A　mathematical　model　for　determining　the　Yin　Yang　nature　of　fruits　was　established　in　our　previous　study.　The　objective　of　this　study　was　to　validate　the　mathematical　model　using　animal　experiments.　One　hundred　and　twenty　Wistar　II　female　rats　were　randomly　divided　into　five　diet　groups:　A,　B,　C,　D　and　E　that　were　administered　with　saline　solution,　hot　Traditional　Chinese　Medicine　(TCM)　tonic,　cold　TCM　tonic,　hot　(Yang)　formulated　mineral　solution　and　cold　(Yin)　formulated　mineral　solution,　respectively.　The　consumption　of　drinking　water　of　rats　in　each　group　was　determined　during　a　feeding　period　of　21　days.　On　the　last　day　of　the　experiment,　the　lingual　superficial　structure　of　the　rats　was　examined,　which　is　a　practice　in　traditional　Chinese　medicine　to　diagnose　Yin-yang　symptoms,　and　blood　samples　were　collected　from　the　rats　to　determine　serum　thrombocytin　(5-HT)　and　thyrotrophic　hormone　(TSH)　and　plasma　noradrenaline　(NE).　The　diet　D　group,　administered　with　the　Yang　mineral　solutions　demonstrated　the　same　trend　as　the　diet　B　group　fed　with　hot　TCM　tonic　(hot　control),　while　the　diet　E　group　administered　with　the　Yin　mineral　solutions　has　the　similar　trend　as　diet　C　group　fed　with　cold　TCM　tonic　(cold　control).　It　was　concluded　that　the　diet　D　had　Yang　nature,　whereas　the　diet　E　had　Yin　nature.　The　results　from　current　study　confirmed　the　findings　from　the　previous　study　that　the　Yin-Yang　nature　of　the　fruits　could　be　determined　by　the　ratio　of　copper,　iron　and　magnesium　content　using　the　mathematical　model.