1、June 2007DEUTSCHE NORM English price group 8No part of this standard may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS 71.040.20!,x|3“9858916www.
2、din.deDDIN ISO 12772Laboratory glassware Disposable microhaematocrit capillary tubes (ISO 12772:1997)English version of DIN ISO 12772:2007-06Laborgerte aus Glas Einmal-Mikrohmatokrit-Kapillaren (ISO 12772:1997)Englische Fassung DIN ISO 12772:2007-06SupersedesDIN 12846:1978-11www.beuth.deDocument com
3、prises 11 pages 09.07DIN ISO 12772:2007-06 2 Contents Page National foreword 3 National Annex NA (informative) Bibliography . 3 1 Scope . 4 2 Normative references . 4 3 Definition . 4 4 Classification . 4 5 Construction 5 5.1 Material 5 5.2 Design 5 5.3 Dimensions 5 5.4 Ring mark 5 5.5 Resistance to
4、 centrifugal force . 5 5.6 Capillarity . 5 5.7 Anticoagulant (Type I only) 5 5.8 Efficacy of heparin coating (Type I only) 6 6 Markings 6 6.1 Labelling 6 6.2 Colour code . 6 Annex A (normative) Resistance to centrifugal force . 7 Annex B (normative) Capillarity . 8 Annex C (normative) Closing the tu
5、be before centrifugation 9 C.1 Sealing . 9 C.2 Melting . 9 Annex D (informative) Efficacy of anti-coagulant coating . 10 D.1 Preparation . 10 D.2 Procedure . 10 Annex E (informative) Test for amount of anti-coagulant in tube and anti-coagulant potency assay 11 E.1 Test for amount of anti-coagulant i
6、n tube: Toluidine blue test 11 E.2 Anti-coagulant potency assay: Chromogenic or fluorogenic substrate test 11 DIN ISO 12772:2007-06 3 National foreword This standard has been prepared by Technical Committee ISO/TC 48 “Laboratory glassware and related apparatus”, Subcommittee SC 1 “Volumetric instrum
7、ents”. The responsible German bodies involved in its preparation were the Normenausschuss Laborgerte und Laboreinrichtungen (Laboratory Equipment Standards Committee), Technical Committee Volumenmess-gerte, and the Normenausschuss Medizin (Medical Standards Committee), Technical Committee Hmatologie
8、. The DIN Standards corresponding to the International Standards referred to in clause 2 of this document are as follows: ISO 719 DIN ISO 719 International Standard ISO 8417 as cited in clause 2 does not exist, since any work on that standards project was stopped following publication of ISO 12772.
9、Amendments This standard differs from DIN 12846: 1978-11 as follows: a) Dimensions and details of marking have been amended. b) Additional and more precise requirements have been specified. c) Five new tests have been included. Previous editions DIN 12846: 1978-11 National Annex NA (informative) Bib
10、liography DIN ISO 719, Glass Hydrolytic resistance of glass grains at 98 C Method of test and classification Laboratory glassware - Disposable microhaematocrit capillary tubes 1 Scope This International Standard provides details for two types of disposable glass capillary tubes, suitable for the mic
11、rohaematocrit test and other analytical tests which include a separation of plasma and cells. The details specified are in conformity with IS0 8417, to the greatest possible extent. WARNING - Capillary tubes can break during the analytical test procedure, resulting in broken sharp glass, blood spill
12、age and aerosols. Appropriate precautions shall be taken to avoid dangerous infections. 2 Normative references The following standards contain provisions which, through references in this text, constitute provisions of this International Standard. At the time of publication, the editions indicated w
13、ere valid. All standards are subject to revision, and parties to agreements based on this International Standard are encouraged to investigate the possibility of applying the most recent editions of the standards indicated below. Members of IEC and IS0 maintain registers of currently valid Internati
14、onal Standards. IS0 719:1985, Glass - Hydrolytic resistance of glass grains at 98 OC - Method of test and classification. IS0 8417:-l, Laboratory volumetric instruments - Principles of design and construction of disposable volumetric articles. 3 Definition For the purposes of this International Stan
15、dard, the following definition applies. 3.1 disposable: Adjective used to describe microhaematocrit capillary tubes which are intended to be used once only and then discarded. NOTE - Such capillary tubes will only be expected to provide their specified performance during the original operation. 4 Cl
16、assification This International Standard describes two types of disposable glass capillary tubes as follows: - Type I: coated with anti-coagulant - Type II: uncoated NOTE - Ammonium heparin, lithium heparin and sodium heparin are the preferred anti-coagulants for the microhaematocrit test. Salts of
