Template for Electronic Submission to ACS Journals · Web viewUV-vis spectra for gold nanoparticle solutions modified with (a) M6EG and (b) 5EG. Figure S1----S2 Images of contact

Properties of Modified Surface for Biosensing Interface

Mutsuo Tanakaa,*, Takahiro Sawaguchia, Yoshiki Hirataa, Osamu Niwab, Keiko Tawac, Chisato

Sasakawac, and Koji Kuraokad

a Health Research Institute, Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1

Higashi, Tsukuba, Ibaraki 305-8566, Japan; b Advanced Science Research Laboratory, Saitama Institute

of Technology, 1690 Fusaiji, Fukaya, Saitama 369-0293, Japan; c School of Science and Technology,

Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337, Japan; d Graduate School of

Maritime Sciences, Kobe University, 5-1-1 Fukaeminami, Higashinada, Kobe, Hyogo 658-0022, Japan

*Corresponding authors. E-mail:[email protected]

Supplementary Material

Contents

UV-vis spectra for gold nanoparticle solutions modified with (a) M6EG and (b) 5EG. Figure S1----S2

Images of contact angle measurement Figure S2-----S2

Images of water layer measurement by AFM Figure S3-----S3

Image of bare chip surface by AFM Figure S4-----S3

Synthesis procedures for surface modification materials-----S4

S1

Figure S1. UV-vis spectra for gold nanoparticle solutions modified with (a) M6EG and (b) 5EG. The

blue, red, and green lines represent spectra without ions, in the presence of PBS or NaCl, respectively.

Figure S2. Images of contact angle measurement.

S2

Figure S3. Image of hydration layer measurement by AFM.

Figure S4. Image of bare chip surface by AFM.

Surface roughness of a LaSFN9 chip covered with a gold film was measured by scanning probe microscopy (SPA-400 and SPI3800N, SII). Mean value of Ra was evaluated as 0.6 nm from five lines drawn randomly.

S3

Synthesis Procedures for Surface Modification Materials

1. Synthesis of M5EG

Synthesis of Triethylene Glycol Methyl Ether Tosylate

To a three-necked flask, triethylene glycol monomethyl ether (8.21 g, 50 mmol), sodium hydroxide

(7.00 g, 175 mmol), THF (35 mL), and water (35 mL) were put, and the mixture was stirred at 0 ˚C. To

the mixture, a THF solution (50 mL) of p-toluenesulfonyl chloride (11.4 g, 60 mmol) was added

dropwise. The reaction mixture was stirred at 0 ˚C for 2h, and then, at room temperature for 12 h. The

reaction mixture was poured to 5wt% aqueous hydrochloric acid, and the product was extracted with

chloroform. After the solvent was evaporated, the obtained crude product by drying under vacuum

condition was used for the subsequent reaction without purification: pale yellow liquid, yield 96%.

1H-NMR (CDCl3, 500MHz) 2.43 (3H, s), 3.35 (3H, s), 3.48~3.68 (10H, m), 4.14 (2H, t, J=4.80 Hz),

7.32 (2H, d, J=8.25 Hz), 7.78 (2H, d, J=8.20 Hz).

Synthesis of Pentaethylene Glycol Monomethyl Ether

Sodium hydride (2.00 g, 60%, 50 mmol) and DMF (120 mL) were put to a three-necked flask, and the

mixture was stirred at room temperature. To the mixture, diethylene glycol (5.30 g, 50 mmol) was

added, and the mixture was heated at 100 ˚C. A DMF solution (20 mL) of triethylene glycol methyl

ether tosylate (3.18 g, 10 mmol) was added dropwise to the mixture, and the reaction mixture was

stirred at 100 ˚C for 3 h. The reaction mixture was allowed to cool at room temperature, and methanol

was added to decompose excess sodium hydride. The solvent was evaporated, and the obtained residue

was poured into 5wt% aqueous hydrochloric acid. The product was extracted with chloroform three

S4

times. The product obtained by solvent evaporation was purified with silica-gel column

chromatography (eluent; chloroform:methanol = 100:0 then 100:2 vol.): pale brown liquid, yield 53%.

1H-NMR (CDCl3, 500MHz) 3.36 (3H, s), 3.51~3.72 (20H, m).

Synthesis of Pentaethylene Glycol Methyl Ether Tosylate

To a three-necked flask, pentaethylene glycol monomethyl ether (2.52 g, 10 mmol), sodium hydroxide



dropwise. The reaction mixture was stirred at 0 ˚C for 2 h, and then, at room temperature for 12 h. The


chloroform twice. The solvent was evaporated, and the product was purified by HPLC (gel permeation

chromatography, GPC): pale brown liquid, yield 83%.

1H-NMR (CDCl3, 500MHz) 2.44 (3H, s), 3.36 (3H, s), 3.51~3.68 (18H, m), 7.32 (2H, d, J=8.25 Hz),

7.79 (2H, d, J=8.25 Hz).

Synthesis of M5EG

Under nitrogen atmosphere, pentaethylene glycol methyl ether tosylate (2.03 g, 5 mmol) and ethanol

(80 mL) were put into a three-necked flask, and the mixture was stirred at room temperature. To the

mixture, thiourea (1.14 g, 15 mmol) was added, and the reaction mixture was stirred and refluxed for 24

h under nitrogen atmosphere. The reaction mixture was allowed to cool at 0 ˚C, and an ethanol solution

(20 mL) of ethylene diamine (7.50 g, 125 mmol) was added dropwise. The reaction mixture was stirred

at 0 ˚C for 10 h under nitrogen atmosphere, and poured into ice water to neutralize with conc. aqueous

hydrochloric acid. The product was extracted with chloroform twice. Purification of the product was

performed with silica-gel column chromatography (eluent; chloroform:methanol = 100:0 then 100:1

vol.): colorless liquid, yield 72%.

1H-NMR (CDCl3, 500MHz) 1.58 (1H, t, J=8.25 Hz), 2.68 (2H, q, J=7.02 Hz), 3.37 (3H, s),

3.51~3.68 (18H, m); 13C-NMR (CDCl3) 23.4, 58.1, 69.3, 69.60, 69.62, 69.69(3C), 69.72, 71.0, 72.0.


