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1
Towards engineering of self-assembled nanostructures
using non-ionic dendritic amphiphiles Bala N. S. Thota,
a Hans v. Berlepsch,
b Christoph Böttcher
b and Rainer Haag
a*
a Institut für Chemie und Biochemie, Freie Universität Berlin, Takustrasse 3, 14195 Berlin, Germany. b Forschungszentrum für Elektronenmikroskopie,and Core facility Biosupramol, Institut für Chemie und Biochemie, Freie Universität
Berlin, Fabeckstrasse 36a, 14195 Berlin, Germany .
Content:
1) General synthetic and Analytic methods
2) Synthetic procedures and Characterization data
3) Critical aggregation concentration (CAC) measurements
4) Dye encapsulation experiments and calculation of transport capacity
5) Dynamic light scattering measurements
6) Cryo-TEM experiments
7) Packing parameter approach
8) Additional cryo-TEM images
9) Representation NMR spectra for final compounds
1) General synthetic and Analytic methods
All commercially available compounds were used as received without further purification. Millipore
water was obtained from a Merck Millipore Milli-Q Integral System. UV/Vis measurements were
performed on a PerkinElmer LAMBDA 950 UV/Vis/NIR spectrophotometer. Standard disposable
PMMA UV/Vis cuvettes with a path length of 1 cm from PLASTIBRAND were used. DLS measurements
were performed on Malvern Zetasizer ZS. Disposable BRAND UV-Cuvettes micro were used. NMR
spectra were recorded on JOEL ECX400, JOEL ECP500 and BRUKER AV500 spectrometers. IR spectra
were recorded on a JASCO FT/IR 4100 spectrometer. Mass analyses were performed on an Agilent 6210
ESI-TOF, Agilent Technologies, Santa Clara, CA, USA. Surface tension measurements were performed
on a Dataphysics OCA 20.
Electronic Supplementary Material (ESI) for ChemComm.This journal is © The Royal Society of Chemistry 2015
2
2) Synthetic procedures and Characterization data
Synthesis of glycerol symmetric esters:
To a solution of Glycerol (1 equiv.) in THF, vinyl ester of the corresponding alkyl chain (2.5 equiv.) was
added followed by addition of the enzyme (20% wt. of the glycerol), Novozyme 435. The reaction
mixture was left stirring at room temperature, without breaking the beads of the enzyme. The progress of
the reaction was followed by TLC. After complete conversion of the starting materials, stopped stirring,
filtered the reaction mixture and washed the beads with excess THF. The filtrate was concentrated, dried
and purified by column chromatography to obtain the pure product as a white solid.
Compound 7: 4.3 g of the product was obtained from 1 g of glycerol. Yield 86%.
1H NMR (500 MHz, CDCl3 ) δ 4.19 (dd, J= 4, 11Hz, 2H, -COOCH2-), 4.14 (dd, J= 6, 11Hz, 2H,
COOCH2-), 4.08 (m, 1H, HOCH(CH2O-)2), 2.35 (t, J=7.5 Hz, 4H, -CH2COO-), 1.63 (m, 4H, -
CH2CH2COO-), 1.33-1.22 (m, 32H, Alkyl chain), 0.88 (t, J= 7Hz, 6H, -CH3). 13
C NMR (126 MHz,
CDCl3) δ 174.08, 77.38, 68.58, 65.20, 34.26, 32.06, 29.75, 29.60, 29.48, 29.40, 29.28, 25.05, 22.83,
14.26. MS (ESI) m/z = 479.3709 [M+Na]+ (calcd. 479.3712). IR: 2911, 2845, 1722,1182.
Compound 8: 5.0 g of product was obtained from 1g of glycerol. Yield 76%.
1H NMR (500 MHz, CDCl3) δ 4.19 (dd, J= 4.5, 11 Hz, 2H, -COOCH2-), 4.14 (dd, J= 6, 11 Hz, 2H,
COOCH2-), 4.08 (m, 1H, HOCH(CH2O-)2), 2.34 (t, J= 7.5 Hz, 4H, -CH2COO-), 1.62 (m, 4H,
Scheme S1. Synthesis of lipophilic azides
3
CH2CH2COO-), 1.32-1.22 (m, 56H, Alkyl chain), 0.88 (t, J= 7 Hz, 6H, -CH3). 13
C NMR (126 MHz,
CDCl3) δ 174.09, 77.36, 68.56, 65.19, 34.26, 32.08, 29.85, 29.83, 29.81, 29.80, 29.75, 29.61, 29.51,
29.40, 29.28, 25.04, 22.84, 14.27. MS (ESI) m/z = 647.5607 [M+Na]+ (calcd. 647.5590). IR: 2915, 2849,
1756,1149.
Synthesis of azido derivatives: To a solution of alcohol (7/8) in dichloromethane, triethyl amine (1.2
equiv.) and methanesulfonyl chloride (1.2 equiv.) were added at 0 °C. Reaction mixture was allowed to
warm-up to room temperature during the progress of the reaction. After complete conversion of the
alcohol, the reaction mixture was quenched by adding water and both the phases were separated.
Compound was extracted from aqueous phase using DCM. The combined organic phase was washed
further with dil. HCl, satd. NaHCO3 and water, then dried over anhyd. Na2SO4. The reaction mixture was
concentrated and subjected for the next step.
The mesylate was dissolved in DMF followed by addition of sodium azide (5 equiv.). The reaction
mixture was left stirring at 90 °C. Progress of the reaction was monitoring by TLC. After completion of
the reaction, the reaction mixture was filtered and DMF was removed under reduced pressure. Reaction
mixture was dissolved in DCM and washed with water, and dried over anhyd. Na2SO4. The crude mixture
was purified by column chromatography using DCM/Hexane.
Compound 9: 3.2 g of product was obtained from 3.1 g of compound 7. Yield 95%.
1H NMR (500 MHz, CDCl3) δ 4.23 (dd, J= 4.5, 11.5 Hz, 2H, -COOCH2-), 4.14 (dd, J= 6.5, 11.5 Hz, 2H,
COOCH2-), 3.87 (m, 1H, N3CH(CH2O)2), 2.35(t, J= 7.5 Hz, 4H, -CH2COO-), 1.63 (m, 4H,
CH2CH2COO-), 1.33-1.25 (m, 32H, Alkyl chain), 0.88 (t, J= 7Hz, 6H, -CH3). 13
C NMR (126 MHz,
CDCl3) δ 173.41, 77.36, 63.15, 58.84, 34.16, 32.05, 29.73, 29.58, 29.47, 29.37, 29.24, 24.96, 22.82,
14.25. MS (ESI) m/z = 504.3781 [M+Na]+ (calcd. for C27H51N3NaO4: 504.3777). IR: 2923, 2853, 2116,
1743.
Compound 10: 5 g of product was obtained from 5.2 g of compound 7. Yield 92%.
1H NMR (500 MHz, CDCl3) δ 4.23 (dd, J= 4.5 Hz, 11.5 Hz, -COOCH2-), 4.14 (dd, J= 7, 11.5 Hz, 2H,
COOCH2-), 3.87 (m, 1H, N3CH(CH2O)2-), 2.35 (t, J= 7.5 Hz, 4H, -CH2COO-), 1.61 (m, 4H,
CH2CH2COO-), 1.33-1.20 (m, 56H, Alkyl chain), 0.88 (t, J= 7 Hz, 6H, -CH3).13
C NMR (126 MHz,
CDCl3) δ 173.42, 63.16, 58.82, 34.16, 32.07, 29.85, 29.82, 29.80, 29.80, 29.74, 29.60, 29.51, 29.38,
29.25, 24.96, 22.84, 14.27. MS (ESI) m/z = 672.5667 [M+Na]+ (calcd. for C39H75N3NaO4: 672.5655). IR:
2917, 2850, 2126, 1741.
