WO1988009655A1

WO1988009655A1 - Method and device for disinfecting biological fluids and container for same

Info

Publication number: WO1988009655A1
Application number: PCT/US1988/001814
Authority: WO
Inventors: Habib Al-Sioufi
Original assignee: Al Sioufi Habib
Priority date: 1987-06-05
Filing date: 1988-06-03
Publication date: 1988-12-15
Also published as: AU1938188A

Abstract

A technique and receptacle (20) for disinfecting biological fluids such as whole blood is described in which the disinfectant (23) is prepositioned in a receptacle (20) for biological fluids utilized for clinical evaluation in an amount which is sufficient to disinfect the fluid without interfering with subsequent clinical evaluation. The amount of disinfectant (23) positioned in the receptacle (20) is adjusted to provide an ultimate concentration in the blood specimen of aldehyde based disinfectant (23) of about 0.001 to 5.0 weight percent and is buffered to a pH of about 7.2 to 8.5, preferably about 7.4. The aldehyde based disinfectant (23) used in accordance with the invention has also been found to facilitate separation of the fluid components of the blood by causing gelling of cellular blood components.

Description

METHOD AND DEVICE FOR DISINFECTING BIOLOGICAL FLUIDS AND CONTAINER FOR SAME

SUMMARY OF THE INVENTION The present invention is directed to a device and method for combating and destroying biological contamination in specimens of biological fluids such as blood intended for medical evaluation without interfering with the integrity of the proposed evaluation. More specifically, the present invention is particularly concerned with disinfecting viral contamination in biological specimens to avoid infecting those coming in contact either with the specimen itself or the receptacles and equipment used to contain and evaluate the specimen. Of particular concern in the present invention is the avoidance of contamination by HTLV-III Virus responsible for Acquired Immune Deficiency Syndrome and Hepatitis Virus which may be present in blood specimens drawn for medical evaluation. Additionally, it has been found that aldehyde based disinfectant compositions used in accordance with the invention as viral disinfectants facilitate separation of the fluid components of the blood by causing gelling and fixing of cellular components.

BACKGROUND OF THE INVENTION The incidence of hospital acquired infections has been increasing in recent years at an alarming rate which has caused great concern among the staffs of hospitals and especially those working in the laboratories. While many disinfection and sterilization techniques have been employed to alleviate this problem in different functional sections of the hospital, these techniques have not consistently provided a safe environment for the staff. Frequently, the disinfection and sterilization techniques which have been used have been employed after overt contamination has taken place from spilling, broken samples, etc. While these techniques have helped to reduce the incidence of laboratory acquired infections, they have not curtailed them. With the increasing incidence of contagious pathogens that can be transmitted by patient's specimens, especially blood and particularly such dangerous contaminants as the AIDS and hepatitis viruses, a new and safe technique for handling laboratory specimens is needed.

Various disinfectants and sterilizing agents have been employed with varying degrees of success, both in hospitals and other environments, Monoaldehydes such as formaldehyde have been used successfully as a disinfectant, however, dialdehydeε, particularly glutaraldehyde, are preferred. Examples of glutaraldehyde-based disinfectants are a dilute sodium phenate-gl taraldehyde solution buffered to pH 7.4, an activated solution which contains 2.0% glutaraldehyde buffered to pH 7.5-8.0 and a disinfectant and sterilizing solution containing 2% glutaraldehyde at pH 7.0-7.5.

The extensive use of glutaraldehyde based compositions as an antiseptic and disinfectant has led to extensive studies of the compound and its activity. Glutaraldehyde has been classified as a chemosterilizer and has been defined by Borick, J. of Pharrα. Sciences, vol. 53, no. 10, October, 1964, as a chemical agent capable of destroying all forms of microbiological life including bacterial and fungus spores, tubercle bacilli and viruses. The compound has in fact been shown to be effective against a wide range of viruses even in the presence of high levels of organic matter which tend to destroy the potency of other disinfectants. The degree of biocidal activity observed in glutaraldehyde solutions is very much dependent on the pH of the solution as enhanced biocidal activity is found in alkaline solutions.