S5

Synthesis of Triethylene Glycol Monobenzyl Ether

Sodium hydride (16.0 g, 60%, 400 mmol) and DMF (500 mL) were put to a three-necked flask, and

the mixture was stirred at room temperature. To the mixture, triethylene glycol (60.0 g, 400 mmol) was

added, and the mixture was heated at 100 ˚C. A DMF solution (80 mL) of benzyl chloride (12.7 g, 100

mmol) was added dropwise to the mixture, and the mixture was stirred at 100 ˚C for 3 h. The reaction

mixture was allowed to cool at room temperature, and methanol was added to decompose excess sodium

hydride. The solvent was evaporated, and the obtained residue was poured to 5wt% aqueous

hydrochloric acid. The product was extracted with chloroform. The organic layer was rinsed with

water. The product obtained by solvent evaporation was purified with silica-gel column

chromatography (eluent; chloroform:methanol = 100:0 then 100:2 vol.): pale brown liquid, yield 86%.

1H-NMR (CDCl3, 500MHz) 3.57~3.76 (12H, m), 4.57 (2H, s), 7.24~7.38 (5H, m).

Synthesis of Hexaethylene Glycol Benzyl Methyl Ether

To a three-necked flask, triethylene glycol monobenzyl ether (2.40 g, 10 mmol), sodium hydride (4.00

g, 60%, 100 mmol), and THF (100 mL) were put, and the mixture was stirred and refluxed. To the

mixture, a THF solution (30 mL) of triethylene glycol methyl ether tosylate (4.78 g, 15 mmol) was

added dropwise, and the mixture was stirred and refluxed for 12 h. The reaction mixture was allowed to

cool at room temperature, and methanol was added to decompose excess sodium hydride. The reaction

mixture was poured into 5wt% aqueous hydrochloric acid, and the product was extracted with

chloroform twice. The product obtained by solvent evaporation was purified with HPLC (GPC): pale

brown liquid, yield 92%.

S6

1H-NMR (CDCl3, 400MHz) 3.38 (3H, s), 3.52~3.74 (24H, m), 4.57 (2H, s), 7.24~7.36 (5H, m).

Synthesis of Hexaethylene Glycol Monomethyl Ether

To an autoclave, hexaethylene glycol benzyl methyl ether (3.86 g, 10 mmol), palladium carbon (5wt

%, 500 mg), and ethanol (100 mL) were put, and the autoclave was sealed. Hydrogen was introduced to

the autoclave, and the reaction mixture was stirred at 100 ˚C for 24 h under 7 atm hydrogen atmosphere.

The autoclave was allowed to cool at room temperature, and depressurized. The reaction mixture was

filtrated to remove palladium carbon. The product obtained by solvent evaporation was dried under

vacuum condition, and used for the subsequent reaction without purification: colorless liquid, yield

quantitative.


Synthesis of Hexaethylene Glycol Mesyl Methyl Ether

To a three-necked flask, hexaethylene glycol monomethyl ether (593 mg, 2 mmol), triethyl amine

(1.01 g, 10 mmol), and dichloromethane (50 mL) were put, and the mixture was stirred at 0 ˚C. To the

mixture, a dichloromethane solution (20 mL) of mesyl chloride (1.15 g, 10 mmol) was added. The

reaction mixture was allowed to warm at room temperature, and stirred for 12 h. The reaction mixture

was poured to 5wt% aqueous hydrochloric acid, and the product was extracted with chloroform twice.

The solvent was evaporated, and the product was purified with silica-gel column chromatography

(eluent; chloroform:methanol = 100:1 then 100:3 vol.): colorless liquid, yield 93%.

1H-NMR (CDCl3, 500MHz) 3.08 (3H, s), 3.38 (3H, s), 3.52~3.78 (22H, m), 4.38 (2H, t, J=4.50 Hz).

Synthesis of M6EG

Under nitrogen atmosphere, hexaethylene glycol methyl ether mesylate (374 mg, 1 mmol) and ethanol


mixture, thiourea (228 mg, 3 mmol) was added, and the reaction mixture was stirred and refluxed for 24

h under nitrogen atmosphere. The reaction mixture was allowed to cool at 0 ˚C, and an ethanol solution



S7

hydrochloric acid. The product was extracted with chloroform twice. Purification of the product was

performed with silica-gel column chromatography (eluent; chloroform:methanol = 100:0 then 100:2

vol.): colorless liquid, yield 69%.


3.53~3.69 (24H, m); 13C-NMR (CDCl3) 24.7, 59.4, 70.65, 70.93, 70.95, 71.00 (5C), 71.05, 72.3, 73.3.


Synthesis of Heptaethylene Glycol Monomethyl Ether

Sodium hydride (2.00 g, 60%, 50 mmol) and THF (120 mL) were put to a three-necked flask, and the

mixture was stirred at room temperature. To the mixture, tetraethylene glycol (9.71 g, 50 mmol) was

added, and the mixture was refluxed. A THF solution (20 mL) of triethylene glycol methyl ether

tosylate (3.18 g, 10 mmol) was added dropwise to the mixture, and the reaction mixture stirred and

refluxed for 12 h. The reaction mixture was allowed to cool at room temperature, and methanol was

added to decompose excess sodium hydride. The solvent was evaporated, and the obtained residue was

poured to 5wt% aqueous hydrochloric acid. The product was extracted with chloroform three times.

The product obtained by solvent evaporation was purified with silica-gel column chromatography

(eluent; chloroform:methanol = 100:0 then 100:2 vol.): pale brown liquid, yield 48%.


Synthesis of Heptaethylene Glycol Methyl Ether Tosylate

To a three-necked flask, heptaethylene glycol monomethyl ether (3.40 g, 10 mmol), sodium hydroxide



S8

dropwise. The reaction mixture was stirred at 0 ˚C for 2 h, and then, at room temperature for 12 h. The


chloroform twice. The solvent was evaporated, and the product was purified with silica-gel column

chromatography (eluent; chloroform:methanol = 100:0 then 100:1 vol.): colorless liquid, yield 52%.

