Blood buffer control

The pH of blood is controlled in part by a buffer composed of carbonic acid, H2CO3, and the hydrogen carbonate ion, HCO3. ... 

The pH of blood is controlled by naturally occurring buffers in the blood. 

A carbonic acid/bicarbonate (H2C03/HC0j ) buffer controls the pH of blood. An irrqtortant feature of this buffer is that carbonic acid decomposes to CO2 and H2O, as shown below ... 

Biomedical Applications. TRIS AMINO is used for a number of purposes in its pure form, it is an acidimetric standard the USP grade can be utilized intraveneously for therapeutic control of blood acidosis TRIS AMINO also is useful in genetic engineering as a buffering agent for enzyme systems, industrial protein purification, and electrophoresis. AMP has found use as a reagent in enzyme-linked immunoassays. The primary appHcation is for alkaline phosphatase assays. 

The most important type of mixed solution is a buffer, a solution in which the pH resists change when small amounts of strong acids or bases are added. Buffers are used to calibrate pH meters, to culture bacteria, and to control the pH of solutions in which chemical reactions are taking place. They are also administered intravenously to hospital patients. Human blood plasma is buffered to pH = 7.4 the ocean is buffered to about pH = 8.4 by a complex buffering process that depends on the presence of hydrogen carbonates and silicates. A buffer consists of an aqueous solution of a weak acid and its conjugate base supplied as a salt, or a weak base and its conjugate acid supplied as a salt. Examples are a solution of acetic acid and sodium acetate and a solution of ammonia and ammonium chloride. 

The temporal correlation between in vitro and in vivo release of insulin was quite good. To induce diabetes, two groups of rats were administered 65 mg/kg of streptozotocin in 0.1 M citrate buffer, pH 4.5. Within several days, the animals had become diabetic, as evidenced by blood glucose levels of approximately 400 mg/dl, and substantial output of glucose in their urine. One group of these animals was then injected subcutaneously with 40-50 mg of 15% insulin-loaded PCPP-SA 50 50 microspheres, 850-1000 pm in diameter. A third group of animals receiving no treatment served as a control. 

Human blood contains a variety of acids and bases that maintain the pH very close to 7.4 at all times. Close control of blood pH Is critical because death results if the pH of human blood drops below 7.0 orrises above 7.8. This narrow pH range corresponds to only a fivefold change in the concentration of hydronium ions. Chemical equilibria work in the blood to hold the pH within this narrow window. Close control of pH is achieved by a buffer solution, so called because it protects, or buffers, the solution against pH variations. 

Figure 8.6 Positive ion LD TOF mass spectra of P. falciparum parasite sample (upper trace), and a control (uninfected blood) sample (lower trace). Protocol D is used for sample preparation. Both samples—in vitro cultured P. falciparum parasites in whole blood, and the whole blood control—are diluted to 5% hematocrit (10-fold) in PBS buffer. In the infected sample the estimated number of deposited parasites per sample well is approximately 100. A commercial LD TOF system is used, and both spectra are normalized to the same (40 mV) detector response value. Each trace represents the average of one hundred single laser shot spectra obtained from linear scanning of an individual well (no data smoothing). The characteristic fingerprint ions of detected heme in the upper trace are denoted.

Moreover, several buffer systems exist in the body, such as proteins, phosphates, and bicarbonates. Proteins are the most important buffers in the body. Protein molecules contain multiple acidic and basic groups that make protein solution a buffer that covers a wide pH range. Phosphate buffers (HPO T /H2P07) are mainly intracellular. The pK of this system is 6.8 so that it is moderately efficient at a physiological pH of 7.4. The concentration of phosphate is low in the extracellular fluid but the phosphate buffer system is an important urinary buffer. Bicarbonate (H2C03/HC0 3) is also involved in pH control but it is not an important buffer system because normal blood pH 7.4 is too far from its pK 6.1 [144],... 

