EN XM XM XM XM XM XM XM XM XM XM XM XM XM Official Journal of the European Communities L 37/4

PROTOCOL TO THE EUROPEAN AGREEMENT

on the Exchange of Therapeutic Substances of Human Origin

PART I

GENERAL PROVISIONS

A. LABELLING

A label printed in English and French, based on the appropriate model to be found in Annexes 2 to 10 to the Protocol, shall be affixed to each container or giving-set.

B. PACKING AND DISPATCH

Whole human blood shall be dispatched in containers in which a temperature of 4 to 6oC is maintained throughout the period of transport.

This condition is not required for the derivatives mentioned in the Protocol.

C. PRODUCTS AND APPARATUS

The products and apparatus referred to in Part II of this Protocol shall be sterile, non-pyrogenic and non-toxic.

It is recommended that the giving-set, as well as the solvents required for the dried products, be sent with each consignment.

D. FREEDOM FROM TOXICITY OF PLASTIC BLOOD TRANSFUSION EQUIPMENT

Equipment shall comply with the provisions set out in Annex 11 to this Protocol.

PART II

SPECIFIC PROVISIONS

1. WHOLE HUMAN BLOOD

Whole human blood is blood which has been mixed with a suitable anticoagulant, after collection from a human subject in normal health.

The blood shall not be obtained from a human subject:

(a) who is known to be suffering from or to have suffered from syphilis; or

(b) whose blood has not been tested with negative results for evidence of syphilitic infection; or

(c) who is not, as far as can be ascertained after medical examination and the study of his antecedents, free from disease transmissible by blood transfusion.

The blood shall be withdrawn aseptically through a closed system of sterile tubing into a sterile container in which the anticoagulant solution has been placed before the container is sterilized. The equipment used must be pyrogen-free. When withdrawal is complete the container shall be immediately sealed and cooled to 4 to 6oC and not opened thereafter until immediately before the blood is to be used.

The blood will be collected into a citrate solution of acid reaction containing dextrose. No antiseptic or bacteriostatic substance shall be added. The volume of the anticoagulant solution must not exceed 220 ml per litre of the whole human blood and the haemoglobin concentration must not less than 97 grams per litre.

Blood group

The blood group under the AB0 system shall have been determined by examination of both corpuscles and serum and that under the Rh system by examination of the corpuscles, using a separate sample of the donor's blood. When there is a national standard, or nationally recommended technique of blood grouping, that technique shall be used.

The term Rh negative is only to be used when specific tests have shown the absence of the antigens C, D, Du and E. All other blood must be labelled Rh positive.

Blood exchange under this agreement should only be used for recipients of the corresponding AB0 group.

Storage

Whole human blood shall be kept in a sterile container sealed so as to exclude micro-organisms and stored at a temperature of 4 to 6oC until required for use, except during any period necessary for examination and transport at higher temperatures, any such period not to exceed 30 minutes after which the blood must immediately be cooled again to 4 to 6 oC.

Labelling

The label on the container shall give all the information shown on the model label (Annex 2). The Rhesus group shall be written as ‘Positive’ or ‘Negative’ or, in abbreviated form, ‘POS’ or ‘NEG’.

1a. HUMAN RED CELL CONCENTRATE

A human red cell concentrate is a unit of whole human blood from which most of the plasma has been removed.

It contains most of the red cells of the unit from which it has been prepared; other cell components may be present or may have been partially removed.

The liquid content of the concentrate will consist either of the residual plasma, or of an appropriate isotonic artificial aqueous solution added after the plasma was removed. The volume of red cells should constitute between 65 and 75 % of the total volume of the product, but if a greater red cell concentration is applied the approximate percentage of erythrocyte volume (haematocrit) shall be indicated on the label.

All operations required in the preparation shall be carried out under aseptic conditions: decantation shall be carried out using a sterile, closed system and by compression only. No antiseptic or bacteriostatic agents should be added.

Blood group and storage

as for whole human blood.

Labelling

The label on the container shall give all the information shown on the model label (Annex 2 a). The Rhesus group shall be written as ‘Positive’ or ‘Negative’ or, in abbreviated form, ‘POS’ or ‘NEC’. If an artificial aqueous solution has been added, the label shall also indicate its volume and composition.