4
Synthesis of dissymmetric azide (11): To a solution of protected glycerol (4.05g, 36.9 mmol) in DCM,
triethylamine (1.2 equiv.) and methanesulfonyl chloride (1.2 equiv.) were added at 0 °C and left stirring
in the ice bath. Reaction mixture was allowed to warm-up to room temperature during the progress of the
reaction and monitored the conversion using TLC. After completion of the reaction, it was quenched by
adding water and both the phases were separated and organic compound was extracted from aqueous
phase using DCM. Combined organic phase was washed with dil. HCl, satd. NaHCO3 solution and water.
Finally the reaction mixture was dried over anhyd. Na2SO4. The reaction mixture was concentrated and
dried, and the crude product was subjected for the next step by dissolving in DMF and adding sodium
azide (5 equiv.). The reaction mixture was left stirring at 90 °C overnight and the reaction mixture was
filtered to remove excess sodium azide. DMF was removed under reduced pressure and the reaction
mixture was dissolved in DCM. This organic phase was washed with water and dried over anhyd.
Na2SO4. The organic phase was concentrated under reduced pressure. To a methanolic solution of this
crude mixture, Dowex-H was added and left stirring at reflux for 5 h. Finally the reaction mixture was
cooled to room temperature and filter to remove Dowex. The filtrate was concentrated and purified by
column chromatography using EtOH/DCM, yield-81%.
1H NMR (400 MHz, CDCl3 ) δ 3.85 (m, 1H, HOCH-), 3.65 (dd, J= 3.6, 12 Hz, 1H, HOCH2-), 3.55 (dd,
J= 6.4, 11.6 Hz, 1H, HOCH2-), 3.37-3.32 (m, 2H, N3CH2-). 13
C NMR (101 MHz, CDCl3) δ 71.13, 64.05,
53.47.
Compound 12: To compound 11 (2.3 g, 19.64 mmol) in THF (50 mL), vinyl stearate (1.2 equiv.) was
added, followed by addition of the enzyme (20% wt.), Novozyme 435. The reaction mixture was left
stirring at room temperature. After complete consumption of the starting material, the reaction mixture
was filtered to remove the enzyme. The filtrate was concentrated under reduced pressure and the crude
mixture was purified by column chromatography (10-25% EtOAc/Hexane) to obtain the compound 12
(5.9 g, yield 79%) as white solid.
1H NMR (400 MHz, CDCl3) δ 4.17 (dd, J= 4.4, 11.6 Hz, 1H, -COOCH2-), 4.12 (dd, J= 6, 11.6 Hz, 1H, -
COOCH2-), 4.01 (m, 1H, HOCH-), 3.4 (m, 2H, N3CH2-), 2.35 (t, J= 7.6 Hz, 2H, -OOCCH2-), 1.62 (m,
2H, -OOCCH2CH2-), 1.29-1.20 (m, 28H, Alkyl chain), 0.87 (t, J= 7.2 Hz, 3H, -CH3)
13C NMR (101 MHz, CDCl3) δ 174.18, 69.27, 65.64, 53.59, 34.22, 32.05, 29.83, 29.80, 29.77, 29.76,
29.73, 29.58, 29.50, 29.48, 29.37, 29.35, 29.24, 25.01, 22.82, 14.26, 14.24. MS (ESI) m/z = 406.3055
[M+Na]+ (calcd. for C21H41N3NaO3: 406.3046).
Compound 13: To a solution of the compound 12 (2.1 g, 5.3 mmol) in DCM, lauric acid (1.2 equiv.) was
added followed by addition of DMAP (0.2 equiv.). The reaction mixture was cooled to 0 °C and added
5
DCC (1.2 equiv.), left stirring in the ice bath for 8 h. The reaction mixture was allowed to warm up to
room temperature during the progress of the reaction. The progress of the reaction was monitored by
TLC. Reaction mixture was filtered to remove the by-products and the precipitate was washed with
DCM/Hexane. The combined organic layer was washed with dil. HCl, satd. NaHCO3 solution, water and
dried over anhyd. Na2SO4. The reaction mixture was concentrated and purified by column
chromategraphy to obtain the compound 13 (2.2 g, yield 74%) as a white solid.
1H NMR (500 MHz, CDCl3) δ 5.17 (m, 1H. –COOCH-), 4.29 (dd, J= 4.5, 12 Hz, 1H, -COOCH2-), 4.15
(dd, J= 6, 12 Hz, 1H, -COOCH2-), 3.46 (m, 2H, -CH2N3), 2.35 (t, J=7.5 Hz, 2H, -CH2COO-), 2.31 (t, J=
7.5 Hz, 2H, -CH2COO-), 1.62 (m, 4H, -CH2CH2COO-), 1.32-1.22 (m, 44H, Alkyl chain), 0.88 (t, J= 7
Hz, 6H, -CH3). 13
C NMR (126 MHz, CDCl3) δ 173.38, 173.01, 70.01, 62.43, 51.03, 34.33, 34.19, 32.07,
32.05, 29.85, 29.83, 29.81, 29.77, 29.75, 29.62, 29.61, 29.51, 29.48, 29.47, 29.41, 29.40, 29.26, 29.21,
25.01, 24.96, 22.83, 14.26. MS (ESI) m/z = 588.4730 [M+Na]+ (calcd. for C33H63N3NaO4: 588.4716). IR:
2922, 2852, 2101, 1743, 1152.
Scheme S2. Synthesis of protected dendritic amphiphiles
6
Synthesis of Propargyl ethers of dendrons (14/15): All the dendrons were synthesized by a literature
reported procedure.1 To a solution of protected dendron in THF, sodium hydride (5 equiv.) was added and
left stirring at 50 °C for 1h. To this reaction mixture, propargyl bromide was added slowly and left
stirring at the same temperature for 8 h. The progress of the reaction was checked by TLC and excess of
NaH was quenched by drop wise addition of water, keeping the reaction flask in ice bath. The reaction
mixture was concentrated under reduced pressure and diluted with water. The compound was extracted
into DCM. The combined organic layer was washed with water, dried over anhyd. Na2SO4. The reaction
mixture was concentrated and purified by column chromatography to obtain a pale yellow liquid (yield
80-85%).
Compound 14:1H NMR (400 MHz, Methanol-d4) δ 4.33 (m, 2H, CHCCH2O-), 4.28-4.23 (m, 4H, CH
acetonide), 4.08-4.03 (m, 4H, CH2 acetonide), 3.85-3.49 (m, 27H, dendron), 2.86 (m, 1H, CHCCH2-),
1.39 (m, 12H, -CH3), 1.33 (m, 12H, -CH3). 13
C NMR (101 MHz, Methanol-d4) δ 110.45, 81.29, 79.90,
79.86, 79.83, 78.47, 78.17, 77.87, 76.22, 76.10, 75.84, 75.78, 75.73, 73.40, 72.52, 72.45, 72.41, 72.38,
72.36, 71.15, 67.76, 67.74, 67.59, 67.56, 58.32, 49.64, 49.49, 49.43, 49.27, 49.21, 49.06, 49.00, 48.82,
48.79, 48.62, 48.57, 48.36, 27.18, 27.13, 27.12, 27.05, 26.13, 25.72, 25.71. MS (ESI) m/z = 757.3997
[M+Na]+ (calcd. for C36H62NaO15: 757.3986).
Compound 15: 1H NMR (500 MHz, Acetone-d6) δ 4.35 (m, 2H, CHCCH2O-), 4.24-4.17 (m, 8H, CH
acetonide), 4.05-4.01 (m, 8H, CH acetonide), 3.75-3.45 (m, 59H, dendron), 2.86 (m, 1H, CHCCH2-),
1.35 (m, 24H, -CH3), 1.29 (m, 24H, -CH3). 13
C NMR (126 MHz, Acetone-d6) δ 109.49, 109.42, 79.56,
79.31, 75.59, 75.55, 75.46, 73.07, 73.05, 72.05, 70.74, 67.51, 67.35, 67.33, 67.31, 67.30, 57.90, 27.20,
27.18, 27.15, 27.13, 27.11, 27.10, 25.76, 25.74, 25.73. MS (ESI) m/z = 1509.8199 [M+Na]+ (calcd. for
C72H126NaO31: 1509.8181).
