Anticoagulant Solution Research Articles

Case Report: Chronic Lymphocytic Leukemia with a Monoclonal Immunoglobulin M λ Lymphocyte Agglutinin

A case of chronic lymphocytic leukemia is described in which peripheral blood films showed lymphocyte agglutination. A serum factor responsible for the agglutination was demonstrated. The factor was dependent upon the presence of anticoagulant solutions and was more active at room temperature than at 37 degrees C. It could be identified as a monoclonal immunoglobulin M. This mechanism has not been previously described in lymphocyte agglutination.

An update on solutions for red cell storage

Anticoagulant and nutrient solutions allow red blood cells to be stored and transported, enabling modern blood banking. The development of these solutions has been slow, covering 90 years, and the reasons for past formulations are best understood in a historical context. Modern red cell storage solutions work well for blood banks, allowing 5-7-week storage, which means more than 90% of collected units find a recipient. Improved scientific understanding of the red cell storage lesion has shown a way to make even better storage solutions, which maintain red cell metabolism and reduce membrane loss.

Catheter lock solutions influence staphylococcal biofilm formation on abiotic surfaces

Microbial biofilms form on central venous catheters and may be associated with systemic infections as well as decreased dialysis efficiency due to catheter thrombosis. The most widely used anticoagulant catheter lock solution in the US is sodium heparin. We have previously shown that sodium heparin in clinically relevant concentrations enhances Staphylococcus aureus biofilm formation. In the present study, we examine the effect of several alternative catheter lock solutions on in vitro biofilm formation by laboratory and clinical isolates of S. aureus and coagulase-negative staphylococci (CNS). Lepirudin, low molecular weight heparin, tissue plasminogen activator, sodium citrate, sodium citrate with gentamicin and sodium ethylene diamine tetra-acetic acid (EDTA) were assessed for their effect on biofilm formation on polystyrene, polyurethane and silicon elastomer. Sodium citrate at concentrations above 0.5% efficiently inhibits biofilm formation and cell growth of S. aureus and Staphylococcus epidermidis. Subinhibitory concentrations of sodium citrate significantly stimulate biofilm formation in most tested S. aureus strains, but not in CNS strains. Sodium EDTA was effective in prevention of biofilm formation as was a combination of sodium citrate and gentamicin. Low molecular weight heparin stimulated biofilm formation of S. aureus, while lepirudin and tissue plasminogen activator had little effect on S. aureus biofilm formation. This in vitro study demonstrates that heparin alternatives, sodium citrate and sodium EDTA, can prevent the formation of S. aureus biofilms, suggesting that they may reduce the risk of biofilm-associated complications in indwelling catheters. This finding suggests a biological mechanism for the observed improvement in catheter-related outcomes in recent clinical comparisons of heparin and trisodium citrate as catheter locking solutions. A novel and potential clinically relevant finding of the present study is the observation that citrate at low levels strongly stimulates biofilm formation by S. aureus.

Detection of blood coagulation and clot formation using quantitative ultrasonic parameters

The ultrasonic parameters of sound velocity, attenuation and integrated backscatter were applied to detect the process of coagulation and clot formation in porcine blood. Fresh porcine blood containing 15% anticoagulant solution was collected. Blood samples with a hematocrit of 45% were obtained by reconstituting the packed erythrocytes with the separated plasma for ultrasound measurements performed with a 10-MHz focused transducer. A 24-mL aliquot of blood was placed in a container and 12 mL of 0.2 mol/L CaCl 2 solution was added to induce clot formation. In each measurement, radio-frequency signals of the blood digitized at 100 MHz were collected for 50 min at a temporal resolution of 1 A-line per s. Results showed that all of the parameters increased within the initial 3 min and, then, immediately decreased dramatically as the CaCl 2 solution was added. Subsequently, the sound velocity gradually increased with time and the integrated backscatter and attenuation increased in accordance with blood coagulation until approximately 500 and 2600 s, respectively. The integrated backscatter, attenuation and sound velocity can be divided into different stages, including red cell aggregation, reduction in hematocrit, blood coagulation and clot formation, corresponding to variations in the physical and chemical properties of the blood. The integrated backscatter, attenuation and sound velocity increased because of the changes in blood properties during the process of coagulation and clot formation: by 8.2 dB, 0.65 dB/cm, and 0.6%, respectively. These results provide a feasibility for further applying ultrasonic parameters to in vivo monitor the progress of clotting and thrombosis research. (E-mail: shyhhau@cycu.edu.tw)

