| 吉素娜,王 芳,张盼望,陈 越,杨长伟,王 冰.LC-MS/MS法检测猪内脏和肌肉中N-乙酰神经氨酸、N-羟乙酰氨酸和脱氨神经氨酸浓度的研究[J].现代生物医学进展英文版,2017,17(4):610-614. |
| LC-MS/MS法检测猪内脏和肌肉中N-乙酰神经氨酸、N-羟乙酰氨酸和脱氨神经氨酸浓度的研究 |
| Determination for the Contents of Neu5Ac, Neu5Gc and KDN in Organ Meats and Muscle of Pigs by LC-MS/MS |
| Received:April 26, 2016 Revised:May 21, 2016 |
| DOI：10.13241/j.cnki.pmb.2017.04.003 |
| 中文关键词: 液相色谱-三重四级杆质谱联用 N-羟乙酰神经氨酸 N-乙酰神经氨酸 脱氨神经氨酸 猪内脏组织 |
| 英文关键词: LC-MS/MS Neu5Ac Neu5Gc KDN Pigs organ |
| 基金项目:国家自然科学基金项目（31271038） |
|
| Hits: 522 |
| Download times: 260 |
| 中文摘要: |
| 摘要 目的：建立液相色谱-三重四级杆质谱联用法（LC-MS/MS）测定猪内脏和肌肉组织中唾液酸N-乙酰神经氨酸(Neu5Ac)、N-羟乙酰神经氨酸(Neu5Gc)和脱氨神经氨酸（KDN）浓度的分析方法。方法：将内脏肉组织匀浆后，利用三氟乙酸（TFA）将结合态的唾液酸从糖脂或糖蛋白链上解离下来后进样分析。色谱条件：色谱柱为ZORBAX Eclipse Plus C18（5 μm，4.6×250 mm，Waters），流动相为0.1 % 醋酸铵-100 %乙腈（9:1），柱温：25℃，流速800 μL?min-1，进样量20 μL。质谱条件：离子源为电喷雾化离子源（ESI），扫描方式为多重反应监测（MRM），监测离子对：Neu5Ac：307.7→87.0，Neu5Gc：323.3→116.0，KDN：266.7→87.0，13C3Neu5Ac内标品：310.9→90.0。结果：Neu5Ac、Neu5Gc和KDN分别在0.1~20.0、0.05~10.00和0.005~1.000 μmol?L-1的浓度范围内和对照品与内标峰面积比值线性关系良好，重复性平均RSD为1.2 %，稳定性平均RSD为1.9 %，日间和日内精密度试验RSD均小于6.7 %，平均回收率为92.9~106.4 %。结论：本方法简便、快速、灵敏度高，可广泛运用于组织和体液中唾液酸的测定。 |
| 英文摘要: |
| ABSTRACT Objective: To establish an LC-MS/MS method for the detection of N-acetylneuraminic acid (Neu5Ac), N-glycolylneuraminic acid (Neu5Gc) and 2-keto-3-deoxy-D-glycero-D-galactonononic acid (KDN) in organ meats and muscle of pigs. Methods: Tissues were acid hydrolyzed with trifluoroacetic acid (TFA) to get ganglioside-bound and glycoprotein-bound sialic acid, followed by injected and analyzed. The analysis was performed on an ZORBAX Eclipse Plus C18 (5 μm, 4.6×250 mm, Waters) column, and the mobile phase consisted of 0.1 % ammonium-100 % acetonitrile (9:1), the flow rate was 800 μL/min, the temperature was maintained at 25 ℃, injection volume was set up 20 μL. Negative electrospray ionization (ESI) and multiple reaction monitoring (MRM) mode was used, MRM: Neu5Ac: 307.7→87.0, Neu5Gc: 323.3→116.0, KDN: 266.7→87.0, 13C3Neu5Ac (IS): 310.9→90.0. Results: The ratio of internal with standard sample peak area and content of Neu5Ac, Neu5Gc and KDN showed good linearity in the range of 0.1~20.0, 0.05~10.00, and 0.005~1.000 μmol/L, RSDs of the repeatability and stability were 1.2 % and 1.9 %, Intra- and inter-day RSDs were all lower than 6.7%, the recoveries of sialic acid were 92.9~106.4%. Conclusion: The established method is simple, accurate and rapid, higher sensitivity and can be applied to determination of Neu5Gc, Neu5Ac and KDN in tissues and fluid of animals and human. |
|
View Full Text
View/Add Comment Download reader |
| Close |
|
|
|
