The eleventh perek of masechet Zevachim discusses the Torah law that if the blood from a chatat (sin offering) hits the kohen's clothing, then it must be laundered within the azarah (Temple courtyard). The law is found in the pasuk: "…and its blood is sprinkled upon a garment, whatever it has been sprinkled upon you shall wash in a holy place." (Vayikra 6:20) The second Mishnah establishes that this laws only applies to a valid korban while the next Mishnah adds that the blood must have been fit for offering on the mizbeach.
The Mishnah continues explaining that if the blood was thrown on the corner of the mizbeach as required and then splashed back on to the clothing of the kohen, this law would not apply. It can be understood that once this blood has reached the mizbeach, the mitzvah is performed and that blood that splashes back is no longer fit to be placed on the mizbeach. Similarly if it splashed back from the base, the clothing would not require laundering. Recall that the blood poured at the base is the blood remaining after the required bloods have been placed on the mizbeach – the shirayim. Importantly, the Bartenura explains that even if the shirayim was not yet poured at the base and it stained the kohen's clothing, it would not require laundering. The basis for all the above laws is that the above cited pasuk states "is sprinkled" (asher yaze) which excludes blood that has already been sprinkled or is not fit for sprinkling. In the case of the shirayim, since the blood for the korban has already been sprinkled, the shirayim no longer qualifies for this law.
Recall that the blood of a chatat must be placed on the four corners of the mizbeach. It is important to also remember that if the blood was only placed on one corner, then be'dieved (after the fact) the korban is valid and an atonement has been achieved. What would the law be, if the blood was placed on one corner of the mizbeach and then some blood spilled on the clothing of the kohen? Would it require laundering? On the one hand, the base requirement has been satisfied, yet on the other hand, the blood is still fit for placement on the other three corners.
The Tosfot R' Akiva Eiger (commenting on the Tosfot Yom Tov) and the Tifferet Yaakov (commenting on the Tifferet Yisrael) both assert that kohen's clothing would require laundering in the azarah, with the latter citing the Gemara (38b) that rules accordingly. Perhaps addressing another question might give us an insight into this ruling.
When explaining the Mishnah's ruling that the blood that ricocheted off the corner does not require laundering, Rashi (38b) explains that this is also based on the above pasuk – since the blood has already been sprinkled it is not covered by the pasuk. The Griz (stencil s.v. "min hakeren") questions the necessity for the pasuk. Once the mitzvah has been performed with the blood it is considered chulin – regular blood – and no longer has any sanctity. Consequently, why would anyone think that this blood is covered by the pasuk?
The Griz explains that this law has nothing to do with the sanctity of the blood. The Torah simply states that a bloodstain originating for a korban chatat requires laundering. Consequently, were it not for the pasuk stating "asher yaze" one might have thought that even the blood that splashed off the corner of the mizbeach would require laundering. "Asher yaze" defines the scope of the law to only blood that is fit for sprinkling.
With the Griz's explanation we can understanding the law in our case. Whether the base mitzvah has been performed or not is irrelevant. "Asher yaze" is what is important. Consequently, since the blood in the cup after the first sprinkling is fit for the other three corners, its staining would require laundering.
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