B-001 牛群妊娠期血清中 25-羟基维生素 D 和 C-3 表聚体的浓度

IF 7.1 2区医学 Q1 MEDICAL LABORATORY TECHNOLOGY

Clinical chemistry Pub Date : 2024-10-02 DOI:10.1093/clinchem/hvae106.365

J P Buchweitz, S Velasquez Rivertte, J A Zyskowski, A Abuelo Sebio

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Methods In the current study, we validated an LC-MS/MS method to measure the mono-hydroxyvitamin D metabolites 25-hydroxyvitamin D2 (25(OH)D2), 25-hydroxyvitamin D3 (25(OH)D3), and 3-epi-25(OH)D3, in bovine serum. Serum was collected from dairy cows at six stages of pregnancy (n=60 (10 per group), pre-breeding heifers, 30-40 days pregnant, 70-90 days, 120-180 days, 210-260 days, and 30-45 days post-calving). The 25(OH)D metabolites were extracted from serum by supported liquid extraction (SLE) and the eluate was derivatized with 2-Nitrosopyridine. Derivatized samples were introduced to the LC-MS/MS, ionized by electrospray ionization in positive-ion mode, and detected and quantified by multiple-reaction monitoring. Results The LC-MS/MS method was linear in the concentration range of 0.25 ng/mL to 100 ng/mL with an r2 &gt; 0.99 for each analyte. 3-epi-25(OH)D3 and total serum 25(OH)D concentrations were calculated for each stage of pregnancy. 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引用次数: 0

摘要

背景在大多数动物物种中，血清 25-羟基维生素 D（25(OH)D）是维生素 D 状态的指标。最近发现了它的 C-3 表聚体，即 3-epi-25-hydroxyvitamin D3（3-epi-25(OH)D3），这在诊断上仍有混淆。据报道，孕妇和婴儿血清中都出现了这种表聚物；然而，由于这种表聚物是最近才被发现的，其生理作用、生化调节和总体生物重要性尚未完全阐明。鉴于3-epi-25(OH)D3在孕妇中的普遍存在，我们假设3-epi-25(OH)D3可作为一种新的预测动物妊娠的生物标志物。方法在本研究中，我们验证了一种 LC-MS/MS 方法来测量牛血清中的单羟维生素 D 代谢物 25- 羟维生素 D2 (25(OH)D2)、25-羟维生素 D3 (25(OH)D3) 和 3-表-25(OH)D3。从奶牛的六个怀孕阶段（n=60（每组 10 头）、配种前母牛、怀孕 30-40 天、70-90 天、120-180 天、210-260 天和产后 30-45 天）采集血清。通过支撑液体萃取（SLE）从血清中提取 25（OH）D 代谢物，并用 2-亚硝基吡啶对洗脱液进行衍生处理。衍生化后的样品进入 LC-MS/MS，在正离子模式下通过电喷雾电离，并通过多重反应监测进行检测和定量。结果 LC-MS/MS 方法在 0.25 ng/mL 至 100 ng/mL 的浓度范围内线性关系良好，各分析物的 r2 > 均为 0.99。计算了每个妊娠阶段的 3-epi-25(OH)D3 和血清总 25(OH)D 浓度。育成前小母牛的血清 25（OH）D 浓度范围为 65 - 85 纳克/毫升，在妊娠期间呈非显著增加趋势，平均值接近 100 纳克/毫升。有趣的是，3-epi-25（OH）D3 在最初的 90 天内保持在基线附近（1.3 - 1.9 纳克/毫升），之后则升高 3-4 倍。结论这项研究证实，25（OH）D3 的表聚是一个跨物种的保守生化过程。循环中 3-epi-25(OH)D3 浓度的增加虽然不能预测妊娠本身，但与妊娠中期至晚期观察到的奶牛雌激素浓度的增加是一致的。未来的研究将探索妊娠期雌激素增加与表观化之间的潜在联系。

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B-001 Serum 25-hydroxyvitamin D and C-3 epimer concentrations throughout gestation in a bovine dairy herd

Background Serum 25-hydroxyvitamin D (25(OH)D) serves as an indicator of vitamin D status in most animal species. The recent identification of its C-3 epimer, 3-epi-25-hydroxyvitamin D3 (3-epi-25(OH)D3) remains diagnostically confounding. The appearance of this epimer in blood serum has been reported for both pregnant women and infants; however, because of its recent discovery, its physiologic role, biochemical regulation, and overall biologic importance have yet to be fully elucidated. Given its prevalence in pregnant women, it was hypothesized that 3-epi-25(OH)D3 may serve as a novel predictive biomarker of pregnancy in animals. Methods In the current study, we validated an LC-MS/MS method to measure the mono-hydroxyvitamin D metabolites 25-hydroxyvitamin D2 (25(OH)D2), 25-hydroxyvitamin D3 (25(OH)D3), and 3-epi-25(OH)D3, in bovine serum. Serum was collected from dairy cows at six stages of pregnancy (n=60 (10 per group), pre-breeding heifers, 30-40 days pregnant, 70-90 days, 120-180 days, 210-260 days, and 30-45 days post-calving). The 25(OH)D metabolites were extracted from serum by supported liquid extraction (SLE) and the eluate was derivatized with 2-Nitrosopyridine. Derivatized samples were introduced to the LC-MS/MS, ionized by electrospray ionization in positive-ion mode, and detected and quantified by multiple-reaction monitoring. Results The LC-MS/MS method was linear in the concentration range of 0.25 ng/mL to 100 ng/mL with an r2 > 0.99 for each analyte. 3-epi-25(OH)D3 and total serum 25(OH)D concentrations were calculated for each stage of pregnancy. Pre-bred heifers had serum 25(OH)D concentrations ranging from 65 - 85 ng/mL with trends toward non-significant increases with mean values approaching 100 ng/mL during pregnancy. Interestingly, 3-epi-25(OH)D3 remained near baseline (1.3 - 1.9 ng/mL) for the first 90 days and elevated 3- to 4-fold thereafter. Conclusions This study confirms that epimerization of 25(OH)D3 is a conserved biochemical process across species. While not predictive of pregnancy itself, the increase in circulating 3-epi-25(OH)D3 concentrations was consistent with mid- to late-gestational increases in estrogen concentration observed for dairy cattle. Future studies will explore the potential link between increases in gestational estrogen and epimerization.

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来源期刊

Clinical chemistry 医学-医学实验技术

CiteScore

11.30

自引率

4.30%

发文量

212

审稿时长

1.7 months

期刊介绍： Clinical Chemistry is a peer-reviewed scientific journal that is the premier publication for the science and practice of clinical laboratory medicine. It was established in 1955 and is associated with the Association for Diagnostics & Laboratory Medicine (ADLM). The journal focuses on laboratory diagnosis and management of patients, and has expanded to include other clinical laboratory disciplines such as genomics, hematology, microbiology, and toxicology. It also publishes articles relevant to clinical specialties including cardiology, endocrinology, gastroenterology, genetics, immunology, infectious diseases, maternal-fetal medicine, neurology, nutrition, oncology, and pediatrics. In addition to original research, editorials, and reviews, Clinical Chemistry features recurring sections such as clinical case studies, perspectives, podcasts, and Q&A articles. It has the highest impact factor among journals of clinical chemistry, laboratory medicine, pathology, analytical chemistry, transfusion medicine, and clinical microbiology. The journal is indexed in databases such as MEDLINE and Web of Science.