Water Quality Assessment of Pansipit River After The Taal Volcano Eruption

Republic of the Philippines
BATANGAS STATE UNIVERSITY

Pablo Borbon Campus I, Batangas City
COLLEGE OF ARTS AND SCIENCES
CASE STUDY:
WATER QUALITY ASSESSMENT OF PANSIPIT RIVER AFTER THE TAAL

VOLCANO ERUPTION
For the
Midterm Requirement in Advance Analytical Chemistry
CHEM 343
Submitted by:
Bauan, Marian
Carandang, Abegail
De Castro, Jia Ruth
Magsino, Ian Deniell
Tong, Jayrex
2020
INTRODUCTION
Taal Lake is a freshwater lake in the province of Batangas, on the island of Luzon in the
Philippines. The lake fills Taal Caldera, a large volcanic caldera formed by massive eruptions
between 100,000 and 500,000 years ago. It has had some of the country’s largest and deadliest
volcanic eruptions: At least six eruptions during the recorded history of Taal since 1572 claimed
fatalities, mostly from strong pyroclastic flows, as well as tsunamis produced in the crater lake.
Furthermore, Taal Lake lies within a national park and a major tourist attraction, which is the
most commonly viewed from Tagaytay Ridge in the north. The Pansipit River drains the lake
into Balayan Bay of the South China Sea through a breach in the cliffs to the southwest.
Last January 12, 2020, Taal Volcano erupts again after a couple of years. This event
affected thousands of families and caused millions-worth of damage just days after the initial
eruption. It is also having a significant impact on agricultural land, livestock wherein it killed
many animals, air quality, and water quality. Thus, residents fear after a vital river system that
runs in the towns suddenly dries. According to Dr. Winchell Sevilla of the Volcano Monitoring
Department of the Philippine Institute of Volcanology and Seismology (PHIVOLCS), explained
that what happened to the Pansipit River is indeed Taal Volcano eruption-related. He said given
magma activity inside the volcano, which the tendency is for the ground to swell because of
magma accumulation. The areas where ground swelling has monitored are Lemery, Taal, and
areas close to Taal Volcano. As a consequence, the natural flow of water at a portion of the
Pansipit River at Barangay Tatlong Maria in Taal was disturbed. Water could no longer flow
because the ground suddenly lifted.
Water quality is defined as the chemical, physical and biological characteristics of water,
usually in respect to its suitability for the designated use. Physico-chemical indicators are the
traditional water quality indicators that most people are familiar with. They include dissolved
oxygen, pH, temperature, salinity, and nutrients such as nitrates and phosphates. It provides
information on its impact on the system. Hence, an assessment of water quality is necessary to be
conducted in the Pansipit River, since eventually, the water came back after days of the eruption.
It is important to check the efficiency of a system to decide what changes should take place.
According to the study conducted by Hopson, R. F. (1991), bodies of water near the volcano may
provide information about the state of the activeness of the volcano and its possibility of a near
eruption. Near eruption may be concluded based on the water quality of the bodies of water
around its location, especially the bodies of water. It states that an increase in water temperature
(hot springs), noticeable variation in the chemical content of water bodies, drying up of springs,
or wells may dictate the activeness of a volcano and a near volcanic eruption.
The water quality in every body of water is crucial in maintaining the equilibrium in all
aspects of biotic and abiotic interactions to attain ecological balance, and it is therefore important
to monitor and control the quality of water. Perturbation in the conditions of these bodies of
water is mainly caused by either human-caused or natural disturbances.
This study aims to devise a sampling plan, sample collection technique, and analysis of
water quality in the Pansipit River and to identify the differences of the water quality before and
after a volcanic eruption.
OBJECTIVES
1. To devise a sampling plan, sample collection, and analysis of samples in a water quality
analysis in Pansipit River.
2. To determine various factors in water quality such as pH, DO (Dissolved Oxygen),
temperature, salinity, nitrate, phosphate, turbidity, and BOD (Biological Oxygen Demand)
before and after a volcanic eruption.
3. To identify the differences of the water quality before and after a volcanic eruption in
terms of:
a. pH,
b. DO (Dissolved Oxygen),
c. temperature,
d. salinity,
e. nitrates,
f. phosphate,
g. turbidity
h. and BOD (Biological Oxygen Demand).
SIGNIFICANCE OF THE STUDY