General protocol for click reaction: To a mixture of the alkyne (1 equiv.) and azide (1.5 equiv.) in THF,
diisopropylethylamine, copper sulphate solution (15% mol, aq. 0.1M) and aq. sodium ascorbate (50%
mol, 0.2M) were added. The reaction mixture was left stirring at 45 °C. After complete consumption of
starting material on TLC, stirring was stopped and both phases were separated from reaction mixture and
organic compound were extracted from aqueous phase using DCM. The combined organic phase was
washed with satd. EDTA solution, water and dried over anhyd. Na2SO4. The reaction mixture was
concentrated and purified by column chromatography to obtain pure product as colourless, viscous liquid.
Compound 16: 2.4 g of compound 16 was obtained starting from 1.9 g of Compound 14 (yield 75%).
7
1H NMR (500 MHz, Acetone-d6): 8.06 (m, 1H, Triazole CH), 5.2 (m, 1H, Triazole-CH(CH2O-)2), 4.79-
4.77 (m, 2H, -OCH2-Triazole-), 4.60-4.57 (m, 4H, -CH2OOC-), 4.23-4.18 (m, 4H, CH acetonide), 4.04 -
4.00 (m, 4H, CH2 acetonide), 3.75-3.46 (m, 27H, dendron), 2.31-2.29 (m, 4H, -CH2COO-), 1.56-1.53 (m,
4H, -CH2CH2COO-), 1.34-1.28 (m, 56H, Alkyl chain and acetonide-CH3), 0.88 (m, 6H, -CH3). 13
C NMR
(126 MHz, Acetone-d6) δ 206.12, 173.22, 123.59, 109.58, 79.40, 75.71, 75.57, 73.16, 72.15, 67.59, 67.41,
64.32, 63.21, 59.41, 34.30, 32.65, 30.38, 30.32, 30.30, 30.23, 30.19, 30.15, 30.10, 30.05, 30.04, 30.01,
29.99, 29.88, 29.86, 29.84, 29.73, 29.71, 29.69, 29.57, 29.53, 29.41, 29.38, 27.19, 25.78, 25.51, 23.34,
14.42, 14.39. MS (ESI) m/z = 1238.7930 [M+Na]+ (calcd. for C63H113N3NaO19: 1238.7866). IR: 2984,
2923, 2855, 1744, 1456, 1370, 1253, 1213.
Compound 17: 2.9 g of compound 17 was obtained from 1.8 g of compound 14 (yield 90%)
1H NMR (500 MHz, Acetone-d6) 7.96 (m, 1H, CH(Triazole)), 5.48 (m, 1H, -CHOOC-), 4.77-4.68 (m,
4H, -CH2-Triazole-CH2-), 4.40 (dd, J=12, 4 Hz, 1H,-CH2OOC-), 4.24-4.18 (m, 4H, CH acetonide), 4.15-
4.11(m, 1H, -CH2OOC-), 4.04-4.01 (m, 4H, CH2 acetonide), 3.75-3.46 (m, 27H, dendron), 2.34 (t, J=7.5
Hz, 2H, -OOCCH2-), 2.29 (t, J=7.5 Hz, 2H, -OOCCH2-), 1.63-1.58 (m, 2H, -OOCCH2CH2-), 1.57-1.53
(m, 2H, -OOCCH2CH2-), 1.41-1.23(m, 68H, Alkyl chain and acetonide-CH3), 0.90-0.86 (m, 6H, -CH3).
13C NMR (126 MHz, Acetone-d6) δ 173.33, 172.84, 124.88, 109.58, 79.40, 79.35, 75.58, 75.57, 73.16,
72.08, 70.44, 67.59, 67.43, 67.41, 64.24, 63.10, 50.55, 34.51, 34.40, 32.68, 32.64, 30.42, 30.39, 30.37,
30.32, 30.30, 30.28, 30.17, 30.15, 30.09, 30.01, 29.99, 29.86, 29.84, 29.81, 29.74, 29.71, 29.69, 29.55,
29.53, 29.39, 29.38, 27.23, 27.18, 25.78, 25.60, 25.53, 23.36, 23.33, 14.40, 14.37. MS (ESI) m/z =
1322.8842 [M+Na]+ (calcd. for C69H125N3NaO19: 1322.8805). IR: 2972, 2853, 1743, 1457.
Compound 18: 3.6 g of compound 18 was obtained from 2.1 g of compound 14 (yield 91%)
1H NMR (500 MHz, Acetone-d6) 8.06 (m, 1H, CH(Triazole)), 5.21 (m, 1H, Triazole-CH(CH2O-)2), 4.78
(t, J= 2.5 Hz, 2H, -OCH2-Triazole-), 4.59 (d, J= 10Hz, 4H, -CH2OOC-), 4.23-4.18 (m, 4H, CH
acetonide), 4.03-4.00 (m, 4H, CH2 acetonide), 3.9-3.46 (m, 27H, dendron), 2.30 (t, J=7.5 Hz, 4H, -
OOCCH2-), 1.55 (m, 4H, -OOCCH2CH2-), 1.34-1.25 (m, 80H, Alkyl chain and acetonide-CH3), 0.88 (t,
J=7.5 Hz, 6H, -CH3). 13
C NMR (126 MHz, Acetone-d6) δ 173.20, 123.57, 109.57, 109.51, 79.41, 75.57,
75.56, 73.16, 72.14, 67.60, 67.43, 67.41, 63.19, 34.30, 30.43, 30.42, 30.38, 30.30, 30.24, 30.15, 30.08,
30.05, 29.99, 29.84, 29.73, 29.69, 29.53, 29.38, 27.22, 27.19, 27.17, 25.79, 25.77, 25.76, 25.74, 23.33.
MS (ESI) m/z = 1406.9755 [M+Na]+ (calcd. for C75H137N3NaO19: 1406.9744). IR: 2984, 2922, 2853,
1744, 1456, 1370, 1253, 1213.
Compound 19: 1.7 g of compound 19 was obtained starting from 1.5 g of compound 15 (yield 86%)
8
1H NMR (500 MHz, Acetone-d6): δ 8.06 (m, 1H, CH(Triazole)), 5.20 (m, 1H, Triazole-CH(CH2O-)2),
4.81 (m, 2H, -OCH2-Triazole-), 4.60-4.58 (m, 4H, -CH2OOC-), 4.25-4.17 (m, 8H, CH acetonide), 4.05-
4.01 (m, 8H, CH2 acetonide), 3.91-3.87 (m, 1H), 3.74-3.35 (m, 59H, dendron), 2.31 (t, J= 7.5 Hz, 4H, -
OOCCH2-), 1.56 (m, 4H, -OOCCH2CH2-), 1.35-1.28 (m, 80H, Alkyl chain and acetonide-CH3), 0.86 (t,
J= 6 Hz, 6H, -CH3). 13
C NMR (126 MHz, Acetone-d6) δ 173.25, 123.58, 109.59, 79.67, 79.36, 75.72,
75.68, 75.59, 75.57, 73.20, 73.19, 72.22, 72.20, 72.12, 72.11, 70.92, 67.64, 67.49, 67.46, 67.45, 67.45,
67.43, 63.70, 63.25, 59.36, 34.32, 32.65, 30.38, 30.30, 30.25, 30.15, 30.10, 30.06, 29.99, 29.84, 29.76,
29.69, 29.54, 29.53, 29.38, 27.29, 27.27, 27.23, 27.21, 25.84, 25.83, 25.83, 25.81, 25.75, 25.53, 23.34,
14.39. MS (ESI) m/z = 1991.2026 [M+Na]+ (calcd. for C99H177N3NaO35: 1991.2060). IR: 2985, 2923,
2854, 1745, 1457, 1370, 1253, 1213.