Boucher et al. , Proc. West Pharmacal Soc. 16, pp.282-288, 1973, postulated that the biocidal activity of gluaraldehyde is controlled by the distance between the aldehyde groups and their tendency to polymerize, thereby allowing free aldehyde groups to interact with the amino groups of the bacterial cell. This agrees with the findings of Rubbo et al., J. Appl. Bacteriol 30, pp.78-87, 1967, that antibacterial activity is due to the two aldehyde groups present on the molecule. After considering these results, Navarro and Monεan, Ann. Microbol 127B, pp.295-307, 1976, concluded that only structures containing two aldehyde groups allow formation of an aldol type polymer at an alkaline pH, and also produces a similar sterilizing effect at acid pHs on increasing concentrations. In other words, while the extent of polymerization is considerable at alkaline pHs , it is negligible in acid solutions unless the concentration is increased. On the other hand, acid solutions at pH3-4 of glutaraldehyde are considerably more stable than alkaline solutions.

The antimicrobal activity in any compound can not be viewed in isolation but must be described with reference to a number of factors including pH, temperature, organic matter present, and concentration. For glutaraldehyde, it has been common to use a 2% solution at room temperature and an alkaline pH of about 7.9. Unfortunately, alkaline solutions of glutaraldehyde are much less stable than acid solutions owning to the polymerization reactions already described, with a corresponding loss of antimicrobiol activity. A reduction in sporicidal activity of activated glutaraldehyde on storage has been observed in reports of Kelsey et al., J. Clin. Pathol. 27, pp.632-638, 1974, Thomas and Russell, J. Appl. Microbiol 28, pp.331-225, 1974b, Gorman and Scott, Int. J. Pharma 4, pp.57-65, 1979a. This reduction in sporicidal activity is directly related to a drop in concentration of the free aldehyde which appears to be essential for biological activity. Borick, Adv. Appl. Microbiol 10, pp.291-312, 1968, has estimated that glutaraldehyde concentration actually falls from 2.1% at pH 8.5 to 1.3% at pH 7.4 over a period of twenty-eight days at ambient temperatures. Accordingly, it has generally been the practice to employ glutaraldehyde as a 2% solution to which an activator is added to bring the pH to approximately 8 at the time of use. Such a solution used at room temperature will, for example, disinfect within 10 minutes and sterilize within 10 hours. However, it has been recommended that this solution be discarded after 14 days because of the significant decrease in activity and free aldehyde concentration. This instability has lead to the development of more stable preparations formulated at lower pHs and some with other potentiators included to increase the otherwise low level of activity observed at lower pH.

The inevitable conditions of clinical use for disinfection and sterilization frequently mean that organic matter is present such as blood and pus. This organic matter can act either by protecting the microbial species from antimicrobial attack or by competing with the microbial cell for active sites on the disinfectant molecules, thus reducing the effective concentration of disinfectant substance. Accordingly, many otherwise effective disinfectants and sterilizing agents may become ineffective where organic material, such as blood, is contacted. Glutaraldehyde, however, has a high resistance to neutralization by organic matter. Borick et al. , J. Pharm. Sci. 53, pp.1273-1275, 1964, for example has reported that the presence of 20% blood serum did not appear to adversely effect the activity of glutaraldehyde while Synder and Cheatle, Am. J. Hosp. Pharm. 22, pp.321-327, 1965, have reported that 1% whole blood did not effect glutaraldehyde activity.

One of the most important considerations in selecting a suitable disinfectant, in addition to its potency and sustained effectiveness as a disinfectant, is the toxicity of the composition to individuals coming in contact with it. Various studies have shown that glutaraldehyde, in moderate effective concentrations, is generally only slightly irritating to the skin, mucous membranes and eyes. Sato and Dobson, Arch. Dermatol 100, pp.564-569, 1969, have found that 5% glutaraldehyde was only irritating if the epidermal barrier was not intact.