1H-NMR (CDCl3, 500MHz) 2.42 (3H,s), 3.35 (3H, s), 3.49~3.68 (26H, m), 4.13 (2H, t, J=4.80 Hz),

7.32 (2H, d, J=8.20 Hz), 7.77 (2H, d, J=8.25 Hz).

Synthesis of M7EG

Under nitrogen atmosphere, hexaethylene glycol methyl ether mesylate (837 mg, 2 mmol) and ethanol


mixture, thiourea (457 mg, 6 mmol) was added, and the reaction mixture was stirred and refluxed for 24

h under nitrogen atmosphere. The reaction mixture was allowed to cool at 0 ˚C, and ethanol solution



hydrochloric acid. The product was extracted with chloroform twice. The solvent was evaporated, and

purification of the product was performed with silica-gel column chromatography (eluent;

chloroform:methanol = 100:0 then 100:1 vol.): colorless liquid, yield 65%.


3.52~3.68 (26H, m); 13C-NMR (CDCl3) 23.5, 58.3, 69.48, 69.77 (2C), 69.82 (7C), 70.10, 71.2, 72.1.

4. Synthesis of 4EG

Synthesis of Tetraethylene Glycol Monotosylate

To a three-necked flask, tetraethylene glycol (18.4 g, 100 mmol), sodium hydroxide (1.20 g, 30

mmol), THF (100 mL), and water (100 mL) were put, and the mixture was stirred at 0 ˚C. To the

mixture, a THF solution (30 mL) of p-toluenesulfonyl chloride (3.81 g, 20 mmol) was added dropwise.

S9

The reaction mixture was stirred at 0 ˚C for 2 h, and then, at room temperature for 20 h. The reaction

mixture was poured to 5wt% aqueous hydrochloric acid, and the product was extracted with chloroform

twice. The solvent was evaporated, and the obtained crude product was purified with silica-gel column


1H-NMR (CDCl3, 500MHz) 2.43 (3H, s), 3.56~3.73 (14H, m), 4.15 (2H, t, J=4.80 Hz), 7.33 (2H, d,

J=7.75 Hz), 7.79 (2H, d, J=8.25 Hz).

Synthesis of 4EG

Under nitrogen atmosphere, tetraethylene glycol monotosylate (1.74 g, 5 mmol) and ethanol (70 mL)

were put into a three-necked flask, and the mixture was stirred at room temperature. To the mixture,

thiourea (1.14 g, 15 mmol) was added, and the reaction mixture was stirred and refluxed for 24 h under

nitrogen atmosphere. The reaction mixture was allowed to cool at 0 ˚C, and an ethanol solution (20 mL)

of ethylene diamine (7.50 g, 125 mmol) was added dropwise. The reaction mixture was stirred at 0 ˚C

for 10 h under nitrogen atmosphere, and poured into ice water to neutralize with conc. aqueous

hydrochloric acid. The product was extracted with chloroform three times. The solvent was

evaporated, and purification of the product was performed with silica-gel column chromatography


1H-NMR (CDCl3, 500MHz) 1.58 (1H, t, J=8.25 Hz), 2.64 (2H, q, J=7.02 Hz), 3.51~3.70 (14H, m);

13C-NMR (CDCl3) 23.5, 61.0, 69.5, 69.6, 69.8, 69.9, 71.8, 72.2.

5. Synthesis of 5EG

Synthesis of Pentaethylene Glycol Monotosylate

To a three-necked flask, pentaethylene glycol (2.38 g, 10 mmol), sodium hydroxide (800 mg, 20

mmol), THF (25 mL), and water (25 mL) were put, and the mixture was stirred at 0 ˚C. To the mixture,

THF solution (20 mL) of p-toluenesulfonyl chloride (2.86 g, 15 mmol) was added dropwise. The

S10

reaction mixture was stirred at 0 ˚C for 2 h, and then, at room temperature for 20 h. The reaction





J=8.25 Hz), 7.79 (2H, d, J=8.25 Hz).

Synthesis of 5EG

Under nitrogen atmosphere, pentaethylene glycol monotosylate (1.96 g, 5 mmol) and ethanol (70 mL)










13C-NMR (CDCl3) 23.3, 60.9, 69.3, 69.5, 69.63, 69.69, 69.71, 69.73, 71.6, 72.0.

6. Synthesis of 6EG

Synthesis of Hexaethylene Glycol Monotosylate

To a three-necked flask, hexaethylene glycol (2.82 g, 10 mmol), sodium hydroxide (880 mg, 22

mmol), THF (25 mL), and water (25 mL) were put, and the mixture was stirred at 0 ˚C. To the mixture,

a THF solution (20 mL) of p-toluenesulfonyl chloride (2.86 g, 15 mmol) was added dropwise. The

S11

reaction mixture was stirred at 0 ˚C for 2 h, and then, at room temperature for 48 h. The reaction





J=8.25 Hz), 7.78 (2H, d, J=8.70 Hz).

Synthesis of 6EG

Under nitrogen atmosphere, hexaethylene glycol monotosylate (2.18 g, 5 mmol) and ethanol (70 mL)










13C-NMR (CDCl3) 23.4, 60.9, 69.3, 69.5, 69.65, 69.69, 69.71 (2C), 69.75, 69.76, 71.6, 72.0.

7. Synthesis of 8C3EG

Synthesis of Triethylene Glycol Mono(8-bromooctyl) Ether

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To a three-necked flask, triethylene glycol (60.1 g, 400 mmol), 1,8-dibromooctane (10.9 g, 40 mmol),

and THF (800 mL) were put, and the mixture was stirred at room temperature. To the mixture,

potassium t-butoxide (4.48 g, 40 mmol) was added. The reaction mixture was stirred and refluxed for 6

h. The reaction mixture was allowed to cool at room temperature, and the solvent was evaporated. The

obtained residue was poured to 5wt% aqueous hydrochloric acid, and the product was extracted with

chloroform. The solvent was evaporated, and the obtained crude product was purified with silica-gel

column chromatography (eluent; chloroform:methanol = 100:0 then 100:0.5 vol.): colorless liquid, yield

41%. The product still contained a byproduct (ca. 10 mol%) via elimination reaction, which was

removed by the following procedures.

