Evaluation of a portable fluorometer for the quantification of vitamin E in blood at key physiological stages of dairy cattle

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Abstract

Vitamin E is essential in mitigating the impact of oxidative stress on periparturient dairy cows and neonatal calves. Therefore, it is essential to measure circulating vitamin E concentrations accurately. Currently, the only reliable method is an expensive and time-consuming procedure using liquid chromatography-mass spectrometry (LC-MS). However, a cheaper and faster method has been developed, which allows the quantification of circulating vitamin E through the use of a handheld fluorometric analyzer (HFA) called the vitamin E iCheck (BioAnalyt GmbH). Our objective was to compare the accuracy of the HFA to the reference LC-MS method for measuring vitamin E in bovine samples. A total of 177 samples collected for other studies were used: 98 newborn calf serum samples from a vitamin E supplementation study (including treated and control animals) and 79 whole-blood samples from cows 1 to 7 d postcalving. Vitamin E concentrations were measured on thawed calf serum and fresh cow EDTA blood using the HFA, following the manufacturer's instructions. Whole blood from cows was then centrifuged to obtain plasma. Vitamin E was also quantified in calf serum and cow plasma at the Michigan State University Veterinary Diagnostic Laboratory using LC-MS. Calf and cow results were analyzed separately because they represent different biological matrices and physiological times. In each dataset, results between the HFA and LC-MS determinations were compared using Passing-Bablok regressions and Bland-Altman plots. The HFA showed a poor linear relationship with LC-MS for calf serum and cow plasma (intercept = 0.33 and 0.67 μg/mL, respectively). The HFA unreliably estimated vitamin E, with a mean bias of −3.2 and 0.6 μg/mL for calves (vitamin E concentration range: 0.28 to 30.75 μg/mL) and cows (0.8 to 5.88 μg/mL), respectively. Moreover, 40.4% of the calf samples read below the linear range of acceptable results for the HFA, making it unsuitable for this age group. Hence, under the conditions of our study, the HFA yielded unreliable results and cannot be recommended for field use.

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对便携式荧光计在奶牛关键生理阶段定量检测血液中维生素 E 的评估
维生素 E 对于减轻氧化应激对围产期奶牛和新生犊牛的影响至关重要。因此,准确测量循环中维生素 E 的浓度至关重要。目前,唯一可靠的方法是使用液相色谱-质谱法(LC-MS),这种方法既昂贵又耗时。不过,一种更便宜、更快速的方法已经开发出来,即使用一种名为维生素 E iCheck(BioAnalyt GmbH)的手持式荧光分析仪(HFA)来定量检测循环中的维生素 E。我们的目的是比较 HFA 和 LC-MS 方法在测量牛样本中维生素 E 方面的准确性。我们总共使用了 177 份为其他研究收集的样本:其中 98 份新生犊牛血清样本来自一项维生素 E 补充剂研究(包括处理过的动物和对照组动物),79 份全血样本来自产后 1 到 7 天的奶牛。按照制造商的说明,使用 HFA 测量解冻的犊牛血清和新鲜奶牛 EDTA 血液中的维生素 E 浓度。然后离心奶牛全血,获得血浆。密歇根州立大学兽医诊断实验室还使用 LC-MS 对犊牛血清和奶牛血浆中的维生素 E 进行了量化。由于犊牛和奶牛代表不同的生物基质和生理时间,因此分别对它们的结果进行了分析。在每个数据集中,使用 Passing-Bablok 回归和 Bland-Altman 图对 HFA 和 LC-MS 测定结果进行比较。对于小牛血清和奶牛血浆,氢氟烷烃与 LC-MS 的线性关系较差(截距分别为 0.33 和 0.67 μg/mL)。氢氟烷烃对维生素 E 的估计不可靠,犊牛(维生素 E 浓度范围:0.28 至 30.75 μg/mL)和奶牛(0.8 至 5.88 μg/mL)的平均偏差分别为-3.2 和 0.6 μg/mL。此外,40.4% 的犊牛样本读数低于 HFA 可接受结果的线性范围,因此不适合该年龄组的犊牛。因此,在我们的研究条件下,氢氟烷烃的结果并不可靠,不能推荐在现场使用。
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JDS communications
JDS communications Animal Science and Zoology
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