Compound 20: 1.8 g of compound 20 was obtained from 1.7 g of compound 15 (yield 76%)
1H NMR (500 MHz, Acetone–d6): δ 7.98 (m, 1H, CH(Triazole)),5.49 (m, 1H, -CHOOC-), 4.79 (m, 2H,
-OCH2-Triazole-), 4.75 (dd, J= 14.5, 4.5, 1H, Triazole-CH2CH-), 4.70 (dd, J= 14.5, 7 Hz, 1H, Triazole-
CH2CH-), 4.41 (dd, J= 12, 4 Hz, 1H, -CH2OOC-), 4.24-4.18 (m, 8H, CH acetonide), 4.13 (dd, J=12, 6
Hz, 1H, -CH2OOC-), 4.05-4.01 (m, 8H, CH2 acetonide), 3.77- 3.46(m, 59H, dendron), 2.35 (t, J= 7.5 Hz,
2H, -OOCCH2-), 2.30 (t, J=7.5 Hz, 2H, -OOCCH2-), 1.63-1.54 (m, 4H, -OOCCH2CH2-), 1.35-1.25 (m,
92H, Alkyl chain and acetonide-CH3), 0.90-0.86 (m, 6H, -CH3). 13
C NMR (101 MHz, Acetone –d6) δ
173.32, 172.85, 146.53, 124.89, 109.58, 109.50, 79.91, 79.65, 79.44, 79.36, 78.76, 75.70, 75.67, 75.56,
73.17, 72.78, 72.39, 72.18, 72.10, 71.18, 70.86, 70.46, 67.62, 67.44, 64.33, 63.13, 50.53, 34.52, 34.41,
32.66, 32.63, 31.33, 30.42, 30.41, 30.28, 30.23, 30.10, 30.04, 29.84, 29.65, 29.46, 29.26, 27.63, 27.27,
27.22, 26.80, 26.22, 25.82, 25.59, 25.54, 25.41, 23.33, 14.39. MS (ESI) m/z = 2075.3006 [M+Na]+ (calcd.
for C105H189N3NaO35: 2075.2999). IR: 2985, 2923, 2855, 1744, 1457, 1370, 1254, 1213.
Compound 21: 1.9 g of compound 21was obtained from 1.85 g of compound 15 (yield 71%)
1H NMR (500 MHz, Acetone–d6): 8.06 (s, 1H, CH(Triazole)), 5.22-5.19 (m, 1H, Triazole-CH(CH2O-)2),
4.81 (t, J=2Hz, 2H, -OCH2-Triazole-), 4.60-4.58 (m, 4H, -CH2OOC-), 4.23-4.18 (m, 8H, CH acetonide),
4.08-3.98 (m, 8H, CH2 acetonide), 3.73-3.46 (m, 59H, dendron), 2.32 (t, J= 7.5 Hz, 4H, -OOCCH2-),
1.56 (m, 4H, -OOCCH2CH2-), 1.35-1.25 (m, 104H, Alkyl chain and acetonide-CH3), 0.88 (t, J= 7 Hz, 6H,
-CH3). 13
C NMR (126 MHz, ACETONE-D6) δ 173.25, 109.60, 75.73, 75.68, 75.59, 75.58, 73.21, 72.19,
72.11, 67.65, 67.49, 67.47, 63.25, 59.34, 34.33, 32.65, 30.44, 30.44, 30.40, 30.38, 30.30, 30.27, 30.15,
30.09, 30.08, 29.99, 29.84, 29.77, 29.69, 29.67, 29.53, 29.51, 29.38, 27.29, 27.27, 27.24, 27.22, 25.84,
25.83, 25.81, 25.54, 23.34, 14.38. MS (ESI) m/z = 2159.3911 [M+Na]+ (calcd. for C111H201N3NaO35:
2159.3938). IR: 2985, 2923, 2854, 1745, 1456, 1370, 1253, 1213.
9
Deprotection of the dendrons: The protected amphiphiles (16-21) were dissolved in MeOH and added
Dowex-H (2 equiv. by weight ) and left stirring at room temperature (the reaction was done in EtOH and
heated to 45 °C in the case of compound 3). Deprotection reaction was monitored using NMR. After
complete deprotection of the amphiphiles, the reaction mixture was filtered using filter paper and Dowex
was washed with excess methanol. The filtrate was concentrated and crude mixture was purified by
reverse phase HPLC using methanol/water (90-95%), yield (70-80%).
Compound 1:
1H NMR (500 MHz, Methanol-d4) δ 8.16 (m, 1H, CH(Triazole)), 5.23-5.18 (m, 1H, Triazole-CH(CH2O-
)2), 4.83-4.79 (2H, m, -OCH2-Triazole-), 4.62-4.55 (m, 4H, -CH2OOC-), 3.80-3.46 (m, 35H, dendron),
2.30 (t, J= 7.5Hz, 4H, -OOCCH2-), 1.55 (m, 4H, -OOCCH2CH2-), 1.34-1.25 (m, 32H, alkylchains), 0.90
(t, J= 7 Hz, 6H, -CH3). 13
C NMR (126 MHz, Methanol-d4) δ 174.52, 124.90, 79.81, 73.95, 72.93, 72.42,
72.28, 72.18, 64.43, 63.44, 34.67, 33.07, 30.75, 30.59, 30.48, 30.40, 30.11, 25.87, 23.74, 14.49. MS (ESI)
m/z = 1078.6623 [M+Na]+ (calcd. for C51H97N3NaO19: 1078.6614). IR: 3396, 2922, 2854, 1743, 1464.
Compound 2:
1H NMR (500 MHz, Methanol-d4) : δ 8.07-8.06 (m, 1H, CH(Triazole)), 5.48 (m, 1H, -CHOOC-), 4.79-
4.66 (m, 4H, -CH2-Triazole-CH2-), 4.44 (dd, J=12, 3.5 Hz, 1H, -CH2OOC-), 4.13-4.09 (m, 1H, -
CH2OOC-), 3.79-3.48 (m, 35 H, dendron), 2.35 (t, J= 7 Hz, 2H, -OOCCH2-), 2.28 (t, J=7.5 Hz, 2H, -
OOCCH2-), 1.62 (m, 2H, -OOCCH2CH2-), 1.54 (m, 2H, -OOCCH2CH2-), 1.36-1.24 (m, 44H), 0.91 (t, J=
6.5 Hz, 3H), 0.90 (t, J= 6.5 Hz, 3H). 13
C NMR (126 MHz, Methanol-d4) δ 174.70, 174.03, 146.70,
126.16, 79.91, 79.87, 79.19, 73.98, 73.95, 72.95, 72.44, 72.29, 72.21, 71.23, 70.99, 64.48, 64.46, 64.43,
64.14, 63.61, 51.22, 49.51, 49.46, 49.34, 49.29, 49.17, 49.12, 49.00, 48.83, 48.66, 48.49, 34.89, 34.82,
33.11, 33.08, 30.83, 30.80, 30.77, 30.68, 30.66, 30.53, 30.48, 30.47, 30.22, 30.14, 25.99, 25.90, 23.77,
23.74, 14.51, 14.47. MS (ESI) m/z = 1162.7564 [M+Na]+ (calcd. for C57H109N3NaO19: 1162.7553). IR:
3390, 2922, 2853, 1742, 1464.
Compound 3:
1H NMR (500 MHz, Methanol-d4) δ 8.16-8.15 (m, 1H, CH(Triazole)), 5.24-5.18 (m, 1H, Triazole-
CH(CH2O-)2), 4.81-4.79 (m, 2H, -OCH2-Triazole-), 4.59-4.56 (m, 4H, -CH2OOC-), 3.80-3.48 (m, 35 H,
dendron), 2.31 (t, J= 7.5 Hz, 4H, -OOCCH2-), 1.58-1.53 (m, 4H, -OOCCH2CH2-), 1.34-1.25 (m, 56H,
alkyl chains), 0.90 (t, J=7.5 Hz, 6H, -CH3). 13
C NMR (126 MHz, Methanol-d4) δ 174.49, 146.63, 124.89,
10
79.94, 79.90, 79.83, 79.30, 74.01, 73.97, 72.98, 72.48, 72.46, 72.32, 72.23, 72.22, 72.20, 71.25, 64.49,
64.45, 64.25, 63.44, 60.32, 60.30, 49.85, 49.63, 49.51, 49.47, 49.42, 49.34, 49.30, 49.29, 49.25, 49.17,
49.13, 49.08, 49.00, 48.89, 48.83, 48.66, 48.50, 48.49, 34.69, 33.09, 30.82, 30.78, 30.77, 30.62, 30.49,
30.42, 30.14, 25.89, 23.75, 14.47. MS (ESI) m/z = 1246.8552 [M+Na]+ (calcd. for C63H121N3NaO19:
1246.8492). IR: 3397, 2916, 2850, 1743, 1467.