Aqueous solutions of glutaraldehyde have been used to treat hyperh drosis and it has been used topically in the treatment of onycho ycosiε. Prevention of dental calculous formation and reduction of dental cavity formation in the mouth has been achieved by using oral compositions incorporating glutaraldehyde. In the cosmetic field, glutaraldehyde has been proposed for disinfection of production equipment and as a preservative. Glutaraldehyde has been used as a disinfectant for control of mastitis.

Accordingly, glutaraldehyde is now a generally accepted disinfectant and is found in a number of commercial preparations for disinfection and sterilization. Babb et al., J. Hosp. Infec. 1, pp.63-75, 1980, for example, have compared nine glutaraldehyde products.

Glutaraldehyde has also been used extensively in various non-microbiological areas including the leather tanning industry and tissue fixation for electromicroscopy. In microbiological areas, glutaraldehyde has been employed principally as a liquid chemical sterilizing agent for medical and surgical material that cannot be sterilized by heat cr irradiation. Compared with other disinfectants, glutaraldehyde has been found to be superior for disinfection of face masks, breathing tubes and other respiratory therapy equipment. Important advantages of glutaraldehyde as a chemosterilizer are: its activity in the presence of organic material, non-corrosive action towards metals, rubber, lenses and most materials, and lack of deleterious effect on cement and lenses of endoscopeε. Further, glutaraldehyde has been recommended for decontamination of dental, surgical instrumentε and working surface where the hepatitus B surface antigen may be present as well as for the treatment of warts.

From the above mentioned studies, testing any biological specimen containing glutaraldehyde will not damage the instrument used in testing. Osterberg, Arch. Pharm. Chemi. Sci. Ed. 6, pp.241-248, 1978, found that damage to leukocytes was apparent only above a 100 microg/ml. glutaraldehyde level. In addition, no erythrocyte damage occurred at the glutaraldehyde concentrations used. The use of aldehydes in electron microscopy was extensively studied and it was found that many cytochemical reactions can be performed on tissue specimens after aldehyde fixation. Glutaraldehyde is effective in preserving both prokaryotes and eukaryotes, including fragile specimens such as marine invertebrateε, embryos, diseased cells and fungi. Glutaraldehyde stabilized blood plasma with little shrinkage of blood clots (Chambers et al. 1968, Arch. Pathol. 85,18.) . Tissue specimens can be left in this fixative for many hours without apparent deterioration. Presently, glutaraldehyde is the most efficient and reliable fixative for preservation of biological specimens for routine electron microscopy and the previously mentioned and available data indicate that proteins are not denaturated to any marked extent by fixation with glutaraldehyde (M.A. Hayat, Fixation for

Electromicroscopy, Academic Press, 1981) . Similarly, glutaraldehyde fixed-erythrocytes remain sensitive to the hemagglutination and hemagglutination inhibition testε for arboviruε antigens and antibodies (Wolff et al. [1977] J. Clin Microbiol. 6.55) . Differential staining of viable and nonviable cells with alcian blue is maintained after fixation with glutaraldehyde (Yip and Auerperg, 1972, In Vitro 7, 323) . From the above mentioned studieε, glutaraldehyde will preserve the biological specimenε without otherwiεe affecting the integrity of the specimen for future evaluation.

As set forth above, the handling of biological specimenε such as blood after sampling, during storage and medical evaluation poses a particular hazard for those coming in contact with the specimens, especially where there is a possibility of AIDS (HTLV-III) or Hepatitis Virus being present. Despite the known effectiveness of disinfectants such as glutaraldehyde in destroying these viruses, their use has essentially been limited to the containers and equipment coming in contact with the fluid, and only after εuch contact has occurred and the fluid disposed of. What remains especially hazardous is the contaminated body fluidε themselves, such as AIDS (HTLV-III) or Hepatitiε infected blood, which are carriers of the infection from the time they are drawn from the donor. Accordingly, what is needed is a technique for destroying εuch viral contamination instantaneously when the sample is taken, but without effecting the specimens for further testing.

DISCUSSION OF THE PRIOR ART

U.S. Patent Number 3,016,328 describes disinfecting with a εporicidal composition containing a C„ to C_fi εaturated dialdehyde, εuch as glutaraldehyde, and an alkalinating agent in either alchoholic or aqueous solution at a pH above 7.4^*.