The use of percutaneous autologous bone marrow transplantation in nonunion and avascular necrosis of bone

During the development of normal bone in the young child, osteoblasts and then haematopoietic stem cells progressively colonise the cartilaginous matrix, resulting in its ossification to bone matrix.[1][1] It is at this time that red bone marrow appears in the skeleton, starting distally in the foot

Loss of Catheter Locking Solution Caused by Fluid Density

The avoidance of clotting in catheter lumina between treatments usually entails locking with an anticoagulant solution such as heparin. In a previous work, it was shown that approximately 20% of locking solution flows from the catheter during instillation of the lock equal in volume to the lumen volume. Furthermore, the locking solution may spill into the blood stream under the influence of gravity. This work investigates the influence of density and viscosity of the locking solution on the volume and speed of locking solution loss from the catheter lumen. A large fraction of the catheter locking solution spills under the influence of gravity if the locking solution's density is higher than the fluid it spills to (blood). Locking solution lost is replaced by blood. Viscosity delays this process, but at 90 minutes after injection, the loss is completed even when highly viscous lock solutions are used. Slow administration of the lock has negligible influence upon the dynamics of the loss.

Washing Stem Cells Cryopreserved in Dimethyl Sulfoxide (DMSO) before Infusion Prevents DMSO-Related Side Effects with Little Loss of CD34 Positive Cells and Prompt Engraftment.

Autologous peripheral blood stem cells are harvested and cryopreserved for weeks to months or even years in advance of the autologous peripheral blood stem cell transplant (PBSCT). The stem cells are cryopreserved in dimethyl sulfoxide (DMSO), a cryoprotectant agent that prevents ice crystals from forming and puncturing the stem cell membranes during the freezing process. Infused with the thawed cells, the DMSO can cause nausea and vomiting, headaches, seizures, renal insufficiency and stem cell death. We report on the results of 103 consecutive PBSCT, in which the DMSO was washed out before reinfusion. Viability, CD 34+ cell recovery as well as clinical endpoints such as time to engraftment (neutrophil and platelet counts) and infusion reactions for the 103 cases were assessed. The cryoprotectant solution used consisted of either 10% DMSO or a combination of 5% DMSO with 6% Hydroxyethyl starch. The stem cells were frozen using a rate-controlled freezer (Cryomed™ 1010) to −180°C and then stored in the liquid phase of liquid nitrogen (kept at approximately −190°C) for up to 4.7 years (mean duration of 42 days) following collection. The sample was thawed rapidly in a 37°C water bath and then washed manually with a washing solution of 0.9% Normal Saline (NS) and 10% anticoagulant citrate dextrose solution. After the sample was resuspended, it was centrifuged (RC-3, Sorvall™) and then suspended again in NS before infusion. Endpoints examined included CD 34+ cell recovery, viability, infusion reactions, and febrile episodes within 24 hours post-infusion, time to engraftment of platelets and neutrophils, highest serum creatinine (Cr) within 4 days following the transplant, and length of hospital stay. Automated WBC differentials were performed on a Beckman Coulter™ AcT Diff Analyzer and manually (when the WBC count was < 1000), using Turk's solution and a hemocytometer. Viability was assessed by exclusion of 2% Trypan blue solution. Mean CD 34+ cell recovery and viability were 85.4% and 79.38%, respectively. The mean serum Cr was 1.1. Time to neutrophil recovery was at day 12.4 (neutrophil count of 500 /uL) and platelet engraftment (defined as a platelet count of 20,000 /uL for three consecutive days without transfusion) was 14.9. Only 21% of patients experienced reactions during infusion, which included chills/rigors (9), fever (6), nausea/emesis (6), diarrhea (1), unusual body tightness or pain (3). There was also a case of mild hypotension during the infusion. 35 patients experienced neutropenic fevers during the 24-hour period post-infusion. All patients engrafted. Mean length of hospital stay was 19.4 days. In this study, fewer than 22% of the cases of PBSCT experienced infusion-related reactions. Most of these reactions were comparatively minor (chills/rigors). The percent CD 34+ cell recovery and viability were only slightly lower than historical infusions - done immediately after thawing, without washing out the DMSO. The data suggest that DMSO can be washed out of the stem cell product without notable loss of CD 34+ cells or a drop in their viability. Many DMSO-related side effects are, thus, eliminated.

