1
Supp
ortin
g in
form
atio
n
A tw
o-st
ep sy
nthe
sis o
f 7,8
-dic
hlor
o-rib
ofla
vin
with
hig
h yi
eld
O
livie
r Cou
rjean
a , Ann
e H
oche
deza , W
ilfrid
Ner
ia , Fré
déric
Lou
ërat
a , Em
ilie
Trem
eya , S
ébas
tien
Gou
nela , S
eiya
Tsu
jimur
ab , Nic
olas
Man
oa*
a U
nive
rsité
de
Bor
deau
x, C
entre
de
Rec
herc
he P
aul P
asca
l, C
NR
S U
PR 8
641,
115
ave
nue
Schw
eitz
er, 3
3600
Pes
sac,
Fra
nce
b Div
isio
n of
Mat
eria
ls S
cien
ce, F
acul
ty o
f Pur
e an
d A
pplie
d Sc
ienc
es, U
nive
rsity
of T
suku
ba1-
1-1
Tenn
odai
, Tsu
kuba
, Iba
raki
, 305
-857
3, Ja
pan
man
o@cr
pp-b
orde
aux.
cnrs
.fr
Exp
erim
enta
l Sec
tion.
Synt
hesi
s of t
he d
ichl
oro-
ribo
flavi
n us
ing
Boc
pro
tect
ion
(Sch
eme
1).
(1a)
: D
ichl
oro-
1,2p
heny
lene
diam
ine
(3.0
0g,
16.9
mm
ol)
was
dis
solv
ed i
n eq
uim
olar
pro
porti
on w
ith s
odiu
m b
icar
bona
te a
nd d
i-ter
t-but
yl
dica
rbon
ate
in 7
5mL
diox
ane
plus
75m
L w
ater
. The
rea
ctio
n w
as c
arrie
d ou
t for
24
hour
s at
roo
m te
mpe
ratu
re a
nd th
e re
actio
n m
ixtu
re w
as
dilu
ted
in 1
50m
L w
ater
. The
n 1a
was
ext
ract
ed tw
ice
with
50m
L di
chlo
rom
etha
ne. T
he o
rgan
ic p
hase
was
firs
t was
hed
with
50m
L sa
tura
ted
sodi
um b
icar
bona
te s
olut
ion,
then
with
100
mL
brin
e. T
he o
rgan
ic p
hase
was
drie
d us
ing
mag
nesi
um s
ulph
ate
and
evap
orat
ed u
nder
vac
uum
.
Solu
tion
(9m
L of
cyc
lohe
xan
and
3mL
of e
thyl
ace
tate
) was
add
ed to
the
brow
n so
lid a
nd fi
ltrat
ed to
giv
e th
e pu
re p
rodu
ct. Y
ield
: 2.7
4g (5
8%).
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
2 M
ass
spec
trom
etry
ana
lysi
s (2
76.6
g m
ol-1
). 1 H
-NM
R (4
00 M
Hz,
DM
SO D
6): δ
= 8
.47
(s, 1
H),
7.53
(s, 1
H),
6.87
(s, 1
H),
5.33
(s, 2
H),
1.46
(s,
9H).
13C
-NM
R (1
00 M
Hz,
DM
SO D
6): δ
= 1
53.2
, 140
.8, 1
25.7
, 123
.9, 1
16.3
, 115
.5, 7
9.5,
66.
4, 2
8.1.
(1b)
: 1
a (1
.10g
, 4m
mol
es),
D-r
ibos
e (3
.58g
, 24
mm
oles
) an
d so
dium
cya
nobo
rohy
drid
e (0
.50g
, 16
mm
oles
) w
ere
diss
olve
d in
100
mL
drie
d
met
hano
l. Th
e re
actio
n w
as p
erfo
rmed
at 6
5°C
, for
48
hour
s. Lo
wer
am
ount
s of D
-rib
ose
and
sodi
um c
yano
boro
hydr
ide
led
to lo
wer
yie
ld o
f 1b.
