高效液相色谱法检测弹性蛋白中锁链素

用高效液相色谱技术对弹性蛋白水解液中锁链素进行了测定。单一锁链素，锁链素、酪氨酸、苯丙氨酸混合液及弹性蛋白水解液经Ｃ１８柱分离，甲醇－水（２８，Ｖ／Ｖ）洗脱，紫外检测器２８０ｎｍ检测。锁链素与酪氨酸峰面积比（ＣＡ）和峰高比（ＣＨ）分别为１．４％和３．４％，标准曲线浓度范围０．１６～０．６４ｍｇ／Ｌ时相关系数为０．９９５６。结果表明：上述色谱条件可定量测定弹性蛋白中锁链素含量，无需将锁链素与水解液中其它氨基酸加以分离，而仅需同其它２７５ｎｍ有吸收的组分区分。

Detection of Desmosine in Elastin by High Performance Liquid Chromatography(HPLC)

An assay of desmosine (DES) in elastin hydrolysate was performed by HPLC. Desmosine alone or its mixture with tyrosine and phenylalanine as well as elastin hydrolysate was separated on a C18 column eluted with a mobile phase of MeOH:H2O (2:8, V/V) and detected at UV-280nm. The results showed that the time of DES eluted in all cases mentioned above was comparable. The ratio of peak area (CA) and peak height (CH) between DES and Tyr is 1.4% and 3.4% resectively. It could be as a reliable measure to evaluate the quantity of DES in elastin. The correlation coefficient r was 0.9956 in the range of 0.16-0.64mg/L for a calibration curve of DES concentration vs peak area. In conclusion, this method could be used as a biochemical indication to detect the degradation level of elastin in the pathological state.