A multi-laboratory validation of the extraction of fat-soluble vitamins from tissue and feed and their analysis by LC-MS/MS

PAR-18-604 VET-LIRN网络能力建设项目 从组织和饲料中提取脂溶维生素及其作用的多实验室验证 LC-MS/MS联用分析 摘要 脂溶维生素A、E和D是所有动物物种必需的饮食成分。虽然是必需的 在少量的情况下，这些化合物在所有阶段都会产生多种代谢和生理效应。 一种动物的生活。因为这些维生素或它们的前体是自然产生的，大多数是自由分布的 动物可以通过它们食用的食物或饲料来维持足够的水平。然而，加工过的谷物 以及用于生产配套、生产和特产的商业饲料的其他配料 物种面临着维生素易降解的挑战。因此，商业饲料 工业用合成形式的这些维生素补充饮食，以提供足够的营养价值。然而， 储存条件和混合误差会对消耗的最终浓度产生不利影响。作为一名 结果，营养维生素浓度可能不足，并导致临床症状缺乏或 过量，并表现出中毒的表现。临床上，缺乏症和中毒症都是可以确定的。 通过测量血清维生素A、E或维生素D代谢物25-羟基维生素来进行死前检查 D(25-羟基-D)。对于死后的动物，通常进行组织维生素分析，肝脏是 选择维生素A和E的样本，肾脏是25-羟基D的最佳样本。临床诊断表明 暗示维生素缺乏或过量通常(但不总是)暗示有潜在的问题 进食。目前解决饲料中脂溶维生素含量的方法有些繁琐、耗时， 并可能使用过时的设备。我们提出了一种提取和分析组织和饲料的方法 这是简单、准确和可在密歇根州CVM Vet-LIRN社区内的实验室之间重现的 州立大学兽医诊断实验室(MSU VDL)，堪萨斯州立大学兽医诊断 田纳西州农业部科德动物健康诊断实验室(TN) 德州农工大学兽医诊断实验室(TVMDL)，加州大学 戴维斯加州动物健康和食品安全(CAHFS)实验室和圭尔夫大学动物健康 实验室(Guelph AHL)配备了液相色谱串联质谱仪。

PAR-18-604 Vet-LIRN Network Capacity-Building Project A multi-laboratory validation of the extraction of fat-soluble vitamins from tissue and feed and their analysis by LC-MS/MS Abstract The fat-soluble vitamins A, E, and D are essential dietary components for all animal species. Although required in small quantities, these compounds exert a multitude of metabolic and physiologic effects through all stages of an animal’s life. Because these vitamins or their precursors are naturally occurring, most free ranging animals can maintain adequate levels through the browse or forage they consume. However, processed grains and other ingredients used in the generation of commercial feeds for companion, production, and specialty species are challenged with the propensity of vitamins toward degradation. Accordingly, the commercial feed industry supplements diets with synthetic forms of these vitamins to provide adequate nutrient value. However, storage conditions and mixing errors can adversely impact the final concentration that is consumed. As a result, nutrient vitamin concentrations may be either inadequate and lead to clinical signs of deficiency or excessive and exhibit manifestations of toxicosis. Clinically, both deficiencies and toxicoses can be established ante-mortem through the measurement of serum vitamin A, E, or the vitamin D metabolite, 25-hydroxy vitamin D (25-OH D). For the post-mortem animal, tissue vitamin analysis is generally performed with liver being the sample of choice for vitamins A and E, and kidney being the best sample for 25-OH D. A clinical diagnosis that is suggestive of vitamin deficiency or excess generally (but not always) implicates a potential problem with feed. Current methods to address fat soluble vitamin content in feed are somewhat onerous, time-consuming, and may utilize outdated equipment. We propose a method of extraction and analysis for both tissues and feed that is simple, accurate, and reproducible across laboratories within the CVM Vet-LIRN community, Michigan State University Veterinary Diagnostic Laboratory (MSU VDL), Kansas State University Veterinary Diagnostic Laboratory (KSU VDL), Tennessee Department of Agriculture Kord Animal Health Diagnostic Laboratory (TN Ag Lab), Texas A&M University Veterinary Medical Diagnostic Laboratory (TVMDL), University of California Davis California Animal Health and Food Safety (CAHFS) lab, and University of Guelph Animal Health Laboratory (Guelph AHL) equipped with liquid chromatography tandem mass spectrometry.