Research Article A Comparison Study of the Blood Component …downloads.hindawi.com/journals/jbt/2013/523539.pdf · Journal of Blood Transfusion 200 mL WB Centrifuged 200 mL ltered

Hindawi Publishing CorporationJournal of Blood TransfusionVolume 2013 Article ID 523539 7 pageshttpdxdoiorg1011552013523539

Research ArticleA Comparison Study of the Blood Component Quality ofWhole Blood Held Overnight at 4∘C or Room Temperature

Shichun Wang Tiantian Wang Yahan Fan Shan HuangZhongmei Yi Ruiqing Li and Shuming Zhao

Department of Blood Transfusion Southwest Hospital TheThird Military Medical University No 30 Street GaotanyanChongqing 400038 China

Correspondence should be addressed to Shuming Zhao shumingzhaoyahoocom

Received 21 February 2013 Accepted 30 July 2013

Academic Editor Erwin Strasser

Copyright copy 2013 Shichun Wang et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Background The use of plasma frozen within 24 hrs is likely to increase Whole blood (WB) and buffy coats (BCs) can be held fora few hrs or overnight before processingMethods Twenty-four bags of WB for plasma and 12 bags for platelet (PLT) concentrateswere collected The fresh frozen plasma (FFP) was prepared within 6 hrs I-FP24 and II-FP24 samples were prepared either fromleukodepletedWB that was held overnight or fromWB that was held overnight before leukodepletionThe PLT concentrates (PCs)were prepared from BCs within 6 hrs (PC1) and within 18 to 24 hrs (PC2) The typical coagulation factors and some biochemicalparameters were determinedResults Compared to the FFP samples the levels of FVII and FVIII in the I-FP24 and II-FP24 samplesdecreased significantly The pH Na+ LDH and FHb levels differed significantly between II-FP24 and FFP Compared to PC1PC2 exhibited lower pH pO

2 and Na+ levels a higher PLT count and increased pCO

2 K+ Lac and CD62P expression levels

Conclusion FP24 is best prepared fromWB that was stored overnight at 4∘C and then leukodepleted and separated within 24 hrsPCs are best produced from BCs derived fromWB that was held overnight at room temperature

1 Introduction

An important step in safeguarding the quality and safety ofthe blood supply is recruiting volunteer donors from low-risk populations and producing qualified blood and bloodcomponents Volunteer donor recruitment is a challengingproposition worldwide In China it was particularly difficultbefore 1998 Traditional Chinese culture believes that theloss of even a small amount of blood has a substantialdetrimental health effect Some people also believe that blooddonation is a disloyal action against onersquos ancestors Oldcultural beliefs combinedwith inadequate efforts tomobilizevolunteer donors have led to a chronic shortage of bloodproducts in some areas of China [1] Therefore many bloodcenters have tried to prepare more blood components suchas platelet (PLT) concentrates (PCs) fromwhole blood (WB)to meet the need for blood products

WB units are generally held at ambient temperatures (20ndash24∘C) when PCs will be prepared within 6 hrs of the bloodcollection The United States Food and Drug Administration

(FDA) guidelines during the early 1980s allowed thepreparation of components at 20 to 24∘C within 8 hrs ofcollection and a 5-day storage period for PCsThe impetus forextending the time to 8 hrs reflected an increased productiondemand for PCs and the limitations of the 6 hr period withrespect to the collection of a large percentage of WB units atmobile drives which were sometimes quite distant from thecomponent production laboratories Currently because ofthe overnight holding practice component manufacturingincluding PLT preparation can be logistically facilitatedwith related cost savings Many countries have converted toovernight-hold blood processing because of these advantages[2] Recently the Canadian Blood Services has adopted theovernight hold approach to prepare PCs from buffy coats [3]

Currently the most commonly used plasma product isfresh frozen plasma (FFP) which can be obtained froma plasmapheresis collection or donated WB FFP must beprepared and frozen within 6 or 8 hrs from the time ofcollection according to the usage of different anticoagulantsfor WB storage These requirements for FFP preparation

2 Journal of Blood Transfusion

200 mL WB

Centrifuged

200 mL filtered WB

Plasma (FFP)

Plasma (I-FP24)

Plasma (II-FP24)

Centrifuged

200 mL WB Centrifuged

RT 400 mL WB

RT

BCs

Plasma P6

RTPCs

RT

BCs RT

PCs

Plasma P24

lt6 hrs

n = 12

n = 12

n = 12

n = 12

n = 12

n = 12

n = 24n = 24

n = 24

200 mL WBn = 12

n = 12

200 mL WBn = 12

200 mL WB

n = 24

4∘C 400 mL WB

18ndash24 hrs filtered

lt6 hrs

4∘C overnight

18ndash24 hrs

18ndash24 hrs

18ndash24 hrs

Overnight

4∘C

Figure 1 Experimental algorithm for the plasma and PCs production validation with WB held overnight (119899 = 2412 units in each arm) Theplasma was frozen as soon as possible after preparation

could limit the number ofWBunits that can be processed intoFFP because of the blood collections from blood-collectingvehiclesWB stored overnight at 4∘C can be used tomanufac-ture plasma and concentrated red blood cells but not for PCsbecause of the deleterious effect of 4∘C storage on PLT qualityPLTs do not tolerate refrigeration and disappear rapidly fromcirculation if subjected to chilling before transplantation [4ndash6] Thus WB used for PC separation must be stored atroom temperature Currently PC preparation according tothe BC removal method is widely practiced [7ndash9] and isassociated with significantly better PLT recoveries and lessPLT activation immediately after preparation [10] Howeverit is also inexpedient for PCs that are prepared during theimmediate processing of freshWB In addition an overnighthold of BC is good for the PC quantity and quality [11]There are numerous operational and economic advantagesto blood services from ambient WB storage within 24 hrsbecause of the reduced restrictions on blood processing forPC separation within 8 hrs of collection

The aim of this study was to compare the quality asmeasured by the coagulation factor levels of plasma preparedfromWB that was processed within 8 hrs after collection andstorage at 4∘C within 18ndash24 hrs of storage at 4∘C or within18ndash24 hrs of storage at ambient temperature and the qualityof PCs from the BCs processed either from freshWB or fromWB that was held overnight at room temperature

