Standardization of herbal is important to meet the WHO parameters. A comparison of methanolic extracts of fruits of Tribulus terrestris Linn roots, fruits and root powder were done with whole plant. Proximate analysis of fruit powder Tribulus terrestris Linn were compared with the reported values of proximate analysis. The whole plant powder extract in HPTLC method indicated that the band with Rf value of 0.65 was observed in each developed and scanned chromatographic plate. The other bands present in Tribulus terrestris Linn methanolic extract of fruits, roots and leaves were found to be different and hence it is possible to differentiate plant parts of Tribulus terrestris Linn. A study of percentage composition of various phyto chemical aspects such as total ash, acid insoluble ash, and nitrate content in Tribulus terrestris Linn was performed. Nitrate content in fruit powder showed linearity curve using Double Beam UV Spectrophotometry and Colorimetry. © 2012 Elsevier B.V., All rights reserved.