\$20 Bonus + 25% OFF

# BCH2011 Laboratory Manual

0 Download2 Pages / 442 Words Add in library Click this icon and make it bookmark in your library to refer it later. GOT IT

## Question:

Students must come to the practical class with written answers to the following QUESTIONS. If NOT completed and submitted before class commences 50% of the total possible marks for the weeks exercise will be deducted. Demonstrators will check these during class and return to students. The Pre-Practical exercise must be submitted with the report’. Both the exercise below and the Assay of Glucose require the preparation of a standard curve. A standard curve, which is determined using a standard solution, relates data for two variables, e.g. absorbance and concentration. These data are connected with a curve of best fit. Consider the data below for a 2.0 mM solution of glucose, which has been diluted to cover a range of concentrations appropriate for a planned experiment.

Tube Number 1 2 3 4 5 6 Volume of 2.0 mM glucose (ml) - 0.2 0.4 0.6 0.8 1.0 Volume of water (ml) 1.0 0.8 0.6 0.4 0.2 - Glucose (µmoles/tube) - 0.4 0.8 1.2 1.6 2.0 Absorbance at 500 nm 0.05 0.30 0.76 1.01 1.23 1.35

A standard curve for a solution consists of a plot of the amount of glucose as the x-axis versus the absorbance at 500 nm as the y-axis. The data are plotted in the figure below. The correct curve of best fit is the unbroken line. The dotted curve simply connects the points and is not a curve of best fit. The curve of best fit is drawn so that the sum of the distances of any points offset from the curve is the same on either side of the curve. Thus, in the simple example shown below, the two offset points are equidistant from the curve.

In this graph, the linear region of the graph where the Beer-Lambert Law is obeyed lies between 0 and 1.2 µmol of glucose per tube. Continued next page Tube Number 1 2 3 4 5 6 Volume of 2.0 mM glucose (ml) - 0.2 0.4 0.6 0.8 1.0 Volume of water (ml) 1.0 0.8 0.6 0.4 0.2 - Glucose (µmoles/tube) - 0.4 0.8 1.2 1.6 2.0 Absorbance at 500 nm 0.05 0.30 0.76 1.01 1.23 1.35 Linear region Pre-Practical Exercise continued The axes of a standard curve need to be labelled, with the units specified, where appropriate.

Thus, in the current example, the x-axis is entitled Glucose and the units are µmoles/tube and the y-axis is entitled Absorbance at 500 nm; no units are specified for the latter as absorbance has no units. Each axis also needs to be marked into divisions (the scale) to allow the data to be plotted and read accurately. These divisions usually involve multiples 1, 2, 5 or 10, or 0.1, 0.2, 0.5 and 1.0, etc. and not 3, 7, etc. which do not allow data to be plotted accurately using the grid of graph paper or read accurately from the graph. Furthermore, the scale should be chosen so that the resulting curve has a gradient of 45° with respect to the x-axis and y-axis; this minimises errors due to scaling.

Exercise The haemoglobin (Hb) content of blood is normally within the following ranges: males 130-180 g per litre females 110-160 g per litre. It can be ascertained by treating a blood sample with a reagent containing an excess of ferricyanide and cyanide, which converts both Hb and oxygenated Hb into cyanomet-Hb. The latter can be determined conveniently from its absorbance at 540 nm by comparison with a standard curve. In order to construct a standard curve Tubes 1-5 (see below) were prepared using a standard solution containing 0.5g Hb per litre. At the same time a 0.01 ml sample of blood was diluted with water and treated with ferricyanide/cyanide reagent (Tube 6).

