发布时间 : 星期六 文章良好的实验动物给药和采血（包括途径和体积）规范指南更新完毕开始阅读
A Good Practice Guide to the Administration of Substances and Removal of Blood,Including Routes and
Volumes
良好的实验动物给药和采血(包括途径和体积)规范指南
Karl-Heinz Diehl1, Robin Hull2, David Morton3, Rudolf Pfister4, Yvon Rabemampianina5, David Smith6,*, Jean-Marc Vidal7 and Cor van de Vorstenbosch 8
1
Aventis, PO Box 1140, D35001 Marburg, Germany
德国马尔堡市35001区1140信箱安万特公司
2
N I B S C, Blanch Lane, South Miimms, Potters Bar, Hertfordshire EN6 3QG
英国赫特福德郡EN6 3QG波特斯巴镇South Miimms布兰奇道英国国家生物制品检定所
3
The University of Birmingham, Medical School, Edgbaston, Birmingham B15 2TT
英国伯明翰市B15 2TT艾吉马斯顿伯明翰大学医学院
4
Novartis Pharma AG, CH-4002 Basel, Switzerland
瑞士巴塞尔CH-4002诺华制药公司
5
Centre de Recherche Pfizer, Etablissement d?Amboise, Z1 Poce′-sur-Cisse-BP 159 37401 Amboise Cedex, France
法国Amboise Cedex Z1 Poce′-sur-Cisse-BP 159 37401 Etablissement d?Amboise 辉瑞研究中心
6
AstraZeneca R&D Charnwood, Bakewell Road, Loughborough, Leics LE11 5RH
英国莱斯特郡LE11 5RH拉夫堡市贝克韦尔路Charnwood阿斯利康研究中心
7
Aventis, 102 Route de Noisy, 95235 Romainville Ce′dex, France
法国Romainville Ce′dex 95235 Noisy路102号安万特公司 8N V Organon, PO Box 20, 5340 BH Oss, Netherlands 荷兰BH Oss5340 20号信箱欧加农公司
Key words: blood volumes; blood removal; administration substances; laboratory animals; refinement. 关键词：血容量；采血；给药；实验动物；简化
This article is the result of an initiative between the European Federation of Pharmaceutical Industries Associations (EFPIA) and the European Centre for the Validation of Alternative Methods (ECVAM).Its objectives are to provide the researcher in the safety evaluation laboratory with an up-to-date, easyto-use set of data sheets to aid in the study design process whilst at the same time affording maximum welfare considerations to the experimental animals.
该文章为欧盟制药工业协会(EFPIA)和欧洲替代动物实验方法验证中心(ECVAM)之间的初步结果。其目的在于为安全性评价实验室的研究者提供最新的易于使用的数据库以帮助研究设计过程，同时最大可能地考虑到实验动物的福利。
Although this article is targeted at researchers in the European Pharmaceutical Industry, it is considered that the principles underpinning the data sets and refinement proposals are equally applicable to all those who use these techniques on animals in their research, whether in research institutes,universities or other sectors of industry. The implications of this article may lead to discussion with regulators, such as those responsible for pharmacopoeial
testing.
尽管该文章针对的是欧洲制药工业界的研究者，但支撑该数据库的基本原理及改进建议同样适用于所有在他们的研究中使用这些动物实验技术的人，不论是研究所、大学或其它行业中的研究者。
There are numerous publications dealing with the administration of test substances and the removal of blood samples, and many laboratories also have their own ?in-house? guidelines that have been developed by custom and practice over many years. Within European Union Directive 86/609EEC1 we have an obligation to refine experiments to cause the minimum amount of stress. We hope that this article will provide background data useful to those responsible for protocol design and review.
