Abstract: Background and Objective: The p-phenylenediamine (PPD) used as a coloring agent in hair colour and it is known to be allergic due to formation of trimer Bandrowski’s base. At favorable condition the p-phenylenediamine molecule convert in to dimer and trimer configuration, trimer configuration is called as a Bandrowski's Base. The objective of study was identification of Bandrowski’s Base through the HPLC estimation. This estimation will helpful to find out the safety aspects of henna based hair colours. Estimate the Bandrowski’s Base formation with various pH and it help to draw a optimum pH range for the Bandrowski’s Base formation and prevent the formation with adjusted the pH range. Therefore, main objective of the study was formulating the safer henna based hair colour without the formation of sensitive potential of Bandrowski’s Base and develop the safer henna based hair colour for the end user. Materials and Methods: Bandrowski’s Base (BB) was synthesized by the method described by Ritter and Schmitz. BB base formation depends upon the pH condition (>10.0), high alkaline and hydrogen peroxide concentration. Various concentrations of standard PPD and BB base injected and identified the retention time. Samples are kept injected at 24 and 48 h time interval interest to monitor the BB base formation and PPD self -coupling. Results: In this combination of standard materials of PPD and BB standard injected and the peaks showed the different standard retention time, PPD eluted at 2.352 min. Retention time and BB base eluted at 4.696 min. Addition of resorcinol plays a crucial role to prevent the BB base formation. Quinone-diimine react with coupler resorcinol to form the indophenol complex (green component) further it under goes coupling with PPD or resorcinol or co-oxidation coupling it forms a poly indophenol complex. Conclusion: Henna based hair color with pH 10-12.0 with 30 min developmental time does not contribute the formation of Bandrowski Base. However, henna based hair colour products pH fall between 7.0-8.0, it further confirms that there is no BB formation and safe to use. No evidence for the formation of Bandrowski base after 24 h with the treatment of higher ammonia and hydrogen peroxide.