Master is a designation given to an individual, usually a magus, who has become one of the formal participants of the Holy Grail War by obtaining Command Spells and forming a contract with a Heroic Spirit. Under ordinary circumstances, a Master is always a spellcaster, usually a Magus with a Magic Circuit, who is selected by the Grail. It is possible for an ordinary person to become a Master, as in the case of Ryuunosuke Uryuu, though this is rare.

Kiritsugu Emiya is the master of Saber and the main protagonist. Born in Fuyuki City, Kiritsugu was raised on Alimango Island, where he lived with his father, Norikata Emiya, the fourth family head of the Emiya family and a magus specializing in time magic.Despite being the true Master of the Einzbern family, Kiritsugu operates mostly in the background during the conflict, ambushing other Masters while they are otherwise occupied.

Kirei Kotomine is the Master of Assassin. Before the Fourth Holy Grail War, Kirei served as a priest under his father, Risei Kotomine. Although Kirei understood the distinction between good and evil, he could only revel in committing sins. Hounded by guilt and sorrow and with no alternative, Kirei prayed that, through marriage to a woman he loved, he could finally achieve respite from his tortured existence. When Kirei discovered marriage could not change him, he was on the brink of suicide; his terminally ill wife, filled with concern, committed suicide to show to Kirei that he could still feel compassion for others and died happy seeing her husband crying for her. However, after her death, Kirei was disgusted to realize he was crying because he had wanted to kill her himself; from that day on, he ceased pursuing salvation altogether.

Master of Archer, and an arrogant and manipulative mage. He arranges for Kirei Kotomine to be his understudy in magic and his support in the Holy Grail War. He thinks highly of Kotomine and has him serve in all manners of affairs,He is skilled in fire manipulation, and employs a large ruby set in the end of a staff as his Mystic Code to help him shape and control flame. While a skilled mage and confident in his abilities, he is actually one of the weakest Tohsaka in history.

The Master of Rider and future Lord El-Melloi II, Waver steals his teacher's artifact early on and uses it to summon Rider. He plans to use the Grail to force the Magus Association to recognize his genius. Despite his difficulties with Rider's overbearing nature, Waver and his Servant form a strong relationship. Though lacking in practical experience in the field of fighting in a war, Waver, mostly due to his Servant, survives the Holy Grail War with little detriment to himself and develops a greater sense of perspective of the world, though he now harbors some bitterness towards Japan.

Kariya is the Master of Berserker.Kariya had previously left the Matō family ten years prior to the 4th Holy Grail War out of disgust for his own family, but returns and makes a deal with Zōken Matō that he will win the Grail War. Due to his lack of formal training, Kariya was implanted with magical worms.This caused a number of side effects as the worms would literally eat away at their host, causing Kariya's health to progressively fail in return for significantly expanding his magic circuits and magical potential. By the time of his summoning of a Servant, Kariya's hair had turned white and the nerves on the left side of his face were dead. In addition, his natural lifespan had been shortened to such an extent that Zōken estimated he would only live on for one additional year. However, despite the Crest Worms and his own accelerated training, Kariya remained severely lacking as a Magus. At Zōken's behest, he compensated for this by summoning the most powerful of the seven Servant classes, the Servant of Mad Enhancement, Berserker. However, the high volume of magical energy required to maintain Berserker causes him frequent, severe pain.

Ryūnosuke is the Master of Caster and a highly deranged serial killer who summons Caster after murdering a family and using their blood as a component in the summoning ritual. He does not seek the Holy Grail, but follows Caster in order to find macabre ways to kill people and relieve his boredom, using a hypnotic charm to lure children to their deaths. However, Ryūnosuke does admit that he does in fact feel that there is something missing from his life and that, on some level, he desires to find it through his atrocities.

The Master of Lancer, Kayneth is a nobleman from the Magus Association, Lord of the House of El-Melloi and a prodigy at magecraft, said to be the most talented participant in the Holy Grail War. His artifact is stolen by his pupil, Waver Velvet, but he manages to obtain another artifact and summon Lancer instead.Despite his skill, Kayneth's overly-traditional approach to combat causes him to underestimate his opponents' capacity for violence, as he believes that the Holy Grail War will be fought with honorable duels and tests of skill rather than quick blitzkriegs like those perpetrated by Kiritsugu and Kotomine. He is confident in his abilities and takes pride in his heritage, but finds the extent of Kiritsugu's untraditional and underhanded tactics both offensive and unexpectedly challenging to overcome.
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