 (m, 6H, Ar-H), 11.90 (s, 1H, OH), 12.20 (s, 1H, NH).

13

C

NMR spectrum: (DMSO-d6, δ ppm): 14.9, 20.7, 110.0, 110.3, 114.1, 114.3, 117.1, 120.1, 127.7, 129.0, 131.8, 141.9, +

+

156.4, 156.7, 159.1, 165.8. HRMS: m/z (ESI) calcd for C16H15FN3OS , [M+H] : 316.0927; found: 316.0924. 6-Fluoro-2-[2-(1-(4-iodophenyl)ethylidene)hydrazino]benzothiazole (7d) -1

1

Yield 65%, m.p. 218-219 °C. IR spectrum (KBr, v, cm ): 3272 (NH). H NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 2.10 (s, 3H, CH3), 6.80-7.70 (m, 7H, Ar-H), 12.00 (s, 1H, NH).

13

C NMR spectrum: (400 MHz, DMSO-d6, δ ppm): 21.5,

96.6, 107.8, 113.9, 128.2, 130.4, 136.5, 137.7, 137.8, 138.1, 144.1, 156.8, 164.3. HRMS: m/z (ESI) calcd for -

-

C15H10FIN3S , [M-H] : 409.9702; found: 409.9777.

Biological testing Antimicrobial and antiquorum-sensing activities Compounds 4a-e, 5a-c, 6a-r and 7a-d were screened for their antibacterial activity against Escherichia coli, Staphylococcus aureus and Bacillus cereus in Luria-Bertani agar medium [7,8] and antifungal activity against Candida albicans in Sabouraud's agar medium, Aspergillus fumigatus 293 in glucose minimal medium [8,9]. In addition, their antipathogenic potential was checked by examining the antiquorum-sensing activity against Chromobacterium violaceum ATCC 12472 in Luria-Bertani agar medium [10]. E. coli, B. cereus, S. aureus and C. albicans were obtained from the culture collection of the Department of Microbiology, Faculty of Pharmacy, Mansoura University, Mansoura, Egypt. A. fumigatus 293 was kindly provided from Keller Lab, Madison, WI, UW, USA. Ch. violaceum ATCC 12472 was kindly provided from Prof. Bob Mclean, Department of Biology, Texas State University, San Marcos, TX, USA. All bacterial strains were propagated in LuriaBertani agar medium (Merck, Germany). C. albicans was grown in Sabouraud's medium (E. Merck, Germany) and A. fumigatus 293 was cultivated in glucose minimal medium (E. Merck, Germany). Ampicillin was obtained from EPICO Co., Egypt. Fluconazole was obtained from Pfizer Co., Egypt. Catechin and DMSO 100% were obtained from SigmaAldrich Co., USA. Antibacterial screening (Determination of inhibition zone diameter (mm)) All the bacterial strains were propagated in Luria-Bertani broth (1% peptone, 0.5% yeast extract, 0.5% NaCl) and 6

solidified with 1.5% agar. Melted Muller Hinton agar (50 mL) at 50 °C were seeded with 50 µL of 1x10 CFU/mL of 18 h culture of tested microorganisms. The inoculated agar was mixed and poured into 15-cm-diameter plates to solidify. Wells were made in agar using cork borer. Tested compounds were dissolved in DMSO 100% in eppendorff tubes for final concentration 5 mg/mL. Aliquots each of 100 µL of each compound were applied into the wells, DMSO was also included as a negative control, and ampicillin in a concentration of 5 mg/mL was used as a reference antibacterial agent. The compounds were allowed to diffuse for 2 h at 4 °C and incubated at 37 °C for 24 h [7]. Inhibition zones were measured

-7-

using Vernier caliper and the activity of the tested compounds was estimated in comparison to ampicillin (Table 1). The inhibition zone diameter of DMSO was subtracted from the antibacterial activity of tested compounds. Antibacterial screening (Determination of minimum inhibitory concentration (MIC, µg/mL )) Minimum inhibitory concentration (MIC, µg/mL) of compounds 6j and 6o (with strong antibacterial activity against E. coli and S. aureus) was performed by a serial dilution technique using 96-multiwell microtiter plates [8]. The investigated compounds were dissolved in DMSO 100% to prepare stock solutions of 5000 µg/mL. Two fold serial dilutions of the dissolved compounds were performed in Luria-Bertani broth providing eight different concentrations (2500, 1250, 625, 312.5, 156.25, 78.125, 39.062 and 19.531 µg/mL). Overnight culture of microorganisms were diluted 6

to 1x10 CFU/mL in Luria-Bertani broth and 20 µL of the diluted cultures were added to the test solutions (100 µL) in the wells (one well per concentration per bacterial strain). The plates were incubated at 37 °C for 24 h [8]. MIC was detected visually as the least concentration inhibiting bacterial growth (no turbidity) and the activity of the tested compounds was estimated in comparison to ampicillin (Table 2). Antifungal screening (Determination of inhibition zone diameter (mm)) 3

Sabouraud's medium (50 mL) inoculated with 50 µL of 1x10 CFU/mL of 24 h culture of C. albicans. Glucose 3

minimal medium (50 mL) was inoculated with 50 µL of 1x10 SFU/mL of A. fumigatus 293. Wells were made in agar using cork borer. The tested compounds were dissolved in DMSO 100% in eppendorff tubes for final concentration 5 mg/mL and 100 µL of the test solution was applied in the wells. The standard antifungal drug (fluconazole) was also added at the same concentration and volume to each plate. In addition, DMSO (control solvent) was added to each plate. Plates were incubated at 37 °C for 48 h [9]. Antifungal activity of the tested compounds was determined by measuring the diameter of the inhibition zone (Table 1). The inhibition zone diameter of DMSO was subtracted from the antifungal acrivity of tested compounds. Antifungal screening (Determination of minimum inhibitory concentration (MIC, µg/mL)) Minimum inhibitory concentration (MIC, µg/mL) of compounds 4b, 4c and 6j (with strong antifungal activity against C. albicans) and compounds 4a, 5a, 6j, 6l, 7c and 7d (with strong antifungal activity against A. fumigatus 293) was determined by broth microdilution method using 96-multiwell microtiter plates. The investigated compounds were dissolved in DMSO 100% to prepare stock solutions of 5000 µg/mL. Two fold serial dilutions of the dissolved compounds were performed in the appropriate medium providing eight different concentrations (2500, 1250, 625, 312.5, 6

156.25, 78.125, 39.062 and 19.531 µg/mL). Overnight culture of C. albicans was diluted to 1x10 CFU/mL in 3

Sabouraud's medium and A. fumigatus 293 was diluted to 1x10 SFU/mL in glucose minimal medium. The diluted cultures (20 µL) were added to the test solutions (100 µL) in the wells (one well per concentration). The plates were incubated at 37 °C for 48 h [8]. MIC was detected visually as the least concentration inhibiting fungal growth (no turbidity) and the activity of the tested compounds was estimated in comparison to fluconazole (Table 2).

-8-

Antiquorum-sensing screening Cultures were prepared by growing Ch. violaceum ATCC 12472 in Luria-Bertani broth and incubated for 16-18 h in an orbital incubator running at 28 °C and 150 rpm. Cultures were then adjusted to 0.5 McFarland standard (Ca. 1x10

6

CFU/mL). Ch. violaceum (100 µL) was inoculated into Luria-Bertani agar (50 mL), poured into plates and solidified. Wells were made in Luria-Bertani agar medium using cork borer. The tested compounds were dissolved in DMSO 100% in eppendorff tubes for final concentration 5 mg/mL and 100 µL of test solution was applied in the wells. The positive control (catechin) was also added at the same concentration and volume to each plate. The positive control (catechin) was also added at the same concentration and volume to each plate. DMSO (control solvent) was added to each plate. Plates were incubated at 30 °C for 48 h to check the inhibition of pigment production around the well. Bacterial growth inhibition would result in a clear halo around the disc, while a positive quorum sensing inhibition is exhibited by a turbid halo harboring pigmentless bacterial cells of Ch. violaceum ATCC 12472 monitor strain [10]. Bacterial growth inhibition by the tested compounds was measured as radius (r1) in mm, while both growth and pigment inhibition was measured as radius (r2) in mm. The pigment inhibition (QS inhibition) was determined by substracting bacterial growth inhibition (r1) from the total radius (r2), thus; QS inhibition = (r2-r1) in mm (Table 1).

