Annals of Tropical Pathology

: 2022  |  Volume : 13  |  Issue : 1  |  Page : 56--59

A comparative study of factor VIII levels in fresh-frozen plasma from whole blood stored at varying temperatures and durations

Ann Abiola Ogbenna1, Kunmi Mathew Oyewole2, Titilope Adenike Adeyemo1, Fikewa Oyedele3, Dorcas Olubunmi Oyelaran2,  
1 Department of Haematology and Blood Transfusion, College of Medicine, University of Lagos/ Lagos University Teaching Hospital, Nigeria
2 Department of Haematology and Blood Transfusion, Lagos University Teaching Hospital, Nigeria
3 Department of Haematology and Blood Transfusion, College of Medicine, University of Lagos, Nigeria

Correspondence Address:
Mr. Kunmi Mathew Oyewole
Department of Haematology, Lagos University Teaching Hospital, Idi Araba, Lagos


Background: In Nigeria, fresh-frozen plasma (FFP) is an important plasma product to provide coagulation factors such as factor VIII (FVIII) in patients with FVIII deficiency and is still the only product available for the treatment of hemophilia A and B in some countries where recombinant factor concentrate is not available. FVIII is a labile coagulation factor and a quality marker of fresh-frozen plasma. In this study, we assessed the effect of storage temperature and time on FVIII yield. Methods: This was a descriptive-analytical study of a random selection of 136 blood donors, randomly divided into four groups. Whole blood (WB) was collected into 450 ml of blood bag from each participant and an additional 4.5 ml was collected into citrated bottle containing 0.5 ml citrate for baseline FVIII assay. WB from donors in Group A and Group B were stored at 20°C–24°C for 6 h or less and 12 h, respectively, while Group C and Group D were stored at 4°C for 12 h and 24 h, respectively. FFP was prepared from the WB after storage and FVIII levels were determined using Sysmex CA-101 based on the principle of turbo-densitometry principle with automatic zero adjustment and magnetic stir bar for homogenizing the test suspension and for increased sensitivity. The level of FVIII in FFP produced at various storage conditions was determined. Percentage yield was calculated as median level of FVIII in FFP per group/median level of FVIII in citrated plasma per group ×100. This is the proportion of FVIII in FFP relative to the level at baseline in the donor unit, whereas the FVIII level is an indication of just the level in the produced FFP. Results: The median FVIII level in FFP from Group A and B was 0.87 IU/ml and 0.82 IU/ml, respectively. Similarly, the median for Group C and D were 0.86 IU/ml and 0.76 IU/ml, respectively. The highest yield was observed in FFP prepared within 6 h or less of blood sample collection and least yield after 24 h of storage at 4°C. Conclusion: The yield of FVIII produced from WB under the varying temperatures and durations of storage studied is above the recommended level of 0.7 IU/ml; hence will be effective for replacement therapy in hemophiliacs. However, the best yield is obtainable from WB stored at 20°C–24°C for 6 h.

How to cite this article:
Ogbenna AA, Oyewole KM, Adeyemo TA, Oyedele F, Oyelaran DO. A comparative study of factor VIII levels in fresh-frozen plasma from whole blood stored at varying temperatures and durations.Ann Trop Pathol 2022;13:56-59

How to cite this URL:
Ogbenna AA, Oyewole KM, Adeyemo TA, Oyedele F, Oyelaran DO. A comparative study of factor VIII levels in fresh-frozen plasma from whole blood stored at varying temperatures and durations. Ann Trop Pathol [serial online] 2022 [cited 2022 Oct 4 ];13:56-59
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The provision of blood components of good quality is the responsibility of the blood bank. Blood component separation allows optimum usage and availability of blood products and also reduces the risk of patient's exposure to transfusion risk related.[1],[2] However, the quality of the blood component prepared can impact positively or negatively on patient care, hence the need to understand the effect of delayed separation from whole blood (WB) to freezing on the factors level. Fresh-frozen plasma (FFP) has been defined as the fluid portion (plasma) of 1 unit of human whole donated blood that has been centrifuged, separated, and frozen solid to -18°C (or colder) within 6 h after blood donation.[3],[4] It is indicated to replace deficiencies of multiple coagulation factors and control protein levels in massive blood loss, liver diseases, and disseminated intravascular coagulation.[5] It may also be used as the replacement fluid in plasma exchange. The ability to freeze transfusable plasma provides advantages to blood operators and transfusionists, freezing plasma preserves coagulation factors and other plasma protein activities and makes possible a long shelf life of the frozen product ranging from 1 to 3 years.[6] The Council of Europe guidelines require FVIII levels to be greater than 0.70 IU/ml and the guideline mandates that plasma must be separated from WB and within 6 h of blood collection.[7] However, blood from donor drives in Nigeria sometimes comes in very late, and hence these plasmas cannot be separated. Currently, those WB are not used for component products and are issued as red cell concentrate. This practice does not allow for efficient use of blood which is scarce. Production of FFP from WB stored at room temperature for up to 12 h or stored at 2°C–6°C for 24 h will greatly increase operational flexibility. It is hence imperative to determine the maximum time at which stored WB either at room temperature or 2°C–6°C can still be of good quality for intended use, especially for the labile factors. This will ensure optimal use of all units of blood donated.


