• 作者： 林仁混；陳長安；王秋華
• 作者服務機構： 國立臺灣大學醫學院生化學研究所; 國立臺灣大學醫學院實驗診斷科
• 中文摘要： Dabsyl Chloride (4-Dimethylaminoazobenzene-4'-Sulfonyl Chloride)是由本研究室首創之新成色試劑。它可和各種氨基酸(Amino acid, a. a.)(22種)反應形成Dabsyl-a. a.之黃色衍生物。它們(10^-11 moles)可在TLC上由肉眼檢查出，而對光甚為安定。其靈敏度較2, 4-Dinitrofluorobenzene約高60倍，而較Dansyl Chloride約低5倍，但其穩定度則較此兩者為優。Dansyl-a. a.之檢查需在暗室進行，較為不便。 Dabsyl-a. a.在熱與氧夾攻下，可在Silica gel plate上產生脫甲基作用，而形成其衍生物Mabsyl-a. a.此種新衍生物亦可作為鑑定各種氨基酸之根據。 Dabsyl Chloride可與蛋白質或Polypeptide形成Dabsyl-蛋白質或Dabsyl-peptide，經水解後可放出Dabsyl-a. a.而鑑定其N-端氨基酸。據文獻記載有14種氨基酸(表一)可發生代謝異常症。在本研究中利用Dabsyl Chloride與此14種a. a·反應，再以二種混合溶媒將此14種dabsyl-a. a.分開(Fig. 2)。此法可應用於檢查各種氨基酸代謝異常之病人。經以五種(Phe, Gly, Val, Ile, Leu)模擬氨基酸尿作試驗，結果在300 n moles/ml之濃度，即可在TLC Plate上檢查出來。 利用本法也檢查到一些癌症病人之尿中有Trp, Gly, Leu，及Ile之存在。正常人的尿中並無此等氨基酸之存在，(表四)本研究室正在篩試更多的癌症病人，以確定此等氨基酸出現於癌症病人尿中之生物意義。
• 英文摘要： Dabsyl chloride (4-dim'ethylaminoazobenzene-4'-sulfonyl chloride) was synthesized by either reaction of sodium 4-dimethylaminoazobenzene-4'-sulfonate with phosphorus pentachloride, or chlorosulfonation of 4-dimelhyl-aminoazobenzene with chlorosulfonic acid. Dabsyl chloride reacted readily with all amino acids to form dabsyl amino acids which were visualized on thin layer plate and found to be photostable. The sensitivity of dabsyl chloride was found to be approximately 60 times higher than that of 2,4-dinitrofluorobenzene and about 5 times less than that of dansyl chloride. The stability of dabsyl amino acids was superior to that of dansyl- and 2,4-dinitrophenyl-amino acids. Dabsyl amino acids were demethylated to 4-monomethylaminoazobenzene-4'-sulfonyl amino acids through the interaction with hot air and silica gel, this provided additional parameters to dabsylation for identifying amino acids. Dabsyl peptide or dabsyl protein was hydrolyzed with 6 N HC1 at 105蚓 in a sealed thick-walled glass tube for 2-3 hours, and the N-terminal amino acid was detected as dabsyl derivative on TLC plate. Fourteen amino acids related to various inborn-errors of metabolism were dabsylated and separated by bidimensional TLC. The procedure was found to be satisfactory for detecting amino acids in the reconstituted amino aciduria. High urinary tryptophan and glycine levels were found in some patients of chronic myeloblastic leukemia, acute myeloblastic leukemia and lymphosarcoma, while high urinary leucine was found in some patients of lymphosarcoma. Further studies on the significance of the elevation of these urinary amino acids in cancer patients are now in progress.
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