U.S. patent number 3,859,049 [Application Number 05/438,866] was granted by the patent office on 1975-01-07 for blood reference standard and process for blood gas test.
Invention is credited to Edward P. Marbach, Arnold G. Ware.
United States Patent |
3,859,049 |
Ware , et al. |
January 7, 1975 |
BLOOD REFERENCE STANDARD AND PROCESS FOR BLOOD GAS TEST
Abstract
A stable blood reference standard and control for blood gas
tests which includes the use of fluoride, citrate and
iodoacetate.
Inventors: |
Ware; Arnold G. (Pasadena,
CA), Marbach; Edward P. (Los Angeles, CA) |
Family
ID: |
27015859 |
Appl.
No.: |
05/438,866 |
Filed: |
February 1, 1974 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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397347 |
Sep 14, 1973 |
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Current U.S.
Class: |
436/11; 436/18;
436/15 |
Current CPC
Class: |
G01N
33/96 (20130101); G01N 2496/70 (20130101); Y10T
436/105831 (20150115); Y10T 436/102499 (20150115); Y10T
436/108331 (20150115); G01N 2496/05 (20130101); G01N
2333/805 (20130101) |
Current International
Class: |
G01N
33/96 (20060101); G01n 033/16 () |
Field of
Search: |
;23/23B ;252/408
;424/11 |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: Serwin; R. E.
Attorney, Agent or Firm: Jessup & Beecher
Parent Case Text
This is a continuation-in-part of Copending application Ser. No.
397,347, filed Sept. 14, 1973, and now abandoned.
Claims
What is claimed is:
1. A reference standard containing blood, blood cells, hemoglobin,
or the like, reconstituted by the addition of a fluoride and an
iodoacetate or a fluoroacetate.
2. A reference standard defined in claim 1, in which the fluoride
and iodoacetate or fluoroacetate are included with a concentration
for the fluoride substantially in a range of 10-100 millimolar, and
with a concentration for the iodoacetate or fluoroacetate
substantially in a range of 1 to 10 millimolar.
3. The reference standard defined in claim 1, and which also
contains a citrate.
4. The reference standard defined in claim 2, and which further
includes a citrate substantially in a range of 3 to 50
millimolar.
5. The reference standard defined in claim 1, in which the fluoride
is sodium fluoride and the iodoacetate or fluoroacetate is
iodoacetic or fluoroacetic acid.
6. The reference standard defined in claim 1, which comprises
hemoglobin and glycerol in proportions to provide a viscosity of
the order of 3.5 centipoise.
7. A process of providing a reference standard which consists of
deriving a reference standard containing blood, blood cells,
hemoglobin, or the like, dissolving sodium fluoride and iodoacetic
or fluroacetic acid in water, and mixing the resulting solution
with the reference standard.
8. The process defined in claim 7, and which further comprises the
step of dissolving citrate in water and mixing the resulting
solution with the reference standard.
9. The process defined in claim 7, in which 25 milligrams of sodium
fluoride and 93 milligrams of iodoacetic acid are dissolved in 10
ml of water and then mixed with 25 ml of the reference standard or
in multiples or sub-multiples thereof.
10. A process for providing a reference standard which consists of
washing fresh red cells, dissolving sodium fluoride and citrate in
an albumen glucose solution, and suspending the red cells in the
solution.
11. The process defined in claim 7, which comprises deriving a unit
of hemoglobin, and adding glycerol to the hemoglobin to increase
the viscosity thereof to the order of 3.5 centipoise.
Description
BACKGROUND OF THE DISCLOSURE
Blood gas tests are performed in most hospital laboratories on an
emergency basis forthe purpose of diagnosing and treating
abnormalities of pulmonary function and acid base balance. The
three parameters are blood pH, pCO.sub.2 and pO.sub.2. In
performing the test, blood is drawn from the patient and introduced
promptly into specialized equipment containing electrodes specific
for each of the three blood-gas componetns. Since the blood sample
deteriorates rapidly, it must be analyzed promptly, or within
several hours if iced. As time progresses, however, the pH of the
blood sample decreases, the pCO.sub.2 increases, and the pO.sub.2
decreases as a result of cellular metabolism. There is no
recognized process in the prior art for preventing these changes
from occurring in stored blood samples.
