l
For apolar lipids, i.e., triacylglycerol (TAG) and phthiocerol
dimycocerosate (PDIM): petroleum ether/diethyl ether 9:1
(v/v) [32].
l
For polyphthienoyl trehalose (PPT) present in
M. tuberculosis: solvent, chloroform/methanol 99:1
(v/v) [47].
l
For phenolic glycolipids (PGL): chloroform/methanol 98:2
or 95/5 (v/v), depending on the nature of saccharide moiety linked to the lipid core. PGL are present in some strains
of the M. tuberculosis complex (Beijing, Canetti) but absent
in H 37 Rv strain [63].
l
For glycerol monomycolate (GroMM): chloroform/methanol 9:1 (v/v) found in all mycobacteria examined so
far [64].
l
For trehalose mycolates found in all mycobacteria, i.e., trehalose dimycolate (TDM): chloroform/methanol 9:1 (v/v)
or chloroform/methanol/water 30/8/1 (v/v/v), and trehalose monomycolate (TMM): chloroform/methanol/
water 65/25/4 (v/v/v).
l
For glycopeptidolipids (GPL), characteristic species- or
type-species glycolipids produced by some species such as
M. smegmatis, M. avium: chloroform/methanol 9:1 (v/v),
chloroform/methanol/water 30/8/1 (v/v/v), or chloroform/methanol/water 65/25/4 (v/v/v).
l
For sulfoglycolipids of M. tuberculosis (SGL): the mixture
chloroform/methanol 85:15 or 80:20 (v/v) allows the separation of the three main SGL, i.e., the tetraacylated
Ac 4 SGL (previously characterized by Goren as SL1), the
triacylated (Ac 3 SGL), and the diacylated (Ac 2 SGL). In
these solvents, Ac 4 SGL co-migrate with triacyltrehalose
(TAT) and the Ac 3 SGL migrate with diacyltrehalose DAT
[29]. Alternatively, SGL1 can be detected in a crude total
lipid extract by 2-D TLC using in the first-dimension chloroform/methanol/water 60:12:1 (v/v/v), and in the second direction, chloroform/methanol/water 75:11:1 (v/v/
v) [65]. The plate has to dry at room temperature between
the two migrations.
l
For phospholipids and aminolipids, i.e., cardiolipin (CL),
phosphatidyl glycerol (PG), phosphatidylethanolamine
(PE), and the phosphatidylinositol mannosides (PIM): analyzed using the reference developing solvent chloroform/
methanol/water 65:25:4 (v/v/v). This solvent is also suitable for the characterization of aminolipids, e.g., the ornithine lipid (OL) characterized in M. canetii and M. bovis
BCG [66].
Lipid and Lipoarabinomannan
143
For apolar lipids, i.e., triacylglycerol (TAG) and phthiocerol
dimycocerosate (PDIM): petroleum ether/diethyl ether 9:1
(v/v) [32].
l
For polyphthienoyl trehalose (PPT) present in
M. tuberculosis: solvent, chloroform/methanol 99:1
(v/v) [47].
l
For phenolic glycolipids (PGL): chloroform/methanol 98:2
or 95/5 (v/v), depending on the nature of saccharide moiety linked to the lipid core. PGL are present in some strains
of the M. tuberculosis complex (Beijing, Canetti) but absent
in H 37 Rv strain [63].
l
For glycerol monomycolate (GroMM): chloroform/methanol 9:1 (v/v) found in all mycobacteria examined so
far [64].
l
For trehalose mycolates found in all mycobacteria, i.e., trehalose dimycolate (TDM): chloroform/methanol 9:1 (v/v)
or chloroform/methanol/water 30/8/1 (v/v/v), and trehalose monomycolate (TMM): chloroform/methanol/
water 65/25/4 (v/v/v).
l
For glycopeptidolipids (GPL), characteristic species- or
type-species glycolipids produced by some species such as
M. smegmatis, M. avium: chloroform/methanol 9:1 (v/v),
chloroform/methanol/water 30/8/1 (v/v/v), or chloroform/methanol/water 65/25/4 (v/v/v).
l
For sulfoglycolipids of M. tuberculosis (SGL): the mixture
chloroform/methanol 85:15 or 80:20 (v/v) allows the separation of the three main SGL, i.e., the tetraacylated
Ac 4 SGL (previously characterized by Goren as SL1), the
triacylated (Ac 3 SGL), and the diacylated (Ac 2 SGL). In
these solvents, Ac 4 SGL co-migrate with triacyltrehalose
(TAT) and the Ac 3 SGL migrate with diacyltrehalose DAT
[29]. Alternatively, SGL1 can be detected in a crude total
lipid extract by 2-D TLC using in the first-dimension chloroform/methanol/water 60:12:1 (v/v/v), and in the second direction, chloroform/methanol/water 75:11:1 (v/v/
v) [65]. The plate has to dry at room temperature between
the two migrations.
l
For phospholipids and aminolipids, i.e., cardiolipin (CL),
phosphatidyl glycerol (PG), phosphatidylethanolamine
(PE), and the phosphatidylinositol mannosides (PIM): analyzed using the reference developing solvent chloroform/
methanol/water 65:25:4 (v/v/v). This solvent is also suitable for the characterization of aminolipids, e.g., the ornithine lipid (OL) characterized in M. canetii and M. bovis
BCG [66].
Lipid and Lipoarabinomannan
143