The study that is entitled Water Quality Assessment of Pansipit River after the Taal
Volcano Eruption would be beneficial for the following:
To the community, the study gives a broad array of information on the quality of the
water they are using by knowing the content and properties of water. It also serves as a basis if
the water quality is at its safe level after a volcanic eruption.
To the environmental sectors, the study helps in the assessment of materials present in
the Pansipit river for monitoring the health of the environment.
To the Local Government Unit (LGU), the study helps in giving information in the
environmental monitoring and serves as an early warning device for volcanic activity and
possible health effects of the water itself.
To the Disaster Risk Reduction Management Council, the study will serve as a
warning device for a near volcanic activity based on water quality.
To the Department of Environmental and Natural Resources (DENR), this study
gives information on how the water quality analysis is executed and what are the qualities of
water and its properties after a volcanic eruption.
To the present researchers, this study aids in the identification of the sampling plan,
sample collection, and sample analysis techniques in water quality analysis and the differences in
the properties of water quality before and after a volcanic eruption.
To the future researchers, the study will serve as a reference for future and in-depth
studies regarding devising a sampling plan, sample collection, and sample analysis techniques
and the difference in the water quality before and after a volcanic eruption.
SAMPLING PLAN
The Sampling Plan outlines the sites, equipment, methodologies, and protocols for data
management that researchers will use to collect water quality data from the Pansipit River after
the Taal Volcano eruption. It also indicates the procedures to be followed, the timeframe for
sampling and the field and laboratory parameters to be measured.
Sampling Site
The sampling site of this study is the Pansipit River located at Luzon Island, Philippines,
in the province of Batangas (Figure 1). The Pansipit River is an 8.2 km channel, and it is
connected to the south basin of Taal Lake and drains water to Balayan Bay. The river in most
areas has an average depth of 4 m and an estimated channel width of 10 to 15 m. For coastal
communities, it is considered an essential freshwater resource. The river is also being considered
a tourist destination. Due to its importance to Taal Lake, the local government and residents
protect it by ensuring that the river is free of obstruction.
Figure 1. Overview of Pansipit River located in Batangas Province of Luzon Island, Philippines
using GIS mapping.
As of January 2020, Taal Volcano suddenly erupted after being quiet for more than four
decades. According to the Philippine Institute of Volcanology and Seismology, Taal Volcano
blasts steam, ash, and pebbles up to nine miles into the atmosphere. The lava spurting from the
volcano dropped into the crater’s surrounding lake, which an event could contaminate water
resources.
The Pansipit River, which is the main focus of this study, flows into Taal Lake. The
researchers want to determine and identify the various factors in the water quality of Pansipit
River, additionally, to test if there is a significant difference between the quality of water of the
Pansipit River before and after the volcanic eruption.
Sampling Technique
The study will use Systematic Sampling in which it represents a group of sampling
techniques that helps researchers to select representative samples from the target sampling site
that they are interested in studying. Systematic sampling measurements are taken at locations
according to a predetermined pattern. The target sampling site for this study is the Pansipit River.
The river will be divided into three layers: upstream (near the Taal Lake), middle stream, and
downstream (near the Balayan Bay) to collect samples from the study site.
Middle Upstream
stream
Downstrea
m
Figure 2. Division of the Pansipit River in various sites: Upstream, Midstream, Downstream.
Systematic sampling and its application to water quality assessment are illustrated with
square grids of sampling points. The samples will be taken at each point of the grid by collecting
samples from the center of grid lines. Sampling in the grid blocks chosen in a genuinely random
way constitutes random sampling.
Figure 3. Systematic sampling plan for collecting samples from the Pansipit River. Each solid
dot represents a sample will be collected within the sampling grid.
General Equipment
Water sampling equipment is used to collect water quality data from the Pansipit River.
This is often done for site compliance and remedial performance monitoring. Equipment to be
used for sample collection should be properly cleaned and decontaminated, field meters should
be calibrated as directed by the manufacturer, and appropriate sample bottles should be prepared.
Chemical elements of interest in subsequent laboratory analysis dictate the type of container to
be used for sampling, sample treatment, and storage of samples. Equipment necessary for the
collection of samples are as follows:
• Field sheets, sample labels, and chain of custody forms
• Sample collection equipment:
- If necessary, equipment for collection of samples from a safe distance

(e.g. sample pole.)
- Container for field measurements
- Container for collecting sample
- Sample bottles for dispatch to laboratory