U.S. Patent Number 3,282,775 describes disinfecting with a sporicidal composition containing a C~ tc C_g εaturated dialdehyde preferably glutaraldehyde and a cationic surface active agent.

U.S. Patent Number 3,708,263 deεcribeε εterilizing at temperatures below 75°C by contacting the equipment to be treated with an aqueouε εolution by pH 2 to 8.5 containing glutaraldehyde and DMSO εimultaneouεly with ultrasonic wave energy.

U.S. Patent Numbers 3,912,450; 3,968,248; and 3,968,250 describe disinfection or sterilization compositions that contain nonionic and anionic εurfactantε with aqueouε or alchoholic glutaraldehyde εolutionε.

U.S. Patent Number 4,093,744 describes sporicidal compositions containing glutaraldehyde at pH 6.5 to 7.4 which may contain a detergent and also a onoaldehyde.

U.S. Patent Number 3,983,252 describes disinfectant compositions that contain a dialdehyde and an alkaline metal salt of a hydrocarbon carboxilic acid in aqueous solution and optionally an alcohol of up to seven carbon atoms or a diol with up to 4 carbon atoms such as ethylene glycol, propylene glycol, butylene glycol and/or a triol glycerol. The ccmpositionε are described as having improved εtability in the pH range of 6 to 7.4.

U.S. Patent Number 4,103,001 describes a sterilizing composition containing glutaraldehyde, a phenol and a metal phenate as active ingredients. The composition may alεo contain a humectant εuch aε glycerol, propylen glycol or diethylene glycol.

U.S. Patent Number 4,436,754 deεcribeε a disinfectant and sterilizing composition having low odor and irritation potential which is an aqueouε solution containing a 2 to 6 carbon atom dialdehyde and may also contain formaldehyde and a diol or mono-substituted diol. Such compositions can be used at pH of 2 to 9.

U.S. Patent Number 3,886,269 describes a formaldehyde based disinfectant formed by passing formaldehyde gas through a solvent such aε dimethyl sulfoxide or dimethyl formamide to form a gel-like polymer. The disinfectant described exhibits disinfection properties against basterial vegetative cells, bacterial spores, and soil organisms.

U.S. Patent Number 4,048,336 describes the use of a combination of glutaraldehyde and a monoaldehyde such as a formaldehyde to kill spores on inεtrumentε.

M.A. Hayat in Fixation for Electromicroscopy, Academic Press, 1981, pages 64 to 147 describes fixative agents for preserving and fixing blood and/or tissue specimens.

Seymour S. Block in Disinfection, Sterilization and Preservation, Lea and Febiger, 1983, Chapters 2, 3, 9 and 22 describes sterilization techniques using glutaraldehyde and phenolic compounds.

CROSS REFERENCE TO RELATED PATENT APPLICATION

Thiε application is a continuation-in-part of Application Serial Number 780,668 filed September 29, 1985.

DESCRIPTION OF THE INVENTION

In accordance with the present invention, a disinfectant for viral and other contamination in biological fluids such aε blood is provided in a container for the biological fluid in an amount which is effective to destroy the contamination without otherwiεe compromising the integrity of the fluid εpeci en with regard tc εubεequent biomedical evaluation. Additionally, the aldehyde baεed diεinfectant compositions used in connection with the invention facilitate εeparation of blood componentε by cauεing gelling of cellular componentε. The preεent invention is particularly adapted for use with evacuated containers into which freshly drawn specimens of blood are introduced and held for subsequent study. Such containers typically consiεt of a cylindrical tube having one open end into which an elastomeric stopper is fitted which iε capable of accepting a hollow εyringe needle to permit introduction of the biological fluid into the tube. Veεsels of this sort are commercially available under the name of Vacutainer Systems from Becton-Dickinson for example and are evacuated to provide a partial vacuum and provided with a hollow syringe needle which is dispoεed εo that blood is drawn from the donor into the tube by the force of the vacuum in the tube.