MTT assay for antitumor screening Compounds 4a-e, 5a-c, 6a-r and 7a-d were screened for cytotoxic activity against cervical cancer (Hela) and kidney fibroblast cancer (COS-7) cell lines adopting MTT assay [11-13] and using doxorubicin as a reference cytotoxic agent. Cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium with 10% fetal bovine serum, penicillin (100 units/mL) and streptomycin (100 µg/mL) at 37 °C in an atmosphere of 5% CO2 were added. 4

Cells were placed in a 96-multiwell microtiter plate (10 cells/well), for 24 h at 37 °C and in an atmosphere of 5% CO2 before treatment with the compounds to allow attachment of the cells to the wall of the plate. The tested compounds were dissolved in DMSO and diluted with phosphate buffer solution (PBS) to obtain different concentrations. Tested compounds of different concentrations were added to each well and cells were incubated with the compounds for 48 h at 37 °C and in an atmosphere of 5 % CO2. All tests were performed in triplicates. The treated cells were washed with PBS and 100 µL of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide solution (MTT) (5 mg/mL MTT stock in PBS diluted to 1 mg/mL with 10% RPMI-1640 medium) was added. The 96-multiwell plates were read by microarray reader Perkinelmer vector 3V multilabel counter model 1420 (Perkinelmer, Boston, MA) for optical density at 490 nm [11-13]. The relative percentage cell viability was calculated from the following equation:

A

% cell viability =

-A

treated cells

A

blank

-A untreated cells

X 100

blank

The relation between surviving fraction and drug concentration is plotted to get the survival curve for cervical cancer (Hela) and kidney fibroblast cancer (COS-7) cell lines. The concentration required for 50% inhibition of cell viability (IC50) was obtained for each compound from the curve fitting using Sigma plot10.

-9-

References [1] P. Yadav, D. Chauhan, N.K. Sharma, S. Singhal, 2-Substituted hydrazino-6-fluoro-1,3-benzothiazole: Synthesis and characterization of new novel antimicrobial agents. Int. J. ChemTech Res. 2 (2010) 1209-1213. [2] A.K. Pareek, P.E. Joseph, D.S. Seth, Novel and efficient synthesis and spectral evaluation of certain new substituted pyrazolones. Orient. J. Chem. 25 (2009) 203-206. [3] C.P. Singh, A.C. Ojha, Synthesis of 1-isonicotinoyl-3-methyl-4-(substitutedazo)pyrazol-5-ones. J. Chin. Chem. Soc. 28 (1981) 51-57. [4] N.G. Ferguson, C. Valant, J. Horne, H. Figler, B.L. Flynn, J. Linden, D.K. Chalmers, P.M. Sexton, A. Christopoulos, P. J. Scammells, 2-Aminothienopyridazines as novel adenosine A1 receptor allosteric modulators and antagonists. J. Med. Chem. 51 (2008) 6165-6172. [5] N. Jagerovic, L. Hernandez-Folgado, I. Alkorta, P. Goya, M.I. Martin, M.T. Dannert, A. Alsasua, J. Frigola, M.R. Cuberes, A. Dordal, J. Holenz, Structural-activity relationship study on C-4 carbon atom of the CB1 antagonist SR141716: synthesis and pharmacological evaluation of 1,2,4-triazole-3-carboxamides. Eur. J. Med. Chem. 41 (2006) 114-120. [6] N. Prasad, A. Sahay, N.P. Singh, Electron impact studies on some ethyl 2,3-dioxobutyrate-2-phenylhydrazones. Pak. J. Sci. Ind. Res. 44 (2001) 215-217. [7] R.D. Pearson, R.T. Steigbigel, H.T. Davis, S.W. Chapmann, Method for reliable determination of minimal lethal antibiotic concentrations. Antimicrob. Agents Chemother. 18 (1980) 699-708. [8] J.M. Andrews, Determination of minimum inhibitory concentrations. J. Antimicrob. Chemother. 48 (2001) 5-16. [9] R.J. Holt, Laboratory tests of antifungal drugs. J. Clin. Pathol. 28 (1975) 767-774. [10] K.H. McClean, M.K. Winson, L. Fish, A. Taylor, S.R. Chhabra, M. Camara, M. Daykin, J.H. Lamb, S. Swift, B.W. Bycroft, G.S. Stewart, P. Williams, Quorum sensing and Chromobacterium violaceum: exploitation of violacein production and inhibition for the detection of N-acyl homoserine lactones. Microbiology 143 (1997) 3703-3711. [11] T. Mosmann, Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immunol. Methods 65 (1983) 55-63. [12] F. Denizot, R. Lang, Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J. Immunol. Methods 89 (1986) 271-277. [13] D. Gerlier, T. Thomasset, Use of MTT colorimetric assay to measure cell activation. J. Immunol. Methods 94 (1986) 57-63.

- 10 -

10-Jun-201317:04:16

%

MOUSTAFA_MG-I-130C_BWANG-ACCU_06102013_ESI-NEG01 102 (1.896) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Cm (94:111) 397.0523 5.47e4 100

398.0583 399.0540

0 100

212.0724

200

278.0791

300

400.0551

400

Elemental Composition Report Single Mass Analysis Tolerance = 5.0 PPM / DBE: min = -1.5, max = 100.0 Element prediction: Off Number of isotope peaks used for i-FIT = 3

500

* 554.2615

600

795.1523818.1132

700

800

900

m/z 1000

Monoisotopic Mass, Odd and Even Electron Ions 4085 formula(e) evaluated with 58 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT 397.0523 397.0538 -1.5 -3.8 0.5 2824.0 397.0511 1.2 3.0 3.5 2466.3 397.0522 0.1 0.3 -1.0 3002.6 397.0526 -0.3 -0.8 11.5 645.2 397.0529 -0.6 -1.5 2.5 812.6 397.0513 1.0 2.5 1.0 1557.1 397.0504 1.9 4.8 12.5 324.8 397.0542 -1.9 -4.8 7.5 410.7 397.0531 -0.8 -2.0 4.0 595.6 397.0504 1.9 4.8 7.0 453.3 397.0526 -0.3 -0.8 6.0 1008.0 397.0515 0.8 2.0 2.5 1077.7 397.0506 1.7 4.3 8.5 314.1 397.0539 -1.6 -4.0 11.0 628.2 397.0528 -0.5 -1.3 7.5 637.3 397.0517 0.6 1.5 12.0 177.9 397.0517 0.6 1.5 6.5 292.3 397.0539 -1.6 -4.0 5.5 969.8 397.0531 -0.8 -2.0 11.5 79.1 397.0520 0.3 0.8 8.0 169.2 397.0542 -1.9 -4.8 7.0 623.5 397.0504 1.9 4.8 6.5 749.4 397.0506 1.7 4.3 16.0 15.3 397.0506 1.7 4.3 10.5 54.5

Formula C H10 N12 O5 F5 S C H10 N14 O8 F S C H14 N11 O6 F3 S2 C10 H10 N12 O F S2 C10 H16 N2 O9 F3 S C10 H20 N O10 F S2 C10 H7 N12 O2 F2 S C11 H12 N6 O5 F3 S C11 H13 N3 O4 F6 S C11 H13 N5 O7 F2 S C11 H16 N5 O6 F S2 C11 H17 N2 O5 F4 S2 C12 H10 N6 O2 F5 S C12 H12 N9 O2 F S2 C12 H13 N6 O F4 S2 C12 H9 N9 O3 F2 S C13 H15 N2 O8 F2 S C13 H18 N2 O7 F S2 C14 H11 N6 O4 F2 S C14 H12 N3 O3 F5 S C14 H15 N3 O2 F4 S2 C14 H16 N2 O4 F3 S2 C15 H8 N9 O2 F S C16 H14 N2 O7 F S