有关供试品给予和采血的出版物众多，且许多实验室在多年的经验和实践基础之上亦发展了它们自己的内部指南。在欧盟化妆品标准86/609EEC中，我们有义务简化实验以最小化动物的紧张程度。我们希望该文能够对那些负责方案设计和审核的研究者提供有用的背景数据。
This guide is based on peer-reviewed publications whenever possible, but where this is not possible we have used ?in-house? data and the experience of those on the working party (as well as helpful comments submitted by the industry) for a final opinion. The guide also addresses the continuing need to refine the techniques associated with the administration of substances and the withdrawal of blood, and suggests ways of doing so. Data-sharing between laboratories should be encouraged to avoid duplication of animal work, as well as sharing practical skills concerning animal welfare and scientific problems caused by ?overdosing? in some way or another. The recommendations in this guide refer to the ?normal? animal, and special consideration is needed, for instance, during pregnancy and lactation.Interpretation of studies may be confounded when large volumes are administered or excessive sampling employed, particularly if anaesthetics are used. Copyright ? 2001 John Wiley & Sons, Ltd.
该文章基于历年所有可能收集到的同行评议出版物，但我们未能够收集到的内部数据和那些工作组的经验(以及行业提交的有用的注释)除外。该指南亦强调了持续性简化与给药和采血有关的技术的必要性，并且建议该如何去进行这方面的工作。应该鼓励实验室间的数据共享以避免重复性动物研究，以及共享在某些方法或其它情况下的“药物过量”所引起的与动物福利有关的实际技术和科学问题。有必要对该指南中涉及到的“正常动物”要求进行特殊考虑，如妊娠和哺乳期间的动物。当给药体积较大或过度采样时对研究结果的诠释可能会令人感到困惑，特别是使用麻醉动物时。
GOOD PRACTICE GUIDE FOR ADMINISTRATION OF SUBSTANCES 良好的给药规范指南 Introduction 引言
Dosing of experimental animals is necessary for a variety of scientific investigations and to meet regulatory demands. The pharmaceutical industry, in particular,has investigated the levels of dosing compatible with animal welfare and valid science.2 In the preclinical stage of the safety evaluation of new drugs it is normal practice to use multiples of the ?effective dose?in order to attempt to establish the necessary safety margins. Where chemicals are of low toxicity or are only poorly soluble in acceptable formulations, a large volume may be required to be given to
individual animals to satisfy both scientific and regulatory requirements.The intended clinical use may also have an impact on the acceptability of larger than usual dose volumes, e.g. imaging agents or plasma expanders for intravenous application.
各种科学研究都需要对实验动物给药以符合药品注册要求。特别是在制药工业领域已研究了与动物福利以及科学性相一致的给药水平。在新药的临床前安全性评价阶段使用多种“有效剂量”以尽量确定必要的安全性范围是一种常规惯例。在使用低毒或溶解性极差的化学药品以一种可接受的制剂形式进行研究时，动物的个体给药体积可能较大以满足科学性和注册的要求。拟用临床剂量可能会使得给药体积较大，超过了动物可接受的正常给药体积，如静脉内注射用的造影剂或血浆增容药物。
The objectives of the Technical Sub group of EFPIA/ECVAM were as follows:
(i) to provide a guide on administration volumes for use in common laboratory species used in toxicity studies required by regulatory authorities;
(ii) to provide consensus dosage levels for routine use that represent good practice in terms of animal welfare and practicality;
(iii) to produce a guide to dosage levels representing the upper limit of common practice, which leaves scope to make the case for special investigations.
EFPIA/ECVAM技术小组的目标如下：
(i)提供一个药品注册当局所要求的毒性研究中常规使用的实验动物的给药体积的指南； (ii)根据动物福利和实用性提供一个在良好规范条件下常规使用的一致性剂量水平； (iii)提供一个代表常规规范上限的剂量水平，以为特殊研究留下一定的剂量扩展余地。 Administration volumes 给药体积
Table 1 presents administration volumes for the commonly employed routes in the most frequently used species. They are consensus figures based on published literature and internal guidelines. The marmoset and minipig are now considered within this category because they are being used increasingly in Europe.
