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We present a new instrument, Hoxotope, for the in situ measurement of H2O and its heavy deuterium isotopologue ͑HDO͒ in the upper troposphere and lower stratosphere aboard the NASA WB-57. Sensitive measurements of ␦D are accomplished through the vacuum UV photolysis of water followed by laser-induced fluorescence detection of the resultant OH and OD photofragments. The photolysis laser-induced fluorescence technique can obtain S / N Ͼ 20 for 1 ppbv HDO and S / N Ͼ 30 for 5 ppmv H2O for 10 s data, providing the sensitivity required for ␦D measurements in the tropopause region. The technique responds rapidly to changing water concentrations due to its inherently small sampling volume, augmented by steps taken to minimize water uptake on instrument plumbing. Data from the summer 2005 Aura Validation Experiment Water Isotope Intercomparison Flights ͑AVE-WIIF͒ out of Houston, TX show agreement for H2O between Hoxotope and the Harvard water vapor instrument and for HDO between Hoxotope and the Harvard ICOS water isotope instrument, to within stated instrument uncertainties. The successful intercomparison validates Hoxotope as a credible source of ␦D data in the upper troposphere and