2 Materials and Methods

21 Study Design The blood component quality of WB thatwas stored overnight was evaluated the frame paragraphof these experiments is shown in Figure 1 Briefly 400mLWB samples were collected from normal volunteer donors

according to standard procedures in the blood donation lawof the Peoplersquos Republic of China (24 bags used for plasmapreparation were stored at 4∘C and 12 bags used for PCspreparation were stored at room temperature) The studyprotocol was approved by the Southwest Hospital HumanResearch and Ethics Committee EachWB sample (400mL plusmn10) was collected into blood collection packs that containeda CPD anticoagulant (Nigale Biomedical Co Ltd SichuanChina) and then equal volumes were divided between 2units (200mLunit) The blood components were processedin a 2-step centrifugation protocol (2600 rpm or 1642timesg for15min followed by 1000 rpm or 263timesg for 6min in a J6-MC centrifuge Beckman Miami FL USA) The plasma wasremoved and the packed RBCs were resuspended in one ofthe additive solutions SAG-M (100mL) (Nigale BiomedicalCo Ltd Sichuan China)The FFP was prepared within 6 hrsfrom 12 (119899 = 12) of the leukodepleted WB with a WBin-line leukocyte blood filter (Nigale Biomedical Co LtdSichuan China) The I-FP24 (I-FP24 6 hrs18ndash24 hrs) wereprepared from leukodepleted blood held overnight (119899 = 12)using a WB in-line leukocyte blood filter (Nigale BiomedicalCo Ltd Sichuan China) from WB within 6 hrs or II-FP24(II-FP24 18ndash24 hrs18ndash24 hrs) from WB held overnight andthen leukodepleted using a WB in-line leukocyte blood filter(Nigale Biomedical Co Ltd Sichuan China) (119899 = 24)One batch of PCs was prepared from BCs (stored overnight)obtained from fresh WB (freshstored 119899 = 12 controlgroup PC1) and the other batch of PCs was prepared fromBCs obtained from WB that was stored overnight at roomtemperature (storedfresh 119899 = 12 experimental group PC2)At the same time plasma from the 2 groups was preparedincluding P6 (119899 = 12) from the PC1 group and P24 (119899 =12) from the PC2 group All plasma samples were stored

Journal of Blood Transfusion 3

in a minus30∘C freezer before testing and all PCs were testedafter separation The typical coagulation factors (FV FVIIFVIII Fib) and some biochemical indicators (K+ Na+ ClminusTP lactic dehydrogenase [LDH] glucose [GLU] FHb pH)were observed in all plasma samples The PLT count and thelactate pCO

2 pO2 HSR pH GLU and CD62P expression

levels were measured in all PCs

22 In Vitro Assays In general the following methods wereused

221 WB Collection and Processing WB was collected indisposable plastic blood bag (110414 Nigale BiomedicalCo Ltd Sichuan China) and then refrigerated in a bloodrefrigerator (HXC-158 Haier Shanghai China) or held atroom temperature The plasma from WB which was held at4∘C was prepared by a 2-step centrifugation (3500 rpm or3128timesg for 10min J6-MC BeckmanMiami USA) frozen infreezer (MDF-U538 Sanyo Osaka Japan) thawed in plasmathawer (KJX III Szmic Suzhou China) at 37∘C PCs wereprepared by a 2-step centrifugation (2600 rpm or 1642timesgfor 15min followed by 1000 rpm or 263timesg for 6min J6-MCBeckman Miami USA)

222 Coagulation Factors The clotting factor activity levelswere determined with an automatic blood coagulation ana-lyzer (CA1500 Sysmex Kobe Japan) and the assay reagentsincluded coagulation factors Fib FVII FVIII APTT and PT(537991 500725A 546529A 537485A 545363 Dade BehringMarburg Marburg Germany) and coagulation factors FV(503587A ChengduXiehe BiotechnologyCo Ltd ChengduChina)

223 Biochemical Parameters The glucose and LDH levelswere measured using dry chemistry on a chemistry ana-lyzer (AU2700 Olympus Shimadzu Tokyo Japan) The FHblevel was measured by a 3-wavelength method (380 nm415 nm 450 nm) on a microplate reader (1500-992 ThermoFisher ScientificWalthamMAUSA)TheTris-HCl solution(624mmolL pH 80) was prepared as the reagent Brieflythe instrument was zeroed and the plasma sample wasdiluted to a 3mL as Tris-HCl solution plasma = 9 1 (2700120583L 300 120583L) and measured the absorbance against the Tris-HCl solution as blank The level of FHb is calculated as theformula FHb (mgL) = [168 times 119860

415minus 084 (119860

380+ 119860450)] times

1000mgL

224 Quantity and Quality of PCs The PLT counts wereobtained using an automated cell analyzer (KX-21N SysmexKobe Japan) The pH pO

2 pCO

2 glucose and lactate levels

were measured with a blood gas analyzer (ABL715 Radiome-ter Copenhagen Denmark) or pH was measured with apH meter (PB-21 Sartorius AG Goettingen Germany) Theglucose and LDH levels were measured using dry chemistrywith a chemistry analyzer (AU2700 Olympus ShimadzuTokyo Japan)The level of HSR and the PAgTwere measuredwith a spectrophotometer (7200 Unic Bern Switzerland)and an adhesionmeter (HY-I BeijingHongrunda instrument

Co Ltd Beijing China)The PLT surface CD62P expressionwas measured with flow cytometry (FACSCalibur BeckmanCoulterMiami FL) and fluorescein isothiocyanate (FITC) orphycoerythrin-labeled monoclonal antibodies (CD41 FITC(Lot 555469) CD62-P PE (Lot 348107) andMouse IgG-1 PE(Lot 555748) BD USA)

23 Statistical Analysis All test unit groups were com-pared with their respective reference groups The resultsare expressed as the mean plusmn standard deviation (SD) andthe median (range) Data were analyzed with the SPSSsoftware program (version 130 SPSS Inc Chicago IL USA)Comparisons were performed with a two-sample 119905-test witha 95 confidence interval (CI) or a one-way analysis ofvariance (ANOVA) and Dunnettrsquos test to identify the differ-ences between each storage method A 119875 value lt 005 wasconsidered to be significant

3 Results

31 Plasma Appearance Observations All frozen plasmasamples were warmed to 37∘C in a water bath on the thirdday after collection and no precipitation emerged

32 Effect of Overnight Holding on In Vitro Plasma QualityTheclotting factor analysis of plasmaprocessed under variousconditions is shown in Table 1 The plasma was processedapproximately 6 hrs after collection or at 18ndash24 hrs in groupsI-FP24 and II-FP24 Compared to FFP I-FP24 decreased by226 2938 3289 and 4071 including significantlylower FV FVII and FVIII levels (119875 lt 005) in II-FP24 thedecreases were 19 1260 1491 and 3286 includingsignificantly lower FVIII levels (119875 lt 005) Compared to FFPand I-FP24 II-FP24 had higher levels of K+ LDH and FHbThe effect of overnight holding of WB at room temperatureon the in vitro plasma quality was also analyzed Comparedto P6 P24 had higher levels of Na+ K+ and FHb and lowerlevels of Clminus pH and FVIII Differences in the pHNa+ LDHand FHb levels were observed between the II-FP24 and P24samples