表1表示最常使用的种属的一般给药途径下的给药体积。它们是根据发表的文献和内部指南综合得到的结果。现在认为绒猴和小型猪在此类常用动物之列，因为它们在欧洲的使用日渐增加。
Two sets of values are shown in each column:values on the left are intended as a guide to ?good practice? dose volumes for single or multiple dosing;values on the right, where given, are the possible maximal values. If maximal values are exceeded, animal welfare or scientific implications may result and reference to the responsible veterinary surgeon should be made. In some instances values are there to accommodate pharmacopoeial requirements.
每一列显示了两组数据，左侧数据拟用来指导在单次或多次给药的“良好规范”中的给药体积；所给出的右侧数据为可能的最大给药值。如果超过了最大值就会涉及到动物福利或科学性，应当参考负责兽医的外科医生的意见。在某些实际运用中，这些数据应符合药典要求。
Some of these suggested maximum values have been obtained from recent literature,3,4 but appear high when compared with ?good practice? values. The need for careful attention to animal welfare and the formulation of
material used at high dose volumes are emphasized,particularly if repeat dosing is intended. Study duration could be restricted and scientific validity compromised by physiological reaction to high dose volumes. It is therefore essential from an ethical standpoint that these issues are fully considered, e.g. by inspectorate or ethical committee, before protocols are finalized and work commences. It is also strongly recommended for ethical as well as scientific reasons that physicochemical compatibility studies (in vitro) and smallscale pilot studies (small groups of animals) are carried out on any new formulation before committing to larger scale studies. Dose volumes should be the minimum compatible with compound formulation and accuracy of administration.
从近来发表的文献中已经得到了这些建议的最大值中的某些数据，但当与“良好规范”数据相比时显得较高。强调了较高给药体积时对动物福利和使用的原料制剂进行仔细关注的必要性，特别是拟进行重复给药时。研究的持续时间和科学有效性应该让步于由于给药体积过高所出现的生理反应。因此在方案定稿以及着手研究之前通过比如监察员或伦理委员会充分地考虑到了这些出版物中的伦理观要素。亦就伦理及科学依据强烈要求对任何新剂型进行物理化学相容性研究(体外)和小型的先导性研究(少量动物组) 以免在大型研究中出现失误。给药体积应该最大限度地与化合物剂型和给药准确性相一致。 Administrative routes 给药途径
Oral route. On occasions, it may be necessary to restrict the animals? food intake before dosing. This factor may affect absorption. Large dose volumes (40 ml kg?1) have been shown to overload the stomach capacity and pass immediately into the small bowel.5Larger volumes may also reflux into the oesophagus.The duration of fasting will depend upon the feeding pattern of the species, the starting time for food restriction,the physiology of the species, the length of time of dosing, diet and the light cycle.6 It is recommended that for accuracy of dosing and to avoid dosing accidents liquids are administered by gavage.
经口给药途径：某些情况下在给药前有必要限制动物的摄食。该因素可能会影响药物吸收。较大的给药体积(40ml/kg)表明超过了胃容量负荷并快速通过胃进入小肠。较大的给药体积亦可以造成食管返流。禁食时间取决于动物种属的饲养方式、禁食的起始时间、种属的生理学特征、给药时间的长短、食物和光照周期。当药液通过灌胃给予时，要求给药必须准确以避免给药意外。
Parenteral routes. For substances administered parenterally,the dose volume used, stability of the formulation before and after administration, pH, viscosity,osmolality, buffering capacity, sterility and biocompatibility of the formulation are factors to consider. This is particularly important for multiple dose studies. These factors are reviewed in some detail by Claassen.7 The smallest needle size should be used, taking into account the dose volume, viscosity of injection material, speed of injection and species.
胃肠外给药途径：对于经胃肠外给予的药物而言，应考虑所采用的制剂的给药体积、给药前和给药后制剂的稳定性、pH、粘度、等渗性、缓冲能力、无菌及生物相容性因素。这对于多次给药研究尤其重要。Claassen总结了这些因素中的某些细节。应使用最小型号的针头、考虑给药体积、注射物粘度、注射速度和动物种属。
Subcutaneous. This route is frequently used. The rate and extent of absorption depend on the formulation. 皮下给药：该途径经常使用。吸收的速度和程度取决于制剂。