Research Proposal

Topic: To analyse the effective result and quality of fresh frozen plasma after
two month storage.

Abstract
It will be a detailed analytical study organized in the blood bank of Sahiwal Medical College
(Seitz R et al., 2010). The study will be conducted upon one hundred and fifty (Male n=131,
female n=18) healthy volunteer blood donors will be register. Whole blood (450± 10%) will
be collected from all donors into triple packs blood bag. Blood components will be prepared
about 8 hours in a 2 step spin protocol (Acker et al., 2016). Whole blood will be centrifuged
at 4° C with spin (2500rpm for 10 minutes) and plasma containing high amount of platelets
will be transferred into second pack of triple bag followed by a high spin for 10 minutes at
3200 rpm. Plasma containing least amount of platelets of second pack and fresh, new
plasma will be stored at --70° C. Factor V111 will be experimented on freshly newly
prepared. This step will be repeated after 60 days of storage using STA Compact
Haemostasis analyser as explained by manufacturer.

Introduction
The aim of the study is to store blood plasma to provide safe and effective blood
components which are advantageous for the recipients (Lamboo et al. 2007). Blood
collection will be done following respective measures authorized by Food and Drug
Administration (FDA). All the processes are required to be screened properly for
quality of all procedures, reagents, equipment and the contents of the components.
Nowadays, fresh frozen plasma is used normally for bleeding disorders and is
collected by plasmapheresis or separated from whole blood (Denise et al. 2012).
Fresh frozen plasma must be prepared and frozen within 6 or 8 hours of collection.
Fresh frozen plasma must be frozen immediately to a temperature required for the
preservation of normal activity of heat coagulation factors (Antunes et al. 2002).
Pakistan is a country with highly saturated of inherited bleeding disorders and the
fresh frozen plasma is mostly used in patients suffering from coagulation factors
deficiencies or bleeding excessively after any wound or injury (Eder AF and Sebook
AM 2007).
Reasons for abnormal bleeding would be routine surgical bleeding, liver disease, an
inherited disorder or a severe accident. They will be highly useful when a patient on
Warfin undergoing a surgery and there is not enough time for vitamin K to reverse
the effect. Fresh frozen plasma has the quality of storing maximum levels of labile
and non-labile coagulation factors (about 1 micro litre per mL) and can be stored for
12 months. Even at -65° C fresh frozen plasma can be stored for up to 7 years.

Materials and Methods

The study will be conducted in pathology department of Sahiwal Medical College. In
this study the efficacy of frozen fresh plasma will be analysed and factor V111 level
would be checked. All of the blood samples will be be collected from normal
volunteer donors according to standard procedures in the blood bank of Sahiwal
Medical College. It will be made sure that each whole blood sample (450mL± 10%)
into triple packs. Packs that will be containing a CPDA-1 anticoagulant. Centrifugation
protocol will be followed in which, blood components will be processed within 4 to 6 hours.
All of the blood will be centrifuges with initial light spin (2000 x rpm) for 5 minutes in a
refrigerated centrifuge. After that, a heavy spin for 10 minute at 32000rpm with temperature
of 4° C will be given. It will be made sure that plasma is removed properly and the
accumulated RBCs are re-suspended in one of the triple blood bag. Plasma will be stored at
18° C. A digital thermometer will be used to check the storage temperature. It is made sure
that an alarm system and a log book is retained.
Coagulation factor will be evaluated by putting five millilitres aliquots of fresh frozen plasma
into water bath at 37 ° C.
Stago STA Compact Haemostasis Analyzer will be used to check the performance of Factor
V111 Mean± standard deviation will be used to present the data.

Discussion
Apheresis collection or whole blood can be used to produce fresh frozen plasma.
Food and Drug Association states that fresh frozen plasma must be frozen for 8
hours of collection. CPD or CPDA-1 will be used as anticoagulant during 6 hours if
ACD is used as preservative. Fresh frozen plasma has the ability to be preserved at
-18° C or lower for 12months or for 7 years if stored at -65° C. Fresh frozen plasma is
needed to have maximum of both stable and labile clotting factors, about 1 international
unit(IU) per milli litre. Fresh frozen plasma has comparatively lower levels of labile factors,
factors V111 and FV. Fresh frozen plasma is assessed by measuring its levels.
Fresh frozen plasma is used today as well a vital blood product to provide coagulation
factors in an acquired and hereditary bleeding disorders in Pakistan due to financial crisis or
absence of essential products.
It is essential to analyze the effect of temperature and time from collection to freezing FV111
activity to utilize the product in a beneficial way. In this study, we will prepare fresh frozen
plasma from whole blood collection from healthy blood donors and keep units of fresh frozen
plasm at -18° C for one month. The effect of temperature variations and storage periods on
the FV 111 activity in fresh frozen plasma will be checked.

Conclusion
This will be an effective research for most patients needing frozen fresh plasma for
various bleeding disorders.

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