Lagos state is the most populated city in Africa.[8] It is located in South-Western Nigeria, bounded in the North-East by Ogun state, in the South by the Coast of Atlantic Ocean and in the West by the Republic of Benin.

The study was carried out in the blood bank of Lagos University Teaching Hospital (LUTH), Idi Araba, Surulere, Lagos state, from November 1, 2019, to December 2019. Tertiary hospital, established in 1962 and is affiliated with the University of Lagos, College of Medicine. LUTH is an 851-bed hospital with a hospital-based blood bank which collects approximately 8000 units of blood per year. Annually, it produces an average of 3420 units of FFP.

A hundred and thirty-six blood donors between the ages of 18 and 65 years who met the hospital donor selection criteria and gave informed consent were recruited.

A simple random technique was used to select eight donors per day. Donors were randomized into four groups through a ballot. Letter A, B, C, and D were written in papers and placed in envelopes. Donors were grouped according to the letter they chose from the envelope.

Four hundred and fifty (450 ± 50) of WB was collected into Axiom triple bags containing 63 mL of CPDA-1 from the participant and 4.5ml of blood was also collected into sodium citrate bottles (1:9 ratios) to determine the baseline levels of factor VIII (FVIII) in all participants.

A total of 136 participants were recruited for this study, 34 for each group. In group A, FFP was prepared within 6 h from WB stored at 20°C–24°C. In Group B, it was prepared at 12 h from WB stored at 20° –24°C; Group C, at 12 h from WB stored at 2-6°C while Group D was prepared at 24 h from WB stored at 2°C–6°C [Figure 1].{Figure 1}

FFP was prepared using Sigma 6-16k Germany cold centrifuge. WB was centrifuged at 4000 g for 15 min at 4°C and the red cells were separated from the plasma with a manual blood component extractor (Labtop platelet extractor). Ten milliliters of plasma produced was expressed into the 3rd bag. This was used for the study to determine the level of FVIII. All blood components were stored at -40°C until test performed.

The coagulation factors FVIII was assay using a coagulation analyzer CA-101 (Siemens AG, Germany). Manufacture instruction and procedure were fully adhered to.

Results were entered into Excel spreadsheet and analyzed using IBM SPSS statistics software, version 25.0 (Armonk, NY, USA). Percentage yield was calculated as follows: mean level of FVIII in FFP per group/mean level of FVIII in citrated plasma (baseline) per group ×100. The mean levels of the FVIII level and yield were compared across groups. P < 0.05 is considered statistically significant.

Ethical approval was obtained from Health Research Ethics Committee (HREC) LUTH to carry out this project. The assigned HREC number is ADM/DCST/HREC/APP/3298. Approval was obtained from the blood donors' clinic for the purpose of this research and informed consent from donors.


The mean age of the participant was 31.58 (± 9.37) years. The majority (78.7%) were within the ages of 21–40 years. The male-to-female ratio is 8.7:1. Most (58.1%) had O positive blood group [Table 1]. The median plasma levels of FVIII in FFP produced over time irrespective of storage form showed a downward trend, this difference was not statistically significant. P > 0.05 [Table 2]. The median FVIII levels produced at all temperatures and storage forms were quite higher than the WHO recommendation of 0.7 IU/ml.{Table 1}{Table 2}

A downward trend in the yield of FVIII was observed with increasing time irrespective of temperature. This difference was statistically significant. The highest yield was observed in FFP produced within 6 h or less of blood donation and the least in FFP produced after 24 h of storage at 4°C [Table 3].{Table 3}

There was a statistical difference in percentage yield of FVIII across the age groups [Table 4]. The difference was between the age groups 31–40 years and 41–50 years (P = 0.024) and 31–40 years/≥50 years (P = 0.007). The highest yield was observed in 31–40 years age groups. There was no statistical difference in percentage yeild of FVIII across blood groups. [Table 5].{Table 4}{Table 5}


The efficiency of FFP therapeutic is determined by the levels of various coagulation factors contained in a unit. Factors which influence the concentration of coagulation factors in FFP may be donor-specific or related to the process of preparation, storage, and thawing. Since FFP is still an important plasma product in the management of hemophilia patients in Nigeria, it is very important to understand the effect of temperature and time from WB collection, separation, and to freezing on FVIII yield in other to optimize it is production in patient management. One of the quality control criteria for FFP production is a FVIII level ≥0.7 IU/ml. The FFPs with FVIII concentration of <0.7 IU/ml are not suitable to be used for patients with hemophilia.[9] The median FVIII levels produced at the various temperatures and storage time in this current study were all >0.7 IU/ml. The lowest median value was 0.76 IU/ml (interquartile range: 0.50 IU/ml), this was observed in FFP produced from WB stored at 4°C for 24 h [Table 2]. This agrees with different studies[10] measuring the FVIII activity of plasma produced from WB stored at 4°C for 20–24 h. Seventy-nine percent yield of FVIII was observed in FFP produced after 24 h of storage at 4°C [Table 3]. Serrano et al. also reported a yield of 78% for FVIII produced after24h storage at 4°C.[10] Dogra et al. in a comparative analysis of factors V and VIII in FFP produced after 24 h of storage also corroborated our findings.[9] They reported a reduction in the activity of 18.4% for FVIII which is equivalent to a yield of 81.6%.