33 Effect of Overnight Holding of WB at Room Temperatureon In Vitro PC Quality All PCs had PLT counts well abovethe current acceptance level of 20 times 1010U Compared to thecontrol group PC2 had a higher PLT count (349 plusmn 119 times1010U versus 278 plusmn 149 times 1010U) which was 2554 higherthan that of PC1 and 7450 higher than that of the currentacceptance levelThe PLT in vitro functional parameters werenot significantly different but differences were observed withrespect to CD62P expression (1193 plusmn 018 versus 1006 plusmn 028119875 lt 005) and some biochemical indicators (lower pH pO

2

and Na+ levels and higher pCO2 K+ and Lac levels Table 2)

4 Discussion

Blood collection processing and storage conditions signifi-cantly influence the quality of blood components and con-sequently these processes are closely regulated to maximize


Table 1 Clotting factor levels and some biochemical parameters of plasma prepared fromWB stored under different conditions (mean plusmn SDmedian and range)

WB stored at 4∘C WB stored at 22∘CWB holdtimecomponentspreparation (hrs)

lt6 lt6 lt618ndash24 18sim2418ndash24 lt6 18ndash24

Plasma FFP 119899 = 12 I-FP24 119899 = 12 II-FP24 119899 = 24 P6 119899 = 12 P24 119899 = 12

Fib (gL)208 plusmn 016

204(188ndash 226)

161 plusmn 043

148(118ndash214)

204 plusmn 040

215(134ndash243)

178 plusmn 059

197(155ndash224)

179 plusmn 059

201(152ndash219)

FV ()8657 plusmn 1873

9239(6125ndash10736)

6114 plusmn 1376

lowast

6064(4498ndash8161)

7566 plusmn 607

7865(6716ndash8161)

8121 plusmn 654

7865(7378ndash9239)

8759 plusmn 884

8396(7578ndash10414)

FVII ()9171 plusmn 2553

9258(6052ndash11300)

6155 plusmn 1145

lowast

6150(4453ndash7519)

7803 plusmn 1908

7196(4945ndash10849)

8032 plusmn 1737

7798(5092ndash11529)

8084 plusmn 1729

8102(4871ndash10849)

FVIII ()9765 plusmn 2599

10760(6381ndash12834)

5790 plusmn 735

lowast

5580(5126ndash7031)

6556 plusmn 1393

998771

6137(4548ndash9265)

7798 plusmn 1160

7859(4895ndash9265)

6630 plusmn 1016

6506(4388ndash8437)

TP (gL)5724 plusmn 081

5740(5630ndash 5810)

5680 plusmn 450

5730(5180ndash6180)

5896 plusmn 317

5900(5390ndash6390)

5891 plusmn 294

5910(5480ndash6640)

5978 plusmn 314

5910(5600ndash6660)

pH721 plusmn 010

717(712ndash738)

722 plusmn 009

720(711ndash734)

724 plusmn 006

725(717ndash736)

736 plusmn 007

735(730ndash753)

718 plusmn 004

718(712ndash724)

Na+ (mmolL)15678 plusmn 088

15700(15550ndash15760)

15494 plusmn 381

15400(15140ndash15900)

15661 plusmn 201

15635(15380ndash16120)

15605 plusmn 277

15620(14970ndash16000)

15925 plusmn 324

15920(15310ndash16350)

K (mmolL)330 plusmn 021

335(302ndash353)

396 plusmn 022

lowast

403(361ndash420)

433 plusmn

028

998771∙ 439(371ndash469)

345 plusmn 021

344(319ndash384)

425 plusmn 027

428(367ndash469)

Clminus (mmolL)7026 plusmn 259

6990(6730ndash7320)

7032 plusmn 372

7020(6580ndash7450)

7092 plusmn 321

7110(6480ndash7570)

7193 plusmn 195

7195(6850ndash7480)

6905 plusmn 223

7020(6510ndash7220)

LDH (IUL)10575 plusmn 2056

9100(9000ndash13400)

100 plusmn 678

9600(9400ndash11000)

14570plusmn2600

998771∙

15100(11000ndash17500)

10358 plusmn 2207

9950(6700ndash15000)

12083 plusmn 2439

995333

10800(8100ndash16100)

GLU (mmolL)2418 plusmn 124

2458(2211ndash2539)

2278 plusmn 061

2255(2212ndash2371)

2272 plusmn 118

2292(2121ndash2438)

2309 plusmn 138

2278(2102ndash2548)

2193 plusmn 148

2179(1989ndash2521)

FHb (mgL)1634 plusmn 788

1002(123ndash2991)

4354 plusmn 1976

lowast

4439(2081ndash7303)

12996plusmn3433

998771∙

12994(7102ndash17717)

2215 plusmn 1951

1371(167ndash6656)

2746 plusmn 2911

995333

1440(389ndash8974)

lowastP lt 005 versus FFP 998771P lt 005 versus FFPP lt 005 versus P6 995333P lt 005 versus II-FP24∙119875 lt 005 versus I-FP24

safety and efficacy The current FDA regulations require thatWBunits held at room temperaturemust be processedwithin8 hrs after collection or alternatively the WB units must berefrigerated and processed within 24 hrs [12] The Council ofEurope (COE) guidelines specify that WB units can be heldfor up to 24 hrs at room temperature provided that the unitsare rapidly cooled to 20ndash22∘C immediately after collectionThe Chinese guidelines require that the WB units are storedat 4∘C as soon as possible after collection and are processedwithin 8 hrs Overnight room temperature WB holdinghas both logistical advantages and some disadvantages that

potentially affect the quality of components particularly redblood cells (RBCs) In recent years Chinese blood centershave tried to improve the national blood supply and safety InChinaWBunits for clinical use are collected at blood centersMore than 400 blood centers are operated at 3 levels provin-cial regional and county Local government health officesoversee blood center operations The WB unit donationmodels in China have changed from paid donors before 1998to employer-organized donors after the new blood donationlaw took effect in 1998 and inmost areas of China donors arenow volunteers [1] The blood donation law bans all paidWB


Table 2 In vitro quality variables of PCs collected under variousconditions (mean plusmn SD median and range)

PC1 (119899 = 12) PC2 (119899 = 12)

PLTs (times1010U) 278 plusmn 149

26 (111ndash658)349 plusmn 119

333 (211ndash560)

pH 712 plusmn 005

713 (703ndash717)690 plusmn 004

lowast

689 (684ndash699)

pCO2(mmHg) 3461 plusmn 301

3545 (2980ndash3920)4919 plusmn 507

lowast

4790 (4380ndash5750)

pO2 (mmHg) 11750 plusmn 1046

11300 (10100ndash13500)9481 plusmn 1750

lowast

9540 (6090ndash11700)

