A highly purified and heat stable cross-linked nonpolymeric tetrameric hemoglobin suitable for use in mammals without causing renal injury and vasoconstriction is provided. A high temperature and short time (HTST) heat processing step is performed to remove undesired dimeric form of hemoglobin, uncross-linked tetrameric hemoglobin, and plasma protein impurities effectively. Addition of N-acetyl cysteine after heat treatment and optionally before heat treatment maintains a low level of met-hemoglobin. The heat stable cross-linked tetrameric hemoglobin can improve and prolong oxygenation in normal and hypoxic tissue. In another aspect, the product is used in the treatment of various types of cancer such as leukemia, colorectal cancer, lung cancer, breast cancer, liver cancer, nasopharyngeal carcinoma and esophageal cancer. Another application is heart preservation in situations where there is a lack of oxygen supply in vivo, such as in heart transplant or oxygen-deprived heart.提供一種高度純化且熱穩定之經交聯的非聚合的四聚體血紅素，其適合應用於哺乳動物中而不導致腎臟損傷與血管收縮。執行高溫與短時間(high temperature and short time,HTST)之熱處理步驟以有效地移除不希望獲得之二聚體形式的血紅素、未經交聯之四聚體血紅素與血漿蛋白質雜質。在熱處理之後與視需要而定在熱處理之前的N-乙醯基半胱氨酸(N-acetyl cysteine)的添加維持一低程度的變性血紅素(met-hemoglobin)。此熱穩定之經交聯的四聚體血紅素可在正常與缺氧組織中增進並延長充氧作用(oxygenation)。在另一方面，將此產物用於各種形式之癌症的治療中，例如白血病、大腸直腸癌、肺癌、乳癌、肝癌、鼻咽癌與食道癌。另一應用為在in vivo缺乏氧氣供應之情況中的心臟的保存，例如在心臟移植或缺氧心臟(oxygen-deprived heart)中。
