Iris, as well as being very useful in the area of identification can provide information about the health status of the organs and tissue in the body. Determination of high cholesterol in the blood, like many other diseases, is concerned with iris analysis. In this study, examined the relationship between high cholesterol and the sodium ring to be formed around the iris by analysis of eye images, which belong to data bank. Firstly, the pupil has been determined with using integral differential operator method on each black and white formatted eye images. Secondly, width of the sodium ring was determined by analysis of color tone. The relationship between the width was determined and cholesterol in the blood was examined for six different eye images. By results is concluded that there is a direct relationship between cholesterol in the blood and width of sodium ring.