2. DRIED HUMAN PLASMA

Dried human plasma is prepared by drying the supernatant fluids which are separated by centrifuging or by sedimentation from quantities of whole human blood.

During preparation no antiseptic or bacteriostatic or other substance shall be added. Dried human plasma shall be obtained by freeze-drying or by any other method which will avoid denaturation of proteins. The dried product shall be readily soluble in a quantity of water equal to the volume of the liquid from which the substance was prepared. The protein concentration of the solution thus obtained must not be less than 45 grams per litre, and must not show visible evidence of the products of haemolysis. The haemaglutinin titre shall not be greater than 1 : 32.

Dried human plasma prepared from one or two donations of blood

Donations shown to contain dangerous levels of isohaemolysins (determined using a sample of fresh serum) or any immune haemaglutinins shall be excluded. Unless the plasma is pooled and frozen within 48 hours of collecting the blood, the sterility of each unit shall be tested by culturing not less than 10 ml.

Dried human plasma prepared from pools of more than two donations

Pools shown to contain dangerous levels of immune haemaglutinins or of isohaemolysins shall be excluded. To avoid untoward effects due to the products of bacterial growth in the plasma no individual donation shall be used if there is any evidance of bacterial contamination, and the sterility of each pool shall be tested by culturing not less than 10 ml. To minimize the risk of transmitting serum hepatitis, plasma should be prepared from pools which should contain not more than 12 donations, or by any other method that has been shown to diminish the risk in comparable manner.

Solubility in water

Add a quantity of water equal to the volume of the liquid from which the sample was prepared; the substance dissolves completely within 10 minutes at 15 to 20 oC.

Identification

Dissolve a known quantity of the product in a volume of water equal to the volume of the liquid from which it was prepared; the solution passes the following tests:

(i) by precipitation tests with specific antisera, it must be shown to contain only human plasma proteins;

(ii) to 1 ml add a suitable amount of thrombin or calcium chloride; coagulation occurs, which can be accelerated by incubation at 37 oC.

Loss of mass on drying

When dried over phosphorus pentoxide at a pressure not exceeding 0,02 mm of mercury for 24 hours, dried human plasma must not lose more than 0,5% of its weight.

Sterility

The final product, after reconstitution, shall be sterile when examined by a suitable bacteriological method.

Storage

Dried human plasma must be kept in an atmosphere of nitrogen or in a vacuum in a sterile container sealed so as to exclude micro-organisms and, as far as possible, moisture, protected from light and stored at a temperature below 20 oC.

Labelling

The label on the container shall give all the information shown on the model label (Annex 3).

3. HUMAN ALBUMIN AND HUMAN PLASMA PROTEIN FRACTION

Human albumin and human plasma protein fraction are preparations of that protein component which forms about 60 % of the total protein mass in the plasma of whole human blood.

The method of preparation used shall be one which produces a material meeting the requirements herein described. Regardless of whether the final product is liquid or dried, the preparation, after the addition of a suitable stabilizing agent or agents, must have been heated in the liquid state in the final container at 60 oC ± 0,5 oC for 10 hours, in order to inactivate the agent causing serum hepatitis. During preparation no antiseptic or bacteriostatic substance shall be added.

In preparations of human albumin, not less than 95% of the mass of the proteins present shall be albumin. In preparations of human plasma protein fraction, not less than 85% of the protein mass shall be albumin. In both preparations, more than 10 milligrams of immunoglobulin G per gram of product shall be present.

When the final product is freeze-dried, it must contain not less than 950 milligrams of protein per gram of product.

When human plasma protein fraction is prepared as a solution it shall have a total protein concentration of between 45 and 50 grams per litre.

When human albumin is prepared as a solution it shall have a total protein concentration of not less than 45 grams per litre.

Solubility of the dried product

Add water to the recommended volume; the dried preparation must be completely soluble.

Stability

By comparison of the solutions before and after heat treatment no evidence of significant denaturation of the proteins in solution shall have been detected as estimated by viscosity and turbidity measurements, ultracentrifugation and electrophoresis. The solution shall be substantially free from visible particles after heating at 57 oC and after agitation in a mechanical shaker for six hours at this temperature.