Compound 4:
1H NMR (500 MHz, Methanol-d4): 8.16-8.15 (m, 1H, CH(Triazole)), 5.22-5.19 (m, 1H, Triazole-
CH(CH2O-)2), 4.82 (s, 2H, -OCH2-Triazole-), 4.6-4.56 (m, 4H, -CH2OOC-), 3.79-3.46 (m, 75H,
dendron), 2.32 (t, J= 7.5 Hz, 4H, -OOCCH2-), 1.56 (m, 4H, -OOCCH2CH2-), 1.33-1.26 (m, 32H,
alkylchains), 0.90 (t, J= 6.5 Hz, 6H, -CH3). 13
C NMR (126 MHz, Methanol-d4) δ 174.53, 124.83, 80.03,
79.82, 79.77, 79.43, 73.99, 73.95, 72.95, 72.40, 72.29, 72.18, 72.16, 71.35, 71.02, 64.49, 64.48, 64.43,
63.46, 60.22, 49.51, 49.34, 49.17, 49.00, 48.83, 48.66, 48.49, 34.68, 33.06, 30.74, 30.59, 30.46, 30.40,
30.12, 25.87, 23.73, 14.49. MS (ESI) m/z = 1670.9556 [M+Na]+ (calcd. for C75H145N3NaO35: 1670.9556).
IR: 3380, 2922, 2855, 1743, 1463.
Compound 5:
1H NMR (500 MHz, Methanol-d4): δ 8.06 (m, 1H, CH(Triazole)),5.48 (m, 1H, -CHOOC-), 4.80 (m, 2H,
-OCH2-Triazole-), 4.74 (dd, J= 14.5, 4.5 Hz, 1H, Triazole-CH2CH-), 4.68 (dd, J= 14.5, 7.5 Hz, 1H,
Triazole-CH2CH-), 4.44 (dd, J= 12, 3.5 Hz, 1H, -CH2OOC-), 4.10 (dd, J= 12, 6Hz, 1H, -CH2OOC-),
3.79-3.45 (m, 75H, dendron), 2.34 (t, J= 7.5 Hz, 2H, -OOCCH2-), 2.28 (t, J= 7.5 Hz, 2H, -OOCCH2-),
1.61 (m, 2H, -OOCCH2CH2-), 1.54 (m, 2H, -OOCCH2CH2-), 1.34-1.26 (m, 44H,alkylchains), 0.90 (t,
J=7 Hz, 3H, -CH3), 0.89 (t, J= 7 Hz, 3H, -CH3). 13
C NMR (126 MHz, Methanol-d4) δ 174.72, 174.06,
146.79, 126.10, 101.28, 79.83, 73.98, 73.95, 72.95, 72.43, 72.41, 72.29, 72.19, 72.17, 71.02, 64.49, 64.48,
64.44, 63.65, 51.21, 49.51, 49.34, 49.30, 49.17, 49.13, 49.00, 48.96, 48.83, 48.66, 48.49, 34.90, 34.83,
33.10, 33.06, 30.82, 30.79, 30.75, 30.68, 30.66, 30.52, 30.47, 30.22, 30.14, 25.99, 25.91, 23.76, 23.73,
14.52, 14.48.
MS (ESI) m/z = 1755.0493 [M+Na]+ (calcd. for C81H157N3NaO35: 1755.0495). IR: 3389, 2921, 2853,
1742, 1464.
Compound 6:
1H NMR (500 MHz, Methanol-d4): 8.16 (m, 1H, CH(Triazole)), 5.21 (m, 1H, Triazole-CH(CH2O-)2),
4.83 (m, 2H, -OCH2-Triazole-), 4.61-4.57 (m, 4H, -CH2OOC-), 3.80-3.46 (m, 75H, dendron), 2.32 (t,
J=7.5 Hz, 4H, -OOCCH2-), 1.57 (m, 4H, -OOCCH2CH2-), 1.36-1.25 (m, 56H, alkyl chains), 0.90 (t, J= 7
11
Hz, 6H, -CH3).13
C NMR (126 MHz, Methanol-d4) δ 174.51, 124.82, 124.81, 80.01, 79.81, 79.76, 73.98,
73.94, 72.94, 72.42, 72.39, 72.28, 72.17, 72.15, 71.01, 64.48, 64.47, 64.43, 64.41, 63.43, 60.14, 49.51,
49.34, 49.30, 49.17, 49.14, 49.13, 49.00, 48.98, 48.96, 48.96, 48.83, 48.78, 48.66, 48.65, 48.49, 34.70,
33.07, 30.81, 30.77, 30.63, 30.44, 30.14, 25.89, 23.74, 14.50. MS (ESI) m/z = 1839.1412 [M+Na]+ (calcd.
for C87H169N3NaO35: 1839.1434). IR: 3376, 2920, 2852, 1743, 1464.
3) Critical aggregation concentration (CAC) measurements
a) Surface tension method: The critical aggregation concentrations of the amphiphiles were determined
by measuring their surface tension in Milli-Q water at different concentrations. The surface tension
measurements were performed on a commercially available contact angle tensiometer OCA 20
(DataPhysics Instruments GmbH, Filderstadt, Germany) using pendant drop method. The samples
were prepared in Milli-Q water 24 h before measurement. All the measurements were done at 25 ±
0.5 °C. Calculation of surface tension was done by using the Young-Laplace equation. The surface
tension was measured trice per minute and the measurement was stopped when the value did not
change by over more than 0.1 mN m-1
over 5 minutes. Equilibration time was generally between 60-
90 min below the CAC and around 30 min above the CAC.
b) Fluorescence method: We have reinvestigated the aggregation behavior of amphiphile 6,
which did not show a clear levelling off in the surface tension on increasing concentration,
using the so-called pyrene fluorescence method. It is known that the CAC of an amphiphile
depends to a certain extent on the method. We tried to confirm at first our CAC value of
amphiphile 5 by the pyrene fluorescence method. Amphiphile 5 showed a well-defined break
point in the surface tension measurements. The breakpoint in the semilogarithimic plot is
Figure S1. Surface tension measurements
12
conventionally interpreted as the CMC.2 The pyrene intensity ratio (I1/I3) shows a similar
declining behavior when plotted against the concentration. The breaking points in the surface
tension vs. concentration plot coincides with the inflection point of the I1/I3 curve. This
allows us to interpret the points of inflection as the apparent cmc. This empirical procedure
gives a value of 2-4 M for amphiphile 6.
Preparation of samples for fluorescence experiments:
A stock solution of pyrene (1mM) was prepared in methanol. 50 L of the methanolic solution of
pyrene was taken in a 250 mL conical flask and the solvent was evaporated under high vacuum.
100 mL of milliQ water was added to this conical and the aqueous solution was sonicated for 1h.
The aqueous pyrene solution (0.5 mM) was filtered through a membrane filter (RC, 0.45 ) and
the filtrate was used for the fluorescence studies. A stock solution of the amphiphile was
prepared by dissolving required amount of the amphiphile in aqueous pyrene solution. The stock
solution was further diluted to required concentrations of the amphiphile using the same aqueous
Figure S2. CAC measurement of amphiphiles (5&6) a) by pyrene fluorescence and b) by surface
tension. Comparision of CAC from fluorescence and surface tension methods c) amphiphile 5 and d)
amphiphile 6.
13
pyrene solution. All the samples were stabilized overnight and recorded their fluorescence
spectra (ex= 335 nm and em = 350-550 nm). The ratio of first and third vibronic bands were
calculated and plotted against the concentration of amphiphile as shown in the figure S2(a).
