Letter to the Editors · Brief an die Herausgeber Dr. Gerda C. Leitner Klinik für Blutgruppenserologie und Transfusionsmedizin Allgemeines Krankenhaus Wien Währinger Gürtel 18–20, 1090 Wien, Austria E-mail gerda.leitner@meduniwien.ac.at © 2005 S. Karger GmbH, Freiburg Accessible online at: www.karger.com/tmh Fax +49 761 4 52 07 14 E-mail Information@Karger.de www.karger.com Transfus Med Hemother 2005;32:255–256 DOI: 10.1159/0000 87944 Received: March 2, 2005 Accepted: March 3, 2005 Published online: September 23, 2005 Collection and Storage of Leukocyte-Depleted Whole Blood in Autologous Blood Predeposit in Elective Surgery Programs Gerda C. Leitner a Isolde Rach a Michaela Horvath a Christoph Buchta a Dieter Zakel a Guenter Weigel b Paul Hoecker a Michael B. Fischer a a Department of Blood Group Serology and Transfusion Medicine, b Department of Cardiothoracic Surgery, Allgemeines Krankenhaus Wien, Vienna, Austria For the production and storage of autologous blood compo- nents it is recommended by the European Guidelines to fol- low the same rules as for the production and storage of ho- mologous blood components [1]. The collected autologous whole blood is plasma- and leukocyte-depleted and stored as packed red blood cells (PRBC). There seems to be evidence that autologous blood, stored and re-transfused as leukocyte- depleted whole blood (LD WB) has no negative impact on the patients’ postoperative course [2]. In compliance with the warranted quality measures [3, 4] preparation of autologous LD WB is a time- and cost-sparing procedure. In order to as- sess the quality of LD WB units stored for 35 days, we ana- lyzed the release of K + , lactate dehydrogenase (LDH), and free hemoglobin (fHb) into the supernatant over time and measured the increase of hemolysis and the decrease of glu- cose and pH till the end of shelf life. Special attention was paid to the intracellular adenine nucleotide content, consider- ing the latter as surrogate markers for red cell quality [5]. 25 CPDA-1-preserved LD WB units (450 ml whole blood, 70 ml CPDA-1) were collected from 25 healthy male donors (median age 40 years, range 22–64 years) using the double blood bag system (MacoPharma, Toucoing, France) with an integrated filter (Leucoflex; MacoPharma). The samples were evaluated for standard metabolic quality parameters (K + , LDH, lactate, fHb in the supernatant) using a Hitachi 917 spectrometer (Boehringer Mannheim, Mannheim, Germany) and for consumption of glucose, which was analyzed enzymat- ically by the hexokinase assay (Olympus System Reagent; Olympus Life and Material Science Europe GmbH, Ham- burg, Germany). Samples were taken immediately after leukocyte depletion and then in weekly intervals until the end of shelf life. The routine quality parameters were extended by measuring ATP, ADP and AMP as described previously [6]. LD WB met the quality requirements warranted by the Euro- pean Council [4]. Leukocyte contamination was < 1 × 10 6 in all units. The metabolic parameters (K + , LDH, lactate, fHb) in- creased over time, but hemolysis was well below the threshold of 0.8% at the end of shelf life. A mean of 0.15 ± 0.11% was measured after 35 days. The consumption of glucose ranged between 52 and 70% (mean ± SEM 60 ± 6%). Lactate levels increased to the 8-fold of initial values accompanied by a sig- nificant decrease of pH from initially 7.0 to 6.56 till the end of shelf life. The intracellular content of ATP rose within 14 days from a mean of 122 ± 26 to 138 ± 33 pmol/10 6 erythrocytes (p < 0.05) and decreased thereafter. At the end of shelf life 70 ± 18% of the initial value was found. ADP and AMP showed also an in- crease until day 14 and day 28, respectively, (p < 0.05) and re- mained elevated until day 35 (fig. 1). During storage erythrocytes have to carry out certain energy- requiring processes in order to maintain membrane stability [6]. This energy is provided mainly by glycolysis and its suffi- ciency can be measured by the concentration of intracellular purines and the consumption of glucose [7]. A satisfying glu- cose reserve remained at this time. 60% of glucose was uti- lized and intracellular ATP values were well maintained. Si- multaneously ADP and AMP levels remained elevated. En- hanced loss of ADP was found in PRBCs stored after irradia- tion which is known to induce membrane damage, indicating a high phosphorylation rate [6]. Taken together, elevated ADP and AMP values, well maintained intracellular ATP levels as well as acceptable hemolysis measured in stored LD WB at the end of shelf life may account for less initial membrane damage by omitting the centrifugation to split LD WB into its components. In conclusion it can be stated that red blood cells stored as LD WB fulfil the requirements for transfusion warranted by the European Council, and adequate quality of erythrocytes