1H-NMR (CDCl3, 500MHz) 1.24~1.62 (10H, m), 1.79~1.88 (2H, m), 3.39 (2H, t, J=6.88 Hz), 3.44

(2H, t, J=6.88 Hz), 3.55~3.75 (12H, m).

Synthesis of Triethylene Glycol Mono(8-mercaptooctyl) Ether

Under nitrogen atmosphere, triethylene glycol mono(8-bromooctyl) ether (3.79 g, 90 %, 10 mmol)

and ethanol (200 mL) were put into a three-necked flask, and the mixture was stirred at room

temperature. To the mixture, thiourea (2.28 g, 30 mmol) was added, and the reaction mixture was

stirred and refluxed for 24 h under nitrogen atmosphere. The reaction mixture was allowed to cool at 0

˚C, and an ethanol solution (50 mL) of ethylene diamine (15.0 g, 250 mmol) was added dropwise. The

reaction mixture was stirred at 0 ˚C for 10 h under nitrogen atmosphere, and poured into ice water to

neutralize with conc. aqueous hydrochloric acid. The product was extracted with chloroform. The

product obtained by solvent evaporation was dried under vacuum condition, and used for the subsequent

reaction without purification: colorless liquid, yield quantitative.

Synthesis of Triethylene Glycol Mono(8-tritylthiooctyl) Ether

Under nitrogen atmosphere, triethylene glycol mono(8-mercaptooctyl) ether (3.27 g, 90 %, 10 mmol),

trityl chloride (2.79 g, 10 mmol), and acetonitrile (200 mL) were put into a three-necked flask, and the

mixture was stirred at room temperature. To the mixture, potassium carbonate (2.07 g, 15 mmol) was

added, and the reaction mixture was stirred and refluxed for 12 h under nitrogen atmosphere. The

S13

reaction mixture was allowed to cool at room temperature. The solvent was evaporated, and the product

was extracted with chloroform. Purification of the product obtained by solvent evaporation was

performed with HPLC (GPC): colorless liquid, yield 82%.

1H-NMR (CDCl3, 500MHz) 1.10~1.61 (12H, m), 2.13 (2H, t, J=7.33 Hz), 3.43 (2H, t, J=6.88 Hz),

3.55~3.75 (12H, m), 7.18~7.44 (15H, m).

Synthesis of 8C3EG

Under nitrogen atmosphere, triethyl silane (2.33 g, 20 mmol), trifluoroacetic acid (30 mL), and

dichloromethane (30 mL) were put into a three-necked flask, and the mixture was stirred at room

temperature. To the mixture, a dichloromethane solution (50 mL) of triethylene glycol mono(8-

tritylthiooctyl) ether (5.37 g, 10 mmol) was added dropwise, and the reaction mixture was stirred at

room temperature for 30 min under nitrogen atmosphere. The reaction mixture was poured to ice water,

and neutralized with potassium carbonate. The product was extracted with chloroform. The organic

layer was rinsed with water, and then, with 5wt% aqueous hydrochloric acid. The solvent was



1H-NMR (CDCl3, 500MHz) 1.22~1.64 (13H, m), 2.51 (2H, q, J=7.32 Hz), 3.44 (2H, t, J=6.65 Hz),

3.55~3.76 (12H, m).

8. Synthesis of 12C4EG

Synthesis of Tetraethylene Glycol Mono(12-bromododecyl) Ether

To a three-necked flask, tetraethylene glycol (77.7 g, 400 mmol), 1,12-dibromododecane (13.1 g, 40

mmol), and THF (800 mL) were put, and the mixture was stirred at room temperature. To the mixture,

S14

potassium t-butoxide (4.48 g, 40 mmol) was added. The reaction mixture was stirred and refluxed for 6

h. The reaction mixture was allowed to cool at room temperature, and the solvent was evaporated. The

obtained residue was poured into 5wt% aqueous hydrochloric acid, and the product was extracted with

chloroform. The solvent was evaporated, and the obtained crude product was purified with silica-gel

column chromatography (eluent; chloroform:hexane = 100:100 then 100:0 vol.): colorless liquid, yield

39%. The product still contained a byproduct (ca. 10 mol%) via elimination reaction, which was

removed by the following procedures.


(2H, t, J=6.65 Hz), 3.55~3.75 (16H, m).

Synthesis of Tetraethylene Glycol Mono(12-mercaptododecyl) Ether

Under nitrogen atmosphere, tetraethylene glycol mono(12-bromododecyl) ether (4.90 g, 90 %, 10

mmol) and ethanol (200 mL) were put into a three-necked flask, and the mixture was stirred at room

temperature. To the mixture, thiourea (2.28 g, 30 mmol) was added, and the reaction mixture was

stirred and refluxed for 24 h under nitrogen atmosphere. The reaction mixture was allowed to cool at 0

˚C, and an ethanol solution (50 mL) of ethylene diamine (15.0 g, 250 mmol) was added dropwise. The

reaction mixture was stirred at 0 ˚C for 10 h under nitrogen atmosphere, and poured into ice water to

neutralize with conc. aqueous hydrochloric acid. The product was extracted with chloroform. The

product obtained by solvent evaporation was dried under reduced pressure, and used for the subsequent

reaction without purification: colorless liquid, yield quantitative.

Synthesis of Tetraethylene Glycol Mono(12-tritylthiododecyl) Ether

Under nitrogen atmosphere, tetraethylene glycol mono(12-mercaptododecyl) ether (4.38 g, 90 %, 10

mmol), trityl chloride (2.79 g, 10 mmol), and acetonitrile (200 mL) were put into a three-necked flask,

and the mixture was stirred at room temperature. To the mixture, potassium carbonate (2.07 g, 15

mmol) was added, and the reaction mixture was stirred and refluxed for 12 h under nitrogen atmosphere.

The reaction mixture was allowed to cool at room temperature. The solvent was evaporated, and the

S15

product was extracted with chloroform. Purification of the product obtained by solvent evaporation was

performed with HPLC (GPC): colorless liquid, yield 82%.


(2H, t, J=6.88 Hz), 3.56~3.75 (16H, m), 7.17~7.44 (15H, m).