4) Dye encapsulation experiments and calculation of transport capacity
In order to test the transport capacity of the synthesized amphiphiles, two hydrophobic dyes (Nile Red
and Nimodipine) were investigated as water insoluble models. A thin film of the dye (0.5 mg) was
prepared in each vial by evaporating a stock solution of the dye in THF at room temperature, and then the
vials were dried under high vacuum for 10-12 h. To these vials, a solution of the amphiphilic molecule
(0.5% wt., 2.5 mL) was added and left stirring at 500 rpm at room temperature. All the samples were
filtered through 0.45 m RC membranes to remove the insoluble dye particles. A known volume of the
filtrate (0.5 mL) was lyophilised from each sample. The lyophilised samples were dissolved in 4 mL of
organic solvent (Methanol for Nile Red and Ethanol for Nimodipine) and recorded their UV-Vis spectra.
The concentration of the dye molecules in the filtrate was calculated using the molar extinction
coefficients of the dye molecules in corresponding solvents (45,000 M-1
cm-1
for Nile Red at 552 nm in
MeOH and 7,200 M-1
cm-1
for Nimodipine at 356 nm in EtOH) and corresponding dilution factor.
5) Dynamic light scattering measurements
Dynamic light scattering studies were conducted using a Zetasizer Nano (Malvern Instruments Ltd.). All
the samples were prepared by dissolving the compound in Milli-Q water by sonication (for 1min, get a
homogeneous dispersion) and vortexing (30 min), and further tempered at 50 °C for 30 min. DLS
measurements were performed before and after tempering the sample. No considerable influence of
tempering was observed. All the samples were filtered through 0.45 m RC membrane prior to
measurement.
6) Cryogenic transmission electron microscopy (cryo-TEM)
The samples for cryogenic transmission electron microscopy (cryo-TEM) were prepared at room
temperature by placing a droplet (6 μL) of the solution on a hydrophilized perforated carbon filmed
Quantifoil grid (60 s Plasma treatment at 8 W using a BALTEC MED 020 device). The excess fluid was
blotted off to create an ultra-thin layer (typical thickness of 100 nm) of the solution spanning the holes of
the carbon film. The grids were immediately vitrified in liquid ethane at its freezing point (-184°C) using
a standard plunging device. Ultra-fast cooling is necessary for an artifact-free thermal fixation
(vitrification) of the aqueous solution avoiding crystallization of the solvent or rearrangement of the
assemblies. The vitrified samples were transferred under liquid nitrogen into a Philips CM12 transmission
14
electron microscope using the Gatan cryoholder and -stage (Model 626). Microscopy was carried out at
-175°C sample temperature using the microscopes low dose mode at a primary magnification of 58300×.
Accelerating voltage was 100 kV and the defocus was chosen to be 1.2 μm.
7) Packing parameter approach
For common single-tail surfactants, the ratio of the volume of the surfactant tail, v0, and its length, l0, is a
constant independent of tail length, equal to 21 Å2.3 [C. Tanford, The hydrophobic Effect: Wiley-
Interscience: New York, 1973]. For double tail surfactants it is twice that value: 2v0/l0 = 42 Å2. Therefore,
the chain length should not affect the molecular packing parameter, (2v0/al0), where a is the area per
molecule, and consequently also not the shape of aggregate. However, if one alkyl chain contains only
2/3 of the carbon atoms of the second chain, as it is valid for the (G2-C12/18) compound 2, the volume-
to-length ratio reduces to 5/6˖(2v0/l0) = 35 Å2. Hence, as compared to a symmetric double tail compounds,
the volume-to-length ratio is reduced to 35 Å2, i.e. the packing parameter is smaller, which favours an
aggregate morphology with larger curvature.
References:
1. M. Wyszogrodzka and R. Haag, Chemistry, 2008, 14, 9202–14.
2. . guiar, . arpena, . . olina- ol var, and . arnero ui , J. Colloid Interface Sci., 2003,
258, 116–122.
3. The Hydrophobic Effect: Formation of Micelles and Biological Membranes, Charles Tanford,
Wiley-Interscience, New York, 1980, 233pp.
15
8) Additional cryo-TEM images
9) Representation NMR spectra for final compounds
Figure S4. Cryo-TEM images of a sample of amphiphile 2 prior (a) and after (b) encapsulation of
Nimodipine. The coexistence of spherical and worm-like micelles persists even after encapsulation of the
guest molecule. (Scale bars 100 nm)
Figure S3. (a) Additional cryo-TEM image of a sample of amphiphile 3 showing large vesicles and (b) Cryo-TEM
image of the amphiphile 2 after tempering at 60 °C for 3 h. It shows mainly worm-like micelles. (Scale bar 30 nm)
a b
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 16: Protected G2C12/12
5.89
56.5
5
3.90
3.81
26.8
7
3.80
3.79
3.76
1.78
0.75
0.67
0.86
0.87
0.87
0.88
0.89
0.89
0.89
1.28
1.29
1.33
1.33
1.34
1.55
2.05
2.05
2.05
2.29
2.29
2.30
2.31
3.52
3.53
3.54
3.55
3.56
3.57
3.58
3.71
3.71
3.72
4.01
4.02
4.02
4.20
4.57
4.58
4.60
4.78
4.78
5.20
5.21
8.06
8.06
8.07
8.08
0102030405060708090100110120130140150160170180190200210220f1 (ppm)
Compound 16: Protected G2C12/12
14.3
914
.42
25.5
129
.53
ACET
ON
E-D
629
.69
ACET
ON
E-D
629
.71
ACET
ON
E-D
629
.73
ACET
ON
E-D
629
.84
ACET
ON
E-D
629
.86
ACET
ON
E-D
629
.88
ACET
ON
E-D
629
.99
ACET
ON
E-D
630
.01
ACET
ON
E-D
630
.04
ACET
ON
E-D
630
.15
ACET
ON
E-D
630
.38
ACET
ON
E-D
632
.65
34.3
0
59.4
163
.21
64.3
267
.41
67.5
972
.15
73.1
675
.57
75.7
179
.40
109.
58
123.
59
173.
22
206.
12
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 17: Protected G2C12/18
6.04
78.4
1
4.00
0.27
27.8
9
8.84
0.70
3.73
0.74
0.57
0.86
50.
870
0.87
90.
884
0.88
90.
893
0.89
81.
091
1.10
31.
281
1.28
91.
296
1.31
71.
336
1.60
72.
041
2.04
62.
050
2.05
42.
058
2.28
92.
328
2.34
33.
472
3.48
23.
492
3.50
43.
522
3.53
13.
541
3.55
13.
562
3.57
43.
585
3.59
73.
607
3.64
63.
658
3.66
83.
693
3.70
73.
710
3.72
33.
737
4.00
84.
022
4.03
84.
189
4.20
04.
212
4.22
44.
758
4.76
34.
768
5.62
47.
965
0102030405060708090100110120130140150160170180190200210f1 (ppm)
Compound 17: Protected G2C12/18
14.3
714
.40
29.5
3 AC
ETO
NE-
D6
29.5
5 AC
ETO
NE-
D6
29.6
9 AC
ETO
NE-
D6
29.7
1 AC
ETO
NE-
D6
29.8
4 AC
ETO
NE-
D6
29.8
6 AC
ETO
NE-
D6
29.9
9 AC
ETO
NE-
D6
30.0
1 AC
ETO
NE-
D6
30.0
930
.15
ACET
ON
E-D
630
.17
ACET
ON
E-D
630
.42
ACET
ON
E-D
6
50.5
5
63.1
064
.24
67.4
167
.43
67.5
970
.44
72.0
873
.16
75.5
775
.58
79.3
579
.40
109.
58
124.
88
172.
8417
3.33
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5f1 (ppm)
Compound 18: Protected G2C18/18
6.29
86.3
6
4.00
2.54
4.06
28.5
2
4.07
4.02
4.05
1.93
0.86
0.92
0.86
70.
881
0.89
00.
894
1.09
21.