397.0519 397.0508 397.0530 397.0533 397.0535 397.0524 397.0524 397.0513 397.0524 397.0513 397.0535 397.0511 397.0538 397.0538 397.0527 397.0538 397.0527 397.0524 397.0540 397.0524 397.0513 397.0537 397.0526 397.0540 397.0515 397.0504 397.0515 397.0504 397.0537 397.0526 397.0529

0.4 1.5 -0.7 -1.0 -1.2 -0.1 -0.1 1.0 -0.1 1.0 -1.2 1.2 -1.5 -1.5 -0.4 -1.5 -0.4 -0.1 -1.7 -0.1 1.0 -1.4 -0.3 -1.7 0.8 1.9 0.8 1.9 -1.4 -0.3 -0.6

1.0 3.8 -1.8 -2.5 -3.0 -0.3 -0.3 2.5 -0.3 2.5 -3.0 3.0 -3.8 -3.8 -1.0 -3.8 -1.0 -0.3 -4.3 -0.3 2.5 -3.5 -0.8 -4.3 2.0 4.8 2.0 4.8 -3.5 -0.8 -1.5

15.5 12.0 11.0 15.0 4.0 0.5 8.5 5.0 3.0 -0.5 -1.5 3.0 0.0 8.0 4.5 2.5 -1.0 8.0 4.0 2.5 -1.0 7.5 4.0 -1.5 8.5 5.0 3.0 -0.5 2.0 -1.5 8.0

8.6 11.6 570.3 45.9 2067.3 2370.6 1715.3 2105.3 2032.7 2435.3 2651.5 1959.5 2043.9 1351.6 1691.9 1653.4 2007.7 1236.3 1332.0 1719.0 1936.6 1069.0 1218.3 1633.0 861.0 1143.9 1080.1 1375.2 1528.2 1699.7 609.0

C17 H10 N6 O3 F S C17 H11 N3 O2 F4 S C17 H14 N3 O F3 S2 C19 H12 N3 O4 F S C2 H10 N15 O2 F3 S2 C2 H11 N12 O F6 S2 C2 H6 N18 O4 F S C2 H7 N15 O3 F4 S C3 H12 N11 O9 F S C3 H13 N8 O8 F4 S C3 H16 N8 O7 F3 S2 C4 H13 N11 O5 F2 S2 C4 H13 N9 O2 F6 S2 C4 H8 N15 O5 F S C4 H9 N12 O4 F4 S C5 H14 N8 O10 F S C5 H15 N5 O9 F4 S C5 H9 N15 O F2 S2 C6 H11 N9 O5 F4 S C6 H15 N8 O6 F2 S2 C6 H16 N5 O5 F5 S2 C7 H11 N12 O2 F2 S2 C7 H12 N9 O F5 S2 C7 H17 N2 O10 F4 S C7 H8 N12 O3 F3 S C7 H9 N9 O2 F6 S C8 H14 N5 O8 F3 S C8 H15 N2 O7 F6 S C8 H17 N5 O7 F2 S2 C8 H18 N2 O6 F5 S2 C9 H10 N9 O4 F3 S

397.0518 397.0540 397.0513

0.5 -1.7 1.0

1.3 -4.3 2.5

4.5 3.5 6.5

843.9 1055.2 1092.5

C9 H11 N6 O3 F6 S C9 H14 N6 O2 F5 S2 C9 H14 N8 O5 F S2

17.14 12.14

132.12 132.08 123.72 123.64 120.25 114.89 109.79 107.77 107.50

139.70

151.96

Current Data Parameters NAME MG−II−63c EXPNO 1 PROCNO 1 F2 − Acquisition Parameters Date_ 20130514 Time 10.55 INSTRUM spect PROBHD 5 mm PABBO BB− PULPROG zgpg30 TD 65536 SOLVENT CDCl3 NS 503 DS 4 SWH 24038.461 Hz FIDRES 0.366798 Hz AQ 1.3631488 sec RG 90.5 DW 20.800 usec DE 6.50 usec TE 298.0 K D1 2.00000000 sec D11 0.03000000 sec TD0 1 ======== CHANNEL f1 ======== SFO1 100.6253441 MHz NUC1 13C P1 9.00 usec PLW1 62.00000000 W ======== CHANNEL f2 ======== SFO2 400.1416006 MHz NUC2 1H CPDPRG[2 waltz16 PCPD2 90.00 usec PLW2 16.00000000 W PLW12 0.36000001 W PLW13 0.29159999 W F2 − Processing parameters SI 32768 SF 100.6152830 MHz WDW EM SSB 0 LB 1.00 Hz GB 0 PC 1.40

200

180

160

140

120

100

80

60

40

20

0

ppm

rinse90%MeOH w 0.1%FA

10:52:03 29-Apr-2013

%

MOUSTAFA_MG-I-9C_BWANG-ACCU_04292013_ESI-POS01 16 (0.296) AM (Cen,4, 80.00, Ar,6000.0,556.28,0.70); Sm (SG, 3x3.00); Cm (1 313.0555 1.17e4 100

411.9809

556.2771

168.9325

0 100

200

300

400


500

600

700

800

900

m/z 1000

Monoisotopic Mass, Odd and Even Electron Ions 2085 formula(e) evaluated with 19 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-4 Minimum: -1.5 Maximum: 5.0 5.0 50.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 313.0555 313.0564 -0.9 -2.9 0.0 413.7 C H12 N9 O7 F S 313.0557 -0.2 -0.6 9.0 12.0 C10 H9 N7 O F2 S 313.0560 -0.5 -1.6 5.0 10.8 C12 H12 N O F5 S 313.0559 -0.4 -1.3 12.5 27.2 C15 H10 N4 O F S 313.0551 0.4 1.3 0.0 476.2 C2 H13 N9 O3 F2 S2 313.0567 -1.2 -3.8 1.5 318.3 C2 H9 N10 O2 F4 S 313.0542 1.3 4.2 0.5 260.9 C4 H12 N6 O5 F3 S 313.0564 -0.9 -2.9 -0.5 427.0 C4 H15 N6 O4 F2 S2 313.0539 1.6 5.1 4.0 310.3 C5 H12 N9 O2 F S2 313.0555 0.0 0.0 5.5 134.0 C5 H8 N10 O F3 S 313.0555 0.0 0.0 0.0 194.1 C6 H14 N3 O6 F3 S 313.0539 1.6 5.1 -1.5 441.0 C6 H18 N2 O7 F S2 313.0569 -1.4 -4.5 5.0 86.3 C7 H10 N7 O2 F3 S 313.0558 -0.3 -1.0 1.5 130.1 C7 H11 N4 O F6 S 313.0553 0.2 0.6 3.5 289.4 C7 H14 N6 O3 F S2 313.0542 1.3 4.2 0.0 306.0 C7 H15 N3 O2 F4 S2 313.0564 -0.9 -2.9 -1.0 732.2 C7 H18 N3 O F3 S3 313.0544 1.1 3.5 4.0 64.9 C9 H13 N3 O5 F2 S 313.0566 -1.1 -3.5 3.0 281.7 C9 H16 N3 O4 F S2

15-May-201312:54:53

MOUSTAFA_MG-I-10C_BWANG-ACCU_05142013_ESI-NEG01 65 (1.207) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Sm (SG, 2x3.00); Cm (61 390.9484 1.13e4 100

%

388.9507

391.9504

389.9538 384.0985

0 384

385

387.5880

386

387

392.9489

388.6009

388

393.9481 395.0416 397.3223

389


390

391

392

393

394

395

396

397

398

398.9305

m/z 399

Monoisotopic Mass, Even Electron Ions 2386 formula(e) evaluated with 11 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-4 Br: 1-5 Minimum: Maximum: Mass 388.9507

Calc. Mass 388.9519 388.9508 388.9515 388.9519 388.9513 388.9491 388.9504 388.9488 388.9501 388.9497 388.9506

