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*Address all correspondence to Sevgi Güngör, sgungor@istanbul.edu.tr
Development and Validation of a Simple and Selective
Chromatographic Method for Quantification of Ceramide-NP
in Skin-Simulating Liposome Formulations
HÜMEYRA ŞAHİN BEKTAY, EMİNE KAHRAMAN AND SEVGİ GÜNGÖR
Department of Pharmaceutical Technology, Istanbul University, Istanbul, Türkiye (H.S.B, E.K, S.G.)
Health Science Institute, Istanbul University, Istanbul, Türkiye (H.S.B)
Department of Pharmaceutical Technology, Bezmialem Vakıf University, Istanbul, Türkiye (H.S.B)
Accepted for publication September 19, 2023.
Synopsis
Numerous topical products containing ceramide to restore the skin barrier function have been available on the
market in recent years. A simple, rapid, and selective analytical method could contribute to the development
of innovative formulations such as skin-simulating liposomes, which contain ceramides as stratum corneum
components. The similarity of the physicochemical structure of ceramide and lipid excipients in liposome
formulation complicates the separative and quantitative analysis. Therefore, this study aims to develop and
validate a selective high-pressure liquid chromatography (HPLC) method for the simultaneous quantification
of lipids in ceramide-containing skin simulating liposome formulations. The separation was performed with
a reversed-phase C18 column (150 mm x 4.6 mm, 5 µm), the mobile phase system consisted of methanol:
acetonitrile (60:40, v/v), the flow rate of 0.5 ml/min, the oven temperature at 45°C, injection volume of 10 µL,
and UV detection at 210 nm. The method was linear (r2 0.99) in the range of concentration from 80 µg/mL
to 480 µg/mL with acceptable precision, accuracy, and selectivity for the liposome formulations. The limit of
detection (LOD) values of ceramide NP (Cer-NP) was 7.90 µg/mL. The limit of quantification (LOQ) values
of Cer-NP was 24.06 µg/mL. The study indicates that the method is efficient and has accuracy and selectivity
for the simultaneous quantification of Cer-NP in skin-simulating liposome formulations.
INTRODUCTION
The formulations consisting of skin components such as ceramides, cholesterol, and other
natural lipids have recently attracted attention as lipid replacement therapy in atopic
dermatitis.1 Numerous products that contain skin lipids such as ceramide, fatty acids, and
cholesterol have been available on the market to restore the skin barrier.2 The pharmaceutical
and cosmeceutical industries have mostly preferred the production of liposome formulations
for skin barrier recovery.3 Hence, a reliable analytical method is still a requirement for the
quantification of ceramides and lipid components in formulation development studies.