In this study, lipid structural change is monitored using Raman spectroscopy during heat treatment, along with the impact of lipid states on the structural and physical properties during the preparation process of the dried and hydrated lipid mixture (LM) similar to intercellular lipid in stratum corneum. The microstructures and thermal behavior of these LMs change depending on the melting of lipid ingredients in the preparation process. It is recognized that variable temperature Raman spectroscopy (VT‐Raman) is a useful and attractive tool for the sensitive in situ monitoring of lipid state changes and lipid melting. The LMs can incorporate D2O into their structures regardless of preparation temperature due to increasing lattice distance by hydration. These results suggest that monitoring lipid structural changes during the heating step is important to precisely prepare target LMs. Practical Applications: This study reveals that VT‐Raman is a useful and attractive tool in in situ monitoring of lipid state change and lipid melting. The monitoring of the preparation process by VT‐Raman is necessary to precisely prepare the target LM similar to intercellular lipid of stratum corneum because the microstructures and thermal properties of these LMs change depending on the melting of lipid ingredients during the preparation process.

Vibrational spectroscopic imaging has become useful analytical tools for quality control of drug products. In this study, we applied microscopic attenuated total reflection (ATR)-IR and confocal Raman microscopy to elucidate microscopic structure of creams and for the formulation design in the development of semi-solid drug products. The model creams were prepared with prednisolone (PRD) and fluconazole (FLC) as active pharmaceutical ingredients and oily solvents such as mineral oil (MO), isopropyl myristate (IPM), benzyl alcohol (BA) and diethyl sebacate (DES). As a result of microscopic ATR-IR imaging, several domains indicating oily internal phase were observed, which had absorption around 1732 and 1734 cm 1 derived from MO, IPM and DES. In addition, domains of BA around 1009 cm 1 were observed at the complemental or similar position in the formulation with MO or DES, respectively. These results suggested that the creams were oil-in-water type and the distribution of domains would reflect the compatibility of the solvents. The contents of PRD and BA were determined quantitatively in each layer after the intentional separation of the creams and the results agreed well with the imaging analysis. Whereas, confocal Raman imaging allowed to visualize the distribution of the components in depth direction as well as two-dimensional plane. In particular, the Raman imaging would ensure the coexistence of FLC and BA as oily phase in the cream. From these results, the feasibility of spectroscopic imaging techniques was successfully demonstrated for the formulation design of semi-solid dosage forms.