MXPA97005713A - Disposable detector of hemoli - Google Patents

Abstract

A method and apparatus for detecting hemolysis from a patient's blood sample, a sealed chamber having a fixed volume for receiving the blood sample is provided, the sealed chamber having an internal pressure resulting from the presence of a amount of fixed air inside the chamber, a volume of fluid that includes the blood sample is received in the sealed chamber, while the sample is received in the sealed chamber, the internal pressure is raised to an increased internal pressure retaining the amount of fixed air inside the sealed chamber when the fluid is received inside the chamber, the increased internal pressure causes the plasma portion of the blood sample in the chamber to penetrate a membrane that forms at least a portion of one side of the chamber. the chamber, a test volume of the plasma portion of the sample is received by means of hemolysis detection after the test volume of the plasma portion has penetrated the membrane, and a hemolysis condition is detected according to a hue associated with the test volume received in the hemolysis detection means

Description

HEAVY DISPOSABLE DETECTOR BACKGROUND OF THE INVENTION r > The present invention relates to systems and methods for detecting hemolysis in the blood of a patient, and in particular to disposable systems which can be used to de-ect hemolysis in non-laboratory environments. Even more particularly, the present invention se->. Refers systems and methods that doctors can use in a patient's room to detect hemolysis. During the treatment of patients by blood circuits outside the body during hemodialysis, hyperthermia, open-heart surgery, immunosorbent therapy, photochemotherapy outside the body, and blood transfusions, there is a risk that hemolysis or rupture may occur. of IOJOS globules. This breakdown of red blood cells is deleterious not only because of the loss of function of these cells, but also because of the release of hemoglobin into the blood plasma that makes you toxic. Currently, hemolysis is typically detected during out-of-body therapies by first taking a sample of a patient's blood and taking it to a laboratory where the sample is placed in a rotating centrifuge to separate the red blood cells in the plasma sample. , and then comparing the plasma colors before and after or during the tr * at. Hemolysis test systems that require the use of a centrifuge and a laboratory are not a factor, because it is possible that hemolysis may occur. to a significant degree during the time that the b test is being conducted. In addition to a centrifuge, another known method for separating the plasma portion of the blood sample involves the use of a myo-orbital membrane device that allows only non-cellular elements of an ip blood sample to pass through a membrane. However, such icroporous membrane devices typically require a relatively high shear rate of the blood sample on the surface of the membrane to prevent the cellular elements of the blood sample from clogging or clogging the pores of b. membrane. The plasma separators of the membrane therefore require Mu systems or complicated to maintain a regime of shear stress on the surface of the membrane that is high enough to promote a good flow of pLasrna through the membrane without any plugging of what < - 20 membrane pores and also low enough for ovitai damage to blood cells that are separated from the plasma by the membrane. As a result, these systems are complex, expensive and typically require a large sample volume to detect hernol. As described above, membrane plasma separators require high shear rates on the surface of the membrane to keep blood cells away from the surface of the membrane and to prevent blood cells from clogging the pores of the membrane. the membrane. such high shear rates, the pores in these plasma separators of the known membrane will be immediately plugged by the blood cells and only a very small amount of plasma will be able to penetrate through the membrane before it is completely covered by the? cell * - * sanguineous. This small amount of? la? -? na e Typically sufficient to wet the membrane and it is more important to determine if hemolysis has occurred unless massive hemolysis has occurred in the sample. Two examples of micropore membrane systems for separating the plasma portion of a blood sample from its Ib cellular elements are shown in the Pat. Of E.U.n. No. J, 705, 100 of Dlatt et al. And the U.S.A. patent. No. 4,191,182 of Popovich et al. As described in the previous paragraph, plasma separation is achieved in these systems creating a high shear rate regime on the surface of the Furthermore, in order to avoid plugging and pore-blocking of the membrane by the cellular elements of the blood, these systems also include means for inducing an ansmembrane pressure across the surface of the membrane. In order to generate the high shear stress regime and 2"High transmembrane pressure required at the surface of the membrane, these prior art systems incorporate special mechanisms to control the velocity of blood flow and pressure at the surface of the membrane, an object of the present invention is providing a system for detecting hemolysis in an out-of-body circuit that can be rapidly performed by a physician in a patient or in a treatment room without the need for a laboratory, an additional object of the present invention is to provide a system to prepare a sample of blood in its plasma and cellular elements that is inexpensive and that does not require external instruments or mechanisms to control the speed and flow of blood on the surface of the membrane. present invention is provide-Ib a system for detecting hemolysis that requires only a small amount of a patient's plasma to penetrate a membrane in order to to detect if hemolysis has occurred. These and other objects of the invention will be apparent upon studying the accompanying drawings and the description of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a perspective view of a hemolysis detector in accordance with a preferred embodiment of the present invention.

The picture? is a perspective view showing the preferred hemolysis detector of Figure 1 in its inv r id position. Figure 2 is a sectional view showing the preferred hemolysis detector * of Figure 2. Figure 4 is a perspective view of a system for collecting and channeling the plasma portion of a blood sample in accordance with an embodiment of the invention. refer to the f > I resented invention. Figure 5 is a top view of the system for collecting and channeling the plasma portion of a blood sample formed in Figure 4. Figure 6 is a sectional view of the system for collecting and channeling the plasma portion of a sample of the blood sample in Figure b. Figure 7 is a diagram illustrating a preferred method for using the hemolysis detector of Figure 1 to detect hemolysis in a patient's blood sample. Figure 8 is a perspective view of a hemolysis detector in accordance with a preferred embodiment of the present invention. Figure 9 is a perspective view showing the preferred hemolysis detector of Figure 8 in its inverted position. Figure 10 is a sectional view showing the preferred hemolysis detector of Figure 8.