• Field parameter meters or test kits (e.g. alkalinity/acidity test kit)
• Powder-less nitrile gloves
• Esky or portable refrigerator
• Personal Protective Equipment, first aid and communication

equipment
Field Measured Parameters and Laboratory Analyzed Parameters

The researchers aim to determine and identify the various factors in the water quality of
the Pansipit River after the Taal Volcano eruption and compare the results obtained from the
study to the water quality of the river before the volcanic eruption. Below are the following
physical and chemical parameters that will be determined throughout the study.
Field Measured Parameters:
• Dissolved Oxygen (DO) - The dissolved oxygen analysis tests the amount of gaseous
oxygen (O2) dissolved in an aqueous solution. Oxygen dissolved in water by absorption from the
ambient air, by aeration (rapid movement), and by photosynthesis. DO is also a field test that
should be conducted on-site. The amount of DO in water depends primarily on the temperature
of the water; the colder water will hold more DO than the warmer water. When in harmony with
the environment, it is said that the water is saturated at this maximum concentration or that the
DO is saturated at 100%.
• pH - A solution’s pH is the concentration, expressed as a negative logarithm, of hydrogen
ions. It reflects the acidity or alkalinity of a solution in the water. Water with a pH of 7 is neutral;
lower pH levels indicate increasing acidity, whereas pH levels above 7 indicate increasingly
alkaline solutions.
• Temperature - A thermometer with a range of 0 to 50°C or an appropriate electronic
thermometer may be used to determine the temperature. The probe (or thermometer) is
positioned for the water to be measured. After the reading has stabilized, the temperature is
measured: this may take several minutes. Since the solubility of DO decreases with rising water
temperatures, the supply of DO for aquatic life is restricted by high water temperatures.
• Salinity- It is the total concentration of the dissolved salts or the total ionic concentration
of dissolved minerals in sample water. It is also called Total Dissolved Solids (TDS). It may
include various cations such as Sodium (Na+ ), Calcium (Ca2+), Potassium (K+), Magnesium
(Mg2+), and anions like Chloride (Cl-), sulfate (SO42-), Carbonate (CO32-), Bicarbonate (HCO3-),
etc. The high salinity of rivers and streams may degrade town water supplies and affect irrigated
agriculture and horticulture. It may also adversely impact riverine ecosystems by lowering crop
yields and degrading stock water supplies.
Laboratory Analyzed Parameters:
• Nitrates - One form of dissolved nitrogen that naturally occurs in water is nitrate. Nitrates
are highly soluble, meaning that they are easily dissolved in water. Nitrates are colorless and
odorless therefore their presence cannot be determined without the use of special testing
equipment. Most natural concentrations of nitrates are not a human health issue, but this may
pose a problem when excess nitrates enter the water.
• Phosphate - In natural waters and wastewaters, phosphorus exists almost entirely as
phosphates. These are known as organically bound phosphates, orthophosphates (PO43-),
condensed phosphates (pyro-, meta-, and other polyphosphates). High levels of phosphorus
and/or other main nutrients can lead to related problems such as nuisance or toxic algal blooms,
although some waterways are naturally eutrophic (nutrient enriched). They occur in solutions, in
particles or detritus, or aquatic species.
• Turbidity – It is the measure of the presence of suspended matters such as soil, biological
solids, decaying organic matter, and particles discharged in wastewater. Tephra, a fragmental
material produced by a volcanic eruption regardless of composition, fragment size, or
emplacement mechanism, is also considered as a suspended matter. A high presence of
suspended matters in water samples, the murkier it is, and the higher in turbidity, v.v.
• Biological Oxygen Demand (BOD) - It is a measure of the amount of organic material
present in the water that is biologically and/or chemically degradable. It shows the amount of
oxygen that could potentially be consumed by aerobic aquatic organisms in the process of
metabolizing all the organic matter available to them. Low levels of dissolved oxygen in affected
waters are the effect of high BOD, resulting in stress on aquatic life and, in extreme cases,
suffocation and death.
Assessment of Water Quality
Water sampling will be implemented to determine the water quality of the Pansipit River
after the Taal Volcano eruption. The sampling will be conducted on November 8, 2020. Samples
will be collected from the target area which is divided into three layers. The first layer is
upstream, near the location of Taal Lake, the second layer is the midstream, and the third layer is
downstream near the Balayan Bay. Each member of the research team, along with the field
personnel, will be designated for specific sampling points. Sampling will be performed at five
sampling points/sites in each layer of Pansipit River, where the samples are selected randomly
using a grid and collected through grab sampling. The study will have randomly selected five (5)
samples with an overall of fifteen (15) representative samples will be collected from the Pansipit
River. The time required for the collection of water samples is around 6:00 AM to 8:00 AM.
Selected physicochemical characteristics of the Pansipit River after the Taal Volcano
eruption will be determined using standard procedures. The physical parameters include
temperature and turbidity, while the chemical parameters include pH, salinity, nitrates,
phosphates, dissolved oxygen (DO) and biological oxygen demand (BOD) will be tested
throughout the study. Measurements and analyses of some parameters of water analysis such as
pH, salinity, DO, and temperature will be conducted on-site/in situ at the surface with 10cm
depth. Hence, the remaining parameters, which are the turbidity, nitrates, phosphates, and BOD
analyses, will be done in the laboratory having various amounts of water samples obtained from
the river. The collected samples are then put in an Esky or ice cooler and will be transported
immediately to the laboratory for analysis. Quantitative data on water quality parameters will be
compared using Single Factor ANOVA at a 95% level of confidence to determine a significant
difference between the water quality of the Pansipit River before and after the Taal Volcano
eruption in various parameters.
Types of Sample
Each water sample may vary within each test or analysis. A sample may be a grab sample
or composite sample. Grab samples are single samples collected at a specific spot at a site over a
short period while composite samples are the combinations of grab samples which should
provide more representative sampling of heterogeneous matrices in which the concentration of
analytes of interest may vary over short periods and space. In this study, grab samples will be
collected as the representative samples of parameters which are analyzed in the laboratory and in
situ sampling for parameters analyzed in the field.
Minimum Amount of Sample