Synthesis of 12C4EG

Under nitrogen atmosphere, triethyl silane (2.33 g, 20 mmol), trifluoroacetic acid (30 mL), and

dichloromethane (30 mL) were put into a three-necked flask, and the mixture was stirred at room

temperature. To the mixture, a dichloromethane solution (50 mL) of tetraethylene glycol mono(12-

tritylthiododecyl) ether (6.37 g, 10 mmol) was added dropwise, and the reaction mixture was stirred at

room temperature for 30 min under nitrogen atmosphere. The reaction mixture was poured to ice water,

and neutralized with potassium carbonate. The product was extracted with chloroform. The organic

layer was rinsed with water, and then, with 5wt% aqueous hydrochloric acid. The solvent was


(eluent; chloroform:methanol = 100:0 then 100:0.5 vol.): colorless liquid, yield 91%.

1H-NMR (CDCl3, 500MHz) 1.18~1.39 (17H, m), 1.50~1.61 (4H, m), 2.49 (2H, q, J=7.33 Hz), 3.41

(2H, t, J=6.88 Hz), 3.52~3.74 (16H, m); 13C-NMR (CDCl3) 24.7, 26.1, 28.4, 29.1, 29.54, 29.57, 29.64

(4C), 34.1, 61.8, 70.1, 70.3, 70.60, 70.62, 70.65, 70.67, 71.6, 72.7.

9. Synthesis of 6CPC

Synthesis of Choline Tosylate

To a three-necked flask, N,N-dimethylaminoethanol (4.46 g, 50 mmol) and THF (200 mL) were put,

and the mixture was stirred at room temperature. To the mixture, a THF solution (50 mL) of methyl

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tosylate (1.02 g, 55 mmol) was added dropwise, and the reaction mixture was stirred at room

temperature for 24 h. The solvent was evaporated, and the product was purified by recrystallization

with acetonitrile: colorless solid, yield 95%.

1H-NMR (CD3OD, 400MHz) 2.37 (3H, s), 3.19 (9H, s), 3.47 (2H, t, J=4.88 Hz), 3.98 (2H, broad),

7.23 (2H, d, J=4.02 Hz), 7.70 (2H, d, J=4.10 Hz).

Synthesis of 6-Hydroxyhexane Tritylthio Ether

Under nitrogen atmosphere, 6-hydroxyhexanethiol (1.34 g, 10 mmol), trityl chloride (2.79 g, 10

mmol), and acetonitrile (200 mL) were put into a three-necked flask, and the mixture was stirred at

room temperature. To the mixture, potassium carbonate (2.07 g, 15 mmol) was added, and the reaction

mixture was stirred and refluxed for 12 h under nitrogen atmosphere. The reaction mixture was allowed

to cool at room temperature. The solvent was evaporated, and the product was extracted with

chloroform. Purification of the product obtained by solvent evaporation was performed with HPLC

(GPC): colorless solid, yield 76%.

1H-NMR (CDCl3, 500MHz) 1.18~1.53 (8H, m), 2.14 (2H, t, J=7.33 Hz), 3.58 (2H, t, J=6.63 Hz),

7.18~7.43 (15H, m).

Synthesis of 6-Phosphorylcholine-Hexane Tritylthio Ether

Phosphoryl chloride (3.06 g, 20 mmol) and dichloromethane (40 mL) were placed in a round-

bottomed flask and stirred at 0 ˚C. A dichloromethane solution (20 mL) of 6-hydroxyhexane tritylthio

ether (753 mg, 2 mmol) with pyridine (158 mg, 2 mmol) was added dropwise to the mixture, and the

reaction mixture was stirred at 0 ˚C for 1 h, and then, at room temperature for 1h. The solvent and

excess phosphoryl chloride were removed under vacuum condition, and pyridine (40 mL) was poured to

the obtained residue. The mixture was cooled at 0 ˚C, and choline tosylate (2.76 g, 10 mmol) was added

to the mixture at once with vigorous stirring. The reaction mixture was allowed to warm at room

temperature, and stirred for 24 h. To the reaction mixture, water (20 mL) was added and the reaction

mixture was stirred for 6 h at room temperature. The solvent was evaporated, and 5wt% aqueous

hydrochloric acid was added to the obtained residue. The product was extracted with chloroform,

S17

concentrated by solvent evaporation, and purified by silica-gel column chromatography (eluent;

methanol:chloroform = 100:100 then 100:0): colorless waxy solid, yield 43%.

1H-NMR (CDCl3, 500MHz) 12~1.53 (8H, m), 2.10 (2H, t, J=7.10 Hz), 3.33 (9H, s), 3.69~3.78

(4H, m), 4.20 (2H, broad), 7.14~7.39 (15H, m).

Synthesis of 6CPC

Triethyl silane (466 mg, 4 mmol), trifluoroacetic acid (20 mL), and dichloromethane (20 mL) were

placed in a round-bottomed flask and stirred at 0 ˚C. A dichloromethane solution (30 mL) of 6-

phosphorylcholine-hexane tritylthio ether (1.08 g, 2 mmol) was added dropwise to the mixture, and the

reaction mixture was stirred at 0 ˚C for 30 min. The solvent was evaporated, and the obtained crude

product was purified by silica-gel column chromatography (eluent; methanol:chloroform = 100:100 then

100:0): colorless solid, yield 66%.

1H-NMR (CDCl3, 500MHz) 1.21~1.38 (5H, m), 1.45~1.57 (4H, m), 2.44 (2H, q, J= 7.33 Hz), 3.33

(9H, s), 3.68~3.78 (4H, m), 4.18 (2H, broad); 13C-NMR (CDCl3) 23.8, 24.6, 27.3, 30.1, 33.1, 53.4

(3C), 58.4, 64.5, 65.4.

10. Synthesis of 12CPC

Synthesis of 12-Hydroxydodecane Tritylthio Ether

Under nitrogen atmosphere, 12-bromododecanol (2.65 g, 10 mmol), trityl thiol (3.32 g, 12 mmol),

and DMF (100 mL) were placed in a three-necked flask and stirred at room temperature. Potassium

carbonate (2.07 g, 15 mmol) was added to the mixture, and the reaction mixture was stirred at room

temperature for 12 h under nitrogen atmosphere. The reaction mixture was poured to 5wt%

hydrochloric acid, and the product was extracted with chloroform. The crude product obtained by

solvent evaporation was purified by HPLC (GPC): colorless solid, yield 85%.