Figure 11 is a diagram illustrating a method intended to use the hemolysis detector of Figure 0 to detect * hemolysis in a patient's blood sample. Figure 12 is a perspective view of a hemolysis detector b in accordance with a further preferred embodiment of the present invention. Figure 13 is a perspective view showing the preferred hemolysis detector of Figure 12 in its inverted position. Figure 14 is a sectional view showing the preferred hemolysis detector of Figure 12. Figure 15 is a diagram illustrating a preferred method for using the hemolysis detector of Figure 12 to detect hemolysis in the sample of blood of a patient. Ib Figure 16 is a top view showing a hemolysis detector in accordance with a further preferred embodiment of the present invention. Figure 17 is a sectional view of the hemolysis detector of Figure 16. Figure 18 is a sectional view showing a hemolysis detector in accordance with another preferred embodiment of the present invention. Figure 19 is a sectional view showing a hemolysis detector according to another embodiment of the present invention. Figure 20 is a sectional view showing a hemolysis detector in accordance with another preferred embodiment of the present invention.

BRIEF DESCRIPTION OF THE INVENTION b The present invention is directed to a method and apparatus for detecting hemysis of a blood sample from a patient. A sealed chamber that has a fine volume is provided to receive the sample of ° angre. The sealed chamber LO has an internal pressure that results from the presence of a fixed amount of air inside the chamber. A volume of fluid that includes the blood sample is received in the sealed chamber. Although the sample is being received in the sealed chamber, the internal pressure of the sealed chamber is raised to L) an increased internal pressure retaining the fi rst amount of air inside the sealed chamber as the fluid is received in the chamber. The increased internal pressure causes the plasma portion of the blood sample in the chamber to penetrate a membrane that forms at least a portion of one side of the chamber. A test volume of the plasma portion of the sample is received by means of hemolysis detection after the test volume of the plasma portion has penetrated the membrane, and a hemolysis condition is detected in accordance with an associated hue with the volume test received in the hemolysis detection means. In accordance with a further aspect of the present invention, a hernol isis detector for detecting hemolysis of a blood sample includes a sealable chamber having means for reciting a volume of fluid including a blood sample in the chamber. Means are also provided to transform the sealable chamber into a sealed chamber having an internal pressure resulting from the presence of a fixed amount of air in the sealed chamber. The means of receiving the volume of fluid in the chamber include means for raising the internal pressure to an increased internal pressure as the fluid is received in the chamber while retaining the fixed amount of air in the chamber to be tied as the fluid volume is received in the chamber. The means for transforming the sealable chamber into a sealed chamber are formed of a membrane that is permeable to a portion of the sample plasma in the sealed chamber when the internal pressure in the chamber is equivalent to the increased internal pressure. Haemolysis detection means located outside the chamber are also provided to receive a test volume of the plasma portion of the sample after it has penetrated the membrane. The hemolysis detection means ect a hemolysis condition in accordance with a hue associated with the test volume of the plasma portion of the sample. In accordance with another aspect of the present invention, a system for detecting a blood constituent of antibodies of a whole blood sample comprises a membrane for separating a portion of plasma from the sample from a cell portion of the sample. The membrane has a first side for receiving the sample of whole blood and a second side for lowering only a portion of plasma of the sample of whole blood. A blood barrier is coupled and located against the first side of the membrane such that the blood barrier defines a perimeter enclosed by the first side of said membrane. An indicator paper that responds to the blood constituent of antibodies is also provided. Paper indicator * is coupled to the membrane a and located with the second side of the membrane. During the operation of this aspect of the invention, the sample of whole blood is placed within the perimeter and in contact with the first side of the membrane. A determination of whether the constituent of interests is present in the whole blood sample after it is done observing the nuance of the indicator paper. In accordance with another aspect of the present invention, a system for detecting a constituent of whole blood of antibodies of a blood sample comprises a membrane to separate a portion of plasma from the sample of a cell portion of the blood sample. The membrane has a first side to receive the sample of whole blood and a second side to pass only the plasma portion of the sample of whole blood. A blood barrier is coupled to and 'i: placed against the first side of the membrane in such a way that the blood barrier defines a perimeter enclosing the first side of the membrane. An indicator paper is coupled to the membrane by plasma channeling means which is positioned against the second side of the membrane. The indicator paper * responds to the constituent of the blood of antibodies, b During the operation of this aspect of the present invention, the whole blood sample is placed inside the pen and in contact with the first side of the membrane. The plasma portion of the sample is collected as it penetrates the membrane and then channeled with the channeling means to the indicator paper. A determination of whether the constituent of interest is present in the whole blood sample is made by observing the nuance of the indicator paper.