105
1.24
81.
252
1.25
51.
262
1.27
81.
282
1.29
31.
312
1.32
41.
334
1.33
71.
535
1.54
91.
563
2.04
12.
045
2.05
02.
054
2.05
82.
286
2.30
12.
316
2.82
33.
463
3.47
43.
483
3.49
13.
494
3.50
53.
518
3.52
53.
530
3.53
33.
537
3.54
43.
554
3.55
73.
564
3.56
83.
574
3.57
83.
581
3.58
93.
596
3.59
93.
601
3.60
83.
611
3.65
13.
654
3.66
33.
673
3.68
43.
696
3.70
83.
712
3.71
83.
725
3.73
03.
734
3.73
83.
746
4.00
44.
009
4.01
74.
021
4.02
54.
034
4.03
84.
191
4.20
24.
213
4.22
24.
225
4.58
34.
595
4.77
84.
783
4.78
88.
064
8.06
9
0102030405060708090100110120130140150160170180190200210220f1 (ppm)
Compound 18: Protected G2C18/18
23.3
3125
.742
25.7
6325
.771
25.7
8827
.171
27.1
8727
.221
29.3
78 A
CETO
NE-
D6
29.5
33 A
CETO
NE-
D6
29.6
87 A
CETO
NE-
D6
29.7
3029
.840
ACE
TON
E-D
629
.994
ACE
TON
E-D
630
.048
30.0
84 A
CETO
NE-
D6
30.1
49 A
CETO
NE-
D6
30.2
40 A
CETO
NE-
D6
30.3
02 A
CETO
NE-
D6
30.3
78 A
CETO
NE-
D6
30.4
2330
.433
34.3
01
63.1
9167
.410
67.4
3267
.596
72.1
4573
.162
75.5
5875
.572
79.4
06
109.
508
109.
575
123.
568
173.
200
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 19: Protected G3C12/12
6.14
5.61
81.4
1
4.07
3.62
4.00
60.1
3
1.09
8.03
8.05
3.96
1.96
0.96
0.31
0.92
0.87
10.
885
0.89
71.
093
1.10
51.
288
1.29
51.
345
1.54
71.
561
1.57
6
2.29
92.
314
2.32
9
2.77
62.
810
3.53
43.
539
3.54
43.
549
3.55
43.
564
3.57
33.
580
3.58
33.
703
3.71
63.
720
4.02
9
4.58
64.
599
4.81
35.
178
5.19
15.
203
5.21
55.
228
5.62
0
8.06
3
0102030405060708090100110120130140150160170180190200210220f1 (ppm)
Compound 19: Protected G3C12/12
14.3
9525
.805
25.8
2625
.828
25.8
3627
.230
29.3
79 A
CETO
NE-
D6
29.5
32 A
CETO
NE-
D6
29.5
43 A
CETO
NE-
D6
29.6
86 A
CETO
NE-
D6
29.8
41 A
CETO
NE-
D6
29.9
94 A
CETO
NE-
D6
30.1
47 A
CETO
NE-
D6
59.3
5763
.249
63.7
0067
.432
67.4
4567
.454
67.4
6467
.486
67.6
3970
.923
72.1
0872
.125
72.1
9672
.220
73.1
9073
.201
75.5
7475
.589
75.6
8575
.719
79.3
6579
.671
109.
594
123.
582
173.
251
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 20: Protected G3C12/18
7.09
109.
47
3.97
4.00
68.8
3
19.6
7
1.15
4.26
0.92
0.89
0.87
30.
880
0.88
71.
288
1.30
11.
319
1.32
41.
344
2.04
12.
045
2.05
02.
054
2.05
82.
349
3.48
63.
496
3.49
83.
506
3.52
93.
532
3.53
53.
540
3.54
23.
546
3.55
13.
554
3.56
03.
563
3.57
13.
577
3.58
13.
590
3.59
73.
601
3.61
03.
669
3.67
93.
687
3.69
03.
698
3.70
23.
715
3.71
93.
731
3.73
74.
017
4.02
04.
030
4.03
34.
036
4.04
64.
048
4.20
64.
218
4.79
47.
978
0102030405060708090100110120130140150160170180190200210220f1 (ppm)
Compound 20: Protected G3C12/18
14.3
925
.82
27.2
227
.27
29.2
6 AC
ETO
NE-
D6
29.4
6 AC
ETO
NE-
D6
29.6
5 AC
ETO
NE-
D6
29.8
4 AC
ETO
NE-
D6
30.0
4 AC
ETO
NE-
D6
30.1
0 AC
ETO
NE-
D6
30.2
3 AC
ETO
NE-
D6
30.2
8 AC
ETO
NE-
D6
30.4
1 AC
ETO
NE-
D6
30.4
2 AC
ETO
NE-
D6
50.5
363
.13
64.3
367
.44
67.6
270
.46
70.8
671
.18
72.1
072
.18
72.3
972
.78
73.1
773
.54
75.5
675
.67
75.7
078
.76
79.3
679
.44
79.6
579
.91
109.
5010
9.58
124.
89
146.
53
172.
8517
3.32
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 21: Protected G3C18/18
6.23
114.
07
3.95
4.04
0.56
63.8
0
8.53
8.50
4.09
1.95
0.86
0.91
0.86
80.
882
0.89
10.
896
1.28
91.
298
1.31
31.
327
1.34
61.
563
1.56
42.
041
2.04
52.
050
2.05
42.
058
2.30
12.
316
2.33
02.
811
3.47
73.
480
3.48
83.
494
3.49
83.
508
3.53
23.
535
3.53
83.
543
3.54
53.
549
3.55
43.
557
3.56
53.
571
3.57
43.
580
3.58
43.
593
3.59
93.
602
3.61
33.
616
3.62
33.
646
3.65
13.
661
3.67
23.
682
3.68
53.
691
3.70
13.
704
3.71
33.
717
3.72
13.
730
3.73
33.
738
3.74
33.
747
3.75
03.
754
3.76
34.
014
4.01
74.
020
4.03
04.
033
4.03
64.
043
4.04
64.
049
4.19
54.
207
4.21
64.
220
4.22
84.
232
4.58
74.
599
4.81
14.
814
4.81
75.
620
8.06
3
0102030405060708090100110120130140150160170180190200210220f1 (ppm)
Compound 21: Protected G3C18/18
14.3
8023
.334
25.5
3625
.811
27.2
1427
.235
29.3
78 A
CETO
NE-
D6
29.5
33 A
CETO
NE-
D6
29.6
86 A
CETO
NE-
D6
29.8
40 A
CETO
NE-
D6
29.9
92 A
CETO
NE-
D6
30.0
7630
.148
ACE
TON
E-D
630
.265
ACE
TON
E-D
630
.301
ACE
TON
E-D
630
.381
30.4
0030
.434
ACE
TON
E-D
6
63.2
3867
.467
67.4
8872
.186
73.1
9875
.572
75.5
87
109.
593
173.
246
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 1: G2C12/12
6.00
0
33.1
50
4.00
0
3.93
8
35.5
57
4.15
8
1.78
8
1.16
8
0.79
2
0.88
80.
902
0.91
5
1.52
11.
534
1.54
81.
562
1.57
7
2.28
92.
304
2.31
9
4.55
54.
568
4.57
34.
579
4.58
64.
597
4.60
94.
620
4.79
34.
801
4.80
44.
812
5.18
55.
196
5.19
85.
209
5.22
05.
223
5.23
3
8.16
3
0102030405060708090100110120130140150160170180190200210f1 (ppm)
Compound 1: G2C12/12
14.4
8523
.738
25.8
6930
.110
30.3
9830
.478
30.5
9330
.749
33.0
7448
.488
MET
HAN
OL-
D3
48.6
58 M
ETH
ANO
L-D
348
.830
MET
HAN
OL-
D3
49.0
00 M
ETH
ANO
L-D
349
.170
MET
HAN
OL-
D3
49.3
40 M
ETH
ANO
L-D
3
63.4
4164
.425
72.1
8472
.278
72.4
2472
.935
73.9
4779
.808
124.
