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Carolina Kunnen, Simon Brown, Brien Holden, Percy Lazon De La Jara, Eric Papas; Effect of Collection Method on the Lipid Composition of Meibum Samples. Invest. Ophthalmol. Vis. Sci. 2013;54(15):6011. doi: https://doi.org/.
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To quantify the effect of various collection techniques on the lipid content of meibum.
Meibum secretion was collected from five non-contact lens wearers aged between 20 and 35 years. The following four meibum expression methods were used: cotton buds without cleaning the eyelid margin (CBn), cotton buds with cleaning the eyelid margin (CB), the Korb meibomian gland evaluator (Korb) and Meibomian Gland Forceps (MGF). Cleaning of the eyelid margin was performed prior to the last three procedures to reduce contamination with lid margin epithelial cells and tears. The order of technique was randomised and all samples were collected at the same time of day and by the same investigator. Lipids were analyzed by chip-based nano-electrospray mass spectrometry (ESI-MS). Using synthesized internal standards, tandem MS allowed us to identify and quantify over 200 lipid species across seven lipid classes. Individual lipid species were grouped as follows; non polar group: cholesteryl ester (CE), free cholesterol (FC), wax ester (WE) and triacylglyceride (TAG); polar group: (O-acyl)-omega-hydroxy fatty acid (OAHFA), phospholiplids (PL) and lysophospholipids (LPC). Comparison between collection methods for each class was made using repeated measures ANOVA.
Due to volume of data, presentation is simplified to show the rank order of lipids returned by each of the various techniques (Table 1). For all classes, except FC there were significant differences between collection techniques (p <.05). While absolute differences were small, overall for polar species, the Korb method returned smaller quantities than the CBn approach. For non polar species differences were more inconsistent.
Collection technique influences the relative amounts of lipid species in samples of meibum. Greater consistency across polar species appears to be offered by the Korb approach.
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