A minimum sample size of 100 mL is needed for each analysis of turbidity, nitrates, and
phosphate, and 1000mL for BOD, respectively. It is advisable to triple the amount of sample
being collected to ensure that the water samples are adequate for the analysis of the parameters
hence, this minimum amount of sample sizes according to standards have already an excess
amount, approximately tripled amount of samples needed for analyses. Consequently, in nitrates,
additional 50 mL of water will be gathered for the sample collection for it to have an
approximately tripled in amount. In terms of field parameters, there is no need for a minimum
sample size due to the analyses done directly in the sample source.
Number Samples
The study aims to identify the water quality of the Pansipit River after the Taal eruption
in terms of eight (8) parameters. Each parameter has five (5) replicates of water samples that are
going to be analyzed. Since the river is too large for being tested as a whole, it will be divided
into three layers: the upstream, midstream, and downstream; for it to represent the whole quality
of water of the river. Hence, the parameters are subdivided into two, in which there are four (4)
fields and also four (4) laboratory parameters where field parameters do not need water sample
collection. By these, a total of 60 water samples will be collected in the whole study.
SAMPLE COLLECTION
The researchers aim to collect appropriate portions of water samples (representative
samples) to analyze specific parameters done in the laboratory in which are nitrates, phosphates,
turbidity, and BOD (Biological/Biochemical Oxygen Demand). In terms of parameters that will
be analyzed in the field, such as pH, temperature, salinity, and Dissolved Oxygen (DO), sample
collection is not required to proceed with such analyses. Consequently, the study focuses on
determining the water quality of Pansipit River after a volcanic eruption of a nearby volcano,
Taal volcano, and upon collecting representative samples from each three (3) layers designated
in the Pansipit River will supply data that supports the goal of the study. The following detailed
information is presented in specific methods of collection of water samples based on various
analyses.
Procedures for preparation of containers