5.0 mDa -1.2 -0.1 -0.8 -1.2 -0.6 1.6 0.3 1.9 0.6 1.0 0.1

5.0 PPM -3.1 -0.3 -2.1 -3.1 -1.5 4.1 0.8 4.9 1.5 2.6 0.3

-1.5 50.0 DBE 9.5 13.5 2.5 -1.5 0.5 1.5 6.5 -0.5 4.5 -1.5 2.5

i-FIT 24.7 4.6 392.6 1435.8 289.5 333.5 201.4 242.3 132.2 160.5 197.9

Formula C12 H8 N4 O2 F2 S Br C15 H7 N4 O F S Br C2 H6 N10 O2 F4 S Br C3 H16 N8 O F S Br2 C4 H12 N6 O4 F2 S2 Br C4 H9 N6 O5 F3 S Br C5 H5 N10 O F3 S Br C6 H15 N2 O7 F S2 Br C7 H11 N6 O3 F S2 Br C7 H19 N2 O2 F S4 Br C7 H8 N4 O F6 S Br

test

Current Data Parameters NAME MG10 EXPNO 1 PROCNO 1 F2 − Acquisition Parameters Date_ 20130124 Time 17.16 INSTRUM spect PROBHD 5 mm PABBO BB− PULPROG zg30 TD 65536 SOLVENT DMSO NS 16 DS 2 SWH 8012.820 Hz FIDRES 0.122266 Hz AQ 4.0894465 sec RG 203 DW 62.400 usec DE 6.50 usec TE 294.9 K D1 1.00000000 sec TD0 1 ======== CHANNEL f1 ======== SFO1 400.1424710 MHz NUC1 1H P1 13.50 usec PLW1 16.00000000 W F2 − Processing parameters SI 65536 SF 400.1400000 MHz WDW EM SSB 0 LB 0.30 Hz GB 0 PC 1.00

10

9

8

7

6

5

4

3

2 24.79

11

67.29

12

1.01 1.93 0.96 1.26 0.98

13

0.90

14

0.88

15

1

0

−1

−2

ppm

133.30 129.20 122.66 119.90 114.83 114.59 113.55 112.23 110.72 109.35

142.03

157.42

165.39

Current Data Parameters NAME MG−I−10c EXPNO 1 PROCNO 1 F2 − Acquisition Parameters Date_ 20130509 Time 11.37 INSTRUM spect PROBHD 5 mm PABBO BB− PULPROG zgpg30 TD 65536 SOLVENT DMSO NS 150 DS 4 SWH 24038.461 Hz FIDRES 0.366798 Hz AQ 1.3631488 sec RG 181 DW 20.800 usec DE 6.50 usec TE 298.3 K D1 2.00000000 sec D11 0.03000000 sec TD0 1 ======== CHANNEL f1 ======== SFO1 100.6253441 MHz NUC1 13C P1 9.00 usec PLW1 62.00000000 W ======== CHANNEL f2 ======== SFO2 400.1416006 MHz NUC2 1H CPDPRG[2 waltz16 PCPD2 90.00 usec PLW2 16.00000000 W PLW12 0.36000001 W PLW13 0.29159999 W F2 − Processing parameters SI 32768 SF 100.6152830 MHz WDW EM SSB 0 LB 1.00 Hz GB 0 PC 1.40

200

180

160

140

120

100

80

60

40

20

0

ppm

03-Jun-201314:30:39

%

MOUTAFA_MG-I-11C_BWANG-ACCU_06032013_ESI-NEG02 87 (1.615) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Sm (Md, 3.00); Cm (80:87 325.0563 4.32e3 100

* 554.2615 326.0582 328.2484 374.2374 358.2317

0 320

340

360

380

555.2675 415.2628 425.2538 465.3808

400

420


440

460

481.3591

480

515.3647 526.3660

500

520

540

557.2233

m/z 560

580

Monoisotopic Mass, Odd and Even Electron Ions 1821 formula(e) evaluated with 21 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 325.0563 325.0564 -0.1 -0.3 6.5 153.6 C H6 N16 O2 F S 325.0553 1.0 3.1 3.0 188.3 C H7 N13 O F4 S 325.0571 -0.8 -2.5 2.0 55.9 C10 H13 N O2 F6 S 325.0566 -0.3 -0.9 4.0 58.8 C10 H16 N3 O4 F S2 325.0557 0.6 1.8 10.0 19.2 C11 H9 N7 O F2 S 325.0571 -0.8 -2.5 9.5 9.6 C13 H11 N4 O2 F2 S 325.0560 0.3 0.9 6.0 18.7 C13 H12 N O F5 S 325.0559 0.4 1.2 13.5 3.1 C16 H10 N4 O F S 325.0573 -1.0 -3.1 13.0 5.1 C18 H12 N O2 F S 325.0564 -0.1 -0.3 1.0 176.9 C2 H12 N9 O7 F S 325.0551 1.2 3.7 1.0 146.9 C3 H13 N9 O3 F2 S2 325.0578 -1.5 -4.6 6.0 121.2 C3 H8 N13 O3 F S 325.0567 -0.4 -1.2 2.5 151.7 C3 H9 N10 O2 F4 S 325.0578 -1.5 -4.6 0.5 143.1 C4 H14 N6 O8 F S 325.0564 -0.1 -0.3 0.5 124.0 C5 H15 N6 O4 F2 S2 325.0555 0.8 2.5 6.5 79.6 C6 H8 N10 O F3 S 325.0555 0.8 2.5 1.0 94.0 C7 H14 N3 O6 F3 S 325.0578 -1.5 -4.6 0.0 105.6 C7 H17 N3 O5 F2 S2 325.0569 -0.6 -1.8 6.0 56.7 C8 H10 N7 O2 F3 S 325.0558 0.5 1.5 2.5 78.3 C8 H11 N4 O F6 S 325.0553 1.0 3.1 4.5 68.0 C8 H14 N6 O3 F S2

15-May-201311:09:18

%

MOUSTAFA_MG-I-68C_BWANG-ACCU_05142013_ESI-NEG01 48 (0.892) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Sm (SG, 2x3.00); Cm (48:62) 260.0293 8.86e3 100

165.9978 167.0044

0 100

261.0319 * 554.2615

262.0277 425.0189

200

300

400

500


600

700

800

900

m/z 1000

Monoisotopic Mass, Odd and Even Electron Ions 867 formula(e) evaluated with 8 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-4 Minimum: -1.5 Maximum: 5.0 5.0 50.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 260.0293 260.0294 -0.1 -0.4 10.5 0.4 C12 H7 N3 O F S 260.0290 0.3 1.2 3.5 215.9 C2 H5 N9 O F3 S 260.0287 0.6 2.3 1.5 266.9 C4 H11 N5 O3 F S2 260.0303 -1.0 -3.8 3.0 157.0 C4 H7 N6 O2 F3 S 260.0292 0.1 0.4 -0.5 212.0 C4 H8 N3 O F6 S 260.0301 -0.8 -3.1 1.0 239.6 C6 H13 N2 O4 F S2 260.0292 0.1 0.4 7.0 47.3 C7 H6 N6 O F2 S 260.0305 -1.2 -4.6 6.5 23.6 C9 H8 N3 O2 F2 S

03-Jun-201314:09:52

%

MOUTAFA_MG-I-41C_BWANG-ACCU_06032013_ESI-NEG01 118 (2.195) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Sm (Md, 3.00); Cm (117 305.0144 8.88e3 100

306.0159 307.0123 342.2294

0 300

320

340

360

* 554.2615

393.9491

380

400


420

440

460

480

500

520

540

560

580

m/z 600

Monoisotopic Mass, Even Electron Ions 1389 formula(e) evaluated with 10 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 305.0144 305.0150 -0.6 -2.0 -1.5 424.7 C H11 N6 O6 F2 S2 305.0145 -0.1 -0.3 11.5 3.7 C12 H6 N4 O3 F S 305.0141 0.3 1.0 4.5 187.7 C2 H4 N10 O3 F3 S 305.0138 0.6 2.0 2.5 270.2 C4 H10 N6 O5 F S2 305.0143 0.1 0.3 0.5 184.3 C4 H7 N4 O3 F6 S 305.0129 1.5 4.9 8.5 64.5 C5 H3 N10 O2 F2 S 305.0152 -0.8 -2.6 7.5 161.6 C5 H6 N10 O F S2 305.0132 1.2 3.9 4.5 62.3 C7 H6 N4 O2 F5 S 305.0154 -1.0 -3.3 3.5 160.1 C7 H9 N4 O F4 S2 305.0156 -1.2 -3.9 7.5 16.8 C9 H7 N4 O4 F2 S