K+ (mmolL) 301 plusmn 021

295 (270ndash330)333 plusmn 024

lowast

340 (300ndash380)

Na+ (mmolL) 13473 plusmn 156

13500 (13100ndash13700)13136 plusmn 211

lowast

13200 (12900ndash13500)

Clminus (mmolL) 7300 plusmn 205

7300 (6900ndash7600)7300 plusmn 173

7200 (7100ndash7600)

GLU (mmolL) 1917 plusmn 113

1900 (1780ndash2140)2014 plusmn 128

2020 (1820ndash2260)

Lac (mmolL) 325 plusmn 070

310 (230ndash470)413 plusmn 041

lowast

420 (360ndash470)CD62Pexpression ()

1006 plusmn 028

1006 (986ndash1025)1193 plusmn 018

lowast

1193 (1180ndash1206)

Adhesion () 4321 plusmn 227

4321 (4161ndash4481)4037 plusmn 366

4037 (3778ndash4295)HSR 101 plusmn 268

Bacterial culture Neg NeglowastP lt 005 versus PC1

donations for clinical use and encourages all Chinese citizensbetween the ages of 18 and 55 years whomeet the health crite-ria for blood donation to donate blood voluntarily Accordingto the law WB units are mostly collected at blood centers ormobile street stations in strategic locations that are run byblood centers One or 2 units (200 or 400mL) of WB canbe drawn from 1 volunteer stored in a refrigerator and trans-ported at 4∘C by a blood vehicle and separated into bloodcomponents at 4∘C within 8 hrs Currently most blood cen-ters report that they have already achieved the goal ofmeetingall of their regionrsquos blood needs with volunteer donations

In early 1999 OrsquoNeill et al studied the quality of FP24by measuring clotting factors in plasma obtained from CPDWB after storage at 22∘C and 4∘C for as long as 24 hrs beforefrozen storage at minus18∘C for 1 month The authors confirmedthat plasma separated from WB that had been stored for upto 24 hrs after collection could be used under all conditionsthat required FFP for transfusion [13] In 2005 Cardiganet al also examined the quality of FFP produced from WBstored at 4∘C overnight [14]The authors suggested that therewas good retention of relevant coagulation factor activity inplasma produced from WB that had been stored at 4∘C for18 to 24 hrs and that this would be an acceptable productfor most patients who required FFP Several other studieshave assessed the stability of FP24 when thawed and storedat 4∘C for up to 5 days [15] The studies discovered that FFPand FP24 contained adequate coagulation factor activities to

maintain hemostatic activity In China the guideline requiresthat the FVIII level in gt75 of FFP must be gt07 IUmLMeanwhile there is no current quality standard for FP24In our study the most variable affective factor during WBstorage was FVIII as might be predicted Compared to FFPthe FVIII level decreased by 3286 in II-FP24 and by 4071in I-FP24 The activity levels of other coagulation factors inI-FP24 and II-FP24 were not significantly different Theseresults showed that with the exception of FVIII FP24 wouldbe an acceptable product for most patients who required FFPKleinman et al conducted a survey in late 2009 to gatherdetailed information in which 40 of the respondents chosethe highest category indicating thatgt75 of their plasmawassupplied as FP24 [16] Interestingly the FV activity increasedsubstantially during the short period of storage which wasalso observed in this study and might reflect activation ofthe contact system during storage [17] Alhumaidan et alalso found that plasma manufactured after a 24 hr roomtemperature hold contains coagulation factors comparable toFFP except for a possible reduction of up to 20 in FVIIIOther clotting factors either were unchanged or showedminimal reduction (lt15) This plasma appears suitable asa transfusable product and extension of liquid storage to 7daysmerits consideration [18] FFP had the best results for themetabolism variables K+ LDH and FHb Compared to theother 2 groups II-FP24 had higher K+ LDH and FHb levelsThis result might be explained by the RBCmetabolism inWBstored overnight at 4∘C and the increased osmotic fragilitythat resulted in RBC lysis increasing some parameters inthe plasma processed from WB The differences in severalbiochemical parameters between the II-FP24 and P24 groupsmight be caused by the holding of WB at different tem-peratures which induced different levels of RBC membranefragility and deformation properties Comprehensively thequality of plasma from theWB that was held overnight at 4∘Cand then leukodepleted ismuch better than that of the plasmafrom leukodepleted WB that was then held overnight Thusonce filtrated plasma should be separated as soon as possible

Although PCs must be prepared within 6 hrs throughfresh WB processing this process is inexpedient becauseof blood collection from mobile vehicles or from a widegeographic areaThus it is important to extend the allowableprocessing time for PCs The BC removal method which isconsidered as a good PC preparationmethod is used inmanycountries Some scientists think that overnight BCs holdingbenefits the quantity and quality of PCs [10] In this study thequantity and quality of PCs from BCs andWB that were heldovernight were compared All PCs had PLT counts well abovethe current acceptance level of 20 times 1010U In particular thePLT counts were much higher in PC2 (2554 higher thanthose in PC1 and 7450 higher than the current acceptancelevel) Other studies have shown levels that were 1860ndash3300 higher in the overnight-held PCs [19 20] We studiedthe effect of BCs holding time before the PLT preparationon the quality of PCs and found lower PLT counts with a0 hr holding time than with 4 or 16 hr holding times Thuswe concluded that PLT recovery was comparatively affectedby immediate BCs processing This result could be explained


by the relatively short rest period of BCs (ie the PLTswere still forming aggregates that were easily removed duringcentrifugation prior to disaggregation) Some biochemicalindicators were different between the PC1 and PC2 groupswhich could be explained by RBC metabolism in WB duringovernight storage and by the lower amount of PLT in thestorage bag thus resulting in a lower total metabolism levelin the bag Although the CD62P expression was significantlydifferent the levels were still lower [21] The PLT quantity isactually improved by overnight room temperature hold andthe quality is not affected

Recently in a special supplement report to the journalof Transfusion scientists in 9 major blood product develop-mental laboratories evaluated the quality of components fromWB that was held overnight at room temperature includ-ing RBCs plasma and PCs The scientists concluded thatovernight holding of WB before processing had no lastingdeleterious effects on the in vitro qualities of the subsequentlyprepared components and that using different RBC additivesolutions did not appear to offer significant advantages interms of final RBC quality regardless of the processingmethod [21 22] Furthermore Dijkstra-Tiekstra and col-leagues analyzed the differences between PCs from fresh andovernight-stored blood Their data showed that PCs are bestprepared after a 20 to 24 hr WB hold [23] Some scientistsconfirmed that storing WB for 22 plusmn 2 hrs at 22∘C beforepreparing the PCs could increase the yield of PLTs in a con-centrate by 43while concurrently improving allmeasures of7-day poststorage PLT viability [24 25] Furthermore Cardi-gan and his colleague who have extensively studied plasmatested the coagulation factor content of plasma fromWB thatwas stored for 24 hrs at an ambient temperature and indicatedthat WB storage at ambient temperature for 24 hrs hadminimal effects on the plasma coagulation activity and thuswas an acceptable alternative to producing plasma on the dayof blood collection [26]There are numerous operational andeconomic advantages of the blood services of ambient WBstorage for 24 hrs because of the removal of the requirementto process blood for PLTs or plasmawithin 6 hrs of collectionEven the partial loss of FVIII in the plasma can be replacedwith a biological FVIII product for clinical use when neces-sary