Afte
r eva
pora
tion
unde
r vac
uum
, 30m
L co
ncen
trate
d aq
ueou
s H
Cl w
as u
sed
to re
act w
ith th
e re
mai
ning
sod
ium
cya
nobo
rohy
drid
e. T
he s
olut
ion
was
neu
traliz
ed w
ith sa
tura
ted
sodi
um b
icar
bona
te so
lutio
n. 1
b w
as e
xtra
cted
twic
e w
ith 5
0mL
ethy
l ace
tate
. The
org
anic
pha
se w
as w
ashe
d w
ith
brin
e, d
ried
usin
g m
agne
sium
sul
phat
e, th
en th
e so
lven
t was
rem
oved
by
evap
orat
ion
unde
r va
cuum
. Pur
ifica
tion
was
mad
e w
ith p
repa
rativ
e
chro
mat
ogra
phy
(sili
ca g
el) u
sing
AcO
Et. Y
ield
: 0.4
6g (2
8%).
Mas
s spe
ctro
met
ry a
naly
sis (
410.
8g m
ol-1
). 1 H
-NM
R (4
00 M
Hz,
DM
SO D
6) : δ
=
8.59
(s, 1
H),7
.42
(s, 1
H),
6.76
(s, 1
H),
5.32
(t, J
=4.8
1Hz,
1H
), 4.
81 (d
, J=5
.50H
z, 1
H),
4.77
(d, J
=5.5
0Hz,
1H
), 4.
68 (d
, J=5
.04H
z, 1
H),
4.42
(t,
J=5.
50H
z, 1
H),
3.78
(m, 1
H),
3.57
(m, 2
H),
3.43
(m, 2
H),
3.27
(m, 1
H),
3.02
(m, 1
H),
1.45
(s, 9
H).
13C
-NM
R (1
00 M
Hz,
DM
SO D
6): δ
= 1
53.6
,
142.
2, 1
27.1
, 125
.2, 1
24.1
, 115
.7, 1
11.2
, 79.
5, 7
3.2,
72.
8, 6
9.9,
63.
3, 4
5.6,
28.
1.
(2) :
1b
(1.1
0g, 2
.7m
mol
es) w
as d
isso
lved
in 5
3mL
of 4
M H
Cl i
n di
oxan
e. T
he re
actio
n w
as p
erfo
rmed
at r
oom
tem
pera
ture
for 5
hou
rs, u
nder
cons
tant
stir
ring.
The
sol
vent
was
rem
oved
by
evap
orat
ion
unde
r va
cuum
and
the
prod
uct w
as d
isso
lved
in w
ater
(10
0mL)
and
was
hed
thre
e
times
usi
ng 2
0mL
ethe
r. A
queo
us s
olut
ion
cont
aini
ng 2
was
free
ze, f
iltra
te a
nd d
ried
with
out f
urth
er p
urifi
catio
n. Y
ield
: m =
0.6
7g (8
0%).
Mas
s
spec
trom
etry
(ES
I) 3
10.9
g m
ol-1
. 1H
-NM
R (
400
MH
z, E
tOD
D6)
: δ
= 6.
75 (
s, 1H
), 6.
60 (
s, 1H
), 5.
42 (
s, 2H
), 3.
95 (
m, 1
H),
3.74
(m
, 2H
),
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
3 3.
64(m
, 2H
), 3.
51 (b
, 5H
), 3.
34 (m
, 1H
), 3.
14 (m
, 1H
). 13
C-N
MR
(100
MH
z, E
tOD
D6)
: δ =
137
.2, 1
35.4
, 121
.0, 1
19.6
, 115
.9, 1
11.8
, 73.
1, 7
0.9,
63.3
, 53.
5, 4
6.4.