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS Ib Referring now to Figures 1-3, perspective and sectional views of a hemolysis detector 100 are shown in accordance with a preferred embodiment of the present invention. The hemolysis detector 100 is formed of a sealable chamber 110 having a sample injection port 120 for receiving a blood sample into the sealed chamber. The sealable chamber 110 is preferably formed of a rigid plastic and has a fixed internal volume. The right plastic to form the camera sealable 110 include acrylic, PVC, polycarbonate or polysulfone. The injection port of the sample 120 is preferably formed of latex rubber or other elastic material. the sealable chamber 110 is mounted on a base 130 of the chamber which is also preferably formed of a clear rigid plastic. A microporous membrane disc 140 (not shown in FIG. 1, but shown in FIG. 3) is provided in and forms the lower end of the ring 110. The microporous membrane disc 140 preferably has a size of pore on the scale of 0.2 to 1.2 microns, and most preferably at 0.45 - 0.80 microns. In the modality Preferred, the membrane disc 140 is only permeable to the plasma portion of a whole blood sample and not permeable to the cell portion of the sample. Channeling means 150 are located immediately downstream of the microporous membrane disc 140. The channeling means Ib 150 collects the plasma portion of a whole blood sample as the plasma portion penetrates through the membrane disc 140 and then that portion of plasma is channeled to the clear capillary tube 160. The hemolysis detector 100 is preferably assembled by placing the channeling means 150 den of the base 130 of the chamber, and then ultrasonically welding the sealable chamber 110 to the base 130 of the chamber with the membrane disc 140 located between the srna,. Referring now to Figure 7, b is shown a diagram illustrating a preferred method for using * a hemolysis detector 100 to detect hemolysis in sample L2 of a patient's blood. As shown in Figure 7, the procedure begins in step 1 by injecting a humectant solution such as saline into the sealable chamber 110. The moistening solution is preferably injected into the sealable chamber 110 with a syringe that has been inserted into the chamber. through the injection port 120 of the sample. In step 2, the hemolysis detector 100 is inverted so that the wetting solution injected during step 1 moistens the wetted membrane disc 140 at the bottom of the sealable chamber 110. This wetting step causes the membrane disc 140 (which has previously been dried and thus permeable to air) becomes air impermeable, thus transforming the sealable chamber 110 to a sealed chamber having a fixed volume of air in the interior. In step 3, a sample of whole blood from a patient is injected into the sealed chamber 110 through a sample injection port 120. Since the injection port of the sample 120 is made of latex or rubber, no air leaks from the sealed chamber 110 during the injection of the whole blood sample towards the chamber. In accordance with the preferred system for the operation of the hemolysis detector 100, the volume of the whole blood sample injected into the sealed chamber 11 (1 in step 3 preferably corresponds to 1-10% of the internal volume of the sealed chamber 110. Since the fluid is added to the sealed chamber 110 in step 3, but no air is allowed to escape from the chamber, the internal pressure in the sealed chamber rises by approximately 8-80 rnrn llg as a result of the injection of the whole blood sample into the sealed chamber of this step.In a further preferred embodiment, the volume of whole blood injected during step 3 is accurately measured in such a way that the internal pressure in the sealable chamber 110 it is elevated by 28-30 rnrn Hg when the whole blood sample is injected into the sealed chamber 110. The change in pressure that results from the injection of a blood sample The whole to the sealed chamber 110 can be easily determined by solving the following equation (l) for the quantity P2: Pi * Vi --- P2 * V2 (1) where i represents the volume of air in the sealed chamber 110 before of the injection of the whole blood sample towards Ib the chamber, Pi represents the air pressure in the sealed chamber 110 before the injection of the whole blood sample into the chamber (this will typically be the ambient air pressure) V represents Vi minus the volume of the sample. of whole blood injected into the sealed chamber 110 during step 3, and P2 represents the air pressure inside the sealed chamber 110 after the whole blood sample has been injected into the chamber. In the preferred embodiment of the detector 100, the membrane disc 140 is used by itself as the reliable means converting the sealable chamber 110 to a sealed chamber because the membrane disc 110 (which was originally in the dry state prior to air) becomes waterproof when wet. In alternative embodiments, other means such as a sealable valve or opening (not shown) located between the inside of the chamber 110 and the outside can be used to transform the chamber 110 into a sealed state. Still referring to FIG. 7, in step 4 of the method, the hemolysis detector 100 is stirred to mix the whole blood sample injected into the sealed chamber 110 during step 3 with a humectant solution which It remains in the sealed chamber 110 of step 1. Next, in step 5, the hemysis detector 100 is inverted and the plasma portion of the whole blood sample previously injected into the chamber then penetrates through the membrane disc 140. The increased internal pressure generated by the injection of the Ib whole blood sample to the sealed chamber 110 in step 3 operates during step 5 as a driving force to push the plasma portion of the whole blood sample through the membrane disc 140. In the preferred embodiment, the pressure internal increase generated by the injection of The whole blood sample to the sealed chamber 110 should be large enough to drive the plasma portion of the blood sample through the membrane disc 140, but not so large as to cause damage to the blood sample. In a preferred embodiment of the hemolysis detector 100, the surface area of the membrane disc 140 may be of the order of 3.14 square centimeters and, in step 5 of Ib Figure 7, from 3.1 to 0.15 ml of pLasrna will penetrate the membrane disc 140. Therefore, in this preferred embodiment, the ratio of