【摘要】 目的:建立适用于检测人脂蛋白单糖组成的液相色谱方法,探讨糖尿病患者与健康人脂蛋白的单糖组成差异。方法:采用柱前衍生液相色谱法检测脂蛋白上的中性和碱性单糖,液相串联质谱法检测唾液酸的含量。结果:健康人和糖尿病患者高密度脂蛋白上的甘露糖(Man)、葡萄糖胺(GlcN)、N-乙酰葡萄糖胺(GlcNAc)、葡萄糖(Glc)、半乳糖(Gal)和N-乙酰神经氨酸(NANA)的含量依次为:(5.88±0.94)、(16.49±4.11)、(1.31±0.33)、(0.87±0.16)、(7.18±1.64)、(2.14±0.12)mmol/(g protein)和(8.68±0.39)、(24.73±5.50)、(1.91±0.54)、(1.23±0.35)、(9.73±2.85)、(3.53±0.27)mmol/(g protein)。健康人和糖尿病患者低密度脂蛋白上的Man、GlcN、GlcNAc、Glc、Gal和NANA的含量依次为:(29.20±3.57)、(50.77±4.72)、(5.28±0.64)、(10.06±1.37)、(28.44±3.96)、(6.86±0.11)mmol/(g protein)和(30.08±3.78)、(38.52±6.38)、(3.79±0.78)、(7.63±1.50)、(20.05±2.63)、(6.45±0.18)mmol/(g protein);健康人和糖尿病患者极低密度脂蛋白上的Man、GlcN、Glc、Gal和NANA的含量依次为:(91.21±4.12)、(27.05±2.34)、(4 230.95±15.83)、(43.40±3.75)、(2.95±0.24)mmol/(g protein)和(82.40±0.51)、(30.16±0.32)、(4 722.73±93.27)、(34.05±2.81)、(4.42±0.15)mmol/(g protein)。结论:本文建立的柱前衍生液相色谱法适用于脂蛋白上的单糖组成分析。糖尿病患者脂蛋白的糖基化修饰与健康人相比存在显著差异。更多还原

【Abstract】 AIM: To establish a liquid chromatography method for determining the monosaccharide composition of human lipoproteins,and to investigate the differences between diabetic patients and healthy participants. METHODS:Liquid chromatography with pre-column derivatization was used to determine the neutral and basic monosaccharides,and liquid chromatography tandem mass spectrometry was applied to quantify N-acetylneuraminic acid content. RESULTS: The contents of mannose,glucosamine,N-acetylglucosamine,glucose,galactose and N-acetylneuraminic acid in high-density lipoprotein from healthy participants and diabetic patients were( 5. 88 ± 0. 94),( 16. 49 ± 4. 11),( 1. 31 ± 0. 33),( 0. 87± 0. 16),( 7. 18 ± 1. 64),( 2. 14 ± 0. 12) mmol /( g protein) and( 8. 68 ± 0. 39),( 24. 73 ± 5. 50),( 1. 91 ± 0. 54),( 1.23 ± 0.35),( 9.73 ± 2.85),( 3.53 ± 0.27) mmol/( g protein),respectively. The contents of mannose,glucosamine,N-acetylglucosamine,glucose,galactose and N-acetylneuraminic acid in low-density lipoprotein from healthy participants and diabetic patients were( 29. 20 ± 3. 57),( 50. 77 ± 4. 72),( 5. 28 ± 0. 64),( 10. 06 ± 1. 37),( 28. 44 ±3. 96),( 6. 86 ± 0. 11) mmol /( g protein) and( 30. 08 ± 3. 78),( 38. 52 ± 6. 38),( 3. 79 ± 0. 78),( 7. 63 ± 1. 50),( 20.05 ±2.63),( 6.45 ±0.18) mmol/( g protein),respectively. The contents of mannose,glucosamine,glucose,galactose and N-acetylneuraminic acid in very-low-density lipoprotein from healthy participants and diabetic patients were( 91.21 ±4.12),( 27.05 ± 2.34),( 4 230.95 ± 15.83),( 43.40 ± 3.75),( 2.95 ± 0.24) mmol/( g protein) and( 82.40 ±0.51),( 30.16 ±0.32),( 4 722.73 ±93.27),( 34.05 ±2.81),( 4.42 ±0.15) mmol/( g protein),respectively. CONCLUSION: Liquid chromatography with pre-column derivatization is suitable for the neutral and basic monosaccharide analysis in human lipoproteins,and the glycosylation of lipoproteins in diabetic patients are significantly changed compared with the healthy controls.更多还原