An important step in starting the sampling procedure is to ensure that the containers are
cleaned and maintained constantly. The following steps must be followed to achieve pre-cleaned
containers: (1) Rinse the containers in tap water (2) Clean with an antibacterial/microbial reagent
like De-Con90 (3) Rinse well using tap water (4) Rinse three times with de-ionized water (5)
Allow to dry. Rinsing allows eliminating all the cleaning solution residues. The equipment is
fully calibrated before starting the sampling. Once all the sampling has been finalized, the
sampling and support equipment is placed at multiple locations. Wearing latex gloves is also a
must.
a. Nitrates
For nitrate determination, samples will directly be transferred into a pre-cleaned sample
bottle (plastic, glass, or fluoropolymer) from the collection vessel. A sample bottle is pre-rinsed
three (3) times with a 20 mL water sample, before the final sample collection. After rinsing, a
minimum volume of 100 mL of the sample will be collected in the pre-cleaned sample bottle and
then stored in a cooler with a temperature that is greater than 0°C and less than or equal to 6°C
(0°C < T ≤ 6°C), above the freezing point of water. The sample fill is just below the shoulder of
the bottle and must be analyzed as soon as possible. Also, this analysis must be done as soon as
possible with maximum storage recommended of 48 hours and regulation of 48 hours, as well.
b. Phosphate
For phosphate determination, the sampling method is similar to nitrate determination. A
sample bottle (glass rinsed with HNO3) is pre-rinsed three (3) times with 20 mL sample water
before the final sample collection, and 100 mL of sample water will be transferred into it. The
maximum storage recommended for the water sample for phosphate determination is only 48
hours before the analysis and regulation of also 48 hours, and it must be stored in a cool place
with a temperature of greater than 0°C and less than or equal to 6°C (0°C < T ≤ 6°C), above the
freezing point of water.
c. Turbidity
The minimum amount of water samples undergoing a turbidity test is 100 mL and is then
directly transferred from the collection vessel into the pre-cleaned sample bottle (plastic, glass,
or fluoropolymer). The sample bottle undergoes cleaning by rinsing it with the water sample for
it to be able to minimize or remove other contaminants. The bottle must be filled to the top, and
no air bubbles are trapped. By preventing too much turbulence near the sampling area, the
presence of air bubbles will be minimized in the water. Afterward, the sample will be taken to
the laboratory within a day for analysis. In the preservation of the sample, the sample bottle is
placed in a dark up to 24 h and has a temperature of greater than 0°C and less than or equal to
6°C (0°C < T ≤ 6°C), above the freezing point of water, to minimize the other effects that may
alter analysis due to contamination. A water sample undergoing a turbidity test has only 24 hours
as maximum storage recommended and 48 hours of regulation. It is a note that the sample for
turbidity must be analyzed as soon as it was gathered for the prevention of the microbiological
decomposition of solids and may give inappropriate results.
d. BOD (Biological Oxygen Demand)
The direct transfer of 1000mL of water sample into each pre-rinsed sample bottle (glass,
plastic, or fluoropolymer) will be done since an unfiltered sample is required for this analysis.
Two separate sample containers will be needed for BOD, not incubated and incubated. It will be
filled to the top to exclude air and avoid air bubbles. The first bottle will be analyzed as soon as
possible with a maximum of 6 hours of recommended storage and 48 h of regulation after sample
collection. For sample preservation, samples are stored at cool and with a temperature of less
than or equal to 6°C ( T ≤ 6°C). While the second sample bottle (incubated) is delivered to the
laboratory and stored in the dark for 5 days at 20°C.
Chain of Custody Procedures

