title: Simultaneous determination of hydrazinocurcumin and phenol red in samples from rat intestinal permeability studies: HPLC method development and validation.
abstract: Hydrazinocurcumin (HZC) is a patented multiactivity compound and is a potent derivative of curcumin which is not yet explored for further development as formulation and requires the determination of biopharmaceutical suitability of the molecule. Intestinal permeability and logP of a compound are two vital biopharmaceutical properties by which, any "hit" molecule proceeds towards NCE (new chemical entity) and govern formulation design of bioactive molecules. In this report, a simple, precise and accurate isocratic reverse phase (RP) liquid chromatography method for simultaneous analysis of HZC and phenol red, for the application in rat in situ single-pass intestinal perfusion (SPIP) was developed and validated using FDA bioanalytical guidelines. RP-HPLC method was developed on C-18 column with UV detection at 332 nm for both the compounds. Isocratic run with the mobile phase consisting of organic phase (methanol:acetonitrile:: 50:20 v/v) and water in the ratio of 80:20 v/v provided a short run time of 7 min with resolution of more then two without interference of blank matrix. The working/expected concentration range for HZC and phenol red were 0.5-50 and 2-200 microg/ml. LOQs for HZC and phenol red of the method was found to be 0.167 and 0.271 microg/ml respectively. The validation parameters indicate that method was suitable for the intended purpose. Permeability, considering water flux with the help of non-permeable marker phenol red was calculated to be 0.34 x 10(-4)cm/s. Along with other descriptors, logP (1.78) and MW (<500) of HZC makes it a potential candidate for oral formulation.
authors.affiliation: Department of Pharmaceutics, National Institute of Pharmaceutical Education & Research, Sector 67, S A S Nagar, Mohali 160062, India.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Rathore
authors.forename: Jay Prakash
authors.initials: JP
authors.lastname: Jain
authors.forename: Amit
authors.initials: A
authors.lastname: Srivastava
authors.forename: S M
authors.initials: SM
authors.lastname: Jachak
authors.forename: Neeraj
authors.initials: N
authors.lastname: Kumar
mesh_terms: D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003474:Curcumin / Q000031:analogs & derivatives* / Q000032:analysis; D006834:Hydrazines / Q000032:analysis*; D007408:Intestinal Absorption; D008297:Male; D010637:Phenolsulfonphthalein / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
chemical_list: D006834:Hydrazines; C475807:hydrazinocurcumin; D010637:Phenolsulfonphthalein; D003474:Curcumin