5. Glass test tubes.
6. Filter paper discs 110 cm diameter.
7. HPLC-grade solvents: Methanol, chloroform, and diethyl
ether (see Note 2).
8. Rotary vacuum evaporator (see Note 3).
9. Air or nitrogen stream.
2.2 Characterization
of Free Lipids by TLC
1. Normal phase silica gel G60, 20 cm  20 cm, 0.25-mm plates.
2. Chromatography tank with saturation paper for TLC
migration.
3. HPLC-grade solvents: Petroleum ether, ether, chloroform, and
methanol (see Note 2).
4. Acetic acid.
5. Spray flasks.
6. Pasteur pipettes or glass syringe.
7. Dittmer and Lester reagent [48] (commercially available for
phospholipids detection).
8. 0.2% anthrone in H 2 SO 4 (for glycolipids detection).
9. 0.1% rhodamine B in ethanol diluted to 0.01% in 0.25 M
NaH 2 PO 4 (general reagent for lipid detection).
10. 10% molybdophosphoric acid in ethanol (general reagent, after
carbonization, for lipid detection).
11. 0.2% ninhydrine in acetone (lipid with free amino group) light
charring.
12. Heater for charring the plates.
13. UV light lamp CAMAG.
2.3 Preparation of FA
Methyl Esters (FAME)
and Mycolic Acid
Methyl Esters (MAME)
1. Glass screw-capped tubes 18 Â 1.5 cm.
2. 50 mL screw-capped glass flask.
3. Pasteur pipettes.
4. Glass test tubes (18 cm  1.5 cm) for transfer ether phases of
extraction.
5. 40% KOH in water and methoxy-ethanol (1:7 v/v).
6. TMS diazomethane (Aldrich).
7. 20% H 2 SO 4 in distilled water or Milli-Ro or Milli-Q grade.
8. pH paper.
9. Distilled water, milli-Ro or Milli-Q grade.
10. HPLC-grade solvents: Methoxy-ethanol, diethyl ether, and
dichloromethane (see Note 2).
11. Air or nitrogen stream.
12. Oven at 110
C.
Lipid and Lipoarabinomannan
117
6. Filter paper discs 110 cm diameter.
7. HPLC-grade solvents: Methanol, chloroform, and diethyl
ether (see Note 2).
8. Rotary vacuum evaporator (see Note 3).
9. Air or nitrogen stream.
2.2 Characterization
of Free Lipids by TLC
1. Normal phase silica gel G60, 20 cm  20 cm, 0.25-mm plates.
2. Chromatography tank with saturation paper for TLC
migration.
3. HPLC-grade solvents: Petroleum ether, ether, chloroform, and
methanol (see Note 2).
4. Acetic acid.
5. Spray flasks.
6. Pasteur pipettes or glass syringe.
7. Dittmer and Lester reagent [48] (commercially available for
phospholipids detection).
8. 0.2% anthrone in H 2 SO 4 (for glycolipids detection).
9. 0.1% rhodamine B in ethanol diluted to 0.01% in 0.25 M
NaH 2 PO 4 (general reagent for lipid detection).
10. 10% molybdophosphoric acid in ethanol (general reagent, after
carbonization, for lipid detection).
11. 0.2% ninhydrine in acetone (lipid with free amino group) light
charring.
12. Heater for charring the plates.
13. UV light lamp CAMAG.
2.3 Preparation of FA
Methyl Esters (FAME)
and Mycolic Acid
Methyl Esters (MAME)
1. Glass screw-capped tubes 18 Â 1.5 cm.
2. 50 mL screw-capped glass flask.
3. Pasteur pipettes.
4. Glass test tubes (18 cm  1.5 cm) for transfer ether phases of
extraction.
5. 40% KOH in water and methoxy-ethanol (1:7 v/v).
6. TMS diazomethane (Aldrich).
7. 20% H 2 SO 4 in distilled water or Milli-Ro or Milli-Q grade.
8. pH paper.
9. Distilled water, milli-Ro or Milli-Q grade.
10. HPLC-grade solvents: Methoxy-ethanol, diethyl ether, and
dichloromethane (see Note 2).
11. Air or nitrogen stream.
12. Oven at 110
C.
Lipid and Lipoarabinomannan
117