In this study, we observed that FVIII levels in FFP produced at 12 h from WB stored at room temperature (20°C–24°C) or at 4°C had a good retention of FVIII activity [Table 3]. There was no significant difference between the activities at these two storage conditions. However, there is a statistical difference between yield at all the storage conditions (P < 0.05) [Table 3]. The median yield at conventional storage time and temperature (≤6 h; 20°C–24°C) was 97.65%, this was the highest. The lowest yield was observed at FFP produced at 24 h after storage at 4°C (79.08%). Meanwhile, Omidkhoda et al. reported contrary to our finding that FVIII activity in FFP produced at 20°C–24°C was lower than that produced at 4°C after 8h and 10 h of storage.[11]

There was a statistically significant difference in the yield of FFP produced from donors within different age groups (P = 0.021) [Table 4]. It was shown in this study that the higher the age of the donor, the better the yield of FVIII. The yield of FFP produced from donors with the ages of 31–40 years was the best. This agrees with other studies in the past.[12],[13]

Although FVIII levels across different temperatures and durations studied were above 0.7 IU/ml, other labile factors were not studied; hence FFP produced after storage at 20°C-24°C or 2°C–6°C for 12 h and those produced after 24 h of storage of WB at 2°C–6°C, can only be used for patients with hemophilia and must be labeled as such. This is a limitation of this study. It is recommended that further studies be done to determine the yield of the other labile factors at these varying storage conditions. Knowledge obtained may help in the optimal use of WB which gets to the blood bank after 6 h of collection, especially in low-middle-income countries where the rate of voluntary blood donations is very low compared to demand.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.


1Josephson CD, Abshire TC. Clinical uses of plasma and plasma fractions: Plasma-derived products for hemophilias A and B, and for von Willebrand disease. Best Pract Res Clin Haematol 2006;19:35-49.
2Erber WN, Perry DJ. Plasma and plasma products in the treatment of massive haemorrhage. Best Pract Res Clin Haematol 2006;19:97-112.
3Cash J. Fresh frozen plasma: Is it farewell? Vox Sang 1994;67 Suppl 3:121-4.
4Makroo RN, Arora B, Bhatia A, Chowdhry M, Luka RN. Clinical significance of antibody specificities to M, N and Lewis blood group system. Asian J Transfus Sci 2014;8:96-9.
5Cardigan R, Van der Meer PF, Pergande C, Cookson P, Baumann-Baretti B, Cancelas JA, et al. Coagulation factor content of plasma produced from whole blood stored for 24 hours at ambient temperature: Results from an international multicenter BEST collaborative study. Transfusion 2011;51 Suppl 1:50S-7S.
6O'Neill EM, Rowley J, Hansson-Wicher M, McCarter S, Ragno G, Valeri CR. Effect of 24-hour whole-blood storage on plasma clotting factors. Transfusion 1999;39:488-91.
7American Association of Blood Banks. The 2009 National Blood Collection and Utilization Survey Report. Available from:–nbcus–report.pdf. [Last accessed on 2021 Mar 06].
9Dogra M, Sidhu M, Vasudev R, Dogra A. Comparative analysis of activity of coagulation factors V and VIII and level of fibrinogen in fresh frozen plasma and frozen plasma. Asian J Transfus Sci 2015;9:6-8.
10Serrano K, Scammell K, Weiss S, Culibrk B, Levin E, Gyöngyössy-Issa M, et al. Plasma and cryoprecipitate manufactured from whole blood held overnight at room temperature meet quality standards. Transfusion 2010;50:344-53.
11Omidkhoda A, Tabatabaei MR, Atarodi K, Karimi K, Froushani AR, Pourfathollah AA. A comparative study of the effects of temperature, time and factor VIII assay type on factor VIII activity in cryoprecipitate in Iran. Blood Transfus 2011;9:394-9.
12Albánez S, Michels A, Sponagle K, Lillicrap D. Age-related increases in plasma factor VIII and Von Willebrand factor in a C57BL/6 mouse model are associated with increased factor VIII and Von Willebrand factor gene expression and reduced expression of the clearance receptor, stabilin-2. Blood 2014;124:4228.
13Orstavik KH, Magnus P, Reisner H, Berg K, Graham JB, Nance W. Factor VIII and factor IX in a twin population. Evidence for a major effect of ABO locus on factor VIII level. Am J Hum Genet 1985;37:89-101.