Departament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona, Barcelona, Spain.
Departament de Bioquímica i Biologia Molecular, Facultat de Veterinària, Universitat Autònoma de Barcelona, Barcelona, Spain.
Vet Clin Pathol. 2023 Jun;52(2):252-260. doi: 10.1111/vcp.13200. Epub 2023 Feb 6.
The determination of iCa and iMg is important in veterinary medicine, but their immediate determination in whole blood is not always possible. Their stability in other sample types and the existence of interferences must be evaluated before its use.
We aimed to analyze the effects of storage time on the stability of iCa, iMg, and other analytes in whole blood, plasma, and serum samples in horses and assess the interference of heparin in these measurements.
Whole blood, heparin-plasma, and serum samples from 10 horses were stored at 4°C and analyzed 1, 2, 3, 4, 5, 6, 7, 8, 24, 48, and 168 hours after sample collection using the Stat Profile Prime Plus Vet equipment (Nova Biomedical, Waltham, MA, USA). Results were analyzed by ANOVA or mixed-effect models.
The concentration of iCa, iMg, total calcium (tCa), total magnesium (tMg), and the ratios iCa/tCa and iMg/tMg did not differ up to 168 hours when compared to the initial time. Total Ca, iMg, and tMg were not significantly different among sample types, but iCa concentrations were slightly but significantly lower in plasma. Freezing at -20°C did not affect iCa, iMg, tCa, and tMg. The pH increased in serum and plasma after 8 hours, and a mild negative correlation existed between plasma iCa concentration and pH. A negative correlation was observed also between the ratios iCa/tCa or iMg/tMg and pH in plasma and serum. A significant decrease in iCa and iMg was detected when comparing homemade syringes at high heparin concentration (~200-300 U heparin/mL) and commercial lithium-heparin tubes (20-30 U/mL).
Samples stored at 4°C can be used to determine iCa and iMg concentrations up to 7 days after collection. Other metabolites are stable for up to 8 hours; heparin interference should be taken into account if using homemade heparin syringes.
在兽医领域,钙离子(iCa)和镁离子(iMg)的检测十分重要,但并非所有情况下都能直接检测全血中的钙离子和镁离子。在将其用于临床检测前,需要评估样本的稳定性以及是否存在干扰。
分析马的全血、肝素血浆和血清样本在不同储存时间下 iCa、iMg 及其他分析物的稳定性,评估肝素对这些测量的干扰。
采集 10 匹马的全血,使用肝素抗凝管和普通采血管,室温下静置 15 分钟后,以 3000r/min 离心 10 分钟,分离得到肝素血浆和血清。将全血、肝素血浆和血清样本于 4℃储存,分别于采血后 1、2、3、4、5、6、7、8、24、48 和 168 小时使用 Nova Biomedical 公司的 Stat Profile Prime Plus Vet 设备进行分析。采用方差分析或混合效应模型对结果进行分析。
与初始时间相比,iCa、iMg、总钙(tCa)、总镁(tMg)、iCa/tCa 和 iMg/tMg 的浓度在 168 小时内均无显著差异。不同样本类型之间的总钙、iMg 和总镁浓度无显著差异,但血浆中的 iCa 浓度略低但有统计学意义。-20℃冻存不影响 iCa、iMg、tCa 和 tMg 的浓度。血清和血浆的 pH 在 8 小时后升高,且血浆 iCa 浓度与 pH 呈轻度负相关。血浆和血清中 iCa/tCa 或 iMg/tMg 与 pH 之间也存在负相关。与市售的含 20-30U/mL 肝素锂的采血管相比,当使用高浓度(~200-300U/mL)肝素的自制肝素注射器时,iCa 和 iMg 的浓度显著降低。
在采集后 7 天内,4℃保存的样本可用于 iCa 和 iMg 浓度的测定。其他代谢物在 8 小时内稳定,若使用自制肝素注射器,应考虑肝素的干扰。