A.M. Lefsay et al. / Chemistry and Physics of Lipids 174 (2013) 55–63
57
+
chromatography on silica gel (ethyl acetate-hexane, 1:4, v/v). The
ESI(+)MS: m/z 297.4 ([M+Na] ); MS/MS (28% CID) of 297.4: m/z
115.1.
1
separated product was a colorless oil (2.6 g, 83%). H NMR (CD Cl )
2
2
ı: 5.25–5.21 (1 H, m), 4.26 (2H, dd, J = 11.9 and 4.3), 4.13 (2H, dd,
J = 11.9 and 6.1), 4.08–4.04 (2H, m), 2.30 (2H, t, J = 7.5), 2.29 (4H,
t, J = 7.6), 1.93–1.85 (2H, m), 1.84–1.76 (2H, m), 1.63–1.56 (8H, m),
2.4.3. Coupling of mono-acyl glycerol with two equivalent fatty
acids
The coupling procedure described in Section 2.4.1 and a 33-h
reaction time were used.
1
.33–1.26 (50H, m), 0.89 (3H, t, J = 7), 0.88 (6H, t, J = 7); 13C NMR
(
CD Cl ) ı: 173.6 (2C), 173.3, 69.3, 66.4, 66.3, 62.6 (2C), 35.4, 35.3,
2 2
3
2
2
4.7, 34.5 (2C), 32.5 (2C), 32.4, 31.1, 30.2 (4C), 30.0 (2C), 29.9 (2C),
9.84 (2C), 29.76, 29.64 (2C), 29.61 (2C), 29.5, 29.4, 27.22, 27.17,
1-(9,10-Dichlorooctadecanoyl)-2,3-didodecanoylglycerol
(13):
coupling of 1-(9,10-dichlorooctadecanoyl)glycerol (11; 1.5 g,
3.5 mmol) and dodecanoic acid (7; 1.4 g, 7.0 mmol) yielded 13 as a
5.4 (2C), 25.3, 23.25 (2C), 23.21, 14.4 (3C); ESI(+)MS (relative inten-
+
◦
1
sity): m/z 813.6 (100, [M+Na] ); MS/MS (30% CID) of m/z 813.6: m/z
waxy solid (2.4 g, 84%). mp 30–33 C; H NMR (CDCl ) ı: 5.28–5.24
3
7
7
8
77.4 (15), 741.4 (100), 613.2 (5), 595.2 (5); MS/MS (40% CID) of m/z
41.4: m/z 541.1 (100), 519.3 (50), 461.1 (20), 439.3 (14); HRMS:
13.5509 (813.5537 calculated for C45H84Cl O Na).
(1H, m), 4.29 (2H, dd, J = 11.9 and 4.3), 4.14 (2H, dd, J = 11.9 and
6.0), 4.04–4.01 (2H, m), 2.31 (6H, td, J = 7.5 and 3.2), 1.93–1.86 (2H,
m), 1.81–1.73 (2H, m), 1.63–1.56 (8H, m), 1.32–1.24 (50H, m), 0.88
2
6
(
9H, t, J = 7); 13C NMR (CDCl ) ı: 173.4, 173.3, 173.0, 69.0, 65.7,
3
2
2
.4. Chlorinated unsymmetrical triacylglycerols (13 and 14)
65.6, 62.2 (2C), 34.53, 34.49, 34.3, 34.2, 34.1, 32.05 (2C), 31.96,
9.84, 29.75 (4C), 29.63, 29.61, 29.52, 29.48 (2C), 29.42, 29.41,
29.34, 29.25, 29.21, 29.17, 29.09, 29.0, 26.9, 26.8, 25.04, 24.99,
2
.4.1. Monoacylisopropylidenes
A solution of 1,2-isopropylideneglycerol (8), fatty acid (1 equiv.
of 2 or 7), EDCI (1 equiv.) and DMAP (0.5 equiv.) in dichloromethane
20 mL) was stirred at room temperature for 24 h. The solvent was
removed by rotary evaporation and crude product was purified
using flash chromatography on silica gel eluted using ethyl acetate-
hexane (2:1, v/v).
24.9, 22.82 (2C), 22.79, 14.3 (3C); ESI(+)MS (relative intensity): m/z
813.5 (100, [M+Na] ); MS/MS (36% CID) of m/z 813.5: m/z 777.5
+
(
(20), 741.5 (100), 613.2 (5); MS/MS (33% CID) of m/z 741.5: m/z
541.3 (100), 519.3 (50), 461.2 (40), 439.1 (25); HRMS: 813.5560
(813.5537 calculated for C45H84Cl O Na).