(3)
: Allo
xan
(0.3
1g, 2
.2 m
mol
es)
and
boric
aci
d (0
.58g
, 9.3
mm
oles
) w
ere
diss
olve
d in
boi
ling
pure
ace
tic a
cid
(25m
L) f
or 1
hou
r, th
en th
e
solu
tion
was
coo
led
to r
oom
tem
pera
ture
. Afte
r ad
ding
0.4
2g o
f 2,
the
reac
tion
was
per
form
ed a
t 80°
C f
or 3
hou
rs a
nd, t
hen
was
left
at r
oom
tem
pera
ture
ove
rnig
ht. W
ater
(50m
L) w
as a
dded
bef
ore
evap
orat
ion
unde
r vac
uum
. Abo
ut 8
0mL
was
eva
pora
ted
and
extra
ctio
n w
as m
ade
twic
e
by 5
0mL
of c
yclo
hexa
n an
d tw
ice
by 5
0mL
by e
thyl
e ac
etat
. The
aqu
eous
pha
se w
as c
oole
d an
d a
prec
ipite
d ap
pear
. Filt
ratio
n ga
ve th
e go
od
prod
uct.
Yie
ld :
m =
0.4
3g (7
7%).
HR
MS
(ESI
) 439
.1 g
mol
-1. 1 H
-NM
R (4
00 M
Hz,
DM
SO D
6) : δ
= 11
.48
(b, 1
H),
8.36
(s, 1
H),
8.30
(s, 1
H),
5.06
(s,
1H),
4.73
(m
, 2H
), 4.
41 (
s, 1H
), 4.
13 (
s, 1H
), 3.
56 (
s, 2H
), 2.
43 (
s, 4H
). 13
C-N
MR
(10
0 M
Hz,
DM
SO D
6) : δ
= 15
9.8,
155
.8, 1
51.5
,
140.
2, 1
37.2
, 134
.5, 1
34.4
, 132
.2, 1
28.5
, 120
.1, 7
3.9,
73.
1, 6
9.1,
63.
8, 4
8.2.
Dir
ect s
ynth
esis
of d
ichl
oro-
ribo
flavi
n (S
chem
e 2)
.
D-r
ibos
e (2
.20g
, 15m
oles
), so
dium
cya
nobo
rohy
drid
e (2
.00g
, 32m
mol
es) a
nd 1
(1.7
7g, 1
0mm
oles
) wer
e di
ssol
ved
in d
ried
met
hano
l (10
0mL)
.
The
reac
tion
was
per
form
ed a
t 65°
C, f
or 4
8 ho
urs.
Low
er a
mou
nts
of D
-rib
ose
and
sodi
um c
yano
boro
hydr
ide
led
to lo
wer
yie
ld o
f 2.
Afte
r
evap
orat
ion
unde
r vac
uum
, 30m
L co
ncen
trate
d aq
ueou
s H
Cl w
as u
sed
to re
act w
ith th
e re
mai
ning
sod
ium
cya
nobo
rohy
drid
e. T
he s
olut
ion
was
neut
raliz
ed w
ith s
atur
ated
sod
ium
bic
arbo
nate
sol
utio
n. 2
was
ext
ract
ed t
wic
e w
ith 5
0mL
ethy
l ac
etat
e. T
he o
rgan
ic p
hase
was
drie
d us
ing
mag
nesi
um s
ulph
ate,
then
the
solv
ent w
as re
mov
ed b
y ev
apor
atio
n un
der v
acuu
m. T
he c
rude
pro
duct
was
par
tially
dis
solv
ed in
dic
hlor
omet
hane
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
4 an
d fil
tere
d on
a n
°5 fr
it. U
nsol
uble
pro
duct
was
par
tially
dis
solv
ed a
gain
with
war
m d
ichl
orom
etha
ne, c
oole
d to
room
tem
pera
ture
and
filte
red.
Yie
ld :
m =
1.0
1g (3
3%).