plasma volume that penetrates the membrane to the membrane surface area is from the • > 0.318 rnl / cm2 of the membrane surface area. In alternative embodiments, the ratio of plasma volume to membrane surface area may vary from 0.1 to 1.0 rnl / crn2 of area of knowledge of mom r * a na. Although in the preterm mode the sample by injection of sample 120 works to elevate the inner chamber 110 of internal pressure as the sample is injected into the sealed chamber by stopping a small amount of air inside the chamber. As the sample received in the chamber, in alternate modes you can also use other means such as a manual pressure pump (not shown) to increase the internal pressure within the frame 11 to 110. Custom-made that the plasma portion of the whole blood sample penetrates the membrane disc 140 in step 5, (1 this portion of plasma is collected by channel means 150 and then channeled to the clear capillary tube 150. In step 6, the detector of hemolysis 100 is inverted again and the mati or plasma dye in the capillary tube 160 is observed with the naked eye or with a column amplifier (not shown) .If the hue of the plasma is amber *, this indicates that the sample of sangr And whole was normal. Alternatively, if the hue of the plasma is Ib pink, this indicates that hemolysis has occurred. Although in the preferred embodiment of the detector 110, which has just been described, the clear capillary tube 160 can be used only to detect if haemolysis has occurred in the sample simply by observing the hue or the plasma in the tube, in alternative embodiments a indicator paper such as guayaco paper (described below together with detector 400) can be used to detect hemolysis of the plasma after it has penetrated through the membrane disc 140. Referring now to figures 4-6, respective and section views of channeling means 150 for collecting and channeling the plasma portion of a blood sample according to a preferred embodiment of the present invention. The channeler means 150 is formed of a plurality of V-connected V-shaped channels 152. Each adjacent pair of channels 152 is attached to a flange 154. The flanges 154 are located against the membrane disc 140 when the detector of Hemolysis 100 is in its assembled state. A collection channel 156 is coupled to each of the channels 152 and to the clear capillary tube 160. During the operation of the hemolysis detector 100 (and in particular during step 5 shown in Figure 7), the plasma that penetrates the through the membrane disc 140 flows first into the channels 152 and then into the collection channel 156. Subsequently, the plasma in the collection channel 156 flows by gravity towards the clear capillary tube 1? 160. Although in the preferred embodiment of the channeling means 150 the channels 152 are V-shaped, in the alternative embodiments said channels may be in the form of b U "Also, in alternative embodiments, the channels 152 may be coupled to a tube. capillary 160 through collection channels. Finally, in a further alternative embodiment (not shown), the channeling means 150 may be fopnados a container in the form of a bowl with a capillary t bo 160 coupled to the lower portion of the bowl, such that the bowl catches plasma as it penetrates through the membrane disc 140 and then channels the plasma by gravity to the capillary tube 1.60. Referring now to figures 8-10, L5 show perspective and sectional views of a hemolysis detector 200 according to a preferred embodiment of L of the present invention. The hemolysis detector 200 is substantially equivalent to the hemolysis detector 100, except that, as explained in more complete form more ahead, the hemolysis detector 200 is configured slightly differently and has its sample injection port 220 located on the longitudinal portion of the sealable chamber 210. In this manner, the hemolysis detector 200 is formed of a sealable chamber. 210 that has a sample injection port 220 for receiving a blood sample in the sealed chamber. In the same way as the adjustable chamber 110, the sealable chamber 220 is preferably formed of a clear rigid plastic and has a fixed internal volume. the sealable chamber 210 is mounted to a chamber base 230 which is also preferably formed of a clear rigid plastic. An icroporous membrane disc 240 (not shown in FIG. 8, but shown in FIG. 10) is provided in and forms one end of the chamber 210. The microporous membrane disc 240 is substantially equivalent to the membrane disc. 140. Channeling means 250 are located immediately adjacent to an icroporous membrane 240. The channeling means 250 collects the plasma portion of a whole blood sample as this portion of plasma penetrates through the membrane disc. 240 and then channels that portion of plasma to a clear capillary * tube 260. Referring now to Figure 11, there is shown a diagram illustrating a preferred method for using a hemolysis detector 200 to detect hemolysis in a blood sample of a patient. As shown in figure 11, the hemolysis detector 200 is used substantially in the same way as the hemolysis detector 100, except that in the hemolysis detector 200 the injection of the saline solution and the whole blood sample in steps 1 and 3 is achieved using a sample injection port located along the longitudinal portion of the chamber 210; Referring now to Figures 12-14, we are show perspective and sectional views of a hemolysis detector 300 in accordance with a further preferred embodiment of the present invention. The hemolysis detector 300 is formed of a sealable chamber 310 having a sample injection screw 320 for receiving a blood sample in the sealed chamber. The sealable chamber 310 is preferably formed of a clear rigid plastic and has a fixed internal volume. A capillary tube cover 330 is secured to the sealable chamber 310. The capillary tube cover 330 is also preferably formed of a clear rigid plastic. A microporous membrane disc 340 (not shown in Figs. 12-13, but shown or in Fig. 14) is provided in and forms a boundary defining a sealable chamber wall 310. Therefore, unlike the systems of 100 and 200 hemolysis detectors that were formed of sealed chambers that were cylindrical in shape, the hemolysis detector 300 is formed of a seal chamber 310 that