Core decompression with marrow stem cells

Association of core decompression with technique of bone graft is frequent and many different techniques have been described. In this series, grafting was done with autologous bone marrow obtained from the iliac crest of patients operated for hips osteonecrosis. The bone marrow was harvested under general anesthesia. A beveled metal trocar of 6 to 8 cm in length and a bore of 1.5 mm was pushed deep into the cancellous bone. A 10-mL syringe that has been flushed with heparin is used to aspirate the marrow. Once the needle has been inserted to the desired depth, the tip is swept around a full circle in 45° steps, with the bevel pointing in different directions at each step. This procedure is continued until a sufficient quantity of bone marrow has been harvested (150 mL). All the marrow aspirated is discharged into a plastic collection bag containing ACD (acid citrate dextrose) anticoagulant solution. It is then filtered, to remove fat aggregates and clots. The aspirated marrow was reduced in volume by concentration and injected in the femoral head after core decompression with a small trocar. To measure the number of progenitor cells transplanted, we used the fibroblast colony forming unit as an indicator of the stromal cell activity and performed in vitro cultures of the fibroblast progenitor cells. The average volume of bone marrow aspiration was 147 mL ± 12 mL per hip. The number of progenitor cells was average 12.4 ± 3.4 per 10 6 bone marrow nucleated cells and the number of nucleated cells was estimated to be 16.4 million cells per milliliter of bone marrow.

The role of depletion of dimethyl sulfoxide before autografting: on hematologic recovery, side effects, and toxicity

Cryopreservation of stem cells after collection from peripheral blood or bone marrow for autologous transplantation necessitates protection with dimethyl sulfoxide (DMSO). Unfortunately, DMSO, when infused with the thawed cell suspension, may induce serious complications and side effects. To assess whether depletion of DMSO before autografting affects safety and efficacy, 56 consenting consecutive patients treated with high-dose chemotherapy and autologous blood stem cell transplantation were assigned to obtain either an untreated or DMSO-depleted autograft. On the day of transplantation, the cryopreserved cells were thawed and infused to the patient either immediately or after washing 3 times in normal saline supplemented with 6% anticoagulant citrate dextrose solution. Cell count with viability, clonogenic assay, and phenotyping were performed before and after thawing and after washing. Hematologic recovery, side effects, and complications were recorded. The in vitro and clinical data on 56 patients show that the depletion of DMSO in vitro before autografting does not induce a significant loss of cell number, viability, colony-forming unit-granulocyte-macrophage activity, or number of CD34 + cells. Furthermore, it leads to a safe and sustained engraftment. The complications and side effects, as recorded by continuous monitoring, were substantially less; however, the procedure takes 3 to 4 hours of laboratory work per patient.