5 Conclusion

In this study in which 400mL WB samples were dividedequally the group data revealed real differences OvernightWBholdingmight be a suitablemethod for blood componentprocessing FP24 is best prepared from WB that was storedovernight at 4∘C followed by leukodepletion and separationwithin 24 hrs PCs are best prepared within 6 hrs from BCthat was derived from WB that was held overnight at roomtemperature

Abbreviations

FFP Fresh frozen plasmaFP24 Plasma frozen within 24 hrs of phlebotomyBC(s) Buffy coat(s)

HSR Hypotonic shock responsePC(s) Platelet concentrate(s)PLT PlateletWB Whole bloodLDH Lactate dehydrogenaseFHb Free hemoglobinhrs HoursTP Total proteinAPTT Activated partial thromboplastin timePT Prothrombin timeGLU GlucoseLac Lactic acid

Conflict of Interests

The authors declare that they have no conflict of interests

Acknowledgments

The authors thank Zerong Wang Ronghua Diao and FangLin for their help with sample collection and Hui Guo forlaboratory measures and analysis This project was in partsupported by the National Natural Science Foundation ofChina (NSFC 81270649) Shichun Wang and Tiantian Wangare co-first authors

References

[1] H Shan J Wang F Ren et al ldquoBlood banking in Chinardquo TheLancet vol 60 no 9347 pp 1770ndash1775 2002

[2] G Moroff J P AuBuchon C Pickard P H Whitley WA Heaton and S Holme ldquoEvaluation of the properties ofcomponents prepared and stored after holding of whole bloodunits for 8 and 24 hours at ambient temperaturerdquo Transfusionvol 51 supplement S1 pp 7Sndash14S 2011

[3] K Serrano K Scammell S Weiss et al ldquoPlasma and cryopre-cipitatemanufactured fromwhole blood held overnight at roomtemperature meet quality standardsrdquo Transfusion vol 50 no 2pp 344ndash353 2010

[4] S Murphy and F H Gardner ldquoEffect of storage temperature onmaintenance of platelet viabilitymdashdeleterious effect of refriger-ated storagerdquoTheNew England Journal ofMedicine vol 280 no20 pp 1094ndash1098 1969

[5] KMHoffmeister TW Felbinger H Falet et al ldquoThe clearancemechanism of chilled blood plateletsrdquoCell vol 112 no 1 pp 87ndash97 2003

[6] V Rumjantseva P K Grewal H H Wandall et al ldquoDual rolesfor hepatic lectin receptors in the clearance of chilled plateletsrdquoNature Medicine vol 15 no 11 pp 1273ndash1280 2009

[7] R R Vassallo and S Murphy ldquoA critical comparison of plateletpreparation methodsrdquo Current Opinion in Hematology vol 13no 5 pp 323ndash330 2006

[8] E Levin B CulibrkM I CGyongyossy-Issa et al ldquoImplemen-tation of buffy coat platelet component production comparisonto platelet-rich plasma platelet productionrdquoTransfusion vol 48no 11 pp 2331ndash2337 2008

[9] W P Sheffield V Bhakta C Jenkins and D V Devine ldquoCon-version to the buffy coat method and quality of frozen plasmaderived from whole blood donations in Canadardquo Transfusionvol 50 no 5 pp 1043ndash1049 2010


[10] W A L Heaton P Rebulla M Pappelettera and W H DzikldquoComparative analysis of different methods for routine bloodcomponent preparationrdquo Transfusion Medicine Reviews vol 11no 2 pp 116ndash129 1997

[11] S Perez-Pujol M Lozano D Perea R Mazzara A Ordinasand G Escolar ldquoEffect of holding buffy coats 4 or 18 hoursbefore preparing pooled filtered PLT concentrates in plasmardquoTransfusion vol 44 no 2 pp 202ndash209 2004

[12] J D Roback M R Combs B J Grossman and C D HillyerEds American Association of Blood Banks Technical ManualAABB Bethesda Md USA 16th edition 2008

[13] E M OrsquoNeill J Rowley M Hansson-Wicher S McCarter GRagno and C R Valeri ldquoEffect of 24-hour whole-blood storageon plasma clotting factorsrdquo Transfusion vol 39 no 5 pp 488ndash491 1999

[14] R Cardigan A S Lawrie I J Mackie and L M WilliamsonldquoThequality of fresh-frozen plasma produced fromwhole bloodstored at 4∘C overnightrdquo Transfusion vol 45 no 8 pp 1342ndash1348 2005

[15] M H Yazer A Cortese-Hassett and D J Triulzi ldquoCoagulationfactor levels in plasma frozen within 24 hours of phlebotomyover 5 days of storage at 1 to 6∘Crdquo Transfusion vol 48 no 12pp 2525ndash2530 2008

[16] S Kleinman B Grossman and P Kopko ldquoA national survey oftransfusion-related acute lung injury risk reduction policies forplatelets and plasma in the United Statesrdquo Transfusion vol 50no 6 pp 1312ndash1321 2010

[17] E A Vogler and C A Siedlecki ldquoContact activation of blood-plasma coagulationrdquo Biomaterials vol 30 no 10 pp 1857ndash18692009

[18] H Alhumaidan T Cheves S Holme and J Sweeney ldquoStabilityof coagulation factors in plasma prepared after a 24-hour roomtemperature holdrdquo Transfusion vol 50 no 9 pp 1934ndash19422010

[19] P F van der Meer J A Cancelas R R Vassallo N RuggM Einarson and J R Hess ldquoEvaluation of the overnighthold of whole blood at room temperature before componentprocessing platelets (PLTs) fromPLT-rich plasmardquoTransfusionvol 51 supplement 1 pp 45Sndash49S 2011

[20] F Q Lu W Kang Y Peng and W M Wang ldquoCharacterizationof blood components separated from donated whole blood afteran overnight holding at room temperature with the buffy coatmethodrdquo Transfusion vol 51 no 10 pp 2199ndash2207 2011