03-Jun-201313:27:28

%


280.2635

296.2567

0 240

312.2566 342.2540 390.2013

260

280

300

320

340

360


380

400

488.0125 519.1009

432.1668

420

440

460

480

500

520

* 554.2615

m/z 540

560

Monoisotopic Mass, Odd and Even Electron Ions 208 formula(e) evaluated with 5 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 259.0446 259.0454 -0.8 -3.1 10.5 12.5 C12 H8 N4 F S 259.0450 -0.4 -1.5 3.5 436.8 C2 H6 N10 F3 S 259.0452 -0.6 -2.3 -0.5 431.3 C4 H9 N4 F6 S 259.0452 -0.6 -2.3 7.0 142.4 C7 H7 N7 F2 S 259.0454 -0.8 -3.1 3.0 140.1 C9 H10 N F5 S

15-May-201312:26:38

%

MOUSTAFA_MG-I-16C_BWANG-ACCU_05142013_ESI-NEG02 82 (1.526) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Sm (SG, 2x3.00); Cm (78:85) 348.9976 8.06e3 100

350.9943

212.0704 347.9673 351.9958 150.0236

0 100

200

* 554.2615

400.8586

300

400

500

Elemental Composition Report Single Mass Analysis Tolerance = 5.0 PPM / DBE: min = -1.5, max = 50.0

600

698.9953 722.9771

700

800

900

m/z 1000

Element prediction: Off Number of isotope peaks used for i-FIT = 3 Monoisotopic Mass, Odd and Even Electron Ions 3175 formula(e) evaluated with 37 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-4 Cl: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 50.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 348.9976 348.9976 0.0 0.0 -1.0 551.7 C H13 N9 O3 F S3 Cl 348.9981 -0.5 -1.4 5.0 506.1 C H5 N13 O4 F S Cl 348.9970 0.6 1.7 1.5 575.1 C H6 N10 O3 F4 S Cl 348.9963 1.3 3.7 1.0 1057.0 C H8 N11 O F3 S Cl2 348.9992 -1.6 -4.6 0.5 540.0 C H9 N10 O2 F3 S2 Cl 348.9992 -1.6 -4.6 3.5 784.4 C10 H13 N2 O3 F2 S Cl2 348.9967 0.9 2.6 8.0 704.6 C11 H10 N5 O F S Cl2 348.9985 -0.9 -2.6 4.0 181.9 C11 H12 N O F4 S2 Cl 348.9974 0.2 0.6 8.5 151.5 C11 H8 N4 O3 F2 S Cl 348.9963 1.3 3.7 5.0 188.7 C11 H9 N O2 F5 S Cl 348.9987 -1.1 -3.2 8.0 118.9 C13 H10 N O4 F2 S Cl 348.9981 -0.5 -1.4 7.5 678.4 C13 H12 N2 O2 F S Cl2 348.9962 1.4 4.0 12.5 66.9 C14 H7 N4 O2 F S Cl 348.9976 0.0 0.0 12.0 46.7 C16 H9 N O3 F S Cl 348.9981 -0.5 -1.4 -0.5 580.4 C2 H11 N6 O9 F S Cl 348.9977 -0.1 -0.3 0.5 1005.0 C3 H10 N8 O2 F3 S Cl2 348.9967 0.9 2.6 -0.5 490.5 C3 H12 N6 O5 F2 S2 Cl 348.9961 1.5 4.3 -1.0 1097.3 C3 H14 N7 O3 F S2 Cl2 348.9989 -1.3 -3.7 -1.5 492.1 C3 H15 N6 O4 F S3 Cl 348.9983 -0.7 -2.0 1.0 503.5 C3 H8 N7 O4 F4 S Cl 348.9981 -0.5 -1.4 4.5 361.8 C4 H8 N10 O F2 S2 Cl

348.9990 348.9981 348.9974 348.9983 348.9967 348.9972 348.9961 348.9965 348.9965 348.9974 348.9979 348.9968 348.9969 348.9985 348.9960 348.9983

-1.4 -0.5 0.2 -0.7 0.9 0.4 1.5 1.1 1.1 0.2 -0.3 0.8 0.7 -0.9 1.6 -0.7

-4.0 -1.4 0.6 -2.0 2.6 1.1 4.3 3.2 3.2 0.6 -0.9 2.3 2.0 -2.6 4.6 -2.0

0.0 -1.0 -1.5 0.5 -1.0 5.0 1.5 4.5 -1.0 1.0 4.0 0.5 3.0 4.5 9.0 8.0

957.9 429.2 1051.2 359.9 381.1 327.2 383.2 858.0 932.4 325.1 818.8 856.1 275.2 275.1 190.4 183.1

C5 C5 C5 C6 C6 C6 C6 C6 C7 C8 C8 C8 C8 C8 C9 C9

H12 N5 O3 F3 S Cl2 H14 N3 O6 F2 S2 Cl H16 N4 O4 F S2 Cl2 H11 N4 O F5 S2 Cl H15 N3 O2 F3 S3 Cl H7 N7 O3 F3 S Cl H8 N4 O2 F6 S Cl H9 N8 O F2 S Cl2 H15 N O6 F2 S Cl2 H10 N O3 F6 S Cl H11 N5 O2 F2 S Cl2 H12 N2 O F5 S Cl2 H13 N3 O5 F S2 Cl H9 N4 O4 F3 S Cl H6 N7 O2 F2 S Cl H9 N7 O F S2 Cl

15-May-201310:02:24

%

MOUSTAFA_MG-I-48C_BWANG-ACCU_05142013_ESI-NEG01 47 (0.876) AM (Med,2, Ht,5000.0,554.26,1.00); Cm (43:53) 300.0620 1.44e3 100

212.0728 301.0655 370.0830 165.9937 213.0754

0 100

200

300

* 554.2615

371.0874

400

Elemental Composition Report Single Mass Analysis Tolerance = 5.0 PPM / DBE: min = -1.5, max = 50.0 Element prediction: Off

500

600

623.0963

700

800

900

m/z 1000

Number of isotope peaks used for i-FIT = 3 Monoisotopic Mass, Even Electron Ions 1713 formula(e) evaluated with 7 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-4 Minimum: -1.5 Maximum: 5.0 5.0 50.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 300.0620 300.0618 0.2 0.7 7.5 4.9 C12 H12 N3 O2 F2 S 300.0607 1.3 4.3 11.5 0.3 C15 H11 N3 O F S 300.0614 0.6 2.0 0.5 61.2 C2 H10 N9 O2 F4 S 300.0625 -0.5 -1.7 -1.5 59.3 C3 H15 N5 O8 F S 300.0612 0.8 2.7 -1.5 56.2 C4 H16 N5 O4 F2 S2 300.0605 1.5 5.0 0.5 33.1 C7 H12 N3 O F6 S 300.0630 -1.0 -3.3 3.5 17.6 C9 H13 N3 O3 F3 S

03-Jun-201313:58:56

%

MOUTAFA_MG-I-49C_BWANG-ACCU_06032013_ESI-NEG02 46 (0.854) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Sm (Md, 3.00); Cm (42:49 300.0609 7.22e3 100