Identification

i) By precipitation tests with specific antisera, both preparations must be shown to contain only human plasma proteins.

ii) By electrophoresis, using the moving boundary technique under acceptable and appropriate conditions, it must be shown that the protein fraction having the mobility of the albumin component of normal human plasma, is not less than 95 % of the protein mass in preparations of human albumin, or not less than 85% of the protein mass in preparations of human plasma protein fraction.

Sodium content and sodium concentration

The sodium content of salt-poor human albumin must not exceed 0,61 millimoles per gram of albumin. In other preparations of human albumin and in human plasma protein fraction, the sodium concentration must not exceed 0,15 moles per litre of solution or reconstituted dried product.

Potassium concentration

The potassium concentration of human plasma protein fraction must not exceed 2 millimoles per litre of solution or reconstituted dried product.

Acidity

The pH of either preparation shall be 6,8 ± 0,2 when measured at a temperature of 15 to 25 oC in a solution diluted to a protein concentration of 10 grams per litre by means of a solution containing 0,15 moles sodium chloride per litre.

Loss of mass on drying

Dried preparations, when dried over phosphorus pentoxide at a pressure not exceeding 0,02 mm of mercury for 24 hours, must not lose more than 0,5% of their weight.

Sterility

The final product shall be sterile when examined by a suitable bacteriological method.

Storage

Dried human albumin must be kept in an atmosphere of nitrogen or in a vacuum in a sterile container, sealed so as to exclude micro-organisms and, as far as possible, moisture, protected from light and stored at a temperature below 20 oC.

Solutions of human albumin and human plasma protein fraction must be kept in sterile containers, sealed so as to exclude micro-organisms, protected from light and stored at a temperature of 4 to 6 oC.

Labelling

The label on the container shall give all the information shown on the appropriate model label (Annex 4). For solutions, the date of preparation is the date of heat treatment in the final container.

4. HUMAN NORMAL IMMUNOGLOBULIN

Human normal immunoglobulin is a preparation of the plasma proteins prepared from whole human blood, containing the antibodies of normal adults. It is obtained from pooled liquid human plasma from not less than 1 000 donors.

The method of preparation used should be one which produces a material meeting the requirements herein prescribed and which prevents the transmission of serum hepatitis by the final product. In addition the method of preparation shall be such that the antibodies contained in the starting material shall be concentrated in an adequate amount in the final product. The procedure shall be shown, for each final preparation, to be satisfactory in this respect by titrating in the starting material and in the final product antibodies to at least one virus and one bacterial toxin. The antibodies chosen shall be those for which there are recognized methods of titration.

During preparation no antiseptic or bacteriostatic substance shall be added; a suitable preservative and a stabilizing agent may be added to the final preparation to maintain bacterial sterility and stability of the final product.

The final product is issued as a solution in which the immunoglobulin concentration shall be between 100 and 170 grams per litre.

Identification

(i) By precipitation tests with specific antisera, it must be shown to contain only human plasma proteins.

(ii) By electrophoresis, using the moving boundary technique under acceptable and appropriate conditions, not less than 90% of the mass of the proteins have the mobility of the gamma component of the globulins of normal human plasma.

Stability

Both before and after heating the final solution at 37 oC for seven days there should be no visible evidence of precipitation or turbidity. It is advisable also to carry out tests using an ultracentrifugation method to determine the extent of degradation of the product to smaller molecular weight components. The method used should be one approved by the national control authority.

Acidity

The pH of the final solution shall be 6,8 ± 0,4 when measured at a temperature of 15 to 25 oC in a solution diluted to a protein concentration of 10 grams per litre by means of a solution containing 0,15 moles sodium chloride per litre.

Sterility

The final product shall be sterile when examined by a suitable bacteriological method.

Storage

Human immunoglobulin solution must be kept in a sterile container, sealed so as to exclude microorganisms, protected from light and stored at a temperature of 4 to 6 oC.

Labelling

The label on the container shall give all the information shown on the model label (Annex 5). The date of preparation is the date of filling the final container.

5. HUMAN SPECIFIC IMMUNOGLOBULINS

Human specific immunoglobulins contain antibodies against designated viral or bacterial agents. Therefore they may be prepared from pools of a limited number of donations.