is essential in the form of an L. For clarity purposes, the inner portion of the detector * of hemolysis 300 occupied by the sealable chamber 310 is indicated in figure 14 by shaded line in parallel. As was the case in the hemolysis detectors 100 and 200, the microporous membrane disc 340 in the hemolysis detector 300 preferably has a pore size in the range of 0.2 to 1.2 microns, and most preferably 0.40 to 0.80 microns. , and is only permeable to the plasma portion of a whole blood sample and not permeable to the cell portion of the sample. Channeling means 350 are located immediately adjacent to an icroporous membrane disc 340. The channeling means 350 collects the plasma portion of a whole blood sample as that portion of plasma penetrates through the membrane disc 340 and then that portion of plasma is channeled to a clear capillary tube 360. The hemolysis detector 300 is preferably assembled by placing the channeling means 350 within the capillary tube cover 330 and then sealingly welding the sealable chamber 310 to the cover 330 with the membrane disc 340 located between them. Referring now to Figure 15, a diagram illustrating a preferred method for using a hemolysis detector 300 for detecting hemolysis in a blood sample of a patient is shown. As shown in Figure Ib, the procedure begins in step 1 by injecting a thinning solution such as a saline solution into the sealable chamber 310 through the sample injection port 320. In step 2, the hemolysis detector 300 is milled to the sides and agitated, thus causing the wetting solution injected during step 1 to wet the membrane disk 340. This passing step causes the membrane disk 340 (which was previously dry and therefore air permeable). ) becoming air impermeable, thereby transforming the sealable chamber into a sealed chamber 310 having a fixed volume of air therein. In step 3, a sample of whole blood from a patient is injected into the sealed chamber 310 by means of a sample injection port 320. Because the injection port 320 is made of latex or rubber, the air does not escape of the sealed chamber 310 during the injection of the whole blood sample into the chamber. The volume of the whole blood sample injected into said chamber 310 in step 3 preferably corresponds to 1-10% of the fixed internal volume of the sealed chamber 310. Co or noted above in relation to the hemolysis detector 100, the addition of this fluid in the sealed chamber causes its internal pressure to rise to approximately 8-80 mrn Hg, and preferably to 28-30 rnrn Hg. Still referring to FIG. 15, in step 4 of the procedure, the hemolysis detector 300 is agitated to mix the whole blood sample injected into the sealed chamber 310 during step 3 without leaving any wetting solution remaining in the sealed chamber. 310 from step 1. After, in step 5, the hemolysis detector 300 is reversed and the plasma portion of the whole blood sample previously injected into the chamber subsequently penetrates through the membrane disc 340. The increased internal pressure generated by the injection of the sample of whole blood in the chamber of the 310 in step 3 and the created downward pressure fior * the weight of the whole blood sample itself works together during step 5 as a force of momentum to push the portion of plasma of the whole blood sample in an upward direction through <1, the membrane disc 140. Because the plasma portion of the entire blood sample penetrates the membrane disc 340 in step 5, this portion of plasma is collected by channeling means 350 and then channeled into a ascending direction inside the clear capillary tube 360. In step 6, the hue or dye of the plasma in the capillary tube 360 is observed either with the naked eye or with a column loupe (not shown). If the shade of the plasma is amber, this indicates that the whole blood sample is normal. Alternatively, if the shade of the plasma is pink, this indicates that haemolysis has occurred. Referring now to Figure 17, a sectional view of a hemolysis detector 400 is shown according to a still more preferred embodiment of the present invention. The hemolysis detector 400 is formed of a microporous membrane disc 410 which relies on separating * the plasma portion of a whole blood sample that comes from the cell portion of the whole blood sample. The membrane disk 410 has a first side 420 for receiving the sample of whole blood which will be separated. In the embodiment shown in Figure 17, the whole blood sample is preferably received on the membrane disc 410 by placing one or more drops of the whole blood sample on the first side 420 of the membrane disc 410. The microporous membrane disc 410 preferably has a pore size in the range of 0.2 to 1.2 microns, and still more preferably 0.45-0.80 microns. Because the membrane disc 410 is permeable only to the plasma portion of a whole blood sample and not permeable to the cell-portion of the sample, only the plasma that comes from the whole blood sample placed on the first side 420 will pass through the second Side 430 of the membrane disc 410. L-1 membrane disc 410 is preferably formed of a hydrophilic membrane such as Therrnopor 800, Verapor 800 or Supor 800 manufactured by Gelman Science. Inc. It will be understood by those skilled in the art that other membranes may be used in the presence of hydrotoxic membranes which require a pre-wetting step to form the membrane disc 410. In addition, in the preferred embodiment, the membrane disc 410 It is 5-10 m in diameter, although you can also use disks that are larger or smaller in diameter. It will also be understood by those skilled in the art that membranes configured in forms other than discs may also be used in place of the membrane disc 410. A blood barrier 440 is coupled to, and posited cloned against, the first side 420 of the disc. membrane 410. The blood barrier is preferably circumferentially defined and defines a perimeter 450 enclosing the first side 420 d L membrane disc 410. In the preferred embodiment, membrane disc 410 and blood barrier 440 are adhered to one another. with another, although these elements can be insured between SL using other fixing means. The blood barrier 440 is preferably formed of a molded plastic material.

