Investigation Into Haemoglobin Levels And The Relationship Between Absorption And Concentration

The concentrations and absorbances of the red and blue dyes were used to find the concentration of the purple dyes. From the graph of the blue dye, the linear equation for absorbance was y = mx + b. From that formula came the equation y = 7.915 x 104 (x) + 0.02489, where y represents absorbance, m is slope, x is concentration/molarity, and b is the constant/y-intercept. The same set up was performed for the red dye, but the equation produced was y = 1.045 x 104 (x) +.001298. The equations found when graphing absorbance vs. concentration were used to find the concentration of the purple dyes. The absorbance for purple dye 3 on the red wavelength of 470 nm equaled 0.149 and 0.818 for the blue wavelength of 635 nm. For purple dye 1

In Exercise 1, diaminofluorene is used to determine the hemoglobin concentration in the daphnids. A higher hemoglobin concentration is indicated by a darker blue color. A spectrophotometer was used to determine the absorbance at 610nm. When measuring the absorbance levels a blank is necessary to have a zero reference, the blank is the “starting point” for the measurements of the sample (re-word). The blank consists of 10µL of diaminofluorene(DAF), 50µL of hydrogen peroxide, and 0.5mL of PBS. The PBS acts as a buffer in this experiment. The cuvette with the sample of daphnids consisted of 10µL of DAF, 50µL of hydrogen peroxide, and 0.5mL of the sample of Daphnia. In Exercise 2, the Pasteur pipette was used to obtain the sample of Daphnia. The depression slide used in this experiment isolated the daphnid, cotton was used to keep the daphnid still while the heartbeat was counted. The ocular micrometer on the microscope allows the tail spine length to be measured accurately, as well as using an ocular magnification table.

Scientists use an instrument called a spectrometer to quantitatively determine the amount of light absorbed by a solution. The primary inner parts of a typical spectrometer are described below. The spectrometer has a light source that emits white light containing a vast mixture of different wavelengths of electromagnetic radiation. The wavelength of interest is then selected using a monochromator (“mono” meaning one and “chromate” meaning color) and an additional exit slit. The separation of white light into different colors (wavelengths) is known as diffraction. The selected light then reaches the sample and depending on how the light interacts with the chemical compound of interest, some of the light is absorbed and some passes straight through. By comparing the amount of light entering the sample (P0) with the amount of light reaching the detector (P), the spectrometer is able to tell how much light is absorbed by the sample.

Using the yellow tube, which included everything but starch, as the blank, each group zeroed their spectrophotometer. This was done so that any absorbance observed depends only on the amount of starch present, not on any other reagents (buffer, IKI). To zero the spectrophotometer, the wavelength was first set at 580nm, using knob 3 (45). Next, the groups made sure that the light next to “transmittance” was lit, and the chamber to be tightly closed. Having the chamber empty & closed tightly provides reference for the darkest condition possible. Using knob 1, the transmittance was turned until it read 0.0 (45). Before the groups used their blank test tube to zero the spectrophotometer, each needed to wipe the tube with kimwipes to ensure a clean reading. Turning knob 2, each group was then instructed to zero the absorbance, 0.000. Upon removing the blank, each trial was inserted into the chamber (46). The

The dyes in the laboratory experiment are made of numerous colors, mainly red and blue, the spectra from each of the dyes corresponded to the wavelengths obtained from each of dye i.e. 620 nm for red and 450 nm for blue.

1.Full blood counts: To assess the Hb for evidence of anemia, WBC for evidence of infection.

We hypothesized that similar proteins would also be present in bovine blood, yet the major band present in this sample was located at much higher molecular weight of approximately 45 kDa. Potential causes in weight differences could be the longer amino acid sequence present in rabbit hemoglobin, 170 amino acids, versus the 145 amino acids present in bovine hemoglobin, yet this would result in bovine being further down on the gel versus what is actually shown (Altschul et al, 1997). Given the data obtained, potential misidentification of hemoglobin appearing as the main protein present or possible error in lane one (rabbit blood), could explain the anomaly in

The purpose of this lab was to observe how light is altered as it goes through a solution containing a visible dye. In doing so, we will understand the relationship between absorbance, concentration and light path length. We first created different concentrations of blue stock solutions through dilution. We made 4 dilutions, which gives us 5 solutions in total. Each diluted cup (A-D) had 20 g of stock solution to start with and then 10, 30, 30, 46 g of distilled water was added respectively.

In this lab five blood tests will be performed: hematocrit determination, erythrocyte sedimentation rate, hemoglobin determination, blood typing, and total cholesterol determination. A “hematocrit refers to the percentage of red blood cells (RBC’s) in a sample of whole blood. Since the function of red blood cells is the transport of oxygen to the cells of the body, the higher the hematocrit, the more red blood cells are available to carry oxygen.” In a normal, healthy male the value for males is 47% and in a normal, healthy female it is 42%. A person with a lower than normal hematocrit would indicate they are anemic, and a higher than normal hematocrit would indicate

Hemoglobin is present in red blood cells and carries oxygen from the lungs to tissues and returns carbon dioxide from tissues back to the lungs. Met Hemoglobin is a form of the oxygen-carrying part of Hemoglobin of which a small amount is present in human blood and produced automatically. However, if produced in large amounts it can cause coma and even death. Increased levels of Methaemoglobin can be caused by the ingestion of chemicals such as Cyanide and can be detected in blood stains at a scene of a crime as enhanced levels of Met Hemoglobin changes blood upon leaving the body from red to a dark brown in

In activity three, a total count for white blood cells was conducted using a sample slide with a blood smear. The slide was observed under microscope with the above data as the result. The normal percentages for each blood count are as follows (zukur):

Investigating haemoglobin (Hb) concentration in blood samples using the haemoglobincyanide method and in foetal haemoglobin samples

An advantage of the test was that the blood samples used are one of the leading analysis preferences chosen to identify biomarkers, and easily obtainable from patients. However, as direct assay was unavailable, Bromocresol green solution was used which changed the electronic structure thus specific whilst binding to albumin. As a result, this generates accurate results with no anomalies for both serum and blood. Moreover, within the expense limitations of University of Chester, a simple assay was used (Rehm, 2006) expands that BCA assay may have been used

Figure 1 illustrates the absorbance values of pH 5.00 and pH 10.00 bromothymol standard solutions that were recorded from 340-800 nm. The graph illustrates the maximum wavelengths observed at each respective standard solution. This maximum wavelength value is important because it is the wavelength with the highest sensitivity and can therefore minimize random error in the absorbance values. Table 1 illustrates the different maximum wavelength values for the two standard solutions. The acidic bromothymol standard solution had a maximum wavelength at a lower value than the basic bromothymol standard solution (Table 1). These wavelength values are therefore associated with the different colors of the standard solutions.

Gel Filtration, SDS-PAGE Objective – The purpose of this lab was to separate a mixture of Hemoglobin, BSA, Blue Dextran, and yellow food coloring by size into fractions using gel filtration. Also simple non-quantitative assay was performed to determine the fractions that contained proteins. Another objective was to analyze the protein content of being BSA or Hemoglobin and estimate the molecular weight of the proteins collected.