Optimizing Lipid Extraction Protocols for Comprehensive Lipidomic Analysis of the Skin
Author
Guled, Amaal Mohamed Hassan
Term
4. term
Publication year
2023
Pages
35
Abstract
This thesis evaluates which solvent most effectively extracts skin lipids for comprehensive lipidomic analysis. Skin lipids are central to barrier function, and efficient extraction underpins reliable mass spectrometric profiling. Samples were collected from a single healthy 24-year-old female volunteer using tape stripping on the proximal forearm and a control sample from the nasal alar crease obtained by scalpel. The solvents tested were chloroform, methanol, a 2:1 chloroform:methanol mixture, and 2-propanol with ammonium acetate; analyses were carried out by MALDI-TOF MS with ceramide standards used for calibration and identification. Tape discs produced a measurable non-zero mass spectral signal even without clearly identifiable ions, while ceramide standards displayed expected sodium adducts. Solvent optimization indicated that the 2:1 chloroform:methanol mixture was the most effective extraction condition among those tested, whereas methanol alone was ineffective; care is advised when applying solvents to tape discs. These findings underscore the importance of solvent selection for skin lipid extraction and show that combining tape stripping with MALDI-TOF MS can support robust lipid profiling in dermatological research, based on samples from one healthy subject.
Denne afhandling undersøger, hvilket opløsningsmiddel der bedst udvinder hudlipider til omfattende lipidomiske analyser. Hudlipider er centrale for hudens barrierefunktion, og effektiv udvinding er afgørende for præcis massespektrometrisk profilering. Hudprøver blev indsamlet fra én sund, 24-årig kvindelig deltager via tape-stripping på den proksimale underarm og en kontrolprøve fra næsens alarfure med skalpel. Følgende opløsningsmidler blev testet: kloroform, metanol, en 2:1-blanding af kloroform:metanol samt 2-propanol med ammoniumacetat; analyser blev udført med MALDI-TOF MS, og ceramidstandarder blev anvendt til kalibrering og identifikation. Tape-strips gav et målbart, ikke-nul signal i massespektret selv uden tydeligt identificerbare ioner, mens ceramidstandarderne viste forventede natriumaddukter. Solventoptimeringen pegede på 2:1 kloroform:metanol som den mest effektive ekstraktionsbetingelse blandt de testede, mens metanol alene var ineffektiv; der bør udvises forsigtighed ved påføring af opløsningsmidler på tape-skiver. Resultaterne fremhæver, at valg af opløsningsmiddel er afgørende for hudlipidudvinding, og at kombinationen af tape-stripping og MALDI-TOF MS kan understøtte mere robuste lipidprofiler i dermatologisk forskning, baseret på prøver fra én sund forsøgsperson.
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