用高效液相色谱法检测2型糖尿病患者血浆游离脂肪酸谱的分析研究

Analysis of the plasma free fatty acid composition in T2DM by high performance liquid chromatography

目的 用高效液相色谱法分析T2DM患者血浆游离脂肪酸谱的变化.方法 收集武汉大学中南医院体检人群和住院患者,其中健康对照组94名、单纯T2DM组101例、T2DM合并高脂血症组77例.取受试者空腹静脉血,分离新鲜血浆,以正十七酸作为内标,以改良Doles法萃取游离脂肪酸,溴苯乙酮作为柱前衍化剂,采用Dionex Summit高效液相色谱法分析系统检测血浆游离脂肪酸谱,内标标准曲线法定量.结果 健康对照组、单纯T2DM组与T2DM合并高脂血症组3组之间的总游离脂肪酸浓度分别为（355.63±100.35）、（421.21±200.83）、（473.04±213.40）μmol/L,3组之间差异有统计学意义（X2=13.08,P〈0.01）,不饱和游离脂肪酸浓度依次为（206.29±61.94）、（218.11±110.28）、（240.94±116.79）μmol/L,3组之间差异无统计学意义（x2=2.17,P〉0.05）.单纯T2DM组与健康对照组比较,总游离脂肪酸升高[（421.21±200.83）μmol/L vs（355.63±100.35）μmol/L,x2=7.22,P〈0.05],不饱和游离脂肪酸浓度和健康对照组差异无统计学意义.T2DM合并高脂血症组与健康对照组比较,总游离脂肪酸浓度升高[（473.04±213.40）μmol/L vs （355.63±100.35）μmol/L,x2=27.93,P〈0.01].日间及日内RSD均〈5%,最低检测浓度为0.05～0.35 μmol/L,高、中、低浓度下的回收率为96.4%～104.8%.结论 T2DM患者血浆总游离脂肪酸浓度升高,以饱和游离脂肪酸升高为主,不饱和游离脂肪酸改变差异无统计学意义.可能与T2MD的发生发展有关,有望成为糖尿病脂代谢紊乱的临床早期监测新指标.

Objective To analyze the plasma free fatty acid （FFA） composition in patients with T2DM. Methods All subjects were from Zhongnan hospital of Wuhan university, and they were divided into three groups： normal control （ n = 94 ）, T2DM （ n = 101 ） and T2DM with hyperlipidemia （ n = 77 ）. Fasting blood samples were taken from the participants, and plasma FFA were separated using a modified Doles method with the bromoacetophenone, pre-column-derivative. The quantitation of FFA was performed on were （355.63 ± 100. 35） μmol/L, （421.21 ± 200. 83 ） μ mol/L, （ 473.04 ± 213.40 ） μmol/L in healthy controls, T2DM group and T2DM with hyperlipidemia group, respectively. The significant differences were observed among the 3 groups（x2 = 13.08, P 〈0.01 ）. However, there was no significant difference of UFA concentrations among the 3 groups [（206.29± 61.94） μ mol/L, （218.11 ± 110.28） μmol/L and （ 240.94 ± 116.79 ） μmol/L, x2 = 2.17, P 〉 0.05]. Compared to normal control [（ 355.63 ± 100.35 ）μmol/L], the FFA concentration[（421.21 ±200.83） μmol/L] in T2DM has significantly increased （x2 =FFA concentrations were higher in T2DM with hyperlipidemia [（473.04 ±213.40） μmol/L] （x2 =27.93,P 〈0.01 ）. The RSD values for intra- and inter-day precision were less than 5%, and the minimal detection limits ranged from 0.05 μmol/L to 0.35 μmol/L The recoveries of high, intermediate and low-level materials were 96.4% -104.8%. Conclusions The total FFA concentration in T2DM has increased, most of which are saturated FFA. The unsaturated FFA has not significantly increased. They seem to be related to the development of T2DM, and might be a new biomarker for clinical monitoring of metabolic disorder of T2DM.