Spectrophotometer Article

Objectives:

1 . To look for the wavelength of maximum compression, Amax, of bromophenol green. 2 . To set up a standard concentration curve of bromophenol green. 3. To look for the concentration in the unknown bromophenol blue alternatives. 4. To determine the concentration of two distinct solutes, bromophenol blue and methyl fruit in a combination. Materials and Methods:

Label Biological Technology practical manual page 5-11.

Results:

Part one particular: Determination of Amax of bromophenol green

The wavelength with maximum absorbance browsing form the graph is 590nm. Part 2: The effect of concentration on absorbance of bromophenol blue answer

Tube

1

2

several

4

your five

6

Unadulterated water (ml)

2 . five

2 . zero

1 . five

1 . 0

0. five

0. 0

Bromophenol green 10 mg/L (ml)

0. 0

zero. 5

1 . 0

1 ) 5

2 . 0

2 . 5

Concentration of bromophenol blue (mg/L)

0. zero

2 . 0

4. 0

6. zero

8. 0

10. zero

Absorbance at 590 nm (Amax of bromophenol blue)

0. 500

0. 135

0. 199

0. 404

0. 596

0. 724

From Figure 2 above, the large molar absorptivity of bromophenol green at 590nm is the lean of the thready regression series which is 0. 0744 L mg-1 cm-1. Part several: Determination in the concentration of the bromophenol blue solutions of unknown attentiveness By plotting on the normal concentration shape from Portion 2, the concentration of bromophenol blue are the following:

By using the formula of Beer-Lambert Law,

Component 4: The result of focus on absorbance of methyl orange solutions

Coming from Figure some above, the molar absorptivity coefficient of methyl orange is the lean of geradlinig regression collection at 460nm which is zero. 0722 D mg-1 cm-1.

Part 5: Determination with the concentrations of two several solutes, bromophenol blue and methyl orange colored, in combination C

The large molar absorptivity agent of bromophenol blue by 460nm is definitely 0. 0036 L mg-1 cm-1.

The molar absorptivity coefficient of methyl orange at 590nm is 0. 0003 M mg-1 cm-1.

According to Beer-Lambert law,

0. 375= 0. 0744 (CBB) + 0. 0003 (CMO)

CBB = …………….. … (1)

0. 308 = 0. 0036 (CBB) + 0. 0722 (CMO) …... (2)

Substitute (1) into (2):

0. 308 = 0. 0036(+ zero. 0722(CMO)

CMO = 4. 039 mg/L

When CMO =4. 039 mg/L:

CBB sama dengan

CBB = five. 024 mg/L

Discussions:

Component 1: Perseverance of Amax of bromophenol blue

Chart of absorbance against the corresponding wavelengths was plotted. Graph plotted displays a bells shape competition. The absorbance of bromophenol blue solution increases slowly but surely as the wavelength boosts until it actually reaches the peak. The utmost absorbance of bromophenol green solution is usually 0. 718 when wavelength is 590nm. However , the absorbance of bromophenol blue solution lessens after achieving the maximum reason for the chart. Part two: The effect of concentration on absorbance of bromophenol blue solution The wavelength was set to 590nm when it was the wavelength with the optimum absorbance. Graph of absorbance against several concentrations of bromophenol blue were plotted. A geradlinig regression range was likewise plotted inside the same chart. From the regression line equation y = 0. 0744x –0. 029, the molar absorptivity coefficient of bromophenol blue can be 0. 0744 Lmg-1cm-1. The graph demonstrate a strong great linear relationship between absorbance and attention of bromophenol blue. In respect to Beer-Lambert Law, when the concentration is usually 0, the absorbance must also be equals to 0. As a result, indicating that those straight line around the graph must intercept the foundation. Therefore , problems might have taken place during the real experiment such as air bubbles trap in the cuvette (1). The stuck air pockets will get in the way the amount of lumination pass through the cuvette. Air bubbles should be removed with a pipette tip to obtain correct readings. Portion 3: Determination of the attentiveness of the bromophenol blue solutions of unknown concentration The absorbance of two different...

References: 1 . Hanna Tools. Technical guidance. http://www.hannacan.com/conseils_colorimetres_en.htm (accessed 6 Drive 2015).

installment payments on your Sausen Silmi. Determining the Concentration of any Solution: Beer 's Law. http://www.scienceteacherprogram.org/chemistry/sausen.html (accessed 6 March 2015).

a few. Oliver Seely. Determination with the Mn Content of Stainlesss steel. http://www.csudh.edu/oliver/che230/labmanual/mangnese.htm (accessed 7 Mar 2015).



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