Before performing the sampling methods, the researchers will prepare a chain of custody
forms to maintain sample integrity from sample collection to analysis of data. The following
procedures provide a summary of the major aspects of the chain of custody for the study.
a. Sample labels: The label for each representative sample will be written in a gummed
paper using a water-resistant ink, which includes the necessary details, including the sample
name or code, the sampling date and time, and the sampling site. It is important to avoid sample
misidentification, especially for the laboratory coordinator.
b. Sample seals: Adhesive paper seals to be used will be prepared by the researchers, which
will contain the sample number, collector’s name, and date, and time of sampling. This will be
attached across the cap and body of the sample container. Before leaving the sampling site, the
person in charge of the sample custody will check the containers, whether they are properly
sealed.
c. Field logbook: The field logbook will also be prepared which contains the information
needed for the sampling and serves as references to carry out the analysis. The contents of the
field logbook are based on the objective of the study and various parameters. The directions and
overview of the site, safety protocols regarding the site, and other specific information in sample
collection and preservation are all included.
d. Chain of custody record: The record will contain information such as the sample
number, the sampling date, the position of sampling site, and the signatures of the collector and
other persons involved in handling the documents, and specific date and time of possession. The
details mentioned on the sample labels are also recorded in the chain of custody record to
provide a backup copy of the sampling procedures done.
e. Sample analysis request sheet: The collector will fill up this sample analysis request
sheet based on the information noted in the logbook. Then, it will be handed to the laboratory
personnel to complete the laboratory portion containing the name of the recipient of the sample,
laboratory number, date of sample receipt, condition of each sample, and the determinations to
be performed.
f. Sample delivery to the laboratory: Once the sampling is completed, the samples must be
transported to the laboratory as soon as possible, preferably the same day or within the maximum
time allotted for each sample to be preserved when the sample is collected. The samples will be
placed in a cooler with crushed ice packed closely, then delivered to the laboratory. A complete
chain of custody and sample analysis request sheets will be enclosed together with the samples to
be delivered.
g. Receipt and logging of sample: To whom and when the sample will be delivered as well
as to whom and when it will be received will be stated by the person handling the sample for it to
be accepted in the laboratory. The person who has custody of the sample will also confirm that
no tampering or irregularity occurred during the transportation of the sample. Once the sample is
received, the custodian will assign the laboratory number, which will be recorded in the
laboratory logbook. The sample will be stored in a refrigerator to keep its temperature between
1-4°C, and in the dark for preservation for it to avoid photosynthesis and microbiological
activity.
h. Assignment of sample for analysis: Once the sample is delivered in the laboratory, it will
be handed to the laboratory supervisor, hence they will be responsible for its care and custody.
The laboratory supervisor will also assign which bottle will be analyzed.
i. Disposal: The disposal of the sample will be done with compliance with the local, state,
and approved methods. The water sample will be reviewed and analyzed whether it is found to
be non-hazardous before it could be disposed of. The disposition of the sample will also be
documented.
Sampling Method
Direct collection of surface water samples in the three (3) layers, upstream, midstream,
and downstream, of Pansipit River will be done by the researchers by using manual sampling,
which is the collection of samples without using any equipment.
Sample Containers
Sample bottles or the sampling containers vary according to each parameter analysis.
Mostly all of the parameters that will be analyzed in the laboratory, which needed sample
collection, such as turbidity, nitrate, and BOD, use containers like plastic (polyethylene or
equivalent), glass, and fluoropolymer (polytetrafluoroethylene (PTFE, Teflon). Hence, in
phosphate, G(A) sample container (glass rinsed with HNO3) is the type of container that will be
used in sample collection.
Sample Volumes
The proposed volumes of samples vary according to each parameter analysis, and it must
be assured that the sample will comply with the amount needed for sample handling, storage,
preservation, preparation, and analyses. The minimum sample size for nitrates, phosphates, and
turbidity determination is 100 mL of water sample according to standards. (Baird, R., Eaton, A.
& Rice, E., 2017) Consequently, 1000mL of water samples will be gathered for BOD. The total
sample size or volumes of the water sample at each layer of the stream (upstream, middle stream,
and downstream) of the Pansipit River, having five (5) replicates for the various parameter are
the following: 1.50L for turbidity; 1.50L for nitrate; 1.50L for phosphate; and 15.0L for BOD
having a total of 19.5 L. Increased amount of sample sizes in each parameter may be done for
possible problems like repeating analysis hence since the standard minimum sample sizes have
already excess amounts, approximately tripled, of representative samples tripling the amount of
water sample will not be done in the study. Consequently, in nitrates, additional 50 mL of water
will be gathered for the sample collection for it to have an approximately tripled in amount
totaling of 20.25L.
Table 1. Summary of the Sample Collections

Total
Sample Maximum
Type of
Parameter Container Collected Preservation Storage Regulatory
Sample
in each Recommended
site(mL)
Analyze as
soon as
Nitrates P, G, FP 150 g, c 48 h 48 h
possible,
Cool, ≤ 6°C
Phosphates G(A) 100 g Cool, ≤ 6oC 48 h 48 h
Analyze in
the same
Turbidity P, G, FP 100 g,c day ; store in 24 h 48 h
the dark
Cool, ≤ 6oC
BOD P, G, FP 1000 g,c Cool, ≤ 6oC 6h 48 h
TOTAL=
1.35L
1.35L (5 replicates) ( 3 layers) = 20.25 L (overall amount of sample size collected in the study)
Legend:
Container: P = plastic, G = Glass, FP = fluoropolymer [polytetrafluoroethylene (PTFE, Teflon)],
G(A) = rinsed with HNO3
Type of Sample: g = grabs sample, c = composite sample
Preservation: Cool, ≤ 6oC = stored at 0°C < T ≤ 6oC (above freezing point of water)
Table 1 shows the summary of the sample collection wherein the total amounts of the
sample size per parameters in each site, how it will be gathered and preserved by using various
sample containers, the storage time it may carry and the overall amount of sample size collected
in the whole study were included.
ANALYSIS OF SAMPLE
This section provides the sample preparation and statistical treatment of both parameters
and standard method of analysis on laboratory analyzed parameters. All standard procedures that
will be utilized in this proposal are obtained from Standard Methods for the Examination of
Water and Wastewater (23rd Edition). This also includes the standard equipment that will be used
in field parameters. In addition, the comparison and contrast between the water quality in the
Pansipit River before and after the Taal volcanic eruption will be discussed.
Field Parameters
The field parameters pH, Dissolved Oxygen (DO), temperature, and salinity will be used
in situ field measurement, specifically the equipment and calibrated Hanna HI9829
multiparameter portable meter will be used combined with HI 76x9829 series probes. This
microprocessor-based multi-sensor probe allows for the measurement of these key parameters.
The procedures that will be made must adhere to the guidelines provided in the manual along
with the equipment.
Laboratory Analyzed parameters