RM
N fo
r com
poun
d 2
as d
escr
ibed
abo
ve.
Allo
xan
(0.3
2g, 2
.2 m
mol
es)
and
boric
aci
d (0
.80g
, 12m
mol
es)
wer
e di
ssol
ved
in b
oilin
g pu
re a
cetic
aci
d (2
5mL)
for
1 h
our
and
then
cool
ed to
50°
C. A
fter a
ddin
g 0.
40g
of 2
, the
reac
tion
was
per
form
ed a
t 80°
C fo
r 3 h
ours
and
then
left
at ro
om te
mpe
ratu
re o
vern
ight
. Ext
ract
ion
and
purif
icat
ion
of 3
was
per
form
ed a
s des
crib
ed a
bove
. Yie
ld :
m =
0.4
1g (7
7%).
RM
N fo
r com
poun
d 3
as d
escr
ibed
abo
ve.
Spec
troe
lect
roch
emic
al e
xper
imen
ts
A th
in la
yer q
uartz
gla
ss s
pect
roel
ectro
chem
ical
cel
l with
a li
ght p
ass
leng
th o
f 1 m
m (A
LS) w
as u
sed
as a
n el
ectro
lysi
s ce
ll. A
Pt m
esh
(80
mes
h, 6
mm
× 7
mm
) was
use
d as
the
wor
king
ele
ctro
de. A
Pt w
ire a
nd A
n A
g|A
gCl|s
at.K
Cl (
ALS
) wer
e us
ed a
s th
e au
xilia
ry e
lect
rode
and
the
refe
renc
e el
ectro
de, r
espe
ctiv
ely.
The
se e
lect
rode
s w
ere
fixed
with
a T
eflo
n ca
p on
the
top
of th
e ce
ll. E
lect
roly
te s
olut
ion
was
0.0
67 M
phos
phat
e bu
ffer
of p
H 7
.0 c
onta
inin
g 0.
67 M
KC
l, an
d th
e to
tal v
olum
e w
as 3
00 μ
L. T
he c
once
ntra
tion
of 3
in th
e el
ectro
lyte
sol
utio
n w
as 0
.33
mg
mL-1
. The
dis
solv
ed o
xyge
n ga
s in
ele
ctro
lyte
sol
utio
n w
as re
mov
ed w
ith A
r gas
for 5
min
prio
r to
the
spec
troel
ectro
chem
ical
mea
sure
men
t
to m
inim
ize
oxyg
en le
vel.
A b
lank
et o
f A
r ga
s w
as m
aint
aine
d ov
er th
e el
ectro
lyte
sol
utio
n du
ring
the
entir
e el
ectro
chem
ical
mea
sure
men
ts.
Arg
on g
as w
as m
aint
aine
d ov
er th
e el
ectro
lyte
sol
utio
n du
ring
elec
troly
sis.
Elec
troly
sis
was
car
ried
out o
n a
pote
ntio
stat
(CH
I 660
) the
rmos
tate
d
at 1
5 ˚C
. Spe
ctru
m c
hang
e of
3 a
t eac
h po
tent
ial (
-0.6
0, -0
.50,
-0.4
5, -0
.40,
-0.3
5, -0
.30,
-0.2
5, -0
.20
and
-0.1
5 V
) was
sim
ulta
neou
sly
mon
itore
d
on a
CA
RY
100
spec
troph
otom
eter
. Th
e po
tent
ial d
epen
denc
e of
the
spec
tral c
hang
e w
as a
naly
zed
at 4
50 n
m a
ccor
ding
to fo
llow
ing
equa
tion.
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
5
−−
+=
O
R'
)(
)(
lnA
EA
EA
AnFRT
EE
equa
tion
(1)
whe
re n
, E, E
º’ ar
e th
e nu
mbe
r of e
lect
rons
, ele
ctro
de p
oten
tial,
and
the
redo
x po
tent
ial o
f 3, r
espe
ctiv
ely.
