Spectrophotometer and Its Function

UNIVERSITI PERUBATAN ANTARABANGSA

INTERNATIONAL MEDICAL UNIVERSITY

MALAYSIA

Bachelor of Science (Honours)

Medical Biotechnology Programme

Fundamental of Biological Sciences and Genetics

Semester 1

2015

Experiment 1 : Spectrophotometer and its function

Name : Darvin A/L Anandaraja

Student ID : 00000011854

Course : Medical Biotechnology 1/15

Date : 13/10/2015

TABLE OF CONTENTS

Objectives | 3 |

Materials and Apparatus | 3 |

Results | 4-12 |

Discussion | 13 |

Reference | 14 |

Objectives

1. To determine the wavelength of maximum absorption, Amax, of bromophenol blue.

2. To construct a standard concentration curve for the bromophenol blue.

3. To determine the concentration of the unknown bromophenol blue solutions.

4. To determine the concentration of two different solutes, bromophenol blue and methyl orange, in a mixture.

Materials and Apparatus

* Bromophenol blue 10 mg/L

* Methyl orange 10 mg/L

* Distilled water

* Solution C (mixture of bromophenol blue solution and methyl orange solution)

* Micropipette

* Cuvette

* Test tubes

* Test tube rack

* Vortex mixer

* Tips

* Spectrophotometer

Results

Part 1:

Wavelength (nm) | Absorbance |

470 | 0.119 |

500 | 0.182 |

530 | 0.433 |

560 | 0.747 |

590 | 1.257 |

620 | 0.388 |

650 | 0.090 |

680 | 0.074 |

Table 1.1

Graph 1.1

Part 2:

Tube | 1 | 2 | 3 | 4 | 5 | 6 | A | B |

Distilled water (ml) | 2.5 | 2.0 | 1.5 | 1.0 | 0.5 | 0 | | |

Bromophenol blue 10mg/L (ml) | 0 | 0.5 | 1.0 | 1.5 | 2.0 | 2.5 | | |

Concentration of bromophenol blue (mg/L) | 0 | 2 | 4 | 6 | 8 | 10 | 3.767 | 2.310 |

Absorbance at 590 nm (Amax of bromophenol blue) | 0.000 | 0.244 | 0.469 | 0.728 | 0.963 | 1.145 | 0.437 | 0.268 |

Absorbance at 460 nm | 0.000 | 0.012 | 0.029 | 0.047...