Present day blood gas equipment is standardized and controlled by
cumbersome procedures using buffers for pH, and gases for pCO.sub.2
and pO.sub.2. Blood specimens of known pH, pCO.sub.2 and pO.sub.2
would be preferable for this purpose. However, the instability of
these components in blood has made this approach impractical in the
prior art.
DETAILED DESCRIPTION OF THE INVENTION
The present invention is directed to a means of stabilizing the
aforesaid components in blood, or a suspension of blood cells in an
appropriate media, so as to facilitate the use of stored blood for
standardizing and controlling the blood gas equipment. The process
of the invention may also be used to stabilize, for example, a
mixture of hemoglobin and buffer salts, which does not involve
blood cells. The stabilizing agents are fluoride, citrate and
iodoacetate. Each of these substances prevents a portion of the
cellular metabolic processes which cause the changes in blood gases
noted previously: fluoride by binding magnesium; citrate and
iodoacetate by inhibiting different pathways in the metabolic
processes.
It has been found that combinations of the above-mentioned
inhibitors when added to blood or a suspension of blood cells, or a
mixture of hemogloblin, stabilize the blood gases for periods of 30
days or longer when stored in closed containers under
refrigeration.
The following example will further illustrate the present invention
although the invention is not limited to this specific example
which is provided by way of illustration and not limitation.
______________________________________ Fluoride -- 25 millimolar
Citrate -- 12 millimolar Iodoacetate -- 5 millimolar (fluoroacetate
may be substituted for iodoacetate)
______________________________________
It has been found that the concentrations listed above have been
most efficient. However, it has also been found that the listed
components may be varied considerably in concentration and in
relation with each other without appreciably varying the blood gas
stabilizing effect. For example, the following ranges of
concentration have been found to be permissible, without
significantly reducing the blood gas stabilizing effect.
______________________________________ Fluoride 10 to 100
millimolar Citrate 3 to 50 millimolar Iodoacetate 1 to 10
millimolar ______________________________________
In carrying out the process of the invention in accordance with one
particular example, the following steps are presented:
Fresh blood is drawn and mixed with a solution containing
concentrates of sodium fluoride, citrate and iodoacetic or
fluoroacetic acid in sufficient quantity to yield the final blood
concentrations mentioned above. For example, 25 milligrams of
sodium fluoride and 93 milligrams of iodoacetic or fluoroacetic
acid are dissolved in 10 ml of water and then mixed with 250 ml of
blood. The citrate is also dissolved in water and mixed with the
blood. These anions may be added in combination with any cation so
long as the pH is adjusted to approximately 7.4 @ 37.degree.C.
Following this the blood is stored in air tight containers ideally
leaving little or no air space. After 2 weeks at refrigerated
temperature the pH, pCO.sub.2 and pO.sub.2 are stabilized and
remain unchanged for 30 days or longer if kept under refrigeration.
In contrast, blood specimens without these preservatives show a
progressive drop in pH and pO.sub.2 and increase in pCO.sub.2. Such
changes are sufficient in magnitude to render untreated blood
unsatisfactory as a blood gas standard or control.
As another example, fresh red cells are washed and suspended in an
albumen glucose solution containing concentrates of sodium fluoride
and citrate in sufficient quantity to yield the concentrations set
forth above.
As another example, a mixture of hemoglobin (not cells) and buffer
salts may be treated in the manner described above to stabilize the
pH at about 7.40, the pCO.sub.2 at about 40 mm, and the pO.sub.2 at
about 80 mm. Glycerol may be added to increase the viscosity of the
mixture to about 3.5 centipoise, that of normal whole blood, and to
act as a preservative. About a 50 percent solution at 37.degree. C
would have a viscosity of 3.5 centipoise, a 75 percent solution
would have better preserving power, but the viscosity would be 9.4
centipoise.
It will be readily apparent to those skilled in the particular art
under consideration that other examples of the invention described
herein may be devised from an understanding of the foregoing
specification without departing from the spirit and scope of the
invention. It is intended in the following claims to cover all such
modifications, variations and adaptations which are included within
the scope of the invention.
* * * * *