All tubes had a total volume of 5ml. The absorbance at 540 nm of Tubes 1-6 was measured and is recorded in the protocol below. Tube Number 1 2 3 4 5 6 (sample) Volume of standard Hb solution (ml) - 1.0 2.0 3.0 4.0 - Volume of cyanide reagent (ml) 1.0 1.0 1.0 1.0 1.0 1.0 Volume of water (ml) 4.0 3.0 2.0 1.0 3.99 Volume of blood (ml) - - - - 0.01 Absorbance at 540 nm 0.025 0.090 0.160 0.230 0.290 0.210 Amount of Hb (mg/tube) Determine the amount of Hb in Tubes 1-5 and enter these data in the protocol. Using the data for Tubes 1-5, construct a standard curve by plotting the absorbance at 540 nm versus the amount of Hb (mg/tube). Answer the following questions about the blood sample: The amount of Hb in Tube 6 = . . . . . . . . . mg The amount of Hb in the 0.01 ml blood sample = . . . . . . . . . mg The concentration of Hb in the blood sample = . . . . . . . . . g per litre Is the Hb content normal? . . . . . . . . .

### Cite This Work

"Laboratory Manual." My Assignment Help, 2020, https://myassignmenthelp.com/free-samples/bch2011-laboratory-manual.

My Assignment Help (2020) Laboratory Manual [Online]. Available from: https://myassignmenthelp.com/free-samples/bch2011-laboratory-manual
[Accessed 17 September 2021].

My Assignment Help. 'Laboratory Manual' (My Assignment Help, 2020) <https://myassignmenthelp.com/free-samples/bch2011-laboratory-manual> accessed 17 September 2021.

My Assignment Help. Laboratory Manual [Internet]. My Assignment Help. 2020 [cited 17 September 2021]. Available from: https://myassignmenthelp.com/free-samples/bch2011-laboratory-manual.

### Latest Other Samples

#### EDUC 7009 Assessment Of Learning

Answer: The lifelong process of managing, learning, processing the work, and transition so as to move towards to the personally determined and evolving preferred future is known as career development. There are many phases of career from where an individual has to pass and it is necessary that he should remain determined regarding the future aspects. Managing career plan and following the same is a very crucial task attached with any individua...

Read More Tags: Australia Pascoe Vale Management University of New South Wales

#### CSE2ISD Information Systems Development

Answer: The system owner is concerned with the business procedures for the system. The main business functions for the systems are: Generation of the sales reports, distribution of the funds to the artists, and sending the sales record and the banks statements to the accountant for handling the financial affairs of the company. 1. Business and Information Systems Users The main users of the system are: Members of the company Manage...

Read More Tags: Australia Abbotsford Computer Science CSE2ISD â€“ Information Systems Development University of Queensland Engineering

#### FIN60003 Business Modelling And Analysis

Answer: In a bid to provide a more empowering and nurturing work environment for employees, certain initiatives have been put in place by the firm.  A 100 sample of employees has been derived from the given population (i.e. all employees employed by the firm) in order to perform various statistical tests to gauge if these have achieved the desired impact. Further, this report also highlights the nature and extent of association between so...

#### QAB 105 Quantitative Analysis For Business

Answer: Number of retailers sold products below the minimum price = 79 A random sample is a population subset where every element has an equal chance or probability of being selected. A random sample can be selected by ensuring that all the retailers be given a unique number and then 200 numbers can be randomly selected from the whole population of numbers through software to avoid any bias. The retailers corresponding to the number selecte...

Read More Tags: Australia Punchbowl Accounting University of New South Wales

#### CMS800003 Construction Law

Answer: It’s very common for participants in construction to enter into a contract. It may be a Subcontractor and General Contractor, Builder and Architect or the Owner and the Contractor. Also, construction contracts are often prone to amendments to suit the demands of the work, so the parties at one time may agree to add, change or remove some terms. With that, modifications may occur at the site where writing would not be available, a...

Read More Tags: Australia Gilberton Management Organizational Development & Change University of New South Wales Masters in Business Administration
Next

### We Can Help!

Are you confident that you will achieve the grade?

### Refer & earn

Just refer 5 friends to earn more than \$2000.

## 5% Cashback

On APP - grab it while it lasts!