301.0659 302.0599 324.2304

0 280

300

320

340

360

394.2303

380

400


410.2276

420

440

484.3546 512.3580

460

480

500

526.3524

520

* 554.2615

540

577.3019

m/z 560

580

Monoisotopic Mass, Odd and Even Electron Ions 1287 formula(e) evaluated with 13 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 300.0609 300.0612 -0.3 -1.0 -1.0 368.0 C H13 N8 O7 F S 300.0605 0.4 1.3 8.0 47.4 C10 H10 N6 O F2 S 300.0618 -0.9 -3.0 7.5 26.7 C12 H12 N3 O2 F2 S 300.0607 0.2 0.7 11.5 1.0 C15 H11 N3 O F S 300.0614 -0.5 -1.7 0.5 311.4 C2 H10 N9 O2 F4 S 300.0598 1.1 3.7 -1.0 346.2 C2 H14 N8 O3 F2 S2 300.0612 -0.3 -1.0 -1.5 303.2 C4 H16 N5 O4 F2 S2 300.0603 0.6 2.0 4.5 171.8 C5 H9 N9 O F3 S 300.0603 0.6 2.0 -1.0 209.3 C6 H15 N2 O6 F3 S 300.0616 -0.7 -2.3 4.0 128.1 C7 H11 N6 O2 F3 S 300.0605 0.4 1.3 0.5 168.9 C7 H12 N3 O F6 S 300.0600 0.9 3.0 2.5 191.3 C7 H15 N5 O3 F S2 300.0614 -0.5 -1.7 2.0 171.4 C9 H17 N2 O4 F S2

03-Jun-201314:03:57

%

MOUTAFA_MG-I-18C_BWANG-ACCU_06032013_ESI-NEG02 108 (2.006) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Cm (108:121) 286.0444 5.86e3 100

287.0476 288.0416

323.2381 392.2126

0 280

300

320

340

360

380

400


* 526.3748 554.2615

406.2003

420

440

460

480

500

520

540

560

m/z 580

Number of isotope peaks used for i-FIT = 3 Monoisotopic Mass, Even Electron Ions 1051 formula(e) evaluated with 7 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 286.0444 286.0458 -1.4 -4.9 0.5 275.8 C H8 N9 O2 F4 S 286.0450 -0.6 -2.1 11.5 1.5 C14 H9 N3 O F S 286.0433 1.1 3.8 -0.5 233.0 C3 H11 N5 O5 F3 S 286.0455 -1.1 -3.8 -1.5 262.7 C3 H14 N5 O4 F2 S2 286.0446 -0.2 -0.7 4.5 153.9 C4 H7 N9 O F3 S 286.0449 -0.5 -1.7 0.5 151.7 C6 H10 N3 O F6 S 286.0444 0.0 0.0 2.5 163.3 C6 H13 N5 O3 F S2

15-May-201311:48:52 MOUSTAFA_MG-I-118C_BWANG-ACCU_05142013_ESI-NEG01 177 (3.301) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Sm (SG, 2x3.00); Cm 349.9592 436 100

%

347.9611

350.9666 351.9503

341.1102

363.0709

0 340

342

344

346

348


350

352

354

356

358

360

362

364

m/z

Number of isotope peaks used for i-FIT = 3 Monoisotopic Mass, Even Electron Ions 419 formula(e) evaluated with 3 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 F: 1-6 S: 1-4 Br: 1-5 Minimum: Maximum: Mass 347.9611

Calc. Mass 347.9606 347.9602 347.9605

5.0 mDa 0.5 0.9 0.6

5.0 PPM 1.4 2.6 1.7

-1.5 50.0 DBE 11.5 4.5 0.5

i-FIT 0.9 10.0 9.8

Formula C14 H8 N3 F S Br C4 H6 N9 F3 S Br C6 H9 N3 F6 S Br

40.61 40.40 39.56 39.35

114.03 113.79 109.05 108.79

134.00 132.32 128.78 123.17

159.31 156.94

167.50

Current Data Parameters NAME MG−I−21c EXPNO 1 PROCNO 1 F2 − Acquisition Parameters Date_ 20130521 Time 16.43 INSTRUM spect PROBHD 5 mm PABBO BB− PULPROG zgpg30 TD 65536 SOLVENT DMSO NS 384 DS 4 SWH 24038.461 Hz FIDRES 0.366798 Hz AQ 1.3631488 sec RG 90.5 DW 20.800 usec DE 6.50 usec TE 298.0 K D1 2.00000000 sec D11 0.03000000 sec TD0 1 ======== CHANNEL f1 ======== SFO1 100.6253441 MHz NUC1 13C P1 9.00 usec PLW1 62.00000000 W ======== CHANNEL f2 ======== SFO2 400.1416006 MHz NUC2 1H CPDPRG[2 waltz16 PCPD2 90.00 usec PLW2 16.00000000 W PLW12 0.36000001 W PLW13 0.29159999 W F2 − Processing parameters SI 32768 SF 100.6152830 MHz WDW EM SSB 0 LB 1.00 Hz GB 0 PC 1.40

200

180

160

140

120

100

80

60

40

20

0

ppm

15-May-201311:28:52

%

MOUSTAFA_MG-I-119C_BWANG-ACCU_05142013_ESI-NEG02 39 (0.725) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Cm (39:44) 302.0395 1.23e4 100

303.0433 300.0263 304.0404

137.0184

0 100

448.0538

212.0663

200

300

400

* 605.0671 554.2615 665.0973

500


600

700

961.0190

800

900

m/z 1000

Monoisotopic Mass, Even Electron Ions 1784 formula(e) evaluated with 10 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-4 Minimum: -1.5 Maximum: 5.0 5.0 50.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 302.0395 302.0407 -1.2 -4.0 0.5 509.4 C H8 N9 O3 F4 S 302.0400 -0.5 -1.7 11.5 1.1 C14 H9 N3 O2 F S 302.0382 1.3 4.3 -0.5 430.1 C3 H11 N5 O6 F3 S 302.0404 -0.9 -3.0 -1.5 547.0 C3 H14 N5 O5 F2 S2 302.0391 0.4 1.3 -1.5 808.6 C4 H15 N5 O F3 S3 302.0396 -0.1 -0.3 4.5 265.3 C4 H7 N9 O2 F3 S 302.0398 -0.3 -1.0 0.5 260.7 C6 H10 N3 O2 F6 S 302.0393 0.2 0.7 2.5 351.5 C6 H13 N5 O4 F S2 302.0384 1.1 3.6 8.5 103.6 C7 H6 N9 O F2 S 302.0387 0.8 2.6 4.5 100.4 C9 H9 N3 O F5 S

17-May-201315:38:33

%


296.0485 212.0725 297.0456 376.0791 213.0769 165.9974

0 100

200

300

400


* 554.2615 577.2307

500

600

680.1317

700

800

900

m/z 1000

Number of isotope peaks used for i-FIT = 3 Monoisotopic Mass, Odd and Even Electron Ions 454 formula(e) evaluated with 6 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 F: 1-6 S: 1-4 Minimum: -1.5 Maximum: 5.0 5.0 50.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 295.0455 295.0452 0.3 1.0 10.0 39.4 C10 H7 N7 F2 S 295.0454 0.1 0.3 6.0 37.8 C12 H10 N F5 S 295.0454 0.1 0.3 13.5 1.7 C15 H8 N4 F S 295.0450 0.5 1.7 6.5 199.4 C5 H6 N10 F3 S 295.0458 -0.3 -1.0 0.0 654.4 C7 H16 N3 F3 S3 295.0452 0.3 1.0 2.5 195.7 C7 H9 N4 F6 S

Current Data Parameters NAME MG148 EXPNO 1 PROCNO 1 F2 − Acquisition Parameters Date_ 20130417 Time 18.05 INSTRUM spect PROBHD 5 mm PABBO BB− PULPROG zg30 TD 65536 SOLVENT DMSO NS 16 DS 2 SWH 8223.685 Hz FIDRES 0.125483 Hz AQ 3.9846387 sec RG 71.8 DW 60.800 usec DE 6.50 usec TE 294.4 K D1 1.00000000 sec ======== CHANNEL f1 ======== NUC1 1H P1 13.50 usec PLW1 16.00000000 W SFO1 400.1424710 MHz F2 − Processing parameters SI 65536 SF 400.1400000 MHz WDW EM SSB 0 LB 0.30 Hz GB 0 PC 1.00

