TY - JOUR
T1 - Combined LC/MS-platform for analysis of all major stratum corneum lipids, and the profiling of skin substitutes
AU - Van Smeden, Jeroen
AU - Boiten, Walter A.
AU - Hankemeier, Thomas
AU - Rissmann, Robert
AU - Bouwstra, Joke A.
AU - Vreeken, Rob J.
N1 - Funding Information:
This research is supported by the Dutch Technology Foundation STW , which is part of the Netherlands Organisation for Scientific Research (NWO) , and which is partly funded by the Ministry of Economic Affairs . The authors thank STW partners Astellas Pharma Inc. and Unilever N.V. for (co)financing this project (No. 10064) as well as Evonik (Essen) for the provision of the synthetic CERs. The project was supported by the Netherlands Metabolomics Centre (NMC) which is part of the Netherlands Genomics Initiative/Netherlands Organisation for Scientific Research and the European Cooperation in Science and Technology (COST) . Finally, the authors would like to thank Hannah Scott and Edward Kaye for their technical assistance, and Varsha Thakoersing for her involvement regarding the HSEs.
PY - 2014
Y1 - 2014
N2 - Ceramides (CERs), cholesterol, and free fatty acids (FFAs) are the main lipid classes in human stratum corneum (SC, outermost skin layer), but no studies report on the detailed analysis of these classes in a single platform. The primary aims of this study were to 1) develop an LC/MS method for (semi-)quantitative analysis of all main lipid classes present in human SC; and 2) use this method to study in detail the lipid profiles of human skin substitutes and compare them to human SC lipids. By applying two injections of 10 μl, the developed method detects all major SC lipids using RPLC and negative ion mode APCI-MS for detection of FFAs, and NPLC using positive ion mode APCI-MS to analyze CERs and cholesterol. Validation showed this lipid platform to be robust, reproducible, sensitive, and fast. The method was successfully applied on ex vivo human SC, human SC obtained from tape strips and human skin substitutes (porcine SC and human skin equivalents). In conjunction with FFA profiles, clear differences in CER profiles were observed between these different SC sources. Human skin equivalents more closely mimic the lipid composition of human stratum corneum than porcine skin does, although noticeable differences are still present. These differences gave biologically relevant information on some of the enzymes that are probably involved in SC lipid processing. For future research, this provides an excellent method for (semi-)quantitative, 'high-throughput' profiling of SC lipids and can be used to advance the understanding of skin lipids and the biological processes involved.
AB - Ceramides (CERs), cholesterol, and free fatty acids (FFAs) are the main lipid classes in human stratum corneum (SC, outermost skin layer), but no studies report on the detailed analysis of these classes in a single platform. The primary aims of this study were to 1) develop an LC/MS method for (semi-)quantitative analysis of all main lipid classes present in human SC; and 2) use this method to study in detail the lipid profiles of human skin substitutes and compare them to human SC lipids. By applying two injections of 10 μl, the developed method detects all major SC lipids using RPLC and negative ion mode APCI-MS for detection of FFAs, and NPLC using positive ion mode APCI-MS to analyze CERs and cholesterol. Validation showed this lipid platform to be robust, reproducible, sensitive, and fast. The method was successfully applied on ex vivo human SC, human SC obtained from tape strips and human skin substitutes (porcine SC and human skin equivalents). In conjunction with FFA profiles, clear differences in CER profiles were observed between these different SC sources. Human skin equivalents more closely mimic the lipid composition of human stratum corneum than porcine skin does, although noticeable differences are still present. These differences gave biologically relevant information on some of the enzymes that are probably involved in SC lipid processing. For future research, this provides an excellent method for (semi-)quantitative, 'high-throughput' profiling of SC lipids and can be used to advance the understanding of skin lipids and the biological processes involved.
KW - Ceramide
KW - Free fatty acid
KW - Human skin equivalent
KW - LC/MS
KW - Porcine skin
KW - Stratum corneum lipid
UR - http://www.scopus.com/inward/record.url?scp=84885811767&partnerID=8YFLogxK
U2 - 10.1016/j.bbalip.2013.10.002
DO - 10.1016/j.bbalip.2013.10.002
M3 - Article
AN - SCOPUS:84885811767
SN - 1388-1981
VL - 1841
SP - 70
EP - 79
JO - Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids
JF - Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids
IS - 1
ER -