According to the invention, the receptacle for receiving and holding the specimen of a biological fluid such aε blood is provided with a disinfectant prior to introduction of the biological fluid in an amount sufficient to destroy viral contamination in the fluid and the receptacle without compromising the integrity of the specimen for medical evaluation. The disinfectant can be a mono or dialdehyde such as either glutaraldehyde or formaldehyde or chlorhexidine, phenols or quaternary ammonium compounds or mixtures thereof. The effective concentration of disinfectant according to the invention is about 0.001 to 5.0 weight percent, based upon the total quantity of biological fluid to be placed in the receptacle. Thus, the actual amount of the disinfectant present in the receptacle before introduction of the fluid will depend on the size of the receptacle and the extent to which it is to be filled with fluid since the fluid iε, in effect, the principal dilutent. Additional aldehydes such aε formaldehyde can also be used in amounts of about 0.001 to 5 percent by weight based on the total biological fluid. Where glutaraldehyde is the diεinfectant employed in accordance with the invention, it is desirable to maintain a slightly alkaline pH of preferably about 7.2 to 8.5 preferably 7.4 in order to achieve maximum effect against viral contaminants.

As demonstrated in the prior art, however, glutaraldehyde undergoes increasing polymerization at alkaline pHs and the glutaraldehyde εhould be maintained at acid pH until juεt before use. While the receptacle can be provided with an alkalinating agent such as sodium bicarbonante, sodium phenate, or lower alkanols, which is isolated from the disinfectant until just before introducing the biological fluid, it is preferred according to the invention to increase the pH of the glutaraldehyde by introduction of the blood specimen itself which has a normal pH of about 7.4. Where buffering to a higher pH is required, suitable amounts of alkalinating agent can be used.

Typical phenolic based and quaternary ammonium based disinfectants which can be used in the present invention are described in Seymour Block' ε Disinfection, Sterilization and Preservation, 3rd Edition, Lea & Febiger, 1983 at chapters 9 and 14 respectively, which is incorporated herein by reference. Suitable phenolic compounds in addition to carbolic acid are halogen substituted phenols especially with the halogen being in the ortho or para position relative to the nuclear hydroxyl group. Also preferred are halogen substituted phenols having aliphatic or aromatic substituents on the benzene nucleus, such as ortho alkyl derivative of p-chlorphenol and o-chlorophenol and para and ortho bromophenols.

Suitable quaternary ammonium compounds for the present invention include pol substituted quaternary ammonium salts such aε alkyldimethyl benzene ammonium saccharinate, and alk ldimethylethylbenzyl ammonium cyclohexyεulfa ate, Biε-quaternary ammonium salts such as 1,10-bis (2methyl-4- aminoquinolinium chloride) -decane, polymeric quaternary ammonium saltε εuch aε poly [oxyethylene (di ethylimino) ethylene (dimethylimino) -ethylene dichloride] , poly[N- [3- (dimethylammonio) propyl] -N¹ - [3- (ethylenoxyethylene-dimethyl- ammonio)propyl] urea dichloride], and -4-[l-tris(2 hydroxyethyl) ammonium chloride-2-butenyl]poly [1-demethyl ammonium chloride-2-butenyl] - -triε (2-hydroxyethyl) ammonium chloride. Alεo uεeful are twin chain quaternary alkyl benzyl ammonium chlorides such as octyl decyl dimethyl ammonium chloride, dioctyl dimethyl ammonium chloride, didecyl dimethyl ammonium chloride and dual quaternary-n-alkyl dimethyl ethyl ammonium chloride and n-alkyl dimethyl ethyl ammonium chloride.

It is also desirable to incorporate into the receptacle of the preεent invention effective amounts of substances to increase the permeability of the cell membrane to allow the disinfectant to reach intracellular pathogens more quickly. Such substanceε are dimethyl εulfoxide, and glycerol, either alone or in combination. Additionally, other εubεtances whose use in connection with sampling and testing of biological fluids, εuch as blood, can be used such as anticoagulants, preservatives and biocidal agents. By employing the various configurations which are embodiments of the present invention, activation of the disinfectant can take place prior to, during or after introduction of the specimen and the disinfectant can be released either before, during or after the specimen is introduced. The present invention will however be more fully appreciated by having reference to the drawings.

