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Background:-Prajasthapana Churna is a combination of herbal drugs having a numerous number of references in Ayurveda classics. But its qualitative data hasn’t been explored thoroughly yet. It is mainly useful to treat gynaecological diseases, infertility, repeated abortion and in diseases where according to Ayurveda Vata and Pitta Doshas are vitiated. Aim:- To develop pharmaceutical profile of Prajasthapana Churna. Material and Method:- Study included preparation of Prajasthapana Churna following all SOPs using raw drugs, which were previously authenticated. Later, Prajasthapana Churna was subjected to physicochemical and high performance thin-layer chromatography (HPTLC) analysis as per standard protocols. Result and Discussion:- Pharmaceutical analysis showed that the loss on drying value was 0.40%, ash value was 9.56%w/w, water soluble extract was 7.24%, methanol soluble was 10.22%, acid insoluble ash 0.76%, and pH is 4.5. Methanolic extract of Prajasthapana Churna shows presence of alkaloids, flavonoids, phenols, tannin, sugar, steroids, saponin, cardiac glycosides and carbohydrates which can be hypothesized for its estrogenic activity. HPTLC fingerprinting profile of Prajasthapana Churna revealed 5 spots at 254 nm and 5 spots at 366nm. Conclusion:- The present investigation will be helpful in assessing the pharmacognostical, phytochemical analysis and laying down pharmacopoeial standards for Prajasthapana Churna.