ＣＰＤ液で採血した血液の全血処理型白血球除去ＷＢＦフイルタークローズドシステムの性能評価

To prevent the post-transfusion reactions in transfusion therapy such as HLA alloimmunization, non-hemolytic febrile transfusion reaction (NHFTR) and Cytomegalovirus transmission, bedside leukocyte reduction has been performed. However, it has become clear that prestorage leukocyte reduction has greater efficiency than poststorage, and prestorage leukocyte reduction is now being considered for implementation. We developed a prestorage leukocyte reduction system that contains CPD anticoagulant solution (PLD-C system). In this study, the performance of the PLD-C system was examined with regard to the leukocyte filtration efficacy and the characteristics of blood during storage by comparison with a pre-storage leukocyte reduction system which uses ACD-A solution (PLD-A system) and blood components collected using CPD solution and prepared by the buffy-coat depleted method (BCPD).It was confirmed that the leukocyte reduction ratio of the PLD-C system (more than-4log) was higher than that of the BCPD (less than-0.4log). Platelet depletion rates of the PLD-C group (average more than 99%) was also higher than in the BCPD group, but the red cell recovery ratio was the same between groups (average 88%). Prestorage filtration showed no adverse effects such as hemolysis, whereas various good effects were observed, such as inhibition of macroaggregate formation and hemolysis during storage. Collecting blood into CPD solution improved red cell quality during storage and increased plasma volume compared to the ACD-A solution.It was confirmed that the PLD-C system has advantages in the production of blood components over the present system. Use of this system can be expected to prevent post transfusion reactions.

Pediatric convective hemofiltration: Normocarb replacement fluid and citrate anticoagulation

Background: Continuous venovenous hemofiltration (CVVH) may be preferable over continuous venovenous hemodialysis (CVVHD) in sepsis; however, CVVH use is limited by the lack of commercially produced bicarbonate replacement solutions in the United States. Anticoagulation used for hemofiltration traditionally has been either heparin or citrate based, but to date, citrate protocols have been used predominately in CVVHD. When CVVH has been used with citrate anticoagulation, replacement solutions have had to be custom made in the pharmacy. Methods: We describe a protocol used in a series of 9 children with sepsis who underwent CVVH with the use of a commercially available bicarbonate solution (Normocarb; Dialysis Solutions Inc, Richmond Hill, Ontario, Canada) and a commercially available citrate solution (Anticoagulant Citrate Dextrose Solution Formula A [ACD-A]; Baxter Health Care, Deerfield, IL). This simplified approach to citrate anticoagulation with CVVH used Normocarb as a prefilter replacement solution under provisions of the 1997 Food and Drug Administration Modernization Act. Results: All patients had acceptable levels of solute and ultrafiltration clearance with this approach. CVVH circuit life averaged 71 hours (range, 37 to 131 hours), influenced in part by the industry recommendation to change the circuit at 72 hours, whereas no complications from this protocol were identified. Conclusion: Similar to CVVHD, this protocol is effective and simple in its components, using industry-produced solutions, avoiding the need for pharmacy-made solutions, and limiting cost and potential risk for compounding errors. Although not the purpose of this report, the patient survival rate was 55%.

Overspill of catheter locking solution: safety and efficacy aspects.

Avoidance of clotting in catheter lumens between treatments usually entails locking with an anticoagulant solution such as heparin. Catheter manufacturers specify internal volume of the catheters, and it is commonly assumed that injection of this precise filling volume is safe and efficient. However, it has been shown that activated partial thromboplastin time (aPTT) increases after instillation of the heparin lock volume specified by the manufacturer. We have investigated this phenomenon in vitro using dye and saline dilution and found that injection of the catheter volume results in spillage of up to 15% of the catheter volume into the patient's blood. Depending on fluid dynamics of the instillation process and catheter compliance, the concentration of the locking solution at the catheter tip is much reduced. As a consequence of this overshoot, the mean concentration at the tip is reduced to approximately 90%. In conclusion, the injection volume must exceed 120% of the catheter lumen for achieving the full strength of the locking solution at the tip. The overshoot may have clinical consequences, (e.g., bleeding if highly concentrated heparin is used).