The use of aldehyde based disinfectants such aε glutaraldehyde in accordance with the invention has also been found to facilitate separation of fluid components of the blood by acting as a fixative and causing gelling of the cellular blood components to occur. The following is an example of this εeparation procedure.

A 10 ml blood sample was drawn and divided into nine 1 ml aliquotε. A 25% glutaraldehyde εolution waε added to each of theεe aliquots in the following amounts: 25, 50, 75, 100, 125, 150, 175, 200 and 250 microliters. No separation of blood components waε obεerved in the aliquots containing 75 or more microliters of glutaraldehyde, however, in the samples containing 25 and 50 microliters the red blood cell componentε formed a gel that remained εeparate from the clear plasma component.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 illustrateε one embodiment of the invention in which a closed tube is used having a stopper in one end and containing a disinfectant and activator;

Figure 2 illustrateε an additional embodiment of the preεent invention whereby both endε of the tube are εtoppered and one stopper is provided with the disinfectant or activator

Figure 3 illustrateε an embodiment of the present invention εimilar to that of Figure 2 in which one εtopper contains anticoagulant;

Figure 4 illustrateε an embodiment of the present invention similar to that of Figure 2 excepr for the presence of an inert barrier material;

Figure 5 illustrates an embodiment of the present invention alεo εimilar to that of Figures 1 and 4 in which the stopper contains activator and disinfectant separated;

Figure 6 illustrates an embodiment similar to that of Figure 5 having an anticoagulant rather than an inert barrier material;

Figure 7 illustrates an embodiment of the present invention having a tube similar to that of Figure 2 in which one stopper contains activator and disinfectant separated from each other and containing anticoagulant;

Figure 8 illustrates an embodiment of the present invention having a tube similar to that of Figure 1 but containing a disinfectant on the wallε of the tube without activator; Figure 9 illustrateε an embodiment of the present invention similar to that of Figure 8 except that activator is contained in the stopper;

Figure 10 illuεtrateε an embodiment of the present invention similar to that of Figure 2 but with disinfectant on the inner walls of the tube;

Figure 11 illustrates an embodiment of the present invention in which a stopper is used which containε disinfectant and having a permeable membrane; and

Figure 12 illustrates an embodiment of the invention similar to that of Figure 2 except that no activator is used in connection with the diεinfectant and a εtopper iε uεed having a permeable membrane.

DETAILED DESCRIPTION OF THE DRAWINGS Directing attention to the drawingε. Figure 1 illuεtrateε an embodiment of the present invention in which a cylindrical tube 20 closed at one end is provided with an elaεtomeric stopper 21 at the other end. As previously noted, closed stopper tubes of similar construction are commonly employed for collecting samples of blood. It iε frequently the caεe that these tubes are provided with a partial vacuum and a double ended hollow syringe needle placed in the stopper end so that the blood εample can be drawn directly from the donor into the tube uεing the vacuum in the tube. Although the details of construction of theεe syringe devices is not herein illustrated since they are well known in the art, it will be understood that they can be used in connection with the present invention. In accordance with the embodiment of the invention shown in Figure 1, a disinfectant material 23 iε prediεposed in the bottom of the tube 20 and a suitable alkaline activator 22 such aε εodium bicarbonate is provided in a cavity 24 of the εtopper 21. The two materialε are thuε kept separate from one another until the blood sample iε introduced through the stopper into the tube whereby the mixing of the disinfectant and activator takeε place. It will be underεtood that the amount of disinfectant present in the bottom of the tube 20 will depend upon the size of the tube and the quantity of blood to be drawn into the tube and should be εufficient to insure a concentration of between .001 and 5.0 diεinfectant once the blood sample is in the tube. The amount of activator present in the εtopper cavity 24 will be sufficient to insure that the specimen and disinfectant have an alkaline pH between 7.2 and 8, preferably about 7.4.