[21] P F van der Meer J A Cancelas R Cardigan et al ldquoEvaluationof overnight hold of whole blood at room temperature beforecomponent processing effect of red blood cell (RBC) additivesolutions on in vitro RBC measuresrdquo Transfusion vol 51supplement 1 pp 15Sndash24S 2011

[22] M F Veale G Healey and R L Sparrow ldquoEffect of additivesolutions on red blood cell (RBC) membrane properties ofstored RBCs prepared from whole blood held for 24 hours atroom temperaturerdquo Transfusion vol 51 supplement 1 pp 25Sndash33S 2011

[23] M J Dijkstra-Tiekstra P F van der Meer R Cardigan et alldquoPlatelet concentrates from fresh or overnight-stored blood aninternational studyrdquo Transfusion vol 51 supplement 1 pp 15Sndash24S 2011

[24] S J Slichter J Corson M K Jones T Christoffel E Pellhamand D Bolgiano ldquoPlatelet concentrates prepared after a 20- to24-hour hold of the whole blood at 22∘Crdquo Transfusion vol 52no 9 pp 2043ndash2048 2012

[25] L McMillian V Hornsey A Morrison O Drummond and CProwse ldquoStorage of whole blood for up to 24H and its effect onplatelet concentratesrdquoTransfusionMedicine vol 19 supplement1 article 20S 2009

[26] R Cardigan P F van derMeer C Pergande et al ldquoCoagulationfactor content of plasma produced from whole blood stored for24 hours at ambient temperature results from an internationalmulticenter BEST Collaborative studyrdquo Transfusion vol 51supplement 1 pp 50Sndash57S 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


200 mL WB

Centrifuged

200 mL filtered WB

Plasma (FFP)

Plasma (I-FP24)

Plasma (II-FP24)

Centrifuged

200 mL WB Centrifuged

RT 400 mL WB

RT

BCs

Plasma P6

RTPCs

RT

BCs RT

PCs

Plasma P24

lt6 hrs

n = 12

n = 12

n = 12

n = 12

n = 12

n = 12

n = 24n = 24

n = 24

200 mL WBn = 12

n = 12

200 mL WBn = 12

200 mL WB

n = 24

4∘C 400 mL WB

18ndash24 hrs filtered

lt6 hrs

4∘C overnight

18ndash24 hrs

18ndash24 hrs

18ndash24 hrs

Overnight

4∘C

Figure 1 Experimental algorithm for the plasma and PCs production validation with WB held overnight (119899 = 2412 units in each arm) Theplasma was frozen as soon as possible after preparation

could limit the number ofWBunits that can be processed intoFFP because of the blood collections from blood-collectingvehiclesWB stored overnight at 4∘C can be used tomanufac-ture plasma and concentrated red blood cells but not for PCsbecause of the deleterious effect of 4∘C storage on PLT qualityPLTs do not tolerate refrigeration and disappear rapidly fromcirculation if subjected to chilling before transplantation [4ndash6] Thus WB used for PC separation must be stored atroom temperature Currently PC preparation according tothe BC removal method is widely practiced [7ndash9] and isassociated with significantly better PLT recoveries and lessPLT activation immediately after preparation [10] Howeverit is also inexpedient for PCs that are prepared during theimmediate processing of freshWB In addition an overnighthold of BC is good for the PC quantity and quality [11]There are numerous operational and economic advantagesto blood services from ambient WB storage within 24 hrsbecause of the reduced restrictions on blood processing forPC separation within 8 hrs of collection

The aim of this study was to compare the quality asmeasured by the coagulation factor levels of plasma preparedfromWB that was processed within 8 hrs after collection andstorage at 4∘C within 18ndash24 hrs of storage at 4∘C or within18ndash24 hrs of storage at ambient temperature and the qualityof PCs from the BCs processed either from freshWB or fromWB that was held overnight at room temperature

2 Materials and Methods

21 Study Design The blood component quality of WB thatwas stored overnight was evaluated the frame paragraphof these experiments is shown in Figure 1 Briefly 400mLWB samples were collected from normal volunteer donors

according to standard procedures in the blood donation lawof the Peoplersquos Republic of China (24 bags used for plasmapreparation were stored at 4∘C and 12 bags used for PCspreparation were stored at room temperature) The studyprotocol was approved by the Southwest Hospital HumanResearch and Ethics Committee EachWB sample (400mL plusmn10) was collected into blood collection packs that containeda CPD anticoagulant (Nigale Biomedical Co Ltd SichuanChina) and then equal volumes were divided between 2units (200mLunit) The blood components were processedin a 2-step centrifugation protocol (2600 rpm or 1642timesg for15min followed by 1000 rpm or 263timesg for 6min in a J6-MC centrifuge Beckman Miami FL USA) The plasma wasremoved and the packed RBCs were resuspended in one ofthe additive solutions SAG-M (100mL) (Nigale BiomedicalCo Ltd Sichuan China)The FFP was prepared within 6 hrsfrom 12 (119899 = 12) of the leukodepleted WB with a WBin-line leukocyte blood filter (Nigale Biomedical Co LtdSichuan China) The I-FP24 (I-FP24 6 hrs18ndash24 hrs) wereprepared from leukodepleted blood held overnight (119899 = 12)using a WB in-line leukocyte blood filter (Nigale BiomedicalCo Ltd Sichuan China) from WB within 6 hrs or II-FP24(II-FP24 18ndash24 hrs18ndash24 hrs) from WB held overnight andthen leukodepleted using a WB in-line leukocyte blood filter(Nigale Biomedical Co Ltd Sichuan China) (119899 = 24)One batch of PCs was prepared from BCs (stored overnight)obtained from fresh WB (freshstored 119899 = 12 controlgroup PC1) and the other batch of PCs was prepared fromBCs obtained from WB that was stored overnight at roomtemperature (storedfresh 119899 = 12 experimental group PC2)At the same time plasma from the 2 groups was preparedincluding P6 (119899 = 12) from the PC1 group and P24 (119899 =12) from the PC2 group All plasma samples were stored









415minus 084 (119860

380+ 119860450)] times

1000mgL


2 pCO





3 Results




2


4 Discussion








204(188ndash 226)

161 plusmn 043

148(118ndash214)

204 plusmn 040

215(134ndash243)

178 plusmn 059

197(155ndash224)

179 plusmn 059

201(152ndash219)

FV ()8657 plusmn 1873

9239(6125ndash10736)

6114 plusmn 1376

lowast

6064(4498ndash8161)

7566 plusmn 607

7865(6716ndash8161)

8121 plusmn 654

7865(7378ndash9239)

8759 plusmn 884

8396(7578ndash10414)


9258(6052ndash11300)

6155 plusmn 1145

lowast

6150(4453ndash7519)

