engleski

HPLC methof for determination of natural moisturizing factor components in the stratum corneum

The levels of natural moisturizing factor (NMF) in the skin can be used as a biomarker of hydration, for studying the effect of skin irritants, or as a biomarker of loss-of-function mutations in the filaggrin gene which are the main risk factor for atopic dermatitis. NMF is a complex mixture of hygroscopic compounds in the uppermost layer of the skin, the stratum corneum (SC) and amino acids and their derivatives contribute more than 50 percent to total content of NMF. Recently, a high-performance liquid chromatography (HPLC) method for the direct determination of four most relevant components of NMF in the SC was proposed based on the extraction with ammonia and subsequent determination by HPLC with mobile phase (MF) containing hydrochloric acid. The main disadvantage of this method is a need to evaporate ammonia to dryness and reconstitute dry content in water, which is time consuming. Another disadvantage is use of hydrochloric acid to sufficiently lower pH of the MF which contribute to corrosion of the chromatographic system. The aim of this study was to explore water as possible substitute for ammonia extraction as well as use of phosphoric acid as substitute of hydrochloric acid. Chromatographic performance, recovery, and precision parameters of the water method was investigated and compared with the extraction using ammonia. In summary, the modified analytical method was shown to be feasible and reliable for the determination of NMF components in the skin.

natural moisturizing factor, extraction, liquid chromatography, method validatio

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