Blood collection

To evaluate chemical and drug safety, different animal species are used. Blood is collected to estimate different parameters of drug toxicity under test. The collection of blood from different sites in test animals, along with specific needle size, has been recommended by regulatory bodies and international agencies (Table 12-4). Based on the size of the laboratory animal(s) and the requirement of the blood needed to generate quality data, blood is withdrawn from each test animal. This is to avoid withdrawal of a large volume of blood and to avoid injury to the test animal (Table 12-5). 

TABLE 12-4 Common Bleeding Sites of Laboratory Animals 

Saphenous Femoral Jugular Ear Tail Needle Size Species Heart Vein Vein Vein Vein Vein (gauge) 

Source Modified from Laboratory Animal Information Service and Centre, National Institute of Nutrition, Indian Council of Medical Research, Government of India, Hyderabad, India. 

TABLE 12-5 Practicable Volume of Blood (mL/kg) Obtainable from Various Animals 

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Primary blood components iaclude plasma, red blood cells (erythrocytes), white blood cells (leukocytes), platelets (thrombocytes), and stem cells. Plasma consists of water dissolved proteias, ie, fibrinogen, albumins, and globulins coagulation factors and nutrients. The principal plasma-derived blood products are siagle-donor plasma (SDP), produced by sedimentation from whole blood donations fresh frozen plasma (FFP), collected both by apheresis and from whole blood collections cryoprecipitate, produced by cryoprecipitation of FFP albumin, collected through apheresis and coagulation factors, produced by fractionation from FFP and by apheresis (see Fractionation, blood-plasma fractionation). 

W. H. Dzik, "Characteristics and Controversies of Blood Collected by Intra Operative Salvage."... 

The pH of blood is a variable, and loss of carbon dioxide and the resultant change in pH can be avoided by leaving the stopper of the blood collection tube in place during processing steps such as centrifugation and storage. 

Various additive mixtures have been proposed for inclusion in blood collection tubes to prevent in vitro platelet activation. One formulation involves the use of a mixture of acid-citrate-dextrose (ACD, 1 5 dilution), 30 p-M acetylsalicylic acid... 

In vitro platelet activation is dependent on the anticoagulant that is used for blood collection. In one study it was demonstrated that PF4 levels in platelet-poor plasma isolated after incubation without any stimuli for 1 hour at 37°C were as follows conventional heparin, 1180 ng/ml hirudin, 469 ng/ml citrate, 440 ng/ml and EDTA, 217 ng/ml (110). EDTA appears to suppress platelet degranulation. PF4 levels obtained with a low-molecular-weight heparin preparation called Frag-min were, however, comparable to those obtained with hirudin (110). 

A plasmin inhibitor such as aprotinin used for blood collection, while effective in inhibiting activation of plasminogen by urokinase, is ineffective against the ac-... 

Measurement of free t-PA in plasma presents challenges in terms of preventing t-PA from complexing to PAI-1 released from platelets after blood collection. To dissociate any preformed t-PA-PAI-1 complex, the anticoagulant pH has to be close to 3.0. Even if blood is collected with an acidic anticoagulant, the blood pH will rise because of the powerful buffering action of hemoglobin. Thus, the pH of plasma has to be adjusted to 3.0 in order to dissociate the t-PA-PAI-I complex (115). 

Molecular methods used to uncover mutations are subject to several variables. The anticoagulants used for blood collection can affect digestion with restriction enzymes and amplification reactions. The type of detergent used in cell lysis can affect amplification of DNA by inhibiting the DNA-amplifying enzyme such as the taq polymerase used in the polymerase chain reaction (116). The control of contamination is crucial in ensuring the quality of results obtained by molecular analysis (117). 

Contant G., Gouault-Heilmann M., Martinoli J. L. Heparin inactivation during blood storage Its prevention by blood collection in citric acid, theophylline, adenosine, dipyridamole—CTAD mixture. Thromb Res 1983 31,365-74. 

Kuhne T., Homstein A., Semple J., et al. Flow cytometric evaluation of platelet activation in blood collected into EDTA vs. Diatube-H, a sodium citrate solution supplemented with theophylline, adenosine and dipyridamole. Am J Hematol 1995 50,40-5. 

Mohler M. A., Refino C. J., Chen S. A., Chen A. B Hotchkiss A. J. D-Phe-Pro-Arg-chloromethylketone Its potential use in inhibiting the formation of in vitro artifacts in blood collected during tissue type plasminogen activator thrombolytic therapy. Thromb Haemost 1986 56, 160-4. 

Kluft C., Verheijen J. H. Leiden Fibrinolysis Working Party. Blood collection and handling procedures for assessment of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1). Fibrinolysis 1990 4(Suppl2) 155-61. 

Longnecker, M. P., Bernstein, L., Bird, C. L., Yancey, A. K. and Peterson, J. C. (1996). Measurement of organochlorine levels in postprandial serum or in blood collected in serum separator tubes . Cancer Epidemiol. Biomarkers Prev., 5(9), 753-755. 

In addition to laboratory blood analyzers and portable point-of-care devices, which require blood collection, continuous monitoring of ion activities in a blood stream via implanted ion-selective electrodes is of great interest. The term biocompatibility refers to the ability of a sensor not to cause toxic or injurious effects while being in contact with living tissue. As dealing with any foreign object introduced into the human body, biocompatibility and hemocompatibility particularly are the most important requirements. 

HOLDER BLOOD COLLECTING TUBE PLAS POLYPROP 2.4381G, 750"ID 10 S 6630012309964 PG 3.48 ... 

TUBE BLOOD COLLECTING VACUUM 7ML W/O ANTICOAGULANT NONSTERILE100 6630001451137 PG 7.50 ... 

Test System Human blood. Collect 30 ml heparinized blood for whole blood and plasma (three tubes) and 30 ml clotted blood for serum (two tubes) from each of six donors. 

Blood Collection. In rodent studies, large numbers of satellite animals (often close to the number used in the main study phase) are usually needed for pharmacokinetic blood sampling, whereas with most nonrodent species, blood samples can be collected from the main study animals without compromising their health status. 

Pharmacokinetic Blood collected at specified times after dosing on Days 1 and 28... 

Urine collection Pretreatment, monthly during treatment, and during Week 4 of reversal Pharmacokinetic samples Blood collected at specified times after dosing on Day 1 and during Weeks six and 12... 

Because most, if not all, study-related activities are conducted in the same dogs, the stress induced by repeated manipulation of dogs for activities such as blood collection, ECG, and physical examinations needs to be taken into consideration. Efforts should be made wherever possible to separate study activities by several days. 

Blood and Urine Collection. About 5-10 ml of blood can be collected from adult ferrets using retroorbital blood collection techniques. Other methods of blood... 

Jackson, R.K., Kieffer, VA., Sauber, J.J. and King, G.L. (1988). A tether-restraint system for blood collection from ferrets. Lab. Anim. Sci. 38 625-628. 

Blood collection from the tail vein is a simple and rapid, nonsurgical method which does not require anesthesia. A relatively large number of serial samples can be obtained within a short period of time. However, this method is limited to relatively small sample volumes (<250 pi per sample). Although larger volumes can be obtained by placing the rat in a wanning chamber, this procedure could significantly influence the disposition of the test compound and therefore is not recommended for routine studies. Blood collected from the cut tail has been shown to provide valid concentration data for numerous compounds. 

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