Improved preservation of coagulation factors after pre-storage leukocyte depletion of whole blood

Plasma and red blood cell quality are affected both by citrate concentration and the levels of extracellular leukocyte and platelet derived substances, accumulated during storage of blood. The effect of leukocyte filtration on the storage stability of whole blood was therefore studied in blood collected in standard CPD and 0.5CPD (CPD with half strength citrate concentration). A total of 52 units, 12 of them with reduced citrate concentration, were leukocyte-filtered with Pall ® whole blood filter (WBF1 or 3). No differences in leukocyte or platelet reduction were observed with the two citrate concentrations. However, with 0.5CPD a significantly longer filtration time and increased complement activation was observed. The effect of pre-storage leukocyte filtration on the plasma quality of whole blood was therefore only studied with standard CPDA1 anticoagulant solution (normal strength citrate concentration). Leukocyte filtration did not affect the von Willebrand factor concentration, while a small reduction (7%, p=0.04) in factor VIII (FVIII) concentration was observed. During storage, however, FVIII decreased more slowly in the filtered than in the unfiltered product, and, from day two, the FVIII content was significantly higher in the filtered product (46% versus 30% at 28 days, p<0.001). Factor V (FV) demonstrated a 16% reduction ( p<0.001) upon filtration, followed by an additional 8% in the next 24 h and only a 4% reduction the next 27 days, while unfiltered products demonstrated a continuous reduction to 26% at 28 days. While the β-thromboglobulin (β-TG) concentration significantly increased (from 836 to 2483 IU/ml, p<0.001) during leukocyte filtration, no further increase was observed during storage. In contrast, unfiltered products demonstrated an increase to 5762 IU/ml ( p<0.001) at 14 days, followed by a slight, not significant, reduction. This indicates platelet activation during filtration and explains a parallel reduction in FV. Filtration induced no increase in prothrombin fragment 1 + 2, while a slight increase was observed in some unfiltered products after 28 days of storage. Pre-storage leukocyte depletion thus improves the coagulation factor content of plasma in stored whole blood.

Simultaneous HPLC quantification of monoamines and metabolites in the blood-free rat cochlea

Monoamine quantification in peripheral sensory receptors, such as the cochlea, is of major interest since monoamines could play a role in neurotransmission. A three-step biochemical protocol was developed to analyze monoamine content within the cochlea. Removal of the blood by aortic perfusion was carried out with an anticoagulant solution prior to the dissection of the cochlea from the temporal bone. The cochlear monoamines and some of their metabolites were then quantified, from homogenated cochlear tissue, by a new application of high performance liquid chromatography coupled to electrochemical detection. This method demonstrated enough sensitivity to detect norepinephrine (NE), dopamine (DA), serotonin (5-HT) and some of their metabolites (3,4-dihydroxyphenylacetic acid, DOPAC; homovanillic acid, HVA; and 5-hydroxyindole-3-acetic acid, 5-HIAA). Furthermore, it enabled the demonstration of noise-induced changes in the cochlear concentrations of NE, DA, DOPAC and HVA. In addition, the aortic perfusion allowed removal of the blood-borne 5-HT from the cochlea without inducing systemic alterations or monoamine degradation, as shown by the absence of effects on NE, DA, DOPAC, HVA or 5-HIAA concentrations. The present methodology may constitute a useful strategy to analyze monoamine turnover in the cochlea and other peripheral sensory receptors.