AR a
nd A
O a
re th
e ab
sorb
ance
at E
= –
0.6
V a
nd –
0.15
V, w
here
3 w
ould
be
fully
redu
ced
and
oxid
ized
, res
pect
ivel
y.
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
6 Pr
oton
NM
R sp
ectr
a of
1
Cl
Cl
NH2
NH2
H2O
DM
SO
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
7 abundance
01.02.03.04.05.06.07.08.09.010.011.012.013.0
X :
parts
per
Mill
ion
: Pro
ton
12.0
11.0
10.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
2.0
1.0
0-1
.0-2
.0
8.478
7.529
6.865
5.335
1.464
DM
SO d
6
9.24
2.00
1.00
0.97
0.92
Cl
Cl
NH
NH2
OO
1a
H2O
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
8
abundance00.010.020.030.040.050.060.070.080.090.1
X :
parts
per
Mill
ion
: Car
bon1
3
220.
021
0.0
200.
019
0.0
180.
017
0.0
160.
015
0.0
140.
013
0.0
120.
011
0.0
100.
090
.080
.070
.060
.050
.040
.030
.020
.010
.00
-10.
0-2
0.0
153.229
140.853
125.741 124.044
116.302 115.539
79.479
66.360
28.079
AcO
EtA
cOEt
AcO
Et
Cl
Cl
NH
NH2
OO
1a
DM
SO
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
9
abundance01.02.03.04.05.06.0
X :
parts
per
Mill
ion
: Pro
ton
12.0
11.0
10.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
2.0
1.0
0-1
.0-2
.0
8.636
7.459
6.804
5.365 4.865 4.851 4.779 4.765 4.725 4.713 4.474 4.461 4.447 3.837 3.824 3.655 3.592 3.524 3.449 3.330 3.284 3.091 3.026
9.47
2.74
2.58 2.15
1.04
0.94
0.93
0.90
0.85
0.84
0.83
0.81
Cl
Cl
NH
NH
OH
OH
OH
OH O
O1b
DM
SO
H2O
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
10
(thousandths)01.02.03.04.05.0
X :
parts
per
Mill
ion
: Car
bon1
3
220.
021
0.0
200.
019
0.0
180.
017
0.0
160.
015
0.0
140.
013
0.0
120.
011
0.0
100.
090
.080
.070
.060
.050
.040
.030
.020
.010
.00
-10.
0-2
0.0
159.791 155.873 151.534
140.160 137.185 134.487 134.096 132.179 128.547 120.070
73.875 73.132 69.070 63.769
48.189
Cl
Cl
NH
NH
OH
OH
OH
OH O
O1b
DM
SO
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
11
EtO
D
EtO
D
EtO
D
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
12
EtO
D
EtO
D
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
13
abundance01.02.03.0
X :
parts
per
Mill
ion
: Pro
ton
11.0
10.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
2.0
1.0
0-1
.0-2
.0-3
.0
10.668
7.548 7.487
4.257 4.024 3.919 3.846 3.812 3.605 3.332 2.750
1.6173.07
2.57
2.13
1.94
1.02
1.00
0.95
0.91
Cl
Cl
N N
N
NH
O
O
OH
OH
OH
OH
3
DM
SO
DM
SO
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012
14
(thousandths)01.02.03.04.05.0
X :
parts
per
Mill
ion
: Car
bon1
3
220.
021
0.0
200.
019
0.0
180.
017
0.0
160.
015
0.0
140.
013
0.0
120.
011
0.0
100.
090
.080
.070
.060
.050
.040
.030
.020
.010
.00
-10.
0-2
0.0
159.791 155.873 151.534
140.160 137.185 134.487 134.096 132.179 128.547 120.070
73.875 73.132 69.070 63.769
48.189
Cl
Cl
N N
N
NH
O
O
OH
OH
OH
OH
3
Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012