7803 plusmn 1908

7196(4945ndash10849)

8032 plusmn 1737

7798(5092ndash11529)

8084 plusmn 1729

8102(4871ndash10849)


10760(6381ndash12834)

5790 plusmn 735

lowast

5580(5126ndash7031)

6556 plusmn 1393

998771

6137(4548ndash9265)

7798 plusmn 1160

7859(4895ndash9265)

6630 plusmn 1016

6506(4388ndash8437)


5740(5630ndash 5810)

5680 plusmn 450

5730(5180ndash6180)

5896 plusmn 317

5900(5390ndash6390)

5891 plusmn 294

5910(5480ndash6640)

5978 plusmn 314

5910(5600ndash6660)

pH721 plusmn 010

717(712ndash738)

722 plusmn 009

720(711ndash734)

724 plusmn 006

725(717ndash736)

736 plusmn 007

735(730ndash753)

718 plusmn 004

718(712ndash724)


15700(15550ndash15760)

15494 plusmn 381

15400(15140ndash15900)

15661 plusmn 201

15635(15380ndash16120)

15605 plusmn 277

15620(14970ndash16000)

15925 plusmn 324

15920(15310ndash16350)


335(302ndash353)

396 plusmn 022

lowast

403(361ndash420)

433 plusmn

028

998771∙ 439(371ndash469)

345 plusmn 021

344(319ndash384)

425 plusmn 027

428(367ndash469)


6990(6730ndash7320)

7032 plusmn 372

7020(6580ndash7450)

7092 plusmn 321

7110(6480ndash7570)

7193 plusmn 195

7195(6850ndash7480)

6905 plusmn 223

7020(6510ndash7220)


9100(9000ndash13400)

100 plusmn 678

9600(9400ndash11000)

14570plusmn2600

998771∙

15100(11000ndash17500)

10358 plusmn 2207

9950(6700ndash15000)

12083 plusmn 2439

995333

10800(8100ndash16100)


2458(2211ndash2539)

2278 plusmn 061

2255(2212ndash2371)

2272 plusmn 118

2292(2121ndash2438)

2309 plusmn 138

2278(2102ndash2548)

2193 plusmn 148

2179(1989ndash2521)


1002(123ndash2991)

4354 plusmn 1976

lowast

4439(2081ndash7303)

12996plusmn3433

998771∙

12994(7102ndash17717)

2215 plusmn 1951

1371(167ndash6656)

2746 plusmn 2911

995333

1440(389ndash8974)






PC1 (119899 = 12) PC2 (119899 = 12)


26 (111ndash658)349 plusmn 119

333 (211ndash560)

pH 712 plusmn 005

713 (703ndash717)690 plusmn 004

lowast

689 (684ndash699)


3545 (2980ndash3920)4919 plusmn 507

lowast

4790 (4380ndash5750)


11300 (10100ndash13500)9481 plusmn 1750

lowast

9540 (6090ndash11700)


295 (270ndash330)333 plusmn 024

lowast

340 (300ndash380)


13500 (13100ndash13700)13136 plusmn 211

lowast

13200 (12900ndash13500)


7300 (6900ndash7600)7300 plusmn 173

7200 (7100ndash7600)


1900 (1780ndash2140)2014 plusmn 128

2020 (1820ndash2260)


310 (230ndash470)413 plusmn 041

lowast


1006 plusmn 028

1006 (986ndash1025)1193 plusmn 018

lowast

1193 (1180ndash1206)


4321 (4161ndash4481)4037 plusmn 366

4037 (3778ndash4295)HSR 101 plusmn 268









5 Conclusion


Abbreviations





Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
























415minus 084 (119860

380+ 119860450)] times

1000mgL


2 pCO





3 Results




2


4 Discussion








204(188ndash 226)

161 plusmn 043

148(118ndash214)

204 plusmn 040

215(134ndash243)

178 plusmn 059

197(155ndash224)

179 plusmn 059

201(152ndash219)

FV ()8657 plusmn 1873

9239(6125ndash10736)

6114 plusmn 1376

lowast

6064(4498ndash8161)

7566 plusmn 607

7865(6716ndash8161)

8121 plusmn 654

7865(7378ndash9239)

8759 plusmn 884

8396(7578ndash10414)


9258(6052ndash11300)

6155 plusmn 1145

lowast

6150(4453ndash7519)

7803 plusmn 1908

7196(4945ndash10849)

8032 plusmn 1737

7798(5092ndash11529)

8084 plusmn 1729

8102(4871ndash10849)


10760(6381ndash12834)

5790 plusmn 735

lowast

5580(5126ndash7031)

6556 plusmn 1393

998771

6137(4548ndash9265)

7798 plusmn 1160

7859(4895ndash9265)

6630 plusmn 1016

6506(4388ndash8437)


5740(5630ndash 5810)

5680 plusmn 450

5730(5180ndash6180)

5896 plusmn 317

5900(5390ndash6390)

5891 plusmn 294

5910(5480ndash6640)

5978 plusmn 314

5910(5600ndash6660)

pH721 plusmn 010

717(712ndash738)

722 plusmn 009

720(711ndash734)

724 plusmn 006

725(717ndash736)

736 plusmn 007

735(730ndash753)

718 plusmn 004

718(712ndash724)


15700(15550ndash15760)

15494 plusmn 381

15400(15140ndash15900)

15661 plusmn 201

15635(15380ndash16120)

15605 plusmn 277

15620(14970ndash16000)

15925 plusmn 324

15920(15310ndash16350)


335(302ndash353)

396 plusmn 022

lowast

403(361ndash420)

433 plusmn

028

998771∙ 439(371ndash469)

345 plusmn 021

344(319ndash384)

425 plusmn 027

428(367ndash469)


6990(6730ndash7320)

7032 plusmn 372

7020(6580ndash7450)

7092 plusmn 321

7110(6480ndash7570)

7193 plusmn 195

7195(6850ndash7480)

6905 plusmn 223

7020(6510ndash7220)


9100(9000ndash13400)

100 plusmn 678

9600(9400ndash11000)

14570plusmn2600

998771∙

15100(11000ndash17500)

10358 plusmn 2207

9950(6700ndash15000)

12083 plusmn 2439

995333

10800(8100ndash16100)


2458(2211ndash2539)

2278 plusmn 061

2255(2212ndash2371)

2272 plusmn 118

2292(2121ndash2438)

2309 plusmn 138

2278(2102ndash2548)

2193 plusmn 148

2179(1989ndash2521)


1002(123ndash2991)

4354 plusmn 1976

lowast

4439(2081ndash7303)

12996plusmn3433

998771∙

12994(7102ndash17717)

2215 plusmn 1951

1371(167ndash6656)

2746 plusmn 2911

995333

1440(389ndash8974)