CATHETER LOCKING SOLUTIONS

Avoidance of clotting in catheter lumens between treatments usually entails locking with an anticoagulant solution such as heparin. Catheter manufacturers specify internal volume of the catheters and it is commonly assumed that injection of this precise volume is safe and efficient. However, it has been shown that aPTT increases after instillation of the heparin lock volume specified by the manufacturer. We have investigated this phenomenon in-vitro, employing dye and saline dilution and found that injection of the catheter volume results in spillage of up to 15 % of the catheter volume into the patient's blood. Depending on fluid dynamics of the instillation process and catheter compliance, the concentration of the locking solution at the catheter tip is much reduced. Side holes contribute to the initial loss of the locking solution during injection is well as to the subsequent loss to flowing blood. Thus, the locking solution composition and instillation methods are relevant safety issues. These aspects are especially important for antimicrobial or antibiotic catheter locking solutions intended to prevent catheter related infection. Antibiotics and antibiotic/anticoagulant mixtures have the potential to promote bacterial resistance. Concentrated citric acid was sold previously as a lock solution but was withdrawn in the US market after a fatal overdose. In order to achieve uniform concentration of the locking solute throughout the catheter lumen, “overfilling” is necessary which is only sate if the systemic effect of the locking solution is negligible.

Structure of a neutral glycosphingolipid recognized by human antibodies in polyagglutinable erythrocytes from the rare NOR phenotype.

NOR is a rare inheritable polyagglutination phenomenon that has been described in two families. Our recent studies on these erythrocytes showed they contained at least two unique neutral glycosphingolipids, and based on their reactivity with Griffonia simplicifolia IB4 (GSL-IB4) isolectin (Kusnierz-Alejska, G., Duk, M., Storry, J. R., Reid, M. E., Wiecek, B., Seyfried, H., and Lisowska, E. (1999) Transfusion 39, 32-38), both oligosaccharide chains terminated with an alpha-galactose residue. The reactivity with GSL-IB4 suggested that these oligosaccharide chains terminated with a Galalpha1-->3Gal- sequence and that anti-NOR agglutinins were common human anti-Galalpha1-->3Gal xenoantibodies. In this report we describe the structure of one NOR component (NOR1) that migrated on thin-layer chromatographic plates in the region of pentaglycosylceramides. Treatment of this sample with alpha-galactosidase and beta-N-acetylhexosaminidase was followed by high-performance thin-layer chromatography with product detection by lectins and the anti-Gb4 monoclonal antibody. The results suggested that NOR1 was an alpha-galactosylated Gb4Cer with a beta-N-acetylhexosaminidase-resistant GalNAc residue. Gas phase disassembly by ion trap mass spectrometry analysis showed the sequence to be Hex1-->4HexN1-->3Hex1-->4Hex1-->4Hex linked to a ceramide composed of C18 sphingosine and a C24 monounsaturated fatty acid. Together these data indicate NOR1 to be a novel Galalpha1-->4GalNAcbeta1-->3Galalpha1-->4Galbeta1-->4 Glc-Cer structure. Additionally it has been shown that NOR glycolipids are recognized by human antibodies that were distinct from the known anti-Galalpha1-->3Gal xenoantibodies.

Blood support for pediatric surgery.

Increasingly complicated surgeries are being performed on neonatal and pediatric patients. Provision of a safe and adequate blood supply is essential for the success of many of these surgeries. Depending on the clinical situation, autologous and/or allogeneic blood may be used. However, in all cases, every attempt should be made to minimize the number of donor exposures to reduce the risk of transfusion transmitted infections. Transfusion of neonatal and pediatric patients requires additional considerations too, such as the risk of graft vs host disease, cytomegalovirus infection, the effects of various preservative anticoagulant solutions, electrolyte levels during blood storage, and wheather or not leukoreduced components are indicated.