The following human specific immunoglobulins are included in these requirements:

— Human immunoglobulin anti-tetanus

— Human immunoglobulin anti-vaccinia.

Other specific immunoglobulins may be developed and when the appropriate international standard is in existence, they should be assayed in relation to that standard and their potency expressed in international units.

Human immunoglobulin anti-vaccinia shall contain not less than 500 IU per ml of vaccinia antibody as determined by a neutralization test on chorio-allantoic membranes or in tissue culture. Human immunoglobulin anti-tetanus shall contain not less than 50 IU per ml of tetanus antitoxin as determined by a neutralization test in animals.

Human specific immunoglobulins must further meet the requirements as described in section 4, Human normal immunoglobulin.

Depending on the antibody content, the immunoglobulin concentration of the final solution may vary between 100 and 170 grams per litre.

Labelling

The label on the container shall give all the information shown on the model label (Annex 5). In addition the label shall state the potency in international units in terms of the appropriate International Standard or International Reference Preparation.

6. DRIED HUMAN FIBRINOGEN

Dried human fibrinogen is a dried preparation which contains the soluble constitutent of liquid human plasma which, on the addition of thrombin, is transformed to fibrin. The method of preparation used should be one which produces a material meeting the requirements herein prescribed and which minimizes the risk of transmitting serum hepatitis. Plasma pools used in the preparation of fibrinogen should contain as few donations as possible.

During preparation no antiseptic or bacteriostatic substance shall be added. The final product shall be freeze-dried.

Solubility

Add water to the recommended volume; the dried preparation must be completely soluble. No precipitation shall occur within 60 minutes of reconstitution.

Identification

(i) By precipitation tests with specific antisera, it must be shown to contain only human plasma proteins.

(ii) The freshly reconstituted product has the property of clotting on the addition of thrombin. When thrombin is added to a solution of human fibrinogen of the same concentration as that in fresh normal plasma, clotting shall occur in not more than twice the time taken for clotting to occur in fresh normal plasma after the addition of thrombin.

(iii) Clottable protein. Not less than 50% of the total protein shall be clottable by thrombin.

Loss of mass on drying

Preparations, when dried over phosphorus pentoxide at a pressure not exceeding 0,02 mm of mercury for 24 hours, must not lose more than 0,3 % of their weight.

Sterility

The final product after reconstitution shall be sterile when examined by a suitable bacteriological method.

Storage

Human fibrinogen shall be kept in an atmosphere of nitrogen or in a vacuum in a sterile container, sealed so as to exclude micro-organisms and, as far as possible, moisture, protected from light and stored at the temperature recommended.

Labelling

The label on the container shall give all the information shown on the model label (Annex 6). The date of preparation is the date of placing into final solution before freeze-drying.

7. DRIED OR FROZEN HUMAN COAGULATION FACTOR VIII

I. Requirements applying to donors

Donors must be in good health and, in particular, free of any communicable disease, in accordance with the criteria adopted for dried human plasma.

II. Requirements applying to preparations

Sterility and atoxicity

The final product must be sterile and pyrogen-free. Where cryoprecipitation is performed in plastic bags, the product must not contain organic solvent or other foreign substances present in the freezing mixture. The passage of such products through the walls of the plastic bag can be prevented by placing the bag in a second impermeable bag during the whole period of immersion. The risk of the plastic bag tearing during storage in the frozen state can be reduced by keeping each bag in a protective box.

Erythrocytes, leukocytes and platelets

Centrifuging should be such as to eliminate the formed elements of the blood as soon and as completely as possible after its collection.

Solubility

The addition of the indicated quantity of appropriate solvent must result in the complete solution of the dry product in less than 30 minutes at 37 oC. Small and easily separable aggregates of fibrinogen may persist.

Stability

The preparation conserved at 20 oC, must not show any sign of precipitation within three hours after it has been dissolved.

Potency

The reconstituted preparation should contain the indicated minimum quantity of factor VIII, one unit corresponding to the potency of 1 ml of average normal fresh plasma, the potency being determined by a method approved by the competent national authority.

Abense of irregular antibodies and, if the preparation is intended for patients of any AB0 group, a titre of anti-A and anti-B antibodies not exceeding 32.