In the embodiment of the invention shown in Figure 2, the cylindrical tube 25 is provided with a stopper at either end. The lower end of the tube 25 is closed by elastomeric stopper 27 having a recesε which contains an activator such as sodium bicarbonate 30 which is separated by thin membrane from the disinfectant 31 which iε disposed freely in the tube. The other end of the tube is closed by stopper 26. A sharp pin 29 having a head 28 is provided for piercing the membrane εeparating the activator and diεinfectant before or once the blood εample haε been introduced into the ether end of the tube 25 through stopper 26.

Figure 3 of the drawings illustrate an embodiment of the invention similar to that of Figure 2 except that the upper end of the tube 25 iε provided with a εtopper 32 having a receεsed area 33 provided with an anticoagulant 34 separated from the disinfectant to maintain the fluidity of the blood sample. Introduction of the blood sample through the stopper 32 releases the anticoagulant by rupturing a barrier to allow it to mix with the blood sample, disinfectant and activator which are released by the means of a pin.

In Figure 4 of the drawings, an embodiment of the invention otherwise similar to that of Figure 1 is illustrated in which an activator 39 is provided in the cavity 38 of stopper 37 in the top of the tube. The disinfectant is however mixed with an inert barrier material and placed at the bottom of the tube 36. In this manner, activation of the disinfectant to the appropriate pH will not occur until the blood sample is centrifuged to produce a separation of the serum.

In Figure 5 of the drawings, the stopper 40 is provided with a recess 43 containing the activator 41 and diεinfectant material 42 which are separated from one another by a thin membrane and from the inside of the tube. Inert barrier material is provided at the bottom of the tube 36.

The embodiment of the invention aε shown in Figure 6 is similar to that of Figure 5 except that the inert barrier material iε replaced with an anticoagulant 45.

Figure 7 of the drawingε illustrateε an additional embodiment of the invention whereby stoppers are provided at both ends of the tube 25. The stopper 27 closing the lower end of the tube iε provided with an activator at 30 and diεinfectant 31 separated from one another by a thin membrane and from the inside of the tube. Anticoagulant 45 is placed in the tube directly over the stopper and disinfectant material. A pin 29 with head 28 iε available to puncture the εeparating membraneε to permit the materialε to mix with the blood introduced through stopper 26 at the other end of the tube.

Figure 8 of the drawingε illustrates a preferred embodiment of the invention in which disinfectant material 50 iε coated on the inside of the tube 20 to provide a layer. The upper end of the stop of the tube 20 is closed by stopper 26 but no additional activator iε provided εince the amount of diεinfectant 50 is adjusted so that its pH will become slightly alkaline with the introduction of blood into the tube which also provides the necesεary dilution to reεult in a concentration of 0.001 to 5% disinfectant.

In Figure 9 of the drawings, an embodiment of the invention iε shown similar to that of Figure 8 in that the disinfectant material iε a coating 50 on the inεide of the tube 20. An activator εuch as sodium bicarbonate is provided and separated from the inside of the tube, however, in cavity 38 of stopper 37 at 39. Figure 10 of the drawings illustrates the embodiment of the invention whereby the cylindrical tube 25 is closed at both ends by respective stoppers 26 and 27. The stopper 27 iε however provided with activator 30 which iε separated from the inside of the tube and released into the tube to interact with the disinfectant 50 by inserting the pin 29 into the stopper 27 to rupture a membrane that separates the activator from the interior of the tube.

In Figure 11 of the invention, either an anticoagulant or activator 51 is provided in the bottom of the tube 44. A porouε material container 54 iε provided on stopper 52 to hold the disinfectant 53 and permit it to diffuse through a permeable membrane into the tube 44 once the fluid specimen haε been introduced into the tube and the tube inverted. In Figure 12 of the drawingε, the disinfectant material 57 is provided in an appropriate cavity in stopper 55 closing one end of the tube while stopper 56 closes the other end of the tube. A membrane prevents the disinfectant from entering the tube itself until blood is introduced, at which time the disinfectant diffuses through the membrane into the specimen.

