Arias, Julia Lourdette; Borja, Patrice Camille; Derecho, Mae Ann
ABSTRACT The significance of monitoring physicochemical parameters in laboratory wastewater had
been a constant concern for the university over the years. The sets of experiments were carried out to analyze ammonia, phosphate and nitrite content using colorimetric methods. The results showed that the concentrations for each are relatively low.
Water is an extremely essential using spectrophotometric method. It is expected
substance for all living organisms. The quality that the data from such analyses will provide an of water is a vital concern for mankind as it is insight to future actions which might be taken by related to human and environmental welfare. the university. Wastewater, especially, is subject to numerous pathogenic microorganisms and high content of METHODOLOGY organic matter as well as inorganic pollutants such as heavy metals. Disposal then of untreated Ammonia analysis by phenate colorimetric or failed treatments to waste into the method. Wastewater sample from the Physical environment will endanger all life forms at the Chemistry Laboratory was collected using long run. Thus, it is of importance to monitor the 500-mL polyethylene bottle and refrigerated physicochemical parameters in order to produce prior to analysis. Analysis of ammonia in the the correct assessment of its potential to affect sample was performed using the organisms and also to provide further treatments spectrophotometric phenate method . Ten needed before discharge to the environment. milliliters each of the sample was pipetted out to Among such parameters analyzed are six different test tubes for replication. A drop nitrite, phosphorus content and ammonia. (0.05 mL) of manganese sulfate solution, Nitrogen and phosphorus are both components MnSO4, was added to each test tube containing critical nutrient in the metabolism of all forms of the sample. The test tubes were then shaken with life. However, it is known that increase in such the addition of 0.50 mL hypochlorous reagent parameters also induce harm to the aquatic and continuously done after the immediate environment. In the case of nitrites and addition of 0.60 mL phenate reagent. Vigorous phosphorus, over-nutrient leads to high algal shaking was observed at this period of the growth and thus causes deprivation of oxygen to method. After 10 minutes, the absorbance of the other life forms. Hence, a competition forms six samples were read using a UV-Vis Double with other living organisms which leads to Beam Spectrophotometer at 630 nm.The color eventual death of the flora and fauna around and formation is stable for at least 24 hours. on the area (eutrophication). Ammonia on the A stock solution was prepared using other hand, becomes toxic once it exceeds the pre-dried NH4Cl diluted to 250-mL. A working natural concentrations. standard was taken from this stock solution and With the number and varied experiments was then used to prepare a series of 5 standard conducted in laboratories and non-observance of solutions of 50-mL each which covers the proper waste management at times practiced by expected concentration range of the sample. The students, the quality of wastewater had been a standard solutions along with the solutions for constant concern of University of San Carlos. In recovery were treated as that of the samples. this paper, a set of experiments which utilizes the indicators of water quality i.e pH, NO2-, Phosphate analysis by direct ascorbic acid NH3, and P levels, of wastewater from the colorimetric method. Into a 125-ml Erlenmeyer Physical Chemistry laboratory were determined flask, 50.0 ml of the water sample was pipetted out and added with a drop of phenolphthalein Table 1. The pH of the sample was taken indicator. If a red coloration develops 5N of beforehand and was determined to be 8.13. sulfuric acid was added dropwise to discharge Calibration curves and calculations are given as the color. Then, 8.00 ml of the combined reagent supplementary materials. was added and mixed thoroughly. Absorbance Analysis of ammonia for the wastewater was measured in a span of 30mins, after samples was done using the phenate colorimetric allowing the solution to react for 10 minutes. method. In this method, ammonia (NH3) reacts Recovery test and quality control of the sample with a hypochlorite to produce monochloramine. were also measured at the end of sample It is further reacted with phenate and sodium analysis. hypochlorite in the which gives the intensely blue compound, indophenol (Park et al., 2009). Nitrite analysis by NED colorimetric method. Using the equation calculated from the Wastewater sample was collected on January 20, calibration graph, the sample concentration was 2017 at 10:30 in the morning from Phychem lab. found to have an average concentration of Prior to this, stock nitrite (NO2-) solution was 3.62E-2 ± 8.47E-4 ppm NH3. The percent prepared and standardized using standardized recovery was determined to be 85%. The potassium permanganate (KMnO4) solution and wastewater was from a laboratory which has not sodium oxalate (Na2C2O4). A color reagent was utilized ammonia water recently and thus there also prepared beforehand and stored in a dark is no expected increase in its levels. bottle inside the refrigerator. Through the entire There are three possible ways to procedure, a determine the phosphate content of a water water blank was carried and necessary sample, the one identified and performed in this corrections were made. experiment is the direct ascorbic acid The pH of the water sample was colorimetric method for phosphates. It follows checked immediately. It was ensured to be in the the principle of reacting the ammonium range of pH 5-9. Fifty mL of the clear sample molybdate and potassium antimonyl tartrate was quantitatively transferred to a 100-mL solution in the acidic medium of the diluted beaker and 2-mL of the color reagent was then sample solutions of orthophosphate to form the added. The absorbance was read after 10 intensely