Still referring to Figures 16 and 17, the hemolysis detector 400 is further formed by a paper indicator paper 460, which is secured within a plastic cover 470 by means of glue 480 or by b any or medium Fixing. The indicator paper disc 460 is preferably positioned directly adjacent to, and in contact with, the second side 430 of the membrane disc 410. The indicator paper disc 460 is also preferably coupled to the membrane disc 410 in a detached manner so that the indicator disc 460 and the membrane disc 410 can be easily separated. The indicator paper disc 460 is preferably formed from a porous paper impregnated with guayaco reams such as He occult paper manufactured by Smit-hKline Diagnostics, Tnc. Such a paper displays a blue * color 5 in the presence of hemoglobin when it is irradiated with a hydrogen peroxide sol. A suitable solution of hydrogen peroxide to be used in conjunction with this aspect of the present invention is the Hemoccult developer manufactured by Srni hKline Diagnostics, Tnc. The indicator paper disc 460 preferably has a larger diameter than the membrane disc 410 and in the preferred embodiment is approximately 20 rn in diameter, since in the preferred embodiment the indicator paper disc 460 is larger than the disc. Membrane disk 410, a 1- portion of the disc of the indicator 460 will rest outside the perimeter 450. However, those experts will be understood as 2!:, In the art, it is also possible to use an indicator paper configured in forms other than discs instead of disc 460. To evaluate the presence of hemoglobin in free form in a whole blood sample using the hemysis detector 400, one or more drops of a blood sample is placed within the perimeter 450 on the first side 420 of the membrane disc 410. If the membrane disc 410 is formed of a hydrophobic or slightly hydrophilic paper, the membrane defect 410 should be pre-formed. moistened with a humidifying solution such as an aqueous solution of iopropyl alcohol or 5-20% ethyl alcohol before the whole blood sample is dripped onto the membrane disc 410. Other organic solvents can also be used as a fluoridating agent, as long as these solvents do not leave a residue on the membrane disc 410 that could interfere with the blood test. The purpose of this pre-wetting step is to hydrolyze the membrane disc. The pre-moistening step can be carried out by simply dropping * one or more drops of the moistening solution onto the membrane disc 410. After being placed one or more drops of the whole blood sample within the perimeter 450 on the first side 420 of the membrane disc 410, the plasma portion of the whole blood sample will wet the membrane disc 410, as well as the indicator paper disc 460 that rests under the membrane disc 410. If the paper disk is displaced indicator 460 spreads beyond the perimeter 450, a drop of developer solution is applied to the moistened portion of the indicator paper disc 460 resting outside the perimeter 450. If a blue color appears in the moistened area after about one minute, this is an indication that hemoglobin exists in free form in the plasma and therefore hemolysis is detected as present. Alternatively, if the wetting of the indicator paper disc * 460 does not propagate beyond the perimeter 450, the membrane disc 410 should be removed from its position on the indicator paper disc 460 and a drop of solution r- The leaner should then be applied to the moistened portion of the indicator paper disc * 460 which previously rests under the membrane disc 410. Again, if a blue color appears in the moistened area after about one minute, this is an indication that hemoglobin exists in free form in the plasma and therefore hemolysis is detected as present. Although in the mode of detector 400 just described above the blood constituent is hemoglobin in free form and an indicator paper impregnated with guaiac resin is used to detect hemoglobin, it is understood by those skilled in the art. the technique that the detector 400 of the present invention can be applied to detect * other constituents of the blood in the plasma by varying the type of indicator paper used to form the disk 460. Referring now to FIG. 18, a view is shown section of a hemolysis detector 500 according to b a still preferred mode of the present invention. The hemolysis detector 500 is formed by a chamber 510, which is closed on all sides except its lowermost side 520. In the upper part of the chamber 510 is provided a sample injection port 530 for receiving a whole blood sample) within the chamber 510. A camera base 540 is also provided. Both the camera 510 with the camera base 540 are preferably formed of molded plastic. A microporous membrane disc 550 and a disc of indicator paper 560 are also provided. The disk of Ib membrane 550 is preferably formed of a hydrophilic membrane such as a hydrophilic membrane such as Thernopor 800, V rapor 800 or Supor 800 manufactured by Geman Science. Tnc. It will be understood by those skilled in the art that other membranous membranes can be used, including hydrophobic membranes. Similarly, the indicator paper disc 560 is preferably formed of a porous paper impregnated with guayaco seeds such as Hemoccult paper manufactured by SmithKline Diagnosfics, Tnc. In contrast to the detector 400 system, the membrane disc 550 and the disc of indicator paper 560 in the detector 500 have the same diameter substantially. twenty During the assembly of the detector 500, the membrane disc 550 and the indicator disc 560 are positioned between the upper edge 570 of the camera base 540 and the lower side 520 of the chamber 510, and the upper edge 570 of the The base of the chamber 540 and the lowermost side 520 of the chamber 510 are secured to each other with both discs between them, preferably by means of final welding. Once the detector 500 is assembled, the preferred application of the device begins with the injection of one or two milliliters of a releasing agent (such as 15% isopropyl alcohol in water) into the chamber 510 through the injection port. sample 530. The injected humidifier solution moistens both the membrane disc 550 and the indicator paper disc * 560, and most of the injection solution passes out of the device within one minute. Next, a predetermined volume of a whole blood sample (preferably 1-10% of the internal volume of the chamber 510) is injected into the chamber 510 through the sample injection port 530. After maintaining the 500 detectoi in its erect position with the sample injection port 530 at the top for about one minute, the detector 500 is then poured 180 degrees and one or two drops of the developer solution are applied to the indicator paper disk * 560. The bottom of the chamber base 540 is preferably open or exposed to the outside to allow the developer to be applied directly on the paper; > q inflator If a blue color appears in the area of the developing solution after approximately 30 seconds, this is an indication that hemoglobin exists in the free form in the plasma and therefore hemolysis is detected as present. b Although in the mode of detector 500 described just above the constituent of blood constituents is hemoglobin in free form and an indicator paper impregnated with guaiac resin is used to detect such hemoglobin, it will be understood by those skilled in the art that the The detector 500 of the present invention can be applied to detect other constituents of the blood in the plasma by varying the type of indicator paper used to form the disc 560. Referring now to FIG. 19, a sectional view of a detector is shown. hemolysis 600 according to a still more preferred embodiment of the present invention; The hemolysis detector 600 is formed by a chamber 610, which is closed on all sides except on its lowermost side 620. At the top of the chamber 610 is provided with a sample injection port 630 to receive a sample of whole blood inside the 610 chamber. A 640 chamber base is also provided. Both the chamber 610 and the camera base 640 ostán preferably formed of molded plastic. A microporous membrane disc 650 is also provided. The membrane disc 650 is the substantial equivalent of the membrane disc 550 described above.