12

11

10

9

8

7

6

5

4

3

2 10.87

13

607.15

14

1.57 1.69 1.54 3.12 1.68 1.08 1.40

15

1.00

16

1

0

−1

−2

ppm

40.47 40.26 40.06 39.85 39.64 39.43 39.22

136.97 134.29 133.95 130.19 126.80 117.84 114.18 113.94 109.79 109.10 108.83

157.12

Current Data Parameters NAME MG148c EXPNO 1 PROCNO 1 F2 − Acquisition Parameters Date_ 20130418 Time 16.11 INSTRUM spect PROBHD 5 mm PABBO BB− PULPROG zgpg30 TD 65536 SOLVENT DMSO NS 269 DS 4 SWH 24038.461 Hz FIDRES 0.366798 Hz AQ 1.3631488 sec RG 161 DW 20.800 usec DE 6.50 usec TE 298.0 K D1 2.00000000 sec D11 0.03000000 sec TD0 1 ======== CHANNEL f1 ======== SFO1 100.6253441 MHz NUC1 13C P1 9.00 usec PLW1 62.00000000 W ======== CHANNEL f2 ======== SFO2 400.1416006 MHz NUC2 1H CPDPRG[2 waltz16 PCPD2 90.00 usec PLW2 16.00000000 W PLW12 0.36000001 W PLW13 0.29159999 W F2 − Processing parameters SI 32768 SF 100.6152830 MHz WDW EM SSB 0 LB 1.00 Hz GB 0 PC 1.40

200

180

160

140

120

100

80

60

40

20

0

ppm

03-Jun-201314:15:33

%

MOUTAFA_MG-I-30C_BWANG-ACCU_06032013_ESI-NEG01 63 (1.173) AM (Med,2, Ar,5000.0,554.26,1.00); Sm (Md, 3.00); Cm (62:69) 313.0922 6.32e3 100

314.0945 327.0729 406.9369

344.2377

0 300

320

340

360

380

400


448.1598

420

440

462.1545

460

478.1033

480

499.3824

500

520

* 554.2615 557.2921

540

560

m/z 580

Monoisotopic Mass, Odd and Even Electron Ions 364 formula(e) evaluated with 6 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 313.0922 313.0922 0.0 0.0 0.5 159.1 C8 H15 N4 F6 S 313.0919 0.3 1.0 4.5 161.1 C6 H12 N10 F3 S 313.0923 -0.1 -0.3 11.5 5.7 C16 H14 N4 F S 313.0924 -0.2 -0.6 4.0 51.6 C13 H16 N F5 S 313.0921 0.1 0.3 8.0 52.6 C11 H13 N7 F2 S 313.0917 0.5 1.6 1.0 334.6 C H11 N13 F4 S

05-Jun-201315:58:28

%

MOUSTAFA_MG-1-32C_BWANG-ACCU_06052013_ESI-NEG01 105 (1.958) AM (Cen,2, 80.00, Ht,5000.0,554.26,1.00); Sm (SG, 2x3.00); Cm 357.1184 1.86e3 100

358.1214 359.1176

0 340

372.2741 379.1025

360

380

400

436.0454

420

440


* 554.2615

478.0638

460

480

500

520

540

560

576.2430

580

m/z

Number of isotope peaks used for i-FIT = 3 Monoisotopic Mass, Even Electron Ions 2651 formula(e) evaluated with 10 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 357.1184 357.1184 0.0 0.0 0.5 32.6 C10 H19 N4 O F6 S 357.1179 0.5 1.4 2.5 29.7 C10 H22 N6 O3 F S2 357.1197 -1.3 -3.6 7.5 4.3 C15 H19 N4 O2 F2 S 357.1185 -0.1 -0.3 11.5 0.6 C18 H18 N4 O F S 357.1177 0.7 2.0 -0.5 86.3 C2 H18 N12 O6 F S 357.1190 -0.6 -1.7 4.5 62.4 C3 H14 N16 O2 F S 357.1193 -0.9 -2.5 0.5 61.7 C5 H17 N10 O2 F4 S 357.1168 1.6 4.5 -0.5 50.1 C7 H20 N6 O5 F3 S 357.1190 -0.6 -1.7 -1.5 51.4 C7 H23 N6 O4 F2 S2 357.1181 0.3 0.8 4.5 33.1 C8 H16 N10 O F3 S

03-Jun-201313:37:28

%


272.0494 * 554.2615 565.0738

281.2678 312.2560 342.2386372.2449

0 250

275

300

325

350

375


400

425

450

475

500

525

550

575

m/z

Monoisotopic Mass, Even Electron Ions 241 formula(e) evaluated with 1 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 271.0466 271.0454 1.2 4.4 11.5 0.5 C13 H8 N4 F S

05-Jun-201315:38:05 MOUSTAFA_MG-1-42C_BWANG-ACCU_06052013_ESI-NEG01 148 (2.740) AM (Cen,2, 80.00, Ar,5000.0,554.26,1.00); Sm (SG, 2x3.00); Cm 338.9676 1.14e4 100

%

340.9643

312.2532

342.9621 343.9655

373.2683

401.2305

431.2489

0 320

340

360

380

400

420


440

465.3612

460

480

513.3729

500

520

* 554.2615 556.2557

540

560

580

m/z

Monoisotopic Mass, Odd and Even Electron Ions 549 formula(e) evaluated with 10 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 F: 1-6 S: 1-2 Cl: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 338.9676 338.9675 0.1 0.3 4.0 86.0 C10 H8 N F5 S Cl2 338.9674 0.2 0.6 11.5 2.8 C13 H6 N4 F S Cl2 338.9673 0.3 0.9 0.5 301.3 C5 H7 N4 F6 S Cl2 338.9672 0.4 1.2 8.0 88.4 C8 H5 N7 F2 S Cl2 338.9670 0.6 1.8 4.5 306.2 C3 H4 N10 F3 S Cl2 338.9686 -1.0 -3.0 4.5 855.0 C H3 N12 F3 S2 Cl 338.9688 -1.2 -3.5 0.5 850.1 C3 H6 N6 F6 S2 Cl 338.9688 -1.2 -3.5 8.0 608.7 C6 H4 N9 F2 S2 Cl 338.9690 -1.4 -4.1 11.5 522.0 C11 H5 N6 F S2 Cl 338.9690 -1.4 -4.1 4.0 607.3 C8 H7 N3 F5 S2 Cl

05-Jun-201316:03:27

%

MOUSTAFA_MG-1-29C_BWANG-ACCU_06052013_ESI-NEG01 60 (1.115) AM (Cen,2, 80.00, Ht,5000.0,554.26,1.00); Sm (SG, 2x3.00); Cm (5 394.0822 536 100

* 554.2615

392.2091 395.0838 390.1958 374.2065

555.2604

410.2126 424.1957

465.3607

484.3458

513.3553

528.3573

556.2704

0 360

380

400

420

440

460


480

500

520

540

560

580

612.4763

600

620

m/z

Monoisotopic Mass, Odd and Even Electron Ions 669 formula(e) evaluated with 10 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 F: 1-6 S: 1-2 Minimum: -1.5 Maximum: 5.0 5.0 100.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 394.0822 394.0814 0.8 2.0 19.5 0.1 C24 H13 N3 F S 394.0813 0.9 2.3 8.5 6.3 C16 H14 N3 F6 S 394.0812 1.0 2.5 16.0 1.3 C19 H12 N6 F2 S 394.0810 1.2 3.0 12.5 6.4 C14 H11 N9 F3 S 394.0808 1.4 3.6 9.0 15.8 C9 H10 N12 F4 S 394.0806 1.6 4.1 5.5 29.5 C4 H9 N15 F5 S 394.0806 1.6 4.1 13.0 15.9 C7 H7 N18 F S 394.0804 1.8 4.6 9.5 29.8 C2 H6 N21 F2 S 394.0840 -1.8 -4.6 0.5 35.6 C H13 N15 F5 S2 394.0840 -1.8 -4.6 8.0 21.1 C4 H11 N18 F S2

132.03 131.39 130.68 129.18 128.70 128.50 127.88 127.39 127.09 126.52 126.17 125.73 125.42 124.73 124.31 123.03 114.08 113.84 109.16

