Generally, some drugs are present in our body for quite long time at nano or femto gram level and accumulate in body tissues, causing many side effects. Therefore, in the absence of the techniques capable to detect at nano or low level the absence of these drug residues is assumed. Besides, small biological samples such as blood of infants, some hormones and enzymes in our body need detection techniques of nano levels. Pharmaceutical analysis is an integral part in pharmacokinetics and pharmacodynamics studies, which needs accurate analyses of drugs. The analytical techniques should be capable to detect drugs and pharmaceutical at nano or low detection limits. The detection at nanogram level is becoming more important and scientists and other regulatory authorities are searching for data on the detection at nanogram level. Therefore, the present article describes state-of-art of nano analysis by using Nano Liquid Chromatography (NLC). Besides, attempts have been made to discuss applications, optimization and comparison of analyses.