During the assembly of the detector 600, the membrane dLsco 650 is positioned between the upper edge 660 of the camera base 640 and the lowermost side 620 of the chamber 610, and the upper edge 660 of the camera base 640 and the side The bottom panels * 620 of the chamber 610 are secured to each other with the membrane disc between them, preferably by ultrasonic welding. Once the detector 600 is assembled, the preferred application of the device begins with the injection of one or two milliliters of a releasing agent * (such as a normal saline solution or 15% isopropyl alcohol in water) into the chamber 610 through the sample injection port 630. The injected humidifier solution moistens the membrane disc 650, and most of the injection solution passes out of the device within one minute. In addition, a predetermined volume of a whole blood sample (preferably 1-10% of the internal volume of the chamber 610) is injected into the chamber 610 through the sample injection port 630., After maintaining the detector 600 in its erect position with the sample injection port 630 at the top-during < about one minute, the detector 600 is then inverted 180 degrees and a strip of indicator paper (such as guaiac paper) is wetted leading to the indicator paper (not shown) in contact with the side 670 of the membrane disc 650. bottom of chamber base 640 is preferably open or exposed to the outside to allow the indicator paper to be brought into direct contact with the membrane disc. One or two drops of developer solution is then applied to the indicator paper strip. If a blue color appears in the area of the developing solution after approximately 30 seconds, this is an indication that hemoglobin is present in free form in the plasma and thus the hemoly is detected as present. Although in detector mode 600 just described above the blood constituent of hemoglobin in free form and an indicator paper impregnated with guaiac resin is used to detect such hemoglobin, it will be understood by those skilled in the art that the detector * 600 of the present invention can be applied to detect other constituents of the blood in the plasma by varying the type of indicator paper used during the operation of the system. Referring now to Figure 20, a sectional view of a hemolysis detector 700 is shown according to a still more preferred embodiment of the present invention. The "hemolysis detector 700 is formed by a chamber 71 0.

At the top of the chamber 710 a sample injection port 720 is provided to receive a sample of whole blood within the chamber 710. A camera base 730 is also provided. Both the camera 710 and the camera base 730 are preferably formed of molded plastic. A microporous membrane slide 740 is also provided on the lowermost side of the chamber 710. The membrane elise 740 is the substantial equivalent of the membrane elyse 550 described above. A half pipeline axis 750 is positioned immediately below < Jel ejisco membrane axis 740. The channeling means 750 is substantially equivalent to the channeling means 150, and thus collects the plasma portion of a whole blood sample by passing that plasma through the membrane disc 740 and then it channels that portion of plasma to a clear capillary udder 760. In contrast to the hemolysis detector 100 in which the lower end of the capillary tube is closed, in the hemolysis detector 700 the lower end 770 of the capillary tube 760 is open. During the assembly of the detector 700, the membrane disc 740 and the channeling means 750 are positioned between the camera base 730 and the camera 710, and the camera shaft base 730 and the camera 710 are secured to each other with the ellipse. of membrane and the channeling medium between them, preferably by ultrasonic welding. Once the detector 700 is assembled, the preferred application of the device begins with the injection of one or two milliliters of a wetting agent (such as a normal saline solution and 15% isopropyl alcohol in water) into the chamber 710 a through the sample injection port 720. The injected humidifier solution passes to the membrane disc 740, and most of the injection solution passes outwardly through the one-minute elentro device. Next, a pre-aligned volume, a sample of whole blood, is injected into the chamber 710 through ele- sample injection shaft port 720. After holding the detector 700 in its erect position with the sample injection port 720 at the top * for about one minute, a strip of indicator paper (such as guaiac paper) is moistened by eμje plasma drops exit end 770 of tube 760 on a strip of indicator paper such as guaiac paper (not shown). One or two drops of r-velor solution are then applied to the anger indicator paper. If a blue color appears in the area of the r-velor solution after approximately 30 seconds, this is an indication that hemoglobin is present in the free form in the plasma and therefore hemolysis is detected as resent. Although in detector mode 700 the indicator paper is applied to the plasma output tube 760 to determine if haemolysis is present, it will be understood by those skilled in the art that an "optical" sensor system (formed from a source) may alternatively be used. of light transmission 780 and a light sensor 790) to analyze the plasma leaving the tube 760 and thus determine if 1 has occurred 1. The present invention can be incorporated into other specific forms without departing from the spirit or essential attributes of the invention. the invention.As a consequence, reference must be made to the claims, rather than to the previous description, all time <;] u these indicate the scope of the nvenei ón.

