1、BRITISH STANDARD BS ISO 12772:1997 Laboratory Glassware Disposable microhaematocrit capillary tubes ICS 11.100; 71.040.20BSISO12772:1997 This British Standard, having been prepared under the directionof the Sector Board forMaterials and Chemicals, waspublished under the authorityof the Standards Boa
2、rdand comes intoeffecton 15August 1997 BSI 09-1999 ISBN 0 580 28409 3 National foreword This British Standard reproduces verbatimISO12772:1997 and implements it as the UK national standard. The UK participation in its preparation was entrusted by Technical Committee LBI/36, Laboratory glassware and
3、related apparatus, to Subcommittee LBI/36/1, Volumetric instruments, which has the responsibility to: aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; mo
4、nitor related international and European developments and promulgate them in the UK. A list of organizations represented on this subcommittee can be obtained on request to its secretary. Cross-references The British Standards which implement international or European publications referred to in this
5、 document may be found in the BSI Standards Catalogue under the section entitled “International Standards Correspondence Index”, or using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of
6、 British Standards are responsible for their correct application. Compliance with a British Standard does not of itself confer immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover, pages i and ii, theISO title page, pages ii to iv, pages 1 to
7、 4 and abackcover. This standard has been updated (see copyright date) and may have had amendments incorporated. This will be indicated in the amendment table on the inside front cover. Amendments issued since publication Amd. No. Date CommentsBSISO12772:1997 BSI 09-1999 i Contents Page National for
8、eword Inside front cover Foreword iii Text of ISO 12772 1ii blankBSISO12772:1997 ii BSI 09-1999 Contents Page Foreword iii 1 Scope 1 2 Normative references 1 3 Definition 1 4 Classification 1 5 Construction 1 6 Markings 2 Annex A (normative) Resistance to centrifugal force 3 Annex B (normative) Capi
9、llarity 3 Annex C (normative) Closing the tube before centrifugation 3 Annex D (informative) Efficacy of anti-coagulant coating 3 Annex E (informative) Test for amount of anti-coagulant in tube and anti-coagulant potency assay 4 Descriptors: Laboratory equipment, disposable equipment, laboratory gla
10、ssware, capillary tubes, classification, specifications, dimensions, marking, colour codes.BSISO12772:1997 BSI 09-1999 iii Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International
11、 Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, a
12、lso take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an Interna
13、tionalStandard requires approval by at least75% of the member bodies casting a vote. This International StandardISO12772 has been prepared by Technical Committee ISO/TC48, Laboratory glassware and related apparatus, Subcommittee SC 1, Volumetric instruments. Annex A,Annex B andAnnex C form an integr
14、al part of this InternationalStandard. Annex D andAnnex E are for information only.iv blankBSISO12772:1997 BSI 09-1999 1 1 Scope This International Standard provides details for two types of disposable glass capillary tubes, suitable for the microhaematocrit test and other analytical tests which inc
15、lude a separation of plasma and cells. The details specified are in conformity withISO8417, to the greatest possible extent. WARNING Capillary tubes can break during the analytical test procedure, resulting in broken sharp glass, blood spillage and aerosols. Appropriate precautions shall be taken to
16、 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 were valid. All standards are subject to revision, and parti
17、es 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 ISO maintain registers of currently valid International Standards. ISO 719:1985, Glass Hydrolytic resistance o
18、f glass grains at98 C Method of test and classification. ISO 8417:, Laboratory volumetric instruments Principles of design and construction of disposable volumetric articles 1) . 3 Definition For the purposes of this International Standard, the following definition applies. 3.1 disposable adjective
19、used to describe microhaematocrit capillary tubes which are intended to be used once only and then discarded NOTESuch capillary tubes will only be expected to provide their specified performance during the original operation. 4 Classification This International Standard describes two types of dispos
20、able glass capillary tubes as follows: Type I: coated with anti-coagulant Type II: uncoated NOTEAmmonium heparin, lithium heparin and sodium heparin are the preferred anti-coagulants for the microhaematocrit test. Salts of EDTA 2)and fluorides (usually sodium or potassium) may be used when capillary
21、 tubes are needed for special purposes. 5 Construction 5.1 Material When tested in accordance with the procedure and classification given inISO719, 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 shal
22、l be free from internal stresses which would impair the performance of the capillary tube. 5.2 Design The tubes shall 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 fol
23、lows: The bore of the tubes shall be uniform and shall not vary by more than 4% of the total over a distance of75mm. NOTEAs the bore variation is critical for the analytical result, smaller bore variations than4% may be marked on the packaging of the capillary tubes. 5.4 Ring mark The tubes may be p
24、rovided with a ring mark to ease filling. Such a ring mark shall be at a distance of60mm max. from one end of the tube. The line thickness shall be between0,5mm and1mm. In the case of colour-coded tubes (see6.2), the colour of the ring mark should preferably be that of the colour code, or may be bla
25、ck. 5.5 Resistance to centrifugal force When the tubes are tested as specified inAnnex A, not more than0,5% shall break. 5.6 Capillarity The tube shall be capable of drawing plasma, serum or human whole blood to a level within20mm from the far end of the tube when tested as specified inAnnex B. NOTE