PC1 (119899 = 12) PC2 (119899 = 12)


26 (111ndash658)349 plusmn 119

333 (211ndash560)

pH 712 plusmn 005

713 (703ndash717)690 plusmn 004

lowast

689 (684ndash699)


3545 (2980ndash3920)4919 plusmn 507

lowast

4790 (4380ndash5750)


11300 (10100ndash13500)9481 plusmn 1750

lowast

9540 (6090ndash11700)


295 (270ndash330)333 plusmn 024

lowast

340 (300ndash380)


13500 (13100ndash13700)13136 plusmn 211

lowast

13200 (12900ndash13500)


7300 (6900ndash7600)7300 plusmn 173

7200 (7100ndash7600)


1900 (1780ndash2140)2014 plusmn 128

2020 (1820ndash2260)


310 (230ndash470)413 plusmn 041

lowast


1006 plusmn 028

1006 (986ndash1025)1193 plusmn 018

lowast

1193 (1180ndash1206)


4321 (4161ndash4481)4037 plusmn 366

4037 (3778ndash4295)HSR 101 plusmn 268









5 Conclusion


Abbreviations





Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of






















204(188ndash 226)

161 plusmn 043

148(118ndash214)

204 plusmn 040

215(134ndash243)

178 plusmn 059

197(155ndash224)

179 plusmn 059

201(152ndash219)

FV ()8657 plusmn 1873

9239(6125ndash10736)

6114 plusmn 1376

lowast

6064(4498ndash8161)

7566 plusmn 607

7865(6716ndash8161)

8121 plusmn 654

7865(7378ndash9239)

8759 plusmn 884

8396(7578ndash10414)


9258(6052ndash11300)

6155 plusmn 1145

lowast

6150(4453ndash7519)

7803 plusmn 1908

7196(4945ndash10849)

8032 plusmn 1737

7798(5092ndash11529)

8084 plusmn 1729

8102(4871ndash10849)


10760(6381ndash12834)

5790 plusmn 735

lowast

5580(5126ndash7031)

6556 plusmn 1393

998771

6137(4548ndash9265)

7798 plusmn 1160

7859(4895ndash9265)

6630 plusmn 1016

6506(4388ndash8437)


5740(5630ndash 5810)

5680 plusmn 450

5730(5180ndash6180)

5896 plusmn 317

5900(5390ndash6390)

5891 plusmn 294

5910(5480ndash6640)

5978 plusmn 314

5910(5600ndash6660)

pH721 plusmn 010

717(712ndash738)

722 plusmn 009

720(711ndash734)

724 plusmn 006

725(717ndash736)

736 plusmn 007

735(730ndash753)

718 plusmn 004

718(712ndash724)


15700(15550ndash15760)

15494 plusmn 381

15400(15140ndash15900)

15661 plusmn 201

15635(15380ndash16120)

15605 plusmn 277

15620(14970ndash16000)

15925 plusmn 324

15920(15310ndash16350)


335(302ndash353)

396 plusmn 022

lowast

403(361ndash420)

433 plusmn

028

998771∙ 439(371ndash469)

345 plusmn 021

344(319ndash384)

425 plusmn 027

428(367ndash469)


6990(6730ndash7320)

7032 plusmn 372

7020(6580ndash7450)

7092 plusmn 321

7110(6480ndash7570)

7193 plusmn 195

7195(6850ndash7480)

6905 plusmn 223

7020(6510ndash7220)


9100(9000ndash13400)

100 plusmn 678

9600(9400ndash11000)

14570plusmn2600

998771∙

15100(11000ndash17500)

10358 plusmn 2207

9950(6700ndash15000)

12083 plusmn 2439

995333

10800(8100ndash16100)


2458(2211ndash2539)

2278 plusmn 061

2255(2212ndash2371)

2272 plusmn 118

2292(2121ndash2438)

2309 plusmn 138

2278(2102ndash2548)

2193 plusmn 148

2179(1989ndash2521)


1002(123ndash2991)

4354 plusmn 1976

lowast

4439(2081ndash7303)

12996plusmn3433

998771∙

12994(7102ndash17717)

2215 plusmn 1951

1371(167ndash6656)

2746 plusmn 2911

995333

1440(389ndash8974)






PC1 (119899 = 12) PC2 (119899 = 12)


26 (111ndash658)349 plusmn 119

333 (211ndash560)

pH 712 plusmn 005

713 (703ndash717)690 plusmn 004

lowast

689 (684ndash699)


3545 (2980ndash3920)4919 plusmn 507

lowast

4790 (4380ndash5750)


11300 (10100ndash13500)9481 plusmn 1750

lowast

9540 (6090ndash11700)


295 (270ndash330)333 plusmn 024

lowast

340 (300ndash380)


13500 (13100ndash13700)13136 plusmn 211

lowast

13200 (12900ndash13500)


7300 (6900ndash7600)7300 plusmn 173

7200 (7100ndash7600)


1900 (1780ndash2140)2014 plusmn 128

2020 (1820ndash2260)


310 (230ndash470)413 plusmn 041

lowast


1006 plusmn 028

1006 (986ndash1025)1193 plusmn 018

lowast

1193 (1180ndash1206)


4321 (4161ndash4481)4037 plusmn 366

4037 (3778ndash4295)HSR 101 plusmn 268









5 Conclusion


Abbreviations





Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















PC1 (119899 = 12) PC2 (119899 = 12)


26 (111ndash658)349 plusmn 119

333 (211ndash560)

pH 712 plusmn 005

713 (703ndash717)690 plusmn 004

lowast

689 (684ndash699)


3545 (2980ndash3920)4919 plusmn 507

lowast

4790 (4380ndash5750)


11300 (10100ndash13500)9481 plusmn 1750

lowast

9540 (6090ndash11700)


295 (270ndash330)333 plusmn 024

lowast

340 (300ndash380)


13500 (13100ndash13700)13136 plusmn 211

lowast

13200 (12900ndash13500)


7300 (6900ndash7600)7300 plusmn 173

7200 (7100ndash7600)


1900 (1780ndash2140)2014 plusmn 128

2020 (1820ndash2260)


310 (230ndash470)413 plusmn 041

lowast


1006 plusmn 028

1006 (986ndash1025)1193 plusmn 018

lowast

1193 (1180ndash1206)


4321 (4161ndash4481)4037 plusmn 366

4037 (3778ndash4295)HSR 101 plusmn 268









5 Conclusion


Abbreviations





Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















5 Conclusion


Abbreviations





Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of







































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Documents

Research Article A Comparison Study of the Blood Component …downloads.hindawi.com/journals/jbt/2013/523539.pdf · Journal of Blood Transfusion 200 mL WB Centrifuged 200 mL ltered