Experiment Analysis of riboflavin content in a vitamin pill by fluorescence spectroscopy

A       IDENTITAS

          Hari/Tanggal    : Selasa /7Juli 2012

          Waktu               : 14.30-17.30

B       KEGIATAN                                        

         Experiment Analysis of riboflavin content in a vitamin pill by fluorescence spectroscopy

C       RINGKASAN KEGIATAN

Materials and equipment                       

1.      Material

Riboflavin tablets

2.      Equipment

Fluorescence spectrometer, Balance

Test tubes: 25 mL x 10                       Volumetric flask: 100mL x 2 (Brown)

Beaker: 50mLx2, 250mL x 2              Pipette: 10mLx1, 2mLx1, 1mLx1

Filter paper: 11cm                               Dropper x 2

Coordinate paper, mortar, funnel, rubber suction bulb, test tube rack, pipette rack, loss rods x 1

Reagents

1.      Sodium sulfite

2.      Sodium hydrosulphite (solid)

3.      Riboflavin standard ( 5μg/mL)

Dissolve 10 mg standard riboflavin in 100 mL acetic acid solution (1.71 mol/L) in a 2 1 flask. Warm, cool and make up to volume. Store the solution in amber bottles in a refrigerator.

Procedure

1.      Weigh accurately the riboflavin tablet and ground it thoroughly in a mortar.

2.      Dissolve the powder in 75 mL ddH2O and mix, transfer to a 100mL volumetric flask and make up the volume with water. While transferring, wash the the conical flask completely to transfer the material without any loss

3.      Filter through medium- fast paper

4.      Take 2 mL solution, add 5 mL 1.70mol/L HAc and make up the volume with water in a 100 mL volumetric flask

5.      Standard curve and riboflavin concentration measurement

Take 10 test tubes, add solutions as follows:

Tube Procedure	Riboflavin Standard						Sample			Blank
	1	2	3	4	5	6	I	II	III	0
Riboflavin Standard solution (mL)	0.25	0.5	1.0	1.5	2.0	2.5
Samples (mL)							10	10	10	10
ddH2O (mL)	9.75	9.5	9.0	8.5	8.0	7.5
Fluorimetric determination	Exciting λ= 440nm, emitted λ= 520nm, number 0 tube as blank
F
Reduction reaction	Add 10mg of Sodium hydrosulphite or sodium sulfite, mix and measure F0 immedietly
F0
Riboflavin Content (μg)

Record the fluorescence intensity of the sample solution and determine the concentration of riboflavin in μg/mL.

Calculation

1.      Draw the standard curve on the coordinate paper. F values as Y-axis, riboflavin content as X-axis from tube 1-6

2.      Calculate the mean F value (F) of sample

3.      Calculate riboflavin content of the testes tubes according to the standard curve

4.      Calculate the riboflavin content of the tablet.

Aku sempat bertanya apakah hanya B2 saja yang bisa dianalisis dengan fluorescence spectroscopy? bagaimana dengan vitamin-vitamin B yang lain? jawabannya untuk B2 mengandung fosfor jadi bisa diliat dengan fluorescent spectroscopy, sedangkan untuk vitamin B yang lain perlu pengamatan lebih lanjut.