156.96

167.51

Current Data Parameters NAME MG−I−29c EXPNO 1 PROCNO 1 F2 − Acquisition Parameters Date_ 20130520 Time 18.00 INSTRUM spect PROBHD 5 mm PABBO BB− PULPROG zgpg30 TD 65536 SOLVENT DMSO NS 353 DS 4 SWH 24038.461 Hz FIDRES 0.366798 Hz AQ 1.3631488 sec RG 114 DW 20.800 usec DE 6.50 usec TE 298.0 K D1 2.00000000 sec D11 0.03000000 sec TD0 1 ======== CHANNEL f1 ======== SFO1 100.6253441 MHz NUC1 13C P1 9.00 usec PLW1 62.00000000 W ======== CHANNEL f2 ======== SFO2 400.1416006 MHz NUC2 1H CPDPRG[2 waltz16 PCPD2 90.00 usec PLW2 16.00000000 W PLW12 0.36000001 W PLW13 0.29159999 W F2 − Processing parameters SI 32768 SF 100.6152830 MHz WDW EM SSB 0 LB 1.00 Hz GB 0 PC 1.40

190 180 170 160 150 140 130 120 110 100

90

80

70

60

50

40

30

ppm

15-May-201312:50:17

%

MOUSTAFA_MG-I-12C_BWANG-ACCU_05142013_ESI-NEG01 56 (1.042) AM (Med,2, Ar,5000.0,554.26,1.00); Sm (SG, 2x3.00); Cm (54:62) 6.88e3 377.9720 379.9691 100

378.9741 380.9725 381.9675 382.9733 384.9376

376.0812

389.8051

0 372

374

376

378


380

382

384

386

388

390

m/z 392

Number of isotope peaks used for i-FIT = 3 Monoisotopic Mass, Odd and Even Electron Ions 1957 formula(e) evaluated with 17 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-4 Br: 1-5 Minimum: Maximum: Mass 377.9720

Calc. Mass 377.9717 377.9710 377.9723 377.9712 377.9703 377.9730 377.9719 377.9730 377.9733 377.9717 377.9708 377.9708 377.9721 377.9710 377.9705 377.9735 377.9719

5.0 mDa 0.3 1.0 -0.3 0.8 1.7 -1.0 0.1 -1.0 -1.3 0.3 1.2 1.2 -0.1 1.0 1.5 -1.5 0.1

5.0 PPM 0.8 2.6 -0.8 2.1 4.5 -2.6 0.3 -2.6 -3.4 0.8 3.2 3.2 -0.3 2.6 4.0 -4.0 0.3

-1.5 50.0 DBE -1.0 8.0 7.5 11.5 -1.0 4.0 0.5 -1.5 0.0 -1.5 4.5 -1.0 4.0 0.5 2.5 3.5 2.0

i-FIT 317.5 31.2 17.0 1.8 249.0 211.3 257.6 263.1 208.9 202.0 133.4 175.3 99.0 131.1 99.5 71.3 72.6

Formula C H12 N8 O7 F S Br C10 H9 N6 O F2 S Br C12 H11 N3 O2 F2 S Br C15 H10 N3 O F S Br C2 H13 N8 O3 F2 S2 Br C2 H8 N12 O3 F S Br C2 H9 N9 O2 F4 S Br C3 H14 N5 O8 F S Br C4 H11 N6 O3 F4 S Br C4 H15 N5 O4 F2 S2 Br C5 H8 N9 O F3 S Br C6 H14 N2 O6 F3 S Br C7 H10 N6 O2 F3 S Br C7 H11 N3 O F6 S Br C7 H14 N5 O3 F S2 Br C9 H12 N3 O3 F3 S Br C9 H16 N2 O4 F S2 Br

15-May-201312:21:19

%


331.0760

150.0251

0 100

212.0717

332.0712

* 554.2615

350.9548

200

300

400

Elemental Composition Report Single Mass Analysis Tolerance = 5.0 PPM / DBE: min = -1.5, max = 50.0

500

600

683.1474

700

800

900

m/z 1000

Element prediction: Off Number of isotope peaks used for i-FIT = 3 Monoisotopic Mass, Odd and Even Electron Ions 2684 formula(e) evaluated with 24 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-4 Minimum: -1.5 Maximum: 5.0 5.0 50.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 330.0722 330.0707 1.5 4.5 1.0 661.1 C H10 N12 O2 F4 S 330.0729 -0.7 -2.1 0.0 646.4 C H13 N12 O F3 S2 330.0713 0.9 2.7 -1.5 1026.7 C H17 N11 O2 F S3 330.0718 0.4 1.2 4.5 546.9 C H9 N15 O3 F S 330.0735 -1.3 -3.9 3.5 150.4 C10 H15 N3 O4 F3 S 330.0719 0.3 0.9 2.0 306.6 C10 H19 N2 O5 F S2 330.0711 1.1 3.3 8.0 73.9 C11 H12 N6 O2 F2 S 330.0733 -1.1 -3.3 7.0 186.4 C11 H15 N6 O F S2 330.0706 1.6 4.8 2.0 636.5 C11 H20 N2 O F2 S3 330.0724 -0.2 -0.6 7.5 38.8 C13 H14 N3 O3 F2 S 330.0713 0.9 2.7 11.5 0.8 C16 H13 N3 O2 F S 330.0718 0.4 1.2 -1.0 641.9 C2 H15 N8 O8 F S 330.0731 -0.9 -2.7 4.0 439.0 C3 H11 N12 O4 F S 330.0720 0.2 0.6 0.5 539.7 C3 H12 N9 O3 F4 S 330.0731 -0.9 -2.7 -1.5 530.2 C4 H17 N5 O9 F S 330.0733 -1.1 -3.3 0.0 433.5 C5 H14 N6 O4 F4 S 330.0717 0.5 1.5 -1.5 535.2 C5 H18 N5 O5 F2 S2 330.0709 1.3 3.9 4.5 290.5 C6 H11 N9 O2 F3 S 330.0731 -0.9 -2.7 3.5 339.4 C6 H14 N9 O F2 S2 330.0709 1.3 3.9 -1.0 358.2 C7 H17 N2 O7 F3 S 330.0722 0.0 0.0 4.0 213.0 C8 H13 N6 O3 F3 S

330.0711 330.0733 330.0706

1.1 -1.1 1.6

3.3 -3.3 4.8

0.5 -0.5 2.5

285.6 335.4 339.0

C8 H14 N3 O2 F6 S C8 H17 N3 O F5 S2 C8 H17 N5 O4 F S2

rinse90%MeOH w 0.1%FA

11:01:23 29-Apr-2013

%

MOUSTAFA_MG-I-8C_BWANG-ACCU_04292013_ESI-POS01 83 (1.543) AM (Cen,4, 80.00, Ar,6000.0,556.28,0.70); Sm (SG, 3x3.00); Cm (7 316.0924 1.15e4 100

356.1158

0 100

556.2771

200

300

400


500

600

700

800

900

m/z 1000

Number of isotope peaks used for i-FIT = 3 Monoisotopic Mass, Even Electron Ions 2188 formula(e) evaluated with 7 results within limits (all results (up to 1000) for each mass) Elements Used: C: 1-150 H: 1-150 N: 1-30 O: 1-60 F: 1-6 S: 1-4 Minimum: -1.5 Maximum: 5.0 5.0 50.0 Mass Calc. Mass mDa PPM DBE i-FIT Formula 316.0924 316.0925 -0.1 -0.3 3.5 266.7 C H11 N15 O2 F S 316.0931 -0.7 -2.2 6.5 9.2 C13 H16 N3 O2 F2 S 316.0920 0.4 1.3 10.5 48.0 C16 H15 N3 O F S 316.0927 -0.3 -0.9 -0.5 260.8 C3 H14 N9 O2 F4 S 316.0916 0.8 2.5 3.5 101.2 C6 H13 N9 O F3 S 316.0918 0.6 1.9 -0.5 97.5 C8 H16 N3 O F6 S 316.0913 1.1 3.5 1.5 295.0 C8 H19 N5 O3 F S2