I. Determination of Nitrates
The practical range of determination for persulfate digestion method and the subsequent
cadmium reduction method is 0.05 to 1.00 mg/L as N and produces low toxicity waste and is less
cumbersome than classical TKN methods. In addition, this method has been proven to be
sensitive and reliable (Ameel et al., 1993). Since nitrates are the focal point of this determination,
the process of digestion can be omitted and can directly proceed to the reduction method. In this
method, nitrates are reduced to nitrites in the presence of cadmium. After reduction, the nitrite
produced will be then determined by diazotizing with sulfanilamide and coupled with NED to
form highly colored azo dye that will be measured colorimetrically.
Preparation of Reagents
1. Copper-cadmium granules
Wash 25 g of new Cd granules with 6M HCl and rinse with water. It will be then
swirled in 100 mL 2% copper sulfate solution for 5 minutes until blue color partially
fades. Decant and repeat with fresh copper sulfate until brown colloidal precipitate begins
to develop. The precipitated copper can then be eliminated through washing of

ammonium chloride-EDTA solution. Store activated cadmium covered with dilute
ammonium chloride-EDTA solution.
2. Sulfanilamide Color Reagent
100 mL of concentrated phosphoric acid will be slowly added to approximately
800 mL deionized water. Thereafter 10.0 g of sulfanilamide will be added and is stirred
until dissolved completely. 1.0 g of N-(1-naphthyl)ethylenediamine dihydrochloride is
added and stirred until complete dissolution. The solution must be diluted up to the 1-L
mark. If the reagent will be prepared days prior to the analysis, it must be stored at 4 oC
in dark. Solutions with dark precipitate must be discarded.
3. Copper Sulfate Solution (2%)
Dissolve 20 g of copper sulfate pentahydrate in 500 mL deionized water and
dilute to 1-mL mark.
4. Ammonium chloride-EDTA Solution
Dissolve 13.0 g of ammonium chloride in 900 mL deionized water. pH of 8.5 is
achieved by addition of concentrated sodium hydroxide solution before bringing to 1-L
volume. This solution is stable for 1 year when refrigerated.
5. Dilute Ammonium chloride-EDTA Solution
Dilute 300 ml of ammonium chloride-EDTA solution to 500mL water. This
reagent will be for cadmium-reduction-column storage solution.
6. Hydrochloric acid
Carefully add 500 mL concentrated hydrochloric acid to 400 mL reagent water
then dilute to 1 L with reagent water.
7. Intermediate Glutamic Acid Solution
Dilute 100 mL stock glutamic acid solution to 1000 mL deionized water. The
solution is then preserved by addition of 2 mL of chloroform.
8. Stock Nitrate Solution A
Dry potassium nitrate in an oven at 103-105oC for 24 hours. 0.7218 g ± 0.0005 g
in deionized water and dilute to 1000 mL mark. Solution is also preserved by addition of
2 mL chloroform.
9. Stock nitrate solution B

