This study was designed to develop crosslinking of $eta$-cyclodextrin ($eta$-CD), and determine the optimum conditions of different factors (mixing time, mixing temperature, and mixing speed) on cholesterol reduction from milk. Crosslinked $eta$-CD was prepared with epichlorohydrin. When milk was treated with different conditions, the cholesterol removal rate was in the range of 79.4 to 83.3% with 1 % crosslinked $eta$-CD addition, which were not significantly different among treatments. After cholesterol removal from milk, the used crosslinked $eta$-CD was washed for cholesterol dissociation and reused. For recycling study, the cholesterol removal rate in first trial was 81.8%, which was mostly same as that using new crosslinked $eta$-CD. With five trials repeatedly using the same sample, the mean cholesterol removal rate was 81.2%. The present study indicated that the optimum conditions on cholesterol removal using crosslinked $eta$-CD were 10 min mixing with 400 rpm speed at $5^{circ}C$ with about 80% cholesterol removal. In addition, crosslinked $eta$-CD resulted in the effective recycling efficiency almost 100%.