<p>Yes. When one washes his hands in middle of the night, and he plans to return to sleep, he is to prepare new water for the morning.<sup id="shulchan-fnref-1" class="shulchan-fnref"><a href="#shulchan-fn-1">1</a></sup></p>
<hr class="shulchan-footnotes-sep"><section class="shulchan-footnotes"><div class="shulchan-footnotes-body"><ol><li id="shulchan-fn-1">As the spirit of impurity will once again reside on his hands upon him returning to sleep before Alos. If however it is already past Alos and one plans to return to sleep then there is no need to prepare new water just like is the law regarding one who sleeps during the day. <a class="shulchan-fn-back" href="#shulchan-fnref-1" aria-label="Back to text">↩</a></li></ol></div></section>

Shulchan Aruch HaRav | If one washed and plans on going back to sleep, must one prepare Negel Vasser again for the next time he wakes up?

Yes. When one washes his hands in middle of the night, and he plans to return to sleep, he is to prepare new water for the morning.1