From the Super Mario Wiki, the Mario encyclopedia
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The Handkerchief is an item in the SNES and PC releases of Mario's Time Machine. It is a long, white piece of cloth. Mario receives it from Anne Hathaway after traveling back in time to Stratford-upon-Avon circa 1601. After Anne gives Mario a Feather for Richard Burbage's costume so that he can participate in a play, Mario then wonders out loud if the Skull in his possession also belongs to a play. Anne recognizes the Skull as being a prop for William Shakespeare's most recent play, Hamlet, and wraps it in the Handkerchief before sending Mario to deliver it to Shakespeare. However, Mario then unwraps the Skull and gives the Handkerchief to a nearby man who was complaining about a stain on his doublet. With the Handkerchief, he thanks Mario and attempts to rub out the stain while shouting "Out, darn spot!" (a quote from Shakespeare's Macbeth in which Lady Macbeth is still haunted by the visions of killing King Duncan and attempts to clean the blood from her hands despite there being no blood). He then bids Mario farewell, still rubbing away the stain.