ANALYSIS OF HYDROXYPROLINE IN COLLAGEN OF PIG SKIN TISSUE BY LOW PRESSURE ION CHROMATOGRAPHY SEPARATION AND CONDUCTIVITY DETECTION

摘要

The Key Laboratory of Leather Chemistry and Engineering of the Ministry of Education, Sichuan University, Chengdu, 610065, China A simplified method has been developed to determine the hydroxyproline concentration in pigskin collagen. The sample was prepared by hydrolyzing pigskin tissue collagen for 24 hours overnight at 110℃ in 6M/Lhydrochloride acid followed by determination by low pressure ion chromatography separation and conductivity detection without derivatization. Hydroxyproline was separated and detected successfully using a 5mm x 100mm long column tilled with 25μm-30μm size and 0.02 mmol/g capacity cationic exchange resin. The elution solution was 0.8mmol/L nitric acid solution and the flow rate was 0.6ml/min. The concentration of the hydroxyproline sample was calculated by the five points work curve of standard stock solution. The result shows that the concentration of hyp in the sample was 3.16 ± 0.28 mmol/L. The RSD of the method was 4.63-8.97%. The recovery percentage of hyp added to the sample was 92.5-105.5%.