26、In view of the possible infection risks, human blood should not be used. 1) To be published. 2) EDTA is an abbreviation for ethylenediaminotetraacetic acid Length: (75 0,5) mm Internal diameter: (1,15 0,08) mm Wall thickness: (0,2 0,025) mmBSISO12772:1997 2 BSI 09-1999 5.7 Anti-coagulant (Type I onl
27、y) The inner surface of Type I tubes shall be evenly coated with an anti-coagulant (see clause4). In the case of heparin, the coating shall show an activity of2,13IU to7,45IU (International Units) per capillary when tested as described inAnnex E. An airtight container should be used in order to prev
28、ent deterioration of the anti-coagulant caused by air moisture. 5.8 Efficacy of heparin coating (Type I only) Coagulation of the sheep plasma shall not be evident when viewed under normal room lighting. This can be tested as described inAnnex D. 6 Markings 6.1 Labelling The outer shipping carton sha
29、ll be clearly marked with the following information: a) manufacturers or vendors name and/or mark; b) product description (e.g. disposable microhaematocrit capillary tubes) and the glass material (e.g. borosilicate glass3.3or soda-lime glass); c) number of tubes in the package; d) batch number or da
30、te of manufacture; e) the number of this International Standard, i.e.ISO12772; and, if applicable (Type I only): f) type of anti-coagulant (e.g. lithium heparin); g) anti-coagulant activity and date of expiration. NOTEAs the expiry date may strongly depend on storage conditions, that information sho
31、uld preferably be given in connection with storage requirements. The smallest packaging unit shall be marked at least with the information stated in a), b), c) and f). 6.2 Colour code 6.2.1 Each Type I tube shall be colour coded to identify the type of anti-coagulant. The colour code for tubes coate
32、d with the following anti-coagulants shall be: At the manufacturers discretion, the tubes may be colour coded at the extreme tip of the tube (seealso5.4). Anti-coagulants containing lithium should not be used if the sample is to be tested for lithium; the same constraints apply to sodium, potassium
33、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. orange for lithium heparin red for sodium heparin green for ammonium heparin yellow for sodium or potassium fluoride purple for sodium or potassium EDTABSISO
34、12772:1997 BSI 09-1999 3 Annex A (normative) Resistance to centrifugal force A.1 Fill the tube to capacity with distilled water or a blood product as described inA.2, sealed or melted (seeAnnex C for details) and place it in a microhaematocrit centrifuge. The centrifuge shall be accelerated until th
35、e bottoms of the tubes are subjected to an RCF of at least11,000. Run the centrifuge at that speed for4min only, then shut it off and allow it to 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 sh
36、ould not be used unless it has been tested and shown to be free of HIV and Hepatitis B and C. Annex B (normative) Capillarity B.1 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 inB.2. The tube shall fill within
37、 a15s time interval. B.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. Annex C (normative) Closing the tube before centrifugation In
38、 order to avoid spilling of blood, the tubes shall be closed at one end before placing them in the centrifuge. This can be done by sealing or melting. In either case, the tube shall not be filled completely to allow for dry space. C.1 Sealing The tube filled with blood, and still held horizontal, sh
39、all be pressed with its “dry” end into the sealant. C.2 Melting The “dry” end of the tube, which 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. NOTEThis melting proces
40、s can be facilitated by an optimization of dimensions of the tube and properties of the glass (e.g. internal diameter1,15mm, wall thickness0,225mm, softening point max.700 C). To attain a rectangular, non-conical shape of the melted bottom, the melting should be performed preferably by turning the c
41、apillary with a small motor. Sometimes anti-coagulants can cause remnants if the capillaries 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 seal
42、ing. Annex D (informative) Efficacy of anti-coagulant coating The heparin-coated tubes may be tested by the following method. D.1 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 afterwa
43、rds recalcified as follows:10ml sheep plasma are added to2ml of a10g/l calcium chloride solution. Mix the sheep plasma and calcium chloride solution well. D.2 Procedure Use as controls samples of both the plain sheep plasma and recalcified sheep plasma in accordance with the following: a) positive c
44、ontrol: a plain (i.e. non-coated) tube filled with recalcified 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 she
45、ep plasma, holding the tubes at such an angle as to facilitate quick filling. Fill the tubes to within5mm 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
46、 end of1h, the tubes containing plasma shall be inspected for evidence of coagulation by carefully snapping off segments of tubing in approximately25mm lengths and placing them on a flat surface (use a black background to facilitate observation and comparison with the controls). Coagulation has occu
47、rred if the sheep plasma becomes opaque or if a fine fibrin thread is noted.BSISO12772:1997 4 BSI 09-1999 Annex E (informative) Test for amount of anti-coagulant in tube and anti-coagulant potency assay E.1 Test for amount of anti-coagulant in tube:Toluidine blue test Heparin reacts like other glyco
48、saminoglycans with toluidine blue to give a purple-coloured reaction product. The absorbance of the purple colour, measured in a spectrophotometer at a wavelength of606nm corresponds to the amount of heparin. The spectrophotometer is calibrated with standard solutions of heparin of known concentrati
49、on or activity. E.2 Anti-coagulant potency assay:Chromogenic 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 of405nm, or the decrease of fluorescence, measured in a fluorometer at an excitation of335nm and an emission of415nm corresponds to the activity of the heparin. The spectrophotometer or the fluorometer is calibrated with