2
6
1,2-Di-(9,10-dichlorooctadecanoyl)-3-dodecanoylglycerol (14):
coupling of 1-dodecanoylglycerol (12; 1.0 g, 3.6 mmol) with 9,10-
dichlorooctadecanoic acid (2.5 g, 7.2 mmol) yielded 14 as an oil
1,2-Isopropylidene-3-(9,10-dichlorooctadecanoyl)glycerol
(9):
coupling of 9,10-dichlorooctadecanoic acid (2; 2.6 g, 7.4 mmol)
and 8 (0.97 g, 7.3 mmol) yielded 9 as an oil (2.9 g 85%). 1H NMR
(2.5 g, 73%). 1H NMR (CDCl ) ı: 5.26–5.21 (1H, m), 4.29–4.25 (2H,
3
(
CD Cl ) ı: 4.30–4.25 (1H, m), 4.14–4.02 (5H, m), 3.72 (1H, dd,
m), 4.11 (2H, dd, J = 11.9 and 6.0), 4.02–3.98 (4H, m), 2.30–2.25
2
2
J = 8.3 and 6.1), 2.35–2.30 (2H, m), 1.98–1.90 (2H, m), 1.85–1.75
(6H, m), 1.92–1.84 (4H, m), 1.79–1.70 (4H, m), 1.61–1.49 (10H,
(
2H, m), 1.64–1.52 (4H, m), 1.40 (3H, s), 1.34 (3H, s), 1.34–1.28
m), 1.31–1.22 (52H, m), 0.86 (3H, t, J = 7), 0.85 (6H, t, J = 7); 13
C
(
18H, m), 0.90 (3H, t, J = 7); 13C NMR (CD Cl ) ı: 173.9, 110.3, 74.4,
NMR (CDCl ) ı: 173.3, 173.2, 172.8, 69.0, 65.7 (2C), 65.6 (2C),
2
2
3
6
7.1, 66.49, 66.45, 65.1, 35.52, 35.49, 34.7, 32.5, 30.1, 29.9, 29.7
62.17, 62.14, 34.5 (4C), 34.2, 34.1, 34.0, 32.0, 31.9 (2C), 29.8, 29.75,
29.71 (2C), 29.6, 29.5, 29.43, 29.36, 29.30, 29.2, 29.1 (4C), 29.04,
29.01, 28.93, 28.91, 26.81 (2C), 26.76 (2C), 24.94, 24.88, 24.85, 22.8,
22.7 (2C), 14.2 (3C); ESI(+)MS (relative intensity): m/z 965. 7 (71,
(
(
2C), 29.6, 29.5, 27.35, 27.30, 27.2, 25.9, 25.5, 23.3, 14.5; ESI(+)MS
relative intensity): m/z 489.3 (100, [M+Na] ): MS/MS (35% CID)
+
of 489: m/z 452.9 (16), 416.9 (100); HRMS: 489.2498 (489.2509
calculated for C24H44Cl O Na).
+
[M+Na] ), 966.8 (44), 967.7 (100); MS/MS (30% CID) of m/z 967.7:
2
4
rac-1,2-Isopropylidene-3-dodecanoylglycerol (10): coupling of
m/z 931.4 (15), 895.5 (100), 893.5 (76), 821.6 (3); MS/MS (30% CID)
of m/z 893.5: m/z 857.5 (26), 821.5 (100); MS/MS (36% CID) of m/z
821.5: m/z 621.3 (64), 599.2 (21), 541.1 (100), 519.3 (35); HRMS:
965.5726 (965.5697 calculated for C51H94Cl O Na).
dodecanoic acid (7; 2.8 g, 14 mmol) and 8 (1.9 g, 14 mmol) yielded
◦
1
0 as a white crystalline solid (3.8 g, 86%). mp < 20 C (lit (Batovska
◦
1
13
et al., 2004) mp 32.5 C); H and C NMR spectra were consistent
4
6
with published data (Batovska et al., 2004); ESI(+)MS: m/z 337.3
+
(
[M+Na] ); MS/MS (35% CID) of 337.3: m/z 279.1.
