He was born in Damascus to an eminent family, and was well educated in his youth. Discontented with the wisdom of the world, he entered monastic life in the monastery of St Theodosius, where he became the lifelong friend and disciple of John Moschos. Together they visited the monasteries and hermitages of Egypt; they later wrote down their discoveries among the holy monks in the classic Spiritual Meadow. After the death of his teacher, St Sophronius traveled to Jerusalem, which had just been liberated from the Persians. He was there to see the Precious Cross returned from Persia by the Emperor Heraclius, who carried it into Jerusalem on his back. A few years later, in 634, St Sophronius was elected Patriarch of Jerusalem, where he served his flock wisely for three years and three months. He was zealous in the defense of Orthodoxy against the Monothelite heresy: He convoked a Council in Jerusalem which condemned it before it was condemned at the Sixth Ecumenical Council. The holy Patriarch even traveled to Constantinople to rebuke the Patriarch Sergius and Emperor Heraclius, who had embraced the Monothelite error.

The years of peace were few for the Holy Land; for just as the Persian Empire was decisively defeated by Heraclius, the followers of Islam erupted out of Arabia, conquering most of North Africa and the Middle East in a few years. The Saint was so grieved by the capture of Jerusalem in 637 by the Caliph Omar that begged God to take him, so that he might not live to see the desecration of the holy places. His prayer was granted, and he reposed in peace less than a year later.

St Sophronios is the author of the Life of Saint Mary of Egypt, appointed to be read in the churches during every Great Lent. He also wrote the service of the Great Blessing of the Waters. Some have attributed the Vesperal hymn "Gladsome Light" to him, but we know that it dates from before the time of St Basil the Great, who mentions it in his writings. It seems though, that St Sophronios supplemented the hymn, and that its present form is due to him.