Rebbi and Rabbi Elazar b'Rabbi Shimon disagree about whether the sanctification of a kohen's hands and feet, performed before Temple service, is nullified each night, requiring repetition the next morning. According to Rabbi Elazar b'Rabbi Shimon, the sanctification remains valid overnight, and there is no need to repeat it.

Ilfa raises a question based on this view: If the sanctification remains valid overnight, is the water in the Temple's basin also unaffected and not disqualified by nightfall? Rabbi Ami quotes Rabbi Yochanan, who reports that Ilfa later answered that the water is indeed not disqualified overnight. However, Rabbi Yitzchak bar Bisna challenges this conclusion.

A Mishna in Yoma describes a device used in the Temple—the muchni—which lowered the water into a well each night to prevent it from becoming disqualified by remaining overnight. The Gemara attempts to use this source to support the possibility that Rabbi Elazar b'Rabbi Shimon holds the water is disqualified overnight. This is based on an earlier Mishna in the same chapter that discusses the location of the bull's slaughter on Yom Kippur, which aligns with Rabbi Elazar b'Rabbi Shimon's opinion: the area between the altar and the ulam (entrance hall), designated for slaughtering kodashei kodashim (most holy offerings). If the earlier Mishna accords with his opinion, it stands to reason that the later Mishna accords with his opinion as well. However, since the passage can also be interpreted in accordance with Rebbi's view, no definitive conclusion is reached.

A Mishna in Yoma describes a device used in the Temple, the muchni, to lower the water into a well each night to prevent it from becoming disqualified overnight. The Gemara attempts to prove that this source aligns with Rabbi Elazar b'Rabbi Shimon's view, proving that he holds the water is disqualified overnight, as an earlier Mishna in the chapter that describes the location of the slaughtering of the bull on Yom Kippur accords with his opinion. This location, between the altar and the ulam, matches Rabbi Elazar b'Rabbi Shimon's opinion regarding the designated area for slaughtering kodashei kodashim (most holy offerings). However, the passage can also be interpreted according to Rebbi's view, so no definitive conclusion is reached.

Rabbi Yochanan rules that a kohen who removes ashes from the altar during the final part of the night sanctifies his hands and feet for the day, despite it still being nighttime. Abaye explains this ruling according to Rebbi, while Rava explains it according to Rabbi Elazar b'Rabbi Shimon. A challenge is raised against Rava's interpretation, but it is ultimately resolved.

Two additional questions are discussed:

1. Does leaving the Temple cancel the sanctification of one's hands and feet? Four sources are brought to address this, but each is rejected, and the question remains unresolved.
2. Does becoming impure cancel the sanctification? Two of the sources cited in the previous discussion are brought in an attempt to answer this question as well.