Colour is the visual perceptual property in the cotton grade classification of Universal Cotton Standards. We undertook an experimental study on the variation in cotton colour using the Non-contact method. Rd (degree of reflectance) and +b (yellowness) are globally recognized colour parameters of cotton, which are measured by HVI (High Volume Instrument). The Non-contact method is used for the evaluation of colour variation within cotton samples. The results obtained from the Non-contact method were compared with those of other conventional methods used globally for the color measurement of cotton fibers: Additionally image analysis is interpreted with the variation in cotton colour. Results confirmed a strong co-relation between the Non-contact method and image analysis for the colour variation of cotton.