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1.
Scheme 3

Scheme 3. From: The Meibomian Puzzle: Combining Pieces Together.

Comparison chart of common chromatographic methods of meibum lipid analyses.

Igor A. Butovich. Prog Retin Eye Res. ;28(6):483-498.
2.
Scheme 4

Scheme 4. From: The Meibomian Puzzle: Combining Pieces Together.

Comparison chart of common spectroscopic methods of meibum lipid analyses

Igor A. Butovich. Prog Retin Eye Res. ;28(6):483-498.
3.
Scheme 5

Scheme 5. From: The Meibomian Puzzle: Combining Pieces Together.

Comparison chart of stand-alone mass spectrometric methods in lipid analysis

Igor A. Butovich. Prog Retin Eye Res. ;28(6):483-498.
4.
Figure 1

Figure 1. From: The Meibomian Puzzle: Combining Pieces Together.

Human meibum collected using the eyelid squeezing method and a spatula. Note meibum solidification at room temperature

Igor A. Butovich. Prog Retin Eye Res. ;28(6):483-498.
5.
Scheme 2

Scheme 2. From: The Meibomian Puzzle: Combining Pieces Together.

Structures of representative polar and amphiphilic lipids relevant to lipidomic analysis of meibum, tear film, and aqueous tears.
Ionizable polar groups are shown in red; nonionizable polar groups – in blue; nonpolar groups – in black.

Igor A. Butovich. Prog Retin Eye Res. ;28(6):483-498.
6.
Scheme 1

Scheme 1. From: The Meibomian Puzzle: Combining Pieces Together.

Structures of representative simple and complex lipids relevant to lipidomic analysis of meibum, tear film, and aqueous tears.
Hydrolyzable bonds are marked with dashed lines. Complex lipids are color-coded to show their continuous non-hydrolizable fragments.

Igor A. Butovich. Prog Retin Eye Res. ;28(6):483-498.
7.
Figure 2

Figure 2. From: The Meibomian Puzzle: Combining Pieces Together.

Partial observation APCI-MS spectra of human meibum recorded in the positive (upper panel) and the negative (lower panel) ion modes in an NP HPLC experiment.
The HPLC-MS experiments were performed as described earlier(Butovich et al., 2007a).

Igor A. Butovich. Prog Retin Eye Res. ;28(6):483-498.
8.
Scheme 6

Scheme 6. From: The Meibomian Puzzle: Combining Pieces Together.

Proposed structure of the tear film (on the left) and the amphiphilic lipid sublayer formed of very long chain (O-acyl)-ω-hydroxy fatty acids and diacyl glycerols (on the right).
*Di- and tri-esters as defined by Nicolaides et al (Nicolaides and Santos, 1985). The locations of double bonds in the structures of the very long chain ω hydroxy fatty acids are tentative (Butovich et al., 2009b).

Igor A. Butovich. Prog Retin Eye Res. ;28(6):483-498.

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