Measurement of the Concentrations of Glucose and Citric Acid in the Aqueous Solution of a Blood Anticoagulant Using near Infrared Spectroscopy

Near infrared (NIR) spectroscopy was employed for the simultaneous prediction of the concentrations of glucose and citric acid in the aqueous solution of a blood anticoagulant solution. The solution was placed in an NIR spectrophotometer and the absorbance at wavelengths between 400 and 2500 nm was measured at 2 nm intervals. Multiple linear regression (MLR) was used to obtain calibration equations relating the NIR spectral data and the glucose and citric acid concentrations of a calibration sample set obtained by enzymatic methods. The value of the multiple correlation coefficient ( R) was 0.993 when using the wavelengths of 2274 and 1674 nm for glucose. The value of the simple correlation coefficient ( r) for citric acid was 0.993 when using the wavelength 1690 nm. To validate the calibration equations obtained, glucose and citric acid concentrations in a prediction sample set were compared with the concentrations measured by the enzymatic methods. Excellent agreement between the results of the enzymatic method and those of NIR spectroscopy was observed for both constituents. The procedure of NIR spectroscopy was simple and the operation time required to predict the concentrations was a few minutes. These results indicate that NIR spectroscopy may be a useful method for monitoring the production process of the blood anticoagulant.

Improved Blood Preservation with 0.5CPD Erythro-Sol. Coagulation Factor VIII Activity and Erythrocyte Quality after Delayed Separation of Blood

Background and objectives: Delay between blood collection and the separation of its components may result in lowered yield of factor VIII (FVIII) and loss of 2,3-biphosphoglycerate (2,3-BPG). This study was to see whether the use of 0.5 CPD resulted in better preservation of FVIII and maintenance of 2,3-BPG. Materials and methods: 55 units of blood were collected in 0.5CPD and 48 in CPD SAG-M. Ten of the collections were paired, so that the same donors were bled in a single session partly in an 0.5CPD system and partly in CPD SAG-M. After collection, the blood was promptly cooled to 20 °C and stored at that temperature for up to 24 h. Results: Preservation of FVIII activity was significantly better in 0.5CPD compared with CPD. The content of von Willebrand factor was stable in the anticoagulant solutions for 24 h at that temperature. Plasma separated from both media had low levels of prothrombin fragment 1+ 2 and complement activation. Paired collections substantiated previous reports that red cell storage is significantly improved in 0.5CPD compared with CPD SAG-M with respect to 2,3-BPG and haemolysis. Conclusions: Red cell metabolism and oxygen-releasing capacity are kept at acceptable levels in 0.5CPD blood for 24 h at 20 °C before component separation. The concentration of red cell 2,3-BPG remained at normal or slightly subnormal levels during further storage in 0.5CPD at 4 °C for 2–4 weeks before gradual decay to an average of 39% at 48 days.

Improved Blood Preservation with 0.5CPD Erythro‐Sol. Coagulation Factor VIII Activity and Erythrocyte Quality after Delayed Separation of Blood

Delay between blood collection and the separation of its components may result in lowered yield of factor VIII (FVIII) and loss of 2,3-biphosphoglycerate (2,3-BPG). This study was to see whether the use of 0.5 CPD resulted in better preservation of FVIII and maintenance of 2,3-BPG. 55 units of blood were collected in 0.5CPD and 48 in CPD SAG-M. Ten of the collections were paired, so that the same donors were bled in a single session partly in an 0.5CPD system and partly in CPD SAG-M. After collection, the blood was promptly cooled to 20 degrees C and stored at that temperature for up to 24 h. Preservation of FVIII activity was significantly better in 0.5CPD compared with CPD. The content of von Willebrand factor was stable in the anticoagulant solutions for 24 h at that temperature. Plasma separated from both media had how levels of prothrombin fragment 1 + 2 and complement activation. Paired collections substantiated previous reports that red cell storage is significantly improved in 0.5CPD compared with CPD SAG-M with respect to 2,3-BPG and haemolysis. Red cell metabolism and oxygen-releasing capacity are kept at acceptable levels in 0.5CPD blood for 24 h at 20 degrees C before component separation. The concentration of red cell 2,3-BPG remained at normal or slightly subnormal levels during further storage in 0.5CPD at 4 degrees C for 2-4 weeks before gradual decay to an average of 39% at 48 days.