17、EDTA 21 and fluorides (usually sodium or potassium) may be used when capillary tubes are needed for special purposes. ) To be published. 2, EDTA is a n abbreviation for ethylenediaminotetraacetic acid 4 DIN ISO 12772:2007-065 Construction 5.1 Material When tested in accordance with the procedure and
18、 classification given in IS0 719, the glass used for the capillary tubes shall at least comply with the requirements of class HGB 3. The glass shall be free from visible defects and shall be free from internal stresses which would impair the performance of the capillary tube. 5.2 Design The tubes sh
19、all be straight and open at both ends without lip or constriction. The tubes may be slightly fire- polished on one or both ends. 5.3 Dimensions The dimensions of the tubes shall be as follows: Length: Internal diameter: Wall thickness: (752 0,5) mm (I,15 + 0,081 mm (0,2+0,025) mm The bore of the tub
20、es shall be uniform and shall not vary by more than 4 % of the total over a distance of 75 mm. NOTE - As the bore variation is packaging of the capillary tubes. critical for the analytical result, smaller bore variations than 4 % may be marked on the 5.4 Ring mark The tubes may be provided with a ri
21、ng mark to ease filling. Such a ring mark shall be at a distance of 60 mm max. from one end of the tube. The line thickness shall be between 0,5 mm and 1 mm. In the case of colour-coded tubes (see 6.2), the colour of the ring mark should preferably be that of the colour code, or may be black. 5.5 Re
22、sistance to centrifugal force When the tubes are tested as specified in annex A, not more than 0,5 % shall break. 5.6 Capillarity The tube shall be capable of drawing plasma, serum or human whole blood to a level within 20 mm from the far end of the tube when tested as specified in annex B. NOTE - I
23、n view of the possible infection risks, human blood should not be used. 5.7 Anti-coagulant (Type I only) The inner surface of Type I tubes shall be evenly coated with an anti-coagulant (see clause 4). In the case of heparin, the coating shall show an activity of 2,13 IU to 7,45 IU (International Uni
24、ts) per capillary when tested as described in annex E. An airtight container should be used in order to prevent deterioration of the anti- coagulant caused by air moisture. 5 DIN ISO 12772:2007-065.8 Efficacy of heparin coating (Type I only) Coagu tested . latr as on of the described sheep in ann pl
25、asm ex D. a shall not be ev ident when viewed under normal room lighting . This can be 6 Markings 6.1 Labelling The outer shipping carton shall be clearly marked with the following information: a) manufacturers or vendors name and/or mark; b) product description (e.g. disposable microhaematocrit cap
26、illary tubes) and the glass material (e.g. borosilicate glass 3.3 or soda-lime glass); c) number of tubes in the package; d) batch number or date of manufacture; e) the number of this International Standard, i.e. IS0 12772; and, if applicable (Type I only): f) type of anti-coagulant (e.g. lithium he
27、parin); g) anti-coagulant activity and date of expiration. NOTE - As the connection with depend expiry date may strongly storage requirements. on storage conditions, that information should preferably be given in The smallest packaging unit shall be marked at least with the information stated in a),
28、 b), c) and f). 6.2 Colour code 6.23 Each Type I tube shall be colour coded to identify the type of anti-coagulant. The colour code for tubes coated with the following anti-coagulants shall be: -orange -red -green -yellow -purple for lithium heparin for sodium heparin for ammonium heparin for sodium
29、 or potassium fluoride for sodium or potassium EDTA At the manufacturers discretion, the tubes may be colour coded at the extreme tip of the tube (see also 5.4). Anti-coagulants containing lithium should not be used if the sample is to be tested for lithium; the same constraints apply to sodium, pot
30、assium and ammonium compounds. 6.2.2 Type II tubes may be colour coded to identify that they are uncoated. If colour coded, the colour shall be blue. 3 6 DIN ISO 12772:2007-06Annex A (normative) Resistance to centrifugal force A.1 Fill the tube to capacity with distilled water or a blood product as
31、described in A.2, sealed or melted (see annex C for details) and place it in a microhaematocrit centrifuge. The centrifuge shall be accelerated until the bottoms of the tubes are subjected to an RCF of at least 11,000. Run the centrifuge at that speed for 4 min only, then shut it off and allow it to
32、 stop without using the brake. A.2 Animal (e.g. bovine, equine) plasma, serum or whole or definibrated blood may be used for the above test. Human blood should not be used unless it has been tested and shown to be free of HIV and Hepatitis B and C. 7 DIN ISO 12772:2007-06Annex B (normative) Capillar