It will be understood that while various preferred embodiments of the present invention have been described herein in order to illustrate and dislose Applicant'ε invention, additional variationε and applicationε of the present invention are considered to fall within the scope thereof.

Claims

WHAT IS CLAIMED: Claim 1. An evacuated receptacle for holding a specimen of biological fluid for clinical evaluation, comprising a vesεel cloεed at one of its ends by an elaεtomeric stopper adapted to be penetrated by means for introducing said specimen therein, said vessel also containing prior to introduction of said specimen about 0.001 to 5.0 eight percent based on the total fluid and diεinfectant of a disinfectant for viral infection present in the specimen which disinfectant is one or more compounds or mixtures thereof selected from the group consisting of glutaraldehyde, formaldehyde, chlorohexidine, phenols and guaternary ammonium compounds, εaid diεinfectant being buffered εubεtantially at the time of said biological fluid is introduced therein at a pH of about 7.2 to 8.5.

Claim 2. The receptacle of claim 1 in which said vessel also containε an activator for εaid disinfectant.

Claim 3. The receptacle of claim 1 wherein both ends of said vessel are closed by elasto eric stoppers.

Claim 4. The receptacle of claim 1 wherein said biological fluid is whole blood.

Claim 5. The receptacle of claim 1 wherein an alkaline buffering agent disposed therein is separated from εaid glutaraldehyde prior to introducing εaid biological fluid, in an amount sufficient to accomplish said buffering when said fluid is introduced into the receptacle.

Claim 6. The receptacle of claim 5 wherein the concentration of said disinfectant is about 0.13 to 2.0 weight percent.

Claim 7. The receptacle of claim 1 wherein said diεinfectant iε coated onto the inεide walls of said vessel.

Claim 8. The receptacle of claim 1 wherein either said disinfectant or said buffering agent is disposed in a cavity in said stepper such that introduction of biological fluid through said stopper causeε εaid diεinfectant or agent to be released into εaid vessel and whichever of the agent or IS disinfectant is not disposed in said stopper is otherwise preεent in said vessel.

Claim 9. The receptacle of claim 3 wherein both of said stopperε are provided with cavitieε adapted to retain material until said biological fluid is introduced into said receptacle, the stopper adapted for penetration by said means for introducing the fluid being also adapted to release said material retained therein on penetration, and the other of said stoppers being provided with separate means to release material contained therein into said receptacle.

Claim 10. The receptacle of claim 9 wherein the material retained in one of the said stopperε is disinfectant.

Claim 11. The receptacle of claim 10 wherein an alkaline buffering agent is also retained in one of said stopperε in an amount sufficient to adjust the pH of the blood in said receptacle to about 7.2 to 8.5.

Claim 12. The receptacle of claim 4 which also contains an effective amount of anticoagulant for said blood.

Claim 13. The receptacle of claim 5 wherein said buffering agent is selected from the group consiεting of εodium bicarbonate, εodium phenate and alkanolε of 2-4 carbonε.

Claim 14. The receptacle of claim 4 which also includes an effective amount of a substance or εubεtances to enhance cell permeability selected from the group consiεting of dimethyl sulfoxide and glycerol.

Claim 15. The receptacle of claim 1 in which said receptacle is evacuated to provide a partial vacuum in the interior thereof.

Claim 16. A method for destroying viral contamination in specimens of biological fluids which comprises providing an evacuated container for said fluids having predispoεed therein about 0.001 to 5.0 weight percent of a disinfectant for said viral contamination selected from the group consisting of glutaraldehyde, formaldehyde, chlorohexidene, phenols and quaternary ammonium compounds or mixtureε thereof, said disinfectant being buffered substantially at the time said biological fluid iε introduced therein to a pH of about 7.2 to 8.5.

Claim 17. A method for εeparating cellular blood components into plasma and a gel which comprises adding to a specimen of whole blood an aldehyde composition selected from the group consisting of glutaraldehyde and formaldehyde in an amount sufficient to cause gelling of said cellular blood component.

Claim 18. The method of claim 17 wherein εaid aldehyde compoεition iε added in an amount of 0.001 to 5 percent by weight baεed on the total of whole blood and aldehyde.