We are developing a non- or minimally-invasive method for detecting and measuring specific drugs and biomolecules in vivo using photoacoustic spectroscopy (PAS). This pilot study investigated the feasibility of detecting the concentration of certain drugs in the vitreous or aqueous of the eye. As a prototype for using PAS for molecular detection in vivo, the technique was applied to the detection in a surrogate eye, of drugs with known optical spectrum such as Trypan Blue, Rose Bengal, and Amphotericin B (AB), at concentrations as low as 1 μg/ml. Chopped CW, or short pulse, Q-switch lasers, were used as pumping sources to generate ultrasonic photoacoustic signals in an ocular phantom containing the drug solutions. In addition to an ultrasonic hydrophone, the photothermal deflection technique (PhDT), a non-contact optical method with high sensitivity and fast response, were used to record the photoacoustic signals. The data from both detectors were compared over a range of drug concentrations. The photoacoustic signal generated from the retina was used as a reference, to measure the attenuation of light through drug solutions of different concentrations in the ocular phantom. The results indicated that photoacoustic spectroscopy is feasible in ocular phantoms incorporating ex vivo ocular tissue. The signals recorded using PAS were to be found to be linearly dependent on drug concentration, as predicted by theory. The photoacoustic method was found to be sensitive to drug concentrations as low as 1 μg/ml, a clinically relevant concentration for many drugs. Future work will be directed at adapting this method for in vivo measurement, and enhancing its sensitivity by using a tunable laser as the pump source.© (2006) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.