Analytical Methodology Development

分析方法开发

• 批准号: 6542069
• 负责人: NADJA REHAK
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6542069
• 关键词: analytical chemistry; analytical method; anticoagulants; blood chemistry; blood tests; centrifugation; chemical stability; chemical structure function; enzyme induction /repression; heparin; human subject; lithium; physical separation; plasma; sample collection; serum; technology /technique development; temperature; tissue /organ preservation; analytical chemistry; analytical method; anticoagulants; blood chemistry; blood tests; centrifugation; chemical stability; chemical structure function; enzyme induction /repression; heparin; human subject; lithium; physical separation; plasma; sample collection; serum; technology /technique development; temperature; tissue /organ preservation

Plasma is often the preferred specimen type when the goal of laboratory is to reduce the turnaround time and clotting occurences of the analytical systems. Recently, a blood collection tube containing Li heparin as anticoagulant and inert gel material to separate the blood cells from plasma, was introduced for routine laboratory use (Becton Dickinson Vacutainer? Plus, # 7961).We evaluated this plasma separator tube (PST) with Hitachi 917 for direct sampling and storage effect on 27 commonly measured analytes. Blood was collected into PST and serum separator tubes (SST) (Becton Dickinson Vacutainer Plus #7983) from volunteers (n=30), and for additional studies, into Li heparin anticoagulated tubes (PT) (Becton Dickinson Vacutainer Plus #7886). All tubes were centrifuged at the routine setting for the SST processing (RCF=2190xg, 5 min) within 30 min of collection. Samples were analyzed in batch mode from the primary tubes with Hitachi 917 fresh, and after storage for 8 hours at room temperature (rt), and up to 5 days at 4 oC. Significance of the mean difference (mdiff) was based on the imprecision of the method. In comparison with fresh serum, fresh plasma collected in PST had significantly different (mdiff1=plasma-serum) concentration of K (mdiff1=-0.23 mmol/L), albumin (mdiff1=0.1 g/dL), total protein (mdiff1=0.13 g/dL) and activity of ALT (mdiff1=-7 U/L) and LD (mdiff1=25 U/L). None of the measured analytes were affected by the storage of samples in PST and SST for 8 hours at rt. The effect of extended storage at 4 oC (mdiff2=stored-fresh) on plasma analytes of PST specimens was ignificant for bicarbonate (-5 mmol/L), K (0.9 mmol/L), Pi (-0.5 mg/dL), glucose (-7 mg/dL) and LD (39 U/L). In comparison, storage of serum in SST affected only LD activity (-10 U/L). The LD activity for fresh PST plasma was comparable to that for fresh PT plasma (mean: 180 and 174 U/Lrespectively). The extended storage of plasma in PST and PT at 4 oC caused an increase in the activity of LD (PST: mdiff2=30 U/L , PT: mdiff2=24 U/L), indicating incomplete separation of platelets by the gel barrier in PST. However, the PST plasma LD isoenzymes were not affected by storage at 4 oC. In addition, the increase in the PST plasma LD activity was not related to the centrifugation time (5, 8, and 10 min), or to the plasma platelet count. However, we observed that all centrifuged PSTs had varying amounts of erythrocytes imbedded in the gel and/or on top of the gel. This strongly suggests that the observed changes in LD and K were due to incomplete separation of plasma from red blood cells. In conclusion, we found PST to be acceptable for blood collection of heparinized plasma when appropriate changes in reference intervals for ALT, LD, and K are implemented. Furthermore, most of the common analytes are stable for up to 8 hours at ambient room temperature. However, the gel barrier does not provide satisfactory separation of plasma and cells and, therefore, the extended storage of plasma in primary tubes at 4 oC is not advisable.

当实验室的目标是减少分析系统的周转时间和凝结的情况时，血浆通常是首选的样品类型。最近，引入了一个含有液肝素的血管，该管子是抗凝剂和惰性凝胶材料，以将血细胞与血浆分开，以供常规实验室使用（Becton Dickinson vacutainer？plus，＃7961）。我们评估了该血浆分离器管（PST）与Hitachi 917的血浆分离器管（PST），以直接采样和储存分析和储存率分析27.27常见的效果。将血液从志愿者（n = 30）（n = 30）中收集到PST和血清分离器管（SST）（SST）（Becton Dickinson Vacutainer Plus＃7983），以及其他研究中，进入LI肝素抗凝管（PT）（Becton Dickinson Vacinson Vacutainer Plus＃7886）。在收集30分钟内，在SST处理的常规设置（RCF = 2190xG，5分钟）的常规设置下，将所有试管离心。从Hitachi 917 Fresh的主要试管中分析样品，并在室温（RT）储存8小时后，最多5天以4 oC为单位。平均差异（MDIFF）的重要性是基于该方法的不精确。 In comparison with fresh serum, fresh plasma collected in PST had significantly different (mdiff1=plasma-serum) concentration of K (mdiff1=-0.23 mmol/L), albumin (mdiff1=0.1 g/dL), total protein (mdiff1=0.13 g/dL) and activity of ALT (mdiff1=-7 U/L) and LD (mdiff1=25 u/l）。在RT时，未通过样品在PST和SST中的样品存储8小时都没有影响所有测量的分析物。对于碳酸氢盐（-5 mmol/l），K（0.9 mmol/L），PI（-0.5 mg/dl），Glucose（-7 mg/dl）和LD（-7 mg/dl）和LD（39 u/l），碳酸氢盐（-5 mmol/L），K（-5 mmol/l），k（0.9 mmol/l），k（-5 mmol/l）和pi（-7 mmol/l）和ld（39 u/l），在4 OC（mdiff2 =储存的Fresh）对PST标本的血浆分析物对PST标本的血浆分析物的影响。相比之下，SST中血清的存储仅影响LD活性（-10 U/L）。新鲜PST等离子体的LD活性与新鲜PT血浆相当（平均：180和174 U/Lesprectity）。 PST中血浆的扩展存储在4 OC中的延长导致LD活性增加（PST：MDIFF2 = 30 U/L，PT：MDIFT2 = 24 U/L），表明PST中的凝胶屏障将血小板分离不完全。但是，PST等离子体LD同工酶不受4 oC的存储影响。另外，PST血浆LD活性的增加与离心时间（5、8和10分钟）或血浆血小板计数无关。但是，我们观察到所有离心的PST的凝胶和/或凝胶顶部嵌入的红细胞含量不同。这强烈表明，LD和K的观察到的变化是由于血浆与红细胞的分离不完全。总之，我们发现当实施ALT，LD和K的参考间隔时，PST可以接受肝素化血浆的血液收集。此外，大多数常见分析物在环境室温下最多可容纳8小时。但是，凝胶屏障不能提供血浆和细胞的令人满意的分离，因此，不建议在4 OC下将血浆扩展在原代管中。