33、ity B.l The tubes shall be tested for capillarity when held at a horizontal level by immersing one end in a drop of a blood product as described in 8.2. The tube shall fill within a 15 s time interval. B.2 Animal (e.g. bovine, equine) plasma, serum or whole or definibrated blood may be used for the
34、above test. Human blood should not be used unless it has been tested and shown to be free of HIV and Hepatitis B and C. 8 DIN ISO 12772:2007-06Annex C (normative) Closing the tube before centrifugation In order to avoid spilling of blood, the tubes shall be closed at one end before placing them in t
35、he centrifuge. This can be done by sealing or melting. In either case, the tube shall not be filled completely to allow for dry space. Cl Sealing The tube filled with blood,.and still held horizontal, shall be pressed with its “dry” end into the sealant. C.2 Melting The “dry” end of the tube, which
36、is filled with blood and still held horizontal, shall be slightly turned in the flame of a microburner. After a flat bottom has formed, a few seconds shall be allowed for cooling. NOTE - This melting process can be facilitated by an optimization of dimensions of the tube and properties of the glass
37、(e.g. internal diameter I,15 mm, wall thickness 0,225 mm, softening point max. 700 “C). To attain a rectangular, non-conica by turning the capillary with a smal I shape motor. of the melted bottom, the melting should be performed preferably Sometimes anti-coagulants can cause remnants if the capilla
38、ries are closed by melting. As remnants effect the analytical result, these capillaries shall be closed by sealing. The manufacturer should mark on the packaging of the capillaries if they shall be closed by sealing. 9 DIN ISO 12772:2007-06Annex D (informative) Efficacy of anti-coagulant coating The
39、 heparin-coated tubes may be tested by the following method. D.l Preparation Carry out the test initially by preparing sheep plasma from whole sheep blood by using citrate, oxalate or EDTA as anti-coagulant. The sheep plasma is afterwards recalcified as follows: 10 ml sheep plasma are added to 2 ml
40、of a 10 g/l calcium chloride solution. Mix the sheep plasma and calcium chloride solution well. 0.2 Procedure Use as controls samples of both the plain sheep plasma and recalcified sheep plasma in accordance with the following: a) positive control: a plain (i.e. non-coated) tube filled with recalcif
41、ied sheep plasma; b) negative control: a coated tube filled with plain sheep plasma. Immediately after the preparation of the recalcified sheep plasma, fill the tubes by immersing the tips in the recalcified sheep plasma and in the plain sheep plasma, holding the tubes at such an angle as to facilit
42、ate quick filling. Fill the tubes to within 5 mm from the other end and immediately agitate them to bring the plasma into contact with the anti-coagulant, whereby avoiding premature coagulation. Then place them in a horizontal position. At the end of 1 h, the tubes containing plasma shall be inspect
43、ed for evidence of coagulation by carefully snapping off segments of tubing in approximately 25 mm lengths and placing them on a flat surface (use a black background to facilitate observation and comparison with the controls). Coagulation has occurred if the sheep plasma becomes opaque or if a fine
44、fibrin thread is noted. 10 DIN ISO 12772:2007-06Annex E (informative) Test for amount of anti-coagulant in tube and anti-coagulant potency assay E.l Test for amount of anti-coagulant in tube: Toluidine blue test Heparin reacts like other glycosaminoglycans with toluidine blue to give a purple-colour
45、ed reaction product. The absorbance of the purple colour, measured in a spectrophotometer at a wavelength of 606 nm corresponds to the amount of heparin. The spectrophotometer is calibrated with standard solutions of heparin of known concentration or activity. E.2 Anti-coagulant potency assay: Chrom
46、ogenic or fluorogenic substrate test Heparin reacts with antithrombin and thrombin to form an additive. The residual thrombin releases a chromophore or a fluorophore from a synthetic peptide. The decrease of absorbance, measured in a spectrophotometer at a wavelength of 405 nm, or the decrease of fl
47、uorescence, measured in a fluorometer at an excitation of 335 nm and an emission of 415 nm corresponds to the activity of the heparin. The spectrophotometer or the fluorometer is calibrated with standard solutions of heparin of known activity. NOTE - Details of tests E.1 and E.2 can be found in NCCLS Document H24-T, Vol. 8 No. 5, Appendix A and Appendix B, pp. 39 - 45. ISSN 0273-3099. Purchase address: NCCLS, 771 E. Lancaster Avenue, Villanova, PA 19085 USA. 11 DIN ISO 12772:2007-06