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1H-NMR (CDCl3, 500MHz) 11~1.39 (18H, m), 1.53~1.60 (2H, m), 2.13 (2H, t, J=7.33 Hz), 3.64

(2H, t, J=6.65 Hz), 7.18~7.43 (15H, m).

Synthesis of 12-Phosphorylcholine-Dodecane Tritylthio Ether

Phosphoryl chloride (3.06 g, 20 mmol) and dichloromethane (40 mL) were placed in a round-

bottomed flask and stirred at 0 ˚C. A dichloromethane solution (20 mL) of 12-hydroxydodecane

tritylthio ether (921 mg, 2 mmol) with pyridine (158 mg, 2 mmol) was added dropwise to the mixture,

and the reaction mixture was stirred at 0 ˚C for 1 h, and then, at room temperature for 1h. The solvent

and excess phosphoryl chloride were removed under vacuum condition, and pyridine (40 mL) was

poured to the obtained residue. The mixture was cooled at 0 ˚C, and choline tosylate (2.76 g, 10 mmol)

was added to the mixture at once with vigorous stirring. The reaction mixture was allowed to warm at

room temperature, and stirred for 24 h. To the reaction mixture, water (20 mL) was added and the

reaction mixture was stirred for 6 h at room temperature. The solvent was evaporated, and 5wt%

aqueous hydrochloric acid was added to the obtained residue. The product was extracted with

chloroform, concentrated by solvent evaporation, and purified by silica-gel column chromatography

(eluent; methanol:chloroform = 100:100 then 100:0): colorless waxy solid, yield 35%.


(9H, s), 3.76~3.85 (4H, m), 4.23~4.30 (2H, broad), 7.17~7.42 (15H, m).

Synthesis of 12CPC


placed in a round-bottomed flask and stirred at 0 ˚C. A dichloromethane solution (30 mL) of 12-

phosphorylcholine-dodecane tritylthio ether (1.25 g, 2 mmol) was added dropwise to the mixture, and

the reaction mixture was stirred at 0 ˚C for 30 min. The solvent was evaporated, and the obtained crude


100:0): colorless solid, yield 79%.

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(9H, s), 3.75~3.86 (4H, m), 4.23~4.29 (2H, broad); 13C-NMR (CDCl3) 24.7, 26.0, 28.4, 29.1, 29.53,

29.57, 29.62, 29.67, 29.70, 31.1, 34.1, 54.3 (3C), 59.2, 65.6, 66.3.

11. Synthesis of 6CCP

Synthesis of 6-Bromohexane Tritylthio Ether

Under nitrogen atmosphere, 1,6-dibromohexane (19.5 g, 80 mmol), trityl thiol (11.1 g, 40 mmol),



temperature for 24 h under nitrogen atmosphere. The reaction mixture was poured to 5wt% aqueous


solvent evaporation was dried under vacuum condition, and used for the subsequent reaction without

purification: pale brown liquid, yield quantitative.


(2H, t, J=6.82 Hz), 7.17~7.44 (15H, m).

Synthesis of Dimethyl 2-Hydroxyethyl 6-Tritylthiohexyl Ammonium Chloride

Under nitrogen atmosphere, crude 6-bromohexane tritylthio ether (27.3 g, ca. 40 mmol), dimethyl

aminoethanol (107 g, 1.2 mol), and dioxane (600 mL) were placed in a three-necked flask. The reaction

mixture was stirred and refluxed for 12 h under nitrogen atmosphere. The reaction mixture was allowed

to cool at room temperature, and the solvent was evaporated. The obtained residue was poured to 5wt%

aqueous hydrochloric acid, and the product was extracted with chloroform. The crude product obtained

S20

by solvent evaporation was purified by silica-gel column chromatography (eluent; methanol:chloroform

= 100:2 then 100:25): pale brown solid, yield 63%.


(9H, s), 3.45 (2H, t, J=8.20 Hz), 3.69 (2H, broad), 4.10 (2H, broad), 7.18~7.7.43 (15H, m).

Synthesis of 6-Inverse Phosphorylcholine-Hexane Tritylthio Ether

Under nitrogen atmosphere, methyl phosphorodichloridate (14.9 g, 100 mmol) and dichloromethane

(400 mL) were placed in a three-necked flask, and the mixture was stirred at 0 ˚C. A dichloromethane

solution (80 mL) of dimethyl 2-hydroxyethyl 6-tritylthiohexyl ammonium chloride (9.36 g, 20 mmol)

with pyridine (3.16 g, 40 mmol) was added dropwise to the mixture, and the reaction mixture was

stirred at 0 ˚C for 2 h, and then, at room temperature for 6 h under nitrogen atmosphere. The solvent

and methyl phosphorodichloridate were evaporated, and chloroform (300 mL) and water (50 mL) were

poured to the obtained residue. The reaction mixture was stirred at room temperature for 12 h under

nitrogen atmosphere, and poured to 5wt% aqueous hydrochloric acid. The product was extracted with

chloroform, and the crude product obtained by solvent evaporation was purified by silica-gel column

chromatography (eluent; methanol:chloroform = 100:10 then 100:100): colorless solid, yield 15%.

1H-NMR (CDCl3, 400MHz) 12~1.41 (6H, m), 1.51~1.67 (2H, m), 2.12 (2H, q, J=6.31 Hz),

3.17~3.40 (8H, m), 3.42 (3H, s), 3.73 (2H, broad), 4.24 (2H, broad), 7.15~7.42 (15H, m).

Synthesis of 6CCP


placed in a round-bottomed flask and stirred at 0 ˚C. A dichloromethane solution (30 mL) of 6-inverse

phosphorylcholine-hexane tritylthio ether (1.08 g, 2 mmol) was added dropwise to the mixture, and the

reaction mixture was stirred at 0 ˚C for 30 min. The solvent was evaporated, and the obtained crude


100:0), and then, HPLC (octadecyl silica-gel, ODS): colorless solid, yield 31%.