Identification

Precipitation tests with specific antisera shall show that the product contains only human plasma proteins.

Loss of mass on drying

Freeze-dried preparations, when dried over phosphorus pentoxide at a pressure not exceeding 0,02 mm of mercury for 24 hours must not lose more than 1,5 % of their weight.

Storage

Human factor VIII shall be stored in the deep-frozen state at a temperature under — 30 oC, and in the freeze-dried state below 5 oC, and protected from light. The dried preparation shall be kept in an atmosphere of nitrogen or in vacuo, in a sterile vial, stoppered so as to exclude all micro-organisms and, as far as possible, all humidity. Storage in the frozen state shall not exceed six months, in the dried state one year, unless the preparation has been retested for minimum required potency.

III. Labelling

The label on the preparation shall give all the information shown on the model label (Annex 7).

8. DRIED HUMAN COAGULATION FACTOR IX

I. Requirements applying to donors

Donors must be in good health and, in particular, free from any communicable disease in accordance with the criteria adopted for dried human plasma.

II. Requirements applying to the concentrate

Sterility and atoxicity

The final product, tested by appropriate methods must be sterile, pyrogen-free and free from undesirable vaso-depressor or respiratory effects. The test for absence of vaso-depressor effects should be performed on a dog or cat.

Solubility

The addition of the indicated quantity of the solvent must result in complete solution in 10 minutes at 37 oC.

Thromboplastin activity and absence of free thrombin

The recalcification time of a normal plasma measured at 37 oC in the presence of an equal volume of various dilutions of the reconstituted product, must not be less than 40 seconds. The reconstituted product, with an equal volume of fibrinogen (3 g/l) added to it, must not coagulate within six hours at 37 oC.

Potency

The reconstituted preparation must contain the indicated minimum quantity of factor IX, one unit corresponding to the potency of 1 ml of average normal fresh plasma, the potency being determined by a method approved by the competent national authority.

Yield and stability in vivo

The method of preparation must be such that the injection of a dose of 50 units per kg body weight, rapidly administered intravenously, using several batches of material given to several patients, shall cause, in 15 minutes, in the absence of a specific inhibitor and in basal conditions, an average rise of not less than 300 units per litre of plasma, and of the persistence, after 24 hours; of an average rise of not less than 60 units per litre of plasma.

Identification

Precipitation tests with specific antisera shall show that the product contains solely human plasma proteins.

Loss of mass on drying

When dried over phosphorus pentoxide at a pressure not exceeding 0,02 mm of mercury for 24 hours, the product must not lose more than 1,5 % of its weight.

Storage

The preparations must be stored dry at a temperature below 5 oC. The period of storage must not exceed two years, unless the potency of the preparation has been retested.

III. Labelling

The label on the preparation shall give all the information shown on the model label (Annex 8).

ANNEXE I AU PROTOCOLE ANNEX I TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN

ANNEXE 2 AU PROTOCOLE ANNEX 2 TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN ANNEXE 2 bis AU PROTOCOLE ANNEX 2a TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN

ANNEXE 3 AU PROTOCOLE ANNEX 3 TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN

ANNEXE 4 AU PROTOCOLE ANNEX 4 TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN ANNEXE 4 (suite 1) ANNEX 4 (continued 1) CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN ANNEXE 4 (suite 2) ANNEX 4 (continued 2) CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN

ANNEXE 5 AU PROTOCOLE ANNEX 5 TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN

ANNEXE 6 AU PROTOCOLE ANNEX 6 TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN

ANNEXE 7 AU PROTOCOLE ANNEX 7 TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN

ANNEXE 8 AU PROTOCOLE ANNEX 8 TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN

ANNEXE 9 AU PROTOCOLE ANNEX 9 TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN

ANNEXE 10 AU PROTOCOLE ANNEX 10 TO THE PROTOCOL CONSEIL DE L'EUROPE COUNCIL OF EUROPE ACCORD EUROPÉEN RELATIF À L'ÉCHANGE DE SUBSTANCES THÉRAPEUTIQUES D'ORIGINE HUMAINE EUROPEAN AGREEMENT ON THE EXCHANGE OF THERAPEUTIC SUBSTANCES OF HUMAN ORIGIN