TP analysis procedure

a. Digestion Process
 Weigh 0.3 gram of dry sample in digestion tube writing each weight in
spreadsheet (for tube number 2 – 10).
 Dispense 4 ml HNO3 and then 2ml of H2O2 into digestion tube. The blank is used
for the tube number 1.
 Put digestion tubes into micro digester and start program. The program used is
plant mpr.
 Once digest process is completed, transfer the digested-solution and blank
solution into the 50 ml volumetric flask.
 Using DI water, wash digest container with approximately 20 ml of DI water and
transfer washed-solution into the 50 ml volumetric flask.
 Top up volumetric flask to 50 ml mark (From this procedure we have blank
solution (A1) and digested-solution (A2).
 Shake each solution by inverting the volumetric flask.
 Filter the solution If the solution is not clear.

b. Colorimeter analysis
Standards and reagents
 Sulphuric acid 2 M (B1): 111 ml concentrated sulphuric acid (18 M) is pipetted into 1
L volumetric flask and slowly added DI water to reach1 L solution.
 Potassium antimony tartrate ( K2Sb2(C4H2O6)2 ) solution (B2): 1.37 g potassium
antimony tartrate heptahydrate is added approximately 300 ml of DI water in 500 ml
beaker and stirred until all has dissolved. The solution then is transferred into 500 ml
volumetric flask and DI water is added to the solution to the mark. This solution then
is stored in the fridge.
 Ammonium molybdate((NH4)2MoO4) solution (4%) (B3): 40 grams of ammonium
molybdite is dissolved with DI water using stirrer in the 1 L beaker glass until all
solution is dissolved. The solution then is transferred into 1 L volumetric flask and
diluted with DI water to the mark.
 Ascorbic acid (C8H8O6) 0.1 M (B4): 15 grams of ascorbic acid is dissolved with DI
water in 250 ml volumetric flask.
 Combined reagent solution (B5): 50 ml sulphuric acid 2 M (B1), 5 ml potassium
antimony tartrate solution (B2), 15 ml of ammonium molybdate solution (B3), 30 ml
ascorbic acid (C8H8O6) 0.1 M (B4) are transferred into beaker glass and mixed. The
solution was added in that order.
  Sodium hydroxide 5M (B6): 20 g NaOH was dissolved with DI water in 100 ml of
volumetric flask. (This solution is used to adjust the pH. The pH in this measurement
shuldbe around 7).
 Standard solution (1,000 mg/L) (B7): 4.394 g KH2PO4 was added into 1,000 ml
volumetric flask and diluted with DI water until the mark. To make 10 mg/L (B8),
pipette 1 ml standard 1000 mg/L and transfer to volumetric flask 100 ml and slowly
add the water until mark.
 The standard solutions then are made up according to expected range of
concentration (0.1 – 2 mg/L) using 10 ml of standard solutions (B8), based on the
table below (This solution is made up in 50 ml volumetric flask).
Range of standard Volume of 10 mg/L-P Di water added (ml)
(mg/L) required (ml)
0.1 0.5 49.5
0.2 1.0 49
0.4 2.0 48
0.8 4.0 46
1.0 5.0 45
1.5 7.5 37.5
2.0 10 35

c. Procedures analysis
 Pipette 5 ml of each standard solution and transfer into 25 ml volumetric flask.
 And then pipette 5 ml of blank solution (A1) into 25 ml volumetric flask.
 1.6 ml of NaOH 5M (B6) is added to each 25 volumetric flask.
 The colour (blue) is developed by adding 5 ml of combined reagent solution (B5)
into each 25 ml volumetric flask.
 DI water is added to the mark
 The standards are analysed using a spectrophotometer set at a wavelength of
880 nm.
 The samples are analysed by adding 5 ml of each digested solution sample (A2).
 1.6 ml of NaOH 5M ) is added to each 25 volumetric flask.
 5 ml combined reagent solution (B5) is added into 25 ml volumetric flask to
develop the cthe blue colour.
 These solutions then are added with DI water to mark.
 These samples were measured using a spectrophotometer set up at wavelength
880 nm.