colored antimonyl phospho- molybdate minutes but before 2 hours at 543 nm using complex. The observed coloration is Milton Roy Spectronic 20. proportional to the phosphate concentration. From the stock NaNO2 solution, 1.018 x The concentration of phosphate present 10-3 mg/mL solution was prepared. From this in the water samples were calculated to be equal new stock solution, a series of standards were to 0.01744 mg P L-1 and 0.01730 mg P L−1 at the prepared into 50-mL volumetric flasks and average absorbance of 0.126 and 0.125, diluted to the mark with deionized water. respectively (refer to Table 1); with a percent Two-mL of color reagent were added to each recovery equal to 90%. From the data calculated, and allowed to equilibrate for more than 10 the method used to determine the amount of minutes before reading. phosphate that is present in the wastewater is Recovery was done to the samples by suitable for the kind of sample analyzed. This is adding a known amount of standard and treating in accordance to the Standard Methods for the the solution with color reagent. Examination of Water and Wastewater that referred ascorbic acid as a method of RESULTS AND DISCUSSION determination used for samples containing phosphates at a range of 0.01 to 6 mg P/L. Results of wastewater analysis for The average concentration of the sample ammonia, phosphate and nitrite using is 6.329 x 10-5 mg N/L and the computed percent spectrophotometric method is summarized in recovery was determined to be 118.75%. Relative to others, nitrite concentration is extremely low as can be shown in figure 1. This is suggested to be due to lack of experiments performed on the sampling day. The low levels of these chemical indicators are to be expected with the kind of wastewater sampled. Most chemicals, if deemed too high, pollutes the environment. These would commonly come from municipal or domestic discharges. Figure 1. Wastewater analysis graph showing ammonia (violet), phosphate (brown) and nitrite (green) concentrations relative to each other. Table 1. Summary of results from spectrophotometric analysis of wastewater
Ammonia Phosphate Nitrite
(mg NH3/ L) ( mg P/ L) (mg NO2—N/ L)
Trial 1 0.03712 0.01744 6.269 E-5
Trial 2 0.03566 0.0173 6.39E-05
Trial 3 0.03520 0.01744 6.51E-05
Trial 4 0.03566 0.01744 6.27E-05
Trial 5 0.03728 0.01744 6.27E-05
Trial 6 0.06504 0.01744 6.27E-05
%Recovery 85% 90% 118.75%
Average 3.6163 E-2 1.7417 E-2 6.3412 E-5
standard deviation 8.4681 E-4 5.7155 E-5 1.0772 E-6
CONCLUSION
The analysis of inorganic components in the is still highly suggested that environmental wastewater sample from Physical Chemistry safety should be kept through proper waste laboratory did not show any alarming results management especially involving chemicals. which may need immediate action. However, it SUPPLEMENTARY MATERIALS
I. Ammonia
a. Methodology
Reagents
● Ammonia-free water ● Hypochlorous acid reagent- add 10 mL 5% NaOCl solution (commercial bleach) to 40 mL ammonia-free water. Adjust pH to 6.5-7.0 ● Manganese sulfate solution- dissolve 25 mg MnSO4.5 H2O in 50 mL ammonia-free water ● Phenate reagent- dissolve 1.25 g NaOH and 5g phenol in 50 mL ammonia-free water. Prepare weekly because this darkens on standing ● Stock ammonium solution - dissolve 381.9 mg anhydrous NH4Cl dried at 100oC in water and dilute to 1-L volumetric flask ● Standard ammonium solution- Dilute 1.00 mL of stock ammonium solution to 250-mL volumetric flask with ammonia-free water.
In a 500ml volumetric flask, 1.3715g of potassium antimonyl tartrate solid was dissolved in 400 ml distilled water and diluted to the mark. In another 500 ml glass stoppered volumetric flask, 20 g of ammonium molybdate solids were dissolved in distilled water, mixed thoroughly and diluted to the mark. For the ascorbic acid solution (0.01 M), 1.76g of ascorbic acid solid was dissolved in 100 ml and stored in a glass stoppered bottle. The following proportions of 50ml 5N sulfuric acid, 5ml potassium antimonyl tartrate solution, 15ml ammonium molybdate solution and 30 ml ascorbic acid were combined and mixed thoroughly forming 100 ml of the combined reagent used for the analysis of phosphate in water. The stability of the reagent is only for 4hrs hence the solutions were combined prior to the analysis. Standard phosphate solution was prepared through dissolving 219.5 mg of anhydrous KH2PO4 and diluted to 1000ml [1.00ml = 50.00 µg PO42−—P]. b. Calibration curve
Figure 3. Calibration curve for phosphate analysis
c. Wastewater sample data
Absorbance Concentration (ppm)
0.126 0.01744
0.125 0.01730
0.126 0.01744
0.126 0.01744
0.126 0.01744
0.126 0.01744
III. Nitrite
a. Methodology Potassium permanganate (0.05 N) was standardized using anhydrous sodium oxalate. About 100-120 mg of Na2C2O4 was dissolved in 100 mL water, added with 10 mL of 1+1 H2SO4 and heated to 90-95 deg. C. This solution was titrated with KMnO4 to a slightly pink endpoint that persisted for at least a minute. 0.025 M Na2C2O4 was standardized using volume ratio with KMnO4.
Sodium nitrite stock solution was prepared by dissolving 0.6196 g commercial reagent grade NaNO2 in deionized water in a 500 mL volumetric flask. For the standardization, the following procedures were followed: Pipet in order 50.00 mL standardized 0.05 N KMnO4 solution, 5.00 mL concentrated sulfuric acid and 50.00 mL stock nitrite solution. The solution was warmed gently on a hot plate while titrated with .025M Na2C2O4 to colorless. The excess oxalate was titrated with 0.05 M KMnO4 to the faint pink endpoint. A water blank was carried through the procedure.
The concentration of NO2—N was calculated using the equation below:
(BxC)−(DxE)x 7 A= F
where:
A= [NO2—N] in stock nitrite solution, mg/mL
B= total volume of standard KMnO4 solution used, mL C= KMnO4 standard solution used, N D= total volume standard Na2C2O4 solution used, mL E= Na2C2O4 standard solution used, N F= volume of stock NaNO2 solution taken for titration Sample calculation for NaNO2 standardization:
((50.50 mL)(0.05127 N KMnO4)− (15.00 mL)(0.05126 N Na2C2O4))x 7