2.5. Hexahalogenated triacylglycerols
2.4.2. Mono-acyl glycerols
2.5.1. Tri-(9,10-dichlorooctadecanoyl)glycerol (16)
Purified 1,2-isopropylidene-3-acylglycerol (9 or 10) was mixed
Reaction of tri-(cis-9-octadecenoyl)glycerol (15, glyceryl tri-
oleate; 0.52 g, 0.59 mmol) with sulfuryl chloride (3 mL, 37 mmol)
as described in Section 2.2 yielded hexachlorinated triacylglycerol
(16) as a colorless oil (0.54 g, 84%). 1H NMR (CD Cl ) ı: 5.25–5.21
+
with ion-exchange resin C-244 (2 g, H form) in methanol (20 mL)
and dichloromethane (2 mL). The mixture was stirred at room tem-
perature and monitored using TLC. When the starting material was
consumed (4 h) the resin was filtered. The filtrate was dried (anhy-
drous sodium sulfate) and removed by rotary evaporation.
2
2
(m, 1H), 4.26 (2H, dd, J = 12.0 and 4.3), 4.13 (2H, dd, J = 11.8 and 6.0),
4.08–4.05 (6H, m), 2.30 (6H, t, J = 7.5), 1.92–1.86 (6H, m), 1.84–1.76
(6H, m), 1.63–1.50 (12H, m), 1.42–1.26 (54H, m), 0.88 (9H, t, J = 7.0);
1
-(9,10-Dichlorooctadecanoyl)glycerol (11): hydrolysis of 1,2-
isopropylidene-3-(9,10-dichlorooctadecanoyl)glycerol (1.62 g,
.5 mmol) yielded 11 as a colorless oil (1.35 g, 92%). H NMR
CD Cl ) ı: 4.18–4.02 (4H, m), 3.91–3.87 (1H, m), 3.68–3.55 (2H,
13
C NMR (CD Cl ) ı: 173.6 (2C), 173.2, 69.4, 66.41 (3C), 66.37 (3C),
2
2
1
3
(
62.6 (2C), 35.4 (6C), 34.7, 34.5 (2C), 32.4 (3C), 30.0 (3C), 29.8 (3C),
29.6 (6C), 29.5 (3C), 29.4 (3C), 27.23 (3C), 27.19 (3C), 25.3 (3C),
23.2 (3C), 14.4 (3C); ESI(+)MS (relative intensity): m/z 1117.5 (60,
2
2
m), 2.34 (2H, t, J = 7.4), 1.99–1.88 (2H, m), 1.85–1.75 (2H, m),
.65–1.51 (4H, m), 1.45–1.28 (18H, m), 0.90 (3H, m); 13C NMR
CD Cl ) ı: 174.6, 71.0, 67.1, 66.5, 65.9, 64.1, 35.5, 35.3, 34.7,
+
1
(
3
[M+Na] ); MS/MS (35% CID) of m/z 1119.5: m/z 1083.3 (24), 1047.4
(100), 1045.6 (40), 1009.6 (23), 973.5 (42); MS/MS (33% CID) of m/z
1047.5: m/z 1011.5 (28), 975.5 (100), 973.7 (69), 901.6 (25); MS/MS
(33% CID) of m/z 973.5: m/z 937.3 (40), 901.5 (100); MS/MS (38%
CID) of m/z 901.6: m/z 621.6 (100), 599.5 (38); HRMS: 1117.5812
2
2
2.5, 30.3, 30.0, 29.8, 29.7, 29.6, 29.5, 27.3, 26.9, 25.5, 23.3, 14.5;
+
ESI(+)MS (relative intensity): m/z 449.2 (100, [M+Na] ); MS/MS
(
(
25% CID) of 449.2: m/z 412.9 (20), 376.9 (100); HRMS: 449.2206
449.2196 calculated for C21H40Cl O Na).
(1117.5856 calculated for C57H104Cl O Na).
2
4
6
6
rac-1-Dodecanoylglycerol
(12):
hydrolysis
of
1,2-
isopropylidene-3-dodecanoylglycerol (1.5 g, 4.8 mmol) yielded 12
2.5.2. Tri-(9,10-dibromooctadecanoyl)glycerol (17)
◦
as white crystals (1.25 g, 93%). mp 60–61 C (lit (Rytczak et al.,
Bromine (1.5 mL, 29 mmol) was added over 20 min to a stirred
solution of tri-(cis-9-octadecenoyl)glycerol (15; 0.20 g, 0.23 mmol)
◦
1
13
2
010) mp 62–63 C); H and C NMR spectra were consistent
◦
with published data (Batovska et al., 2004; Rytczak et al., 2010);
in dichloromethane (5 mL) at −10 C. After 1 h and an additional