Claims (2)

3b NOVELTY OF THE INVENTION CLAIMS

1. - An apparatus for detecting hemolysis from a blood sample comprising: (A) a sealed chamber having an internal pressure ejue results from a presence of a quantity of air within the shaft, eg a sealed chamber; (B) said sealed chamber has the purpose of receiving a volume of fluid within said sealed chamber, said volume of fluid includes said blood sample, said means for receiving said volume of fluid includes means for raising said internal pressure to a pressure internally increased upon receipt of said fluid within said sealed chamber retaining said amount of air in a sealed chamber upon receipt of said fluid volume within said sealed chamber; (Or a membrane to form at least a portion of one side of said sealed chamber, said membrane is permeable to a portion of plasma of said sample in said sealed chamber when said internal pressure is equivalent to said increased internal pressure, said membrane being impermeable to blood cells in said sample; (D) hemolysis detection means posi cloned out of said sealed chamber to receive an axis test volume said plasma portion of said sample after said sample volume of said plasma has penetrated said membrane, and to detect a condition of hemolysis according to a nuance associated with said firing volume of said plasma portion

2. The apparatus according to claim 1, wherein said volume fluid includes only said sample of blood 3.- The confi gning device with the claim, in the eXample seal chamber has a volume and in which said blood sample it has a volume ^ or is equivalent to 1-10% of said volume of said sealed chamber. 4. The apparatus in accordance with the claim 1, wherein said hemolysis defecation means detects said haemolysis condition only if said hue associated with said test volume of said portion of plasma is represented by a predetermined color. L5 5.- The axis device according to the claim 4, wherein said hemolysis detection means are formed by a translucent hollow tube and said hue associated with said test volume axis said portion of plasma is a dye that appears within said translucent hollow tube when Said test volume of said portion of plasma is poled within said translucent hollow tube. 6. The apparatus according to the indication rei 4, in the e-ue said means of detection of hemolysis are formed by an indicator paper. 7. The apparatus according to claim 1, wherein said means for detecting hemolysis are formed by an optical sensor. 8. An apparatus for detecting hemolysis from a blood sample comprises: (A) a sealable chamber, said sealable chamber having means for receiving a fluid volume within said chamber, said fluid volume including said sample axis blood; (B) meelios relies to transform said sealable carnage into a sealed chamber having an internal pressure that results from the presence of a volume of air within said sealed chamber; (C) said means for receiving said volume of fluid within said chamber include means for elevating said internal pressure to an increased internal pressure as said fluid is received inside said chamber retaining an amount of air in said chamber sealed to said chamber. be * received the volume of fluid in said sealed chamber; (D) said means for transforming said sealable chamber into a sealed chamber being formed by a membrane, said membrane being permeable to a portion of plasma of said sample in said sealed chamber when said internal pressure is equivalent to said increased internal pressure; (E) Hemolysis defecation means positioned outside said chamber to receive a test volume of said portion of said sample after said test volume of said plasma has penetrated said membrane, and it will be reliable to detect a hemolysis condition in accordance with a nuance associated with said test volume of said plasma portion. < -) .- The apparatus according to claim ü, in which ejicho fluid axis volume includes only said sample sa san e. 10. The apparatus according to claim 8, wherein said sealed chamber has a volume and wherein the blood sample has a volume equivalent to 1-10% of said volume of said sealed chamber. 11. The device < 1e according to the claim 0, in the μje said hemolysis detection methyls detect said haemolysis condition only if said nuance associated with the volume test the said portion of plasma is represented by a predetermined color *. 12. The apparatus according to claim 11, wherein said means for detecting hemysis are formed by * a translucent hollow tube and said nuance associated with said test volume of said portion of plasma is a dye that appears within ejicho shaft translucent hollow tube said volume test of said portion of plasma is positioned elen ro of said translucent hollow tube. 13.- The axis device according to the claim (1, wherein said means of detecting hemolysis are formed by an indicator paper.) 14. The axis according to claim 8, wherein said haemolysis detection axis means are formed by a optical sensor * 15. The apparatus according to claim jq 8, n the e-ue said membrane is adapted to convert to a sealable chamber in a sealed chamber when said membrane is moistened with a moisturizing solution. 16. A method for detecting hemolysis from a blood sample is comprised of the axis steps: (A) provide a sealed chamber to receive said blood sample, said sealed chamber has an internal pressure that a presence of a quantity of air within said sealed chamber results; () received a fluid axis volume in said sealed chamber, said volume of fluid includes said sample of blood; (C) elevating, during said receiving step, said infernal pressure to an increased internal pressure by retaining said amount of air in said sealed chamber when said volume of fluid is received within said sealed chamber; (D) Ib to push, with said increased internal pressure a portion of plasma of ejicha (ours in said sealed chamber to penetrate a membrane, forming said membrane at least a portion of an axis side said sealed chamber; (E) receive a test volume of said plasma portion of the sample shown in media of Hemolysis detection after said test volume of said plasma portion has penetrated said membrane; and (F) detecting a hemysis condition according to a hue associated with said test volume received within said hemolysis axis detection means. 17. The method according to claim 16, wherein step (A) comprises the steps of: (1) providing a sealable chamber for receiving said blood sample; and (?) transforming a sealable chamber into a sealed chamber by applying a moisturizing solution to said membrane, said chamber having an internal pressure resulting from a presence of a quantity of air within said sealed chamber. 18. The method according to claim 16, wherein said volume of fluid includes only said blood sample. 10 19.- The method according to the claim 16, wherein said sealed chamber has a volume and wherein said blood sample has a volume that is equivalent to 1-10% of said volume of the sealed chamber. 20. - The method in accordance with the claim L5 16, wherein said hemolysis condition is < 1 affected in step (F) only if said hue associated with said signature volume of said plasma portion is represented by a predetermined color *. 21.- The axis method according to the claim 20 20, wherein said hemolysis detection means are formed by a translucent hollow tube and wherein said haemolysis condition is detected in step (F) by observing only a nuance that appears within said translucent hollow tube when said test volume of said portion of The plasma is positioned within said translucent hollow tube. 22. The method according to claim 20, wherein said characters of "hemolysis letection" are formed by an indicator paper and in which the hemolysis condition is executed in step (F) by applying at least one drop of said test volume of said portion of plasma to said indicator paper and observing a hue t- | ue appear on said indicator paper.