Basic procedure (ACW kit) Mix 1500 pL of ACW reagent 1 (diluter) with 1000 pL of ACW reagent 2 (buffer) and 25 pL of photosensitizer reagent (lumi-nol based). Start measurement after brief vortexing. Assayed solution (or control) is added before addition of photosensitizer reagent. Volume of ACW reagent 1 is reduced by the volume of assayed plasma sample. Standard substance ascorbic acid. Duration of measurement 2-3 min. Measured parameter effective lag phase = lag-phase sample - lag-phase blank. Assayed amount of human blood plasma 2 pL. 

Blank, calibrator, control, and patient whole-blood samples (50 /iL) were transferred into 1.5 mL conical test tubes, mixed with 100 /xL of the IS, vortexed for 10 sec, and centrifuged at 13,000 g for 5 min. Twenty-five microliters of supernatant were injected onto a Cohesive Technologies Cyclone polymeric turbulent flow column (50 x 1 mm, 50 /flushed with a mixture of methanol and water (10 90 v/v) at a flow of 5 mL/min. Column switching from the TFC to HPLC systems was via a Cohesive Technologies system. The analytical column was a Phenomenex Phenyl-Hexyl-RP (50 x 2.1 mm, 5 /.mi). The mobile phase consisted of methanol and ammonium acetate buffer (97 3 v/v). The buffer was 10mM ammonium acetate containing 0.1% v/v acetic acid. The flow rate was 0.6 mL/min. 

Citrated blood is diluted 1 10 with enzyme buffer solution, and preservative is added (H19). The buffer is prepared by dissolving 0.2 g of Clarase (Fisher Scientific Co., New York) in 100 ml citrate buffer (5 g potassium citrate monohydrate and 1 g citric acid monohydrate in 1000 ml distilled water, pH 5.6). The solution is incubated for 3 days at 37°. After incubation, it is autoclaved 15 minutes to stop enzymatic action and coagulate proteins. It is filtered, and 1.0, 1.5, and 2.0 ml of the supernatant is added to individual flasks and assayed. Control flasks are included to estimate pantothenic acid contamination of the enzyme. 

Serum albumin is the most abundant protein in blood plasma. Its primary function is to control the colloidal osmotic pressure in blood, but is also important for its buffering capacity and for its ability to transport fatty acids and bilirubin, as well as xenobiotic molecules. The physiological implications of its esterase-like activity are unknown (see Sect. 3.7.5). 

Ammonia buffers protons in the tubules of the kidney to prevent formation of an acidic urine, when the kidney excretes (secretes) protons to control the pH of the blood. 

In humans, the pH of blood is held at a remarkably constant value of 7.4 0.05. hi severe diabetes, the pH can drop to pH 7.0 or below, leading to death from acidotic coma. Death may also occur at pH 7.7 or above, because the blood is unable to release CO2 into the lungs. The pH of blood is normally controlled by a buffer system, within rather narrow limits to maintain life and within even narrower limits to maintain health. 

One of the most important buffer systems in the human body is that which keeps the pH of blood around 7.4. If the pH of blood fall below 6.8 or above 7.8, critical problems and even death can occur. There are three primary buffer systems at work in controlling the pH of blood carbonate, phosphate, and proteins. The primary buffer system in the blood involves carbonic acid, H COj and its conjugate base bicarbonate, HCO3. Carbonic acid is a weak acid that dissociates according to the following reaction ... 

The salts of the buffer pairs responsible for control of the pH of plasma and extracellular fluid involve sodium as the principal cation, while the cellular buffers involve potassium salts. See also Acid-Base Regulation (Blood) and Diuretic Agents. 

The Bohr effect is closely related to the major roles that hemoglobin plays in disposing of the C02 produced in tissues, and in controlling the blood pH. While oxygen is being delivered to the tissues in the venous blood, the C02 is being removed from the tissues (fig. 5.5). The C02 that diffuses into the erythrocytes is rapidly converted into carbonic acid, which in turn dissociates into H+ and HC03. The protons produced by this dissociation would lower the pH and reverse the dissociation if it were not for the buffer-... 

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