Silk microspheres prepared by spray-drying were exposed to humid atmosphere (89% relative humidity, RH). Change in the secondary structure of silk fibroin during preparation of silk microspheres and exposure to high humidity was studied. Scoured silk fiber was dissolved in an aqueous solution of calcium chloride mixed with ethanol. After dialysis against purified water, theophylline was added to the solution as a model drug. Silk microspheres were obtained by spray-drying. Silk fibroin and theophylline were found to be amorphous in the microsphere by means of powder X-ray diffractometry. Fibroin assumed beta-sheet conformation in the scoured silk fiber while it has an irregular structure in the microsphere, according to FTIR and Raman spectra. Fibroin recrystallized and its secondary structure changed to beta-sheet conformation by exposure of the microspheres to an atmosphere of 89% RH. (C) 2003 Elsevier Inc. All rights reserved.