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1H-NMR (CDCl3, 400MHz) 35~1.53 (6H, m), 1.70~1.82 (2H, m), 2.53 (2H, q, J=7.27 Hz), 3.35

(6H, s), 3.51 (2H, t, J=8.26 Hz), 3.58 (3H, d, J=10.6 Hz), 3.81 (2H, broad), 4.31 (2H, broad); 13C-NMR

(CDCl3) 22.7, 24.4, 25.8, 27.8, 51.6 (2C), 58.9, 64.0, 65.4; m/z: 300 (M + H+).

12. Synthesis of 12CCP

Synthesis of 12-Bromododecane Tritylthio Ether

Under nitrogen atmosphere, 1,12-dibromohexane (13.1 g, 40 mmol), trityl thiol (11.1 g, 40 mmol),





solvent evaporation was dried under vacuum condition, and used for the subsequent reaction without

purification: pale brown liquid, yield quantitative (isolated yield: 48%).


(2H, t, J=6.88 Hz), 7.16~7.43 (15H, m).

Synthesis of Dimethyl 2-Hydroxyethyl 12-Tritylthiododecyl Ammonium Chloride

Under nitrogen atmosphere, crude 12-bromododecane tritylthio ether (20.9 g, ca. 40 mmol),

dimethyl aminoethanol (35.6 g, 400 mmol), and dioxane (600 mL) were placed in a three-necked flask.

The reaction mixture was stirred and refluxed for 12 h under nitrogen atmosphere. The reaction mixture

was allowed to cool at room temperature, and the solvent was evaporated. The obtained residue was

poured to 5wt% aqueous hydrochloric acid, and the product was extracted with chloroform. The crude

S22

product obtained by solvent evaporation was purified by silica-gel column chromatography (eluent;

methanol:chloroform = 100:2 then 100:25): pale brown solid, yield 47%.


(6H, s), 3.46~3.56 (2H, m), 3.71 (2H, broad), 4.12 (2H, broad), 7.17~7.44 (15H, m).

Synthesis of 12-Inverse Phosphorylcholine-Dodecane Tritylthio Ether

Under nitrogen atmosphere, methyl phosphorodichloridate (14.9 g, 100 mmol) and dichloromethane

(400 mL) were placed in a three-necked flask, and the mixture was stirred at 0 ˚C. A dichloromethane

solution (80 mL) of dimethyl 2-hydroxyethyl 12-tritylthiododecyl ammonium chloride (11.0 g, 20

mmol) with pyridine (3.16 g, 40 mmol) was added dropwise to the mixture, and the reaction mixture

was stirred at 0 ˚C for 2 h, and then, at room temperature for 6 h under nitrogen atmosphere. The

solvent and methyl phosphorodichloridate were evaporated, and chloroform (300 mL) and water (50

mL) were poured to the obtained residue. The reaction mixture was stirred at room temperature for 12 h

under nitrogen atmosphere, and poured to 5wt% aqueous hydrochloric acid. The product was extracted

with chloroform, and the crude product obtained by solvent evaporation was purified by silica-gel

column chromatography (eluent; methanol:chloroform = 100:10 then 100:100), and then, HPLC (ODS):

colorless solid, yield 9.5%.

1H-NMR (CDCl3, 400MHz) 08~1.43 (18H, m), 1.62~1.78 (2H, m), 2.13 (2H, t, J=7.32 Hz),

3.19~3.50 (8H, m), 3.57 (3H, d, J=10.6 Hz), 3.78 (2H, broad), 4.30 (2H, broad), 7.16~7.43 (15H, m).

Synthesis of 12CCP


placed in a round-bottomed flask and stirred at 0 ˚C. A dichloromethane solution (30 mL) of 12-inverse

phosphorylcholine-dodecane tritylthio ether (1.25 g, 2 mmol) was added dropwise to the mixture, and

the reaction mixture was stirred at 0 ˚C for 30 min. The solvent was evaporated, and the obtained crude


100:0), and then, HPLC (octadecyl silica-gel, ODS): colorless solid, yield 56%.

S23


(6H, s), 3.47 (2H, t, J=8.34 Hz), 3.58 (3H, d, J=10.6 Hz), 3.81 (2H, broad), 4.30 (2H, broad); 13C-NMR

(CDCl3) 22.8, 24.6, 26.3, 28.3, 29.0, 29.2, 29.36, 29.41, 29.43 (2C), 34.0, 51.7 (2C), 52.6, 58.9, 64.1,

65.7; m/z: 384 (M + H+).

13. Synthesis of 6CS

Synthesis of 6-Bromohexane Tritylthio Ether

Under nitrogen atmosphere, 1,6-dibromohexane (2.44 g, 10 mmol), trityl thiol (553 mg, 2 mmol),

and DMF (60 mL) were placed in a three-necked flask, and stirred at room temperature. Potassium

carbonate (552 mg, 4 mmol) was added to the mixture, and the reaction mixture was stirred at room



solvent evaporation was purified by HPLC (GPC): colorless solid, yield 79%.


(2H, t, J=6.84 Hz), 7.17~7.44 (15H, m).

Synthesis of 6-Dimethylaminohexane Tritylthio Ether

Under nitrogen atmosphere, 6-bromohexane tritylthio ether (879 mg, 2 mmol), dimethylamine

hydrochloride (816 mg, 10 mmol), and DMF (150 mL) were placed in a three-necked flask, and stirred

at 50 ˚C. Sodium carbonate (1.59 g, 15 mmol) was added to the mixture, and the reaction mixture was

stirred at 50 ˚C for 12 h under nitrogen atmosphere. The reaction mixture was poured to 5wt% aqueous

sodium carbonate, and the product was extracted with chloroform. The crude product obtained by

S24

solvent evaporation was purified by silica-gel column chromatography (eluent; chloroform:methanol =

100:2 then 100:10): colorless solid, yield 67%.

1H-NMR (CDCl3, 400MHz) 12~1.46 (8H, m), 2.14 (2H, t, J=7.30 Hz), 2.18~2.24 (8H, m),

7.17~7.44 (15H, m).