902
174.
521
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
-100000
0
100000
200000
300000
400000
500000
600000
700000
800000
900000
1000000
1100000
1200000
1300000
1400000Compound 2: G2C12/18
6.00
45.5
41.
871.
83
1.90
1.86
35.7
9
0.85
0.86
3.98
0.87
0.88
0.88
90.
891
0.90
30.
905
0.91
60.
919
1.25
51.
265
1.27
51.
291
1.29
61.
311
1.31
71.
321
1.32
71.
328
1.33
21.
343
2.26
62.
281
2.29
62.
336
2.35
12.
365
3.30
43.
307
3.31
03.
313
3.31
73.
460
3.46
23.
472
3.47
43.
480
3.48
23.
492
3.49
53.
500
3.51
23.
515
3.52
33.
526
3.53
53.
545
3.55
13.
555
3.56
13.
563
3.56
53.
571
3.57
33.
576
3.58
33.
585
3.59
33.
595
3.60
53.
608
3.61
53.
681
3.68
43.
692
3.69
63.
700
3.70
53.
742
3.74
83.
752
3.75
93.
764
3.76
83.
771
3.77
53.
780
4.68
94.
704
4.72
34.
787
4.79
64.
878
8.07
1
0102030405060708090100110120130140150160170180190200210220f1 (ppm)
-300000
-200000
-100000
0
100000
200000
300000
400000
500000
600000
700000
800000
900000
1000000
1100000
1200000
1300000
1400000
1500000
1600000
1700000
1800000
1900000
2000000
2100000Compound 2: G2C12/18
14.4
714
.51
30.4
730
.48
30.6
830
.77
30.8
030
.83
33.0
833
.11
48.6
6 M
ETH
ANO
L-D
348
.83
MET
HAN
OL-
D3
49.0
0 M
ETH
ANO
L-D
349
.17
MET
HAN
OL-
D3
49.3
4 M
ETH
ANO
L-D
3
63.6
164
.46
64.4
8
70.9
971
.23
72.2
172
.29
72.4
472
.95
73.9
573
.98
79.1
979
.87
79.9
1
126.
16
146.
70
174.
0317
4.70
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 3: G2C18/18
6.05
59.7
5
4.00
4.00
2.39
36.7
9
4.02
1.93
0.98
0.92
0.89
0.90
0.90
0.92
1.26
1.27
1.28
1.29
1.31
1.31
1.32
1.33
2.29
2.31
2.32
3.31
3.31
3.31
3.35
3.54
3.54
3.56
3.56
3.56
3.57
3.57
3.57
3.58
3.59
4.57
4.58
4.58
4.80
4.80
4.81
4.86
5.19
5.20
5.22
8.15
8.16
-30-20-100102030405060708090100110120130140150160170180190200210220230f1 (ppm)
Compound 3: G2C18/18
14.4
723
.75
25.8
930
.49
30.6
230
.77
30.7
830
.82
33.0
948
.83
MET
HAN
OL-
D3
49.0
0 M
ETH
ANO
L-D
349
.17
MET
HAN
OL-
D3
49.3
4 M
ETH
ANO
L-D
3
60.3
263
.44
64.4
564
.49
71.2
572
.20
72.2
272
.23
72.3
272
.46
72.4
872
.98
73.9
774
.01
79.3
079
.83
79.9
079
.94
124.
89
146.
63
174.
49
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 4: G3C12/12
6.41
35.7
2
3.99
4.00
82.9
8
5.22
3.34
1.53
0.87
0.89
00.
898
0.90
40.
917
1.26
71.
292
1.31
21.
314
1.31
71.
328
1.54
71.
561
1.57
52.
302
2.31
72.
331
3.53
03.
537
3.54
13.
545
3.54
83.
557
3.56
53.
570
3.57
83.
580
3.58
93.
592
3.59
73.
602
4.57
54.
587
4.82
04.
824
4.82
74.
831
4.86
6
5.19
75.
209
5.22
1
8.15
18.
155
8.15
98.
163
0102030405060708090100110120130140150160170180190200210f1 (ppm)
Compound 4: G3C12/12
14.4
85
23.7
2625
.871
30.1
2230
.403
30.4
6430
.592
30.7
4233
.060
34.6
8248
.659
MET
HAN
OL-
D3
48.8
29 M
ETH
ANO
L-D
349
.000
MET
HAN
OL-
D3
49.1
71 M
ETH
ANO
L-D
349
.340
MET
HAN
OL-
D3
60.2
2163
.461
64.4
3264
.481
64.4
9572
.182
72.2
8973
.988
79.4
2979
.770
79.8
2480
.026
124.
825
174.
534
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 5: G3C12/18
6.00
0
45.0
36
3.41
3
3.60
7
76.4
34
0.72
9
0.71
80.
584
1.74
81.
962
0.77
6
0.83
0
0.88
00.
884
0.89
40.
898
0.90
80.
912
1.26
51.
283
1.28
81.
306
1.31
31.
320
1.33
52.
279
2.33
12.
346
3.29
53.
299
3.30
23.
305
3.30
93.
454
3.45
83.
466
3.47
43.
478
3.48
63.
490
3.49
83.
510
3.52
13.
527
3.53
33.
538
3.54
83.
556
3.56
03.
562
3.56
93.
580
3.59
23.
603
3.61
23.
624
3.63
33.
661
3.66
63.
670
3.67
53.
679
3.68
23.
686
3.69
13.
696
3.70
53.
712
3.73
23.
737
3.74
23.
749
3.75
33.
758
3.76
03.
764
3.76
83.
774
4.80
04.
857
8.06
3
0102030405060708090100110120130140150160170180190200210220f1 (ppm)
Compound 5: G3C12/18
14.4
814
.52
30.4
730
.52
30.6
630
.68
30.7
530
.79
30.8
233
.06
33.1
0
48.6
6 M
ETH
ANO
L-D
348
.83
MET
HAN
OL-
D3
48.9
6 M
ETH
ANO
L-D
349
.00
MET
HAN
OL-
D3
49.1
7 M
ETH
ANO
L-D
349
.34
MET
HAN
OL-
D3
63.6
564
.44
64.4
864
.49
72.1
772
.19
72.2
972
.41
79.8
3
101.
28
126.
10
146.
79
174.
0617
4.72
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
Compound 6: G3C18/18
6.00
56.5
5
4.01
3.89
74.7
1
5.09
1.91
1.07
0.96
0.88
90.
903
0.91
71.
256
1.26
21.
275
1.29
41.
312
1.31
61.
326
1.34
11.
356
1.55
61.
570
1.58
31.
598
2.30
72.
322
2.33
73.
304
3.30
73.
310
3.31
33.
317
3.34
93.
464
3.46
83.
476
3.48
43.
496
3.50
03.
508
3.52
03.
530
3.53
63.
542
3.54
53.
549
3.55
73.
566
3.57
03.
579
3.58
93.
593
3.59
73.
602
3.61
13.
614
3.62
23.
625
3.63
53.
646
3.65
03.
656
3.66
33.
677
3.68
53.
688
3.69
73.
708
3.71
83.
727
3.74
03.
749
3.75
23.
760
3.76
33.
768
3.77
43.
786
3.79
74.
576
4.58
84.
598
4.61
04.
821
4.82
54.
828
4.84
74.
879
4.90
74.
911
5.18
75.
197
5.20
95.
221
8.15
8
0102030405060708090100110120130140150160170180190200210220f1 (ppm)
Compound 6: G3C18/18
14.5
00
23.7
3525
.892
30.1
4430
.440
30.6
2630
.771
30.8
1233
.070
34.6
9948
.660
MET
HAN
OL-
D3
48.8
30 M
ETH
ANO
L-D
349
.002
MET
HAN
OL-
D3
49.1
69 M
ETH
ANO
L-D
349
.341
MET
HAN
OL-
D3
63.4
3464
.410
64.4
2864
.470
64.4
8472
.155
72.1
7373
.979
79.7
6079
.813
80.0
15
124.
808
124.
821
174.
506
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