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In vivo and in vitro studies of differential continuous wave photoacoustic spectroscopy (DCW-PAS) for non-invasive blood glucose monitoring were performed. The DCW-PAS technique utilizes amplitude modulation of dual wavelengths of light to determine changes in glucose concentration. The study compared DCW-PAS measurements with results from invasive blood glucose sensor measurements during oral glucose tolerance tests (OGTTs) of healthy people. The trends in blood glucose levels (BGLs) obtained from invasive sensors and from the photoacoustic signal have good agreement, with the standard error and correlation coefficient against BGLs of 48 mg/dL or less and 0.80, respectively. Our proposed photoacoustic spectroscopy (PAS) method shows high potential for use in a non-invasive BGL sensor.