Synthesis of 6-Sulfobetaine-Hexane Tritylthio Ether

To a three-necked flask, 6-dimethylaminohexane tritylthio ether (807 mg, 2 mmol) and dry acetone

(30 mL) were put, and the mixture was stirred at room temperature. A dry acetone solution (5 mL) of

1,3-propanesultone (366 mg, 3 mmol) was added to the mixture, and the reaction mixture was stirred at

room temperature for 48 h. The solid product obtained by solvent evaporation was rinsed with dry

acetone for purification, and dried under vacuum condition: colorless solid, yield 86%.

1H-NMR (CDCl3, 400MHz) 13~1.42 (6H, m), 1.53~1.68 (2H, m), 2.09~2.23 (4H, m), 3.84 (2H, t,

J=6.78 Hz), 3.10 (6H, s), 3.16 (2H, t, J=8.42 Hz), 3.65 (2H, t, J=8.18 Hz), 7.16~7.42 (15H, m).

Synthesis of 6CS

To a round-bottomed flask, triethyl silane (466 mg, 4 mmol), trifluoroacetic acid (20 mL), and

dichloromethane (20 mL) were put, and the mixture was stirred at 0 ˚C. A dichloromethane solution (30

mL) of 6-sulfobetaine-hexane tritylthio ether (1.05 g, 2 mmol) was added dropwise to the mixture, and

the reaction mixture was stirred at 0 ˚C for 30 min. The product obtained by solvent evaporation was

purified by silica-gel column chromatography (eluent; chloroform:methanol = 100:5 then 100:100):

colorless solid, yield 92%.


(2H, t, J=6.70 Hz), 3.16 (6H, s), 3.27~3.33 (2H, m), 3.68~3.74 (2H, m); 13C-NMR (CDCl3) 19.7, 22.9,

24.7, 26.1, 28.0, 33.9, 48.2, 51.2 (2C), 63.6, 64.7; m/z: 284 (M + H+).

14. Synthesis of 12CS

S25

Synthesis of 12-Bromododecyl Dimethyl Amine

Under nitrogen atmosphere, 1,12-dibromododecane (16.4 g, 50 mmol), dimethylamine

hydrochloride (4.49 g, 55 mmol), and DMF (500 mL) were placed in a three-necked flask, and stirred at

50 ˚C. Sodium carbonate (7.95 g, 75 mmol) was added to the mixture, and the reaction mixture was

stirred at 50 ˚C for 12 h under nitrogen atmosphere. The obtained residue by solvent evaporation was

poured to 5wt% aqueous sodium carbonate, and the product was extracted with chloroform. The

solvent was evaporated, and the product was purified by silica-gel column chromatography (eluent;

chloroform:methanol = 100:0 then 100:5): colorless liquid, yield 30%.

1H-NMR (CDCl3, 500MHz) 12~1.48 (18H, m), 1.71~1.79 (2H, m), 2.20 (6H, s), 2.22 (2H, t,

J=7.80 Hz), 3.52 (2H, t, J=6.85 Hz).

Synthesis of 12-Dimethylaminododecane Tritylthio Ether

Under nitrogen atmosphere, 12-bromododecyl dimethyl amine (585 mg, 2 mmol), trityl thiol (663

mg, 2.4 mmol), and DMF (60 mL) were placed in a three-necked flask, and stirred at room temperature.

Potassium carbonate (415 mg, 3 mmol) was added to the mixture, and the reaction mixture was stirred

at room temperature for 12 h under nitrogen atmosphere. The reaction mixture was poured to 5wt%

aqueous sodium carbonate, and the product was extracted with chloroform. The crude product obtained

by solvent evaporation was purified by silica-gel column chromatography (eluent; chloroform:methanol

= 100:2 then 100:5): colorless liquid, yield 67%.

S26

1H-NMR (CDCl3, 500MHz) 09~1.49 (20H, m), 2.14 (2H, t, J=7.58 Hz), 2.22 (6H, s), 2.25 (2H, t,

J=7.56 Hz), 7.17~7.44 (15H, m).

Synthesis of 12-Sulfobetaine-Dodecane Tritylthio Ether

To a three-necked flask, 12-dimethylaminododecane tritylthio ether (976 mg, 2 mmol) and dry

acetone (30 mL) were put, and the mixture was stirred at room temperature. A dry acetone solution (5

mL) of 1,3-propanesultone (366 mg, 3 mmol) was added to the mixture, and the reaction mixture was

stirred at room temperature for 48 h. The solid product obtained by solvent evaporation was rinsed with

dry acetone for purification, and dried under vacuum condition: colorless solid, yield 60%.


(2H, broad), 2.90 (2H, t, J=6.58 Hz), 3.17 (6H, s), 3.20~3.29 (2H, m), 3.69~3.78 (2H, m), 7.16~7.43

(15H, m).

Synthesis of 12CS

To a round-bottomed flask, triethyl silane (466 mg, 4 mmol), trifluoroacetic acid (20 mL), and

dichloromethane (20 mL) were put, and the mixture was stirred at 0 ˚C. A dichloromethane solution (30

mL) of 12-sulfobetaine-dodecane tritylthio ether (1.22 g, 2 mmol) was added dropwise to the mixture,

and the reaction mixture was stirred at 0 ˚C for 30 min. The product obtained by solvent evaporation

was purified by silica-gel column chromatography (eluent; chloroform:methanol = 100:5 then 100:100):

colorless solid, yield 69%.

1H-NMR (CDCl3, 400MHz) 21~1.43 (19H, m), 1.55~1.78 (4H, m), 2.19~2.30 (2H, m), 2.53 (2H,

q, J=7.37 Hz), 2.92 (2H, t, J=6.62 Hz), 3.20 (6H, s), 3.24~3.32 (2H, m), 3.70~3.78 (2H, m); 13C-NMR

(CDCl3) 19.6, 23.1, 24.9, 26.7, 28.7, 29.4, 29.5, 29.78, 29.81, 29.84 (2C), 34.4, 48.1, 51.1 (2C), 63.8,

65.1; m/z: 368 (M + H+).

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Template for Electronic Submission to ACS Journals · Web viewUV-vis spectra for gold nanoparticle solutions modified with (a) M6EG and (b) 5EG. Figure S1----S2 Images of contact