Obtain a commercial source different from stock nitrate solution A and prepare
the same way as the preparation for stock nitrate solution A.
10. Intermediate Nitrate Solution A
Dilute 100 mL stock nitrate solution to 1000 mL with water. Solution is stable for
at least 6 months and is preserved by addition of 2 mL chloroform.
11. Intermediate Nitrate Solution B
Obtain a commercial source different from intermediate nitrate solution A and
prepare the same way as the preparation for intermediate nitrate solution A.
12. Ammonia-free water nitrate-free water
Ammonia-free water can be prepared by either ion-exchange or distillation. For
this experiment, to eliminate traces of ammonia in distilled water, add 0.1 mL of
concentrated sulfuric acid to 1L distilled water and the solution is redistilled.
Apparatus
1. Reduction column
2. Pipette
3. Spectrophotometer for use at 543 nm that provides a light path of 1 cm or longer.
4. Filter photometer with the light path of 1 cm or longer and a filter with a maximum
transmittance near 540 nm.
5. pH meter
Procedure
For the preparation of the reduction column, insert a glass wool plug into the bottom of
the reduction column and fill with water. Add sufficient Cu-Cd granules to produce a column
18.5 cm long. Maintain water level above Cu-Cd granules to prevent entrapment of air. Wash
column with 200 mL dilute NH4Cl-EDTA solution. Activate column by passing through it, at 7
to 10 mL/min, at least 100 mL of a solution composed of 25% 1.0 mg NO3 -N/L standard and
75% NH4Cl-EDTA solution.
A pH range between 7 and 9 for every sample is required. It is urgent to therefore test
its pH level using pH meter, if the pH of the sample does not satisfy the required value, it can be
adjusted by adding 1N sulfuric acid or 1 N sodium hydroxide depending on the pH value that
will be obtained as it ensures a pH of 8.5 after the addition of ammonium chloride-EDTA
solution. For turbid samples, it must be first filtered to eliminate turbidity.
To a 25 mL of sample 75mL of ammonium chloride-EDTA solution is added and mixed
thoroughly. Pour the solution into the column and collect at a rate of 7-10 mL/min. The first 25
mL that will be eluted must be discarded then collect the remaining eluted solution. There is no
need to wash the columns between samples.
Color measurement must be done as soon as after conducting the reduction method. To a
50 mL of reduced sample, add 2.0 mL color reagent and mix. And the absorbance of mixed
samples is measured at 543 nm against a distilled water-reagent blank. The measurement of
absorbance must be conducted 10-120 minutes after mixing with color reagent. If the nitrate
concentration exceeds the standard curve range which is 1 mg/L,use the remainder of the
reduced sample to make an appropriate dilution and analyze again. All of the procedure will be
done in 5 replicates.
Using an intermediate nitrate solution, prepare standard in the range 0.05 to 1.0 mg of
nitrate/L by dilution of 0.5, 1.0, 2.0, 5.0, and 10 mL to 100 ml in volumetric flask. The procedure
for the reduction method for this standard is exactly the same as for the samples. For the
verification of column efficiency, at least one reduced standard nitrate will be compared to a
standard nitrite at the same concentration. If the efficiency of the copper cadmium granules falls
below 75%, the granules must be reactivated. A standard curve can be obtained by plotting the
absorbance of standards against the nitrate concentration. The sample concentration can be then
computed directly from the standard curve and must be reported as milligrams oxidized N/L.
II. Determination of Phosphates

There are three common standard methods known for the determination of
phosphates: (1) Vanadomolybdophosphoric acid method is most useful for routine analysis in
the range of 1 to 20 mg P/L. The stannous chloride method or the ascorbic acid method is more
suited for the range of 0.01 to 6 mg P/L. The ascorbic acid method will be used in the
determination of phosphate in this analysis.
Apparatus
1. Spectrophotometer with infrared phototube for use at 880 nm that provides a light path of
2.5 cm or longer.
2. Acid-washed glassware
1. Antimony potassium tartrate solution.
Dissolve 1.3715 g K(SBO)C4H4O6 • 1/2H2O in approximately 400 mL distilled water and
dilute to 500-mLvolumetric flask. the solution must bestored at 4oC in a dark,glass-stoppered
bottle.
2. Ammonium molybdate solution
Dissolve 20.0 g of ammonium heptamolybdate tetrahydrate in500 mL distilled water. It
must be stored in a plastic bottle at 4oC.
3. 0.1 M ascorbic acid
Dissolve 1.76g ascorbic acid in 100 mL distilled water.
4. Combined Reagent
This is prepared by mixing 50mL 5 N sulfuric acid, 5 mLantimony potassium tartrate
solution, 15 mL ammonium molybdate solution and 30 mL ascorbic acid solution. It is
recommended to mix the solution after the addition of each reagent. If the solution is
turbid, shake it and stand for a few minutes until turbidity disappears before proceeding.
5. Sulfuric acid solution
Slowly add 70 mL of concentrated sulfuric acid to 500 mLdistilled water. When the
solution has cooled, dilute it to 1L
6. Stock phosphate solution
Dry 439.3 mg anhydrous KH2PO4 for 1 hour at 105oC and dilute it to 1000 mL.
7. Intermediate phosphate solutions
Dilute 100.0 mL stock phosphate solution to 1000 mL with distilled water.
8. Standard phosphate solutions
Prepare a suitable series of standards by diluting appropriate volumes of intermediate
phosphate solution.
Procedure
Pipet 50.0 mL sample into a clean, 125-mL erlenmeyer flask. 1 drop (0.05 mL)
phenolphthalein indicator is added into the flask. If red color develops in the solution upon
addition of the indicator, add 5N sulfuric acid solution to eliminate the appearance of the color
thereafter addition of combined reagent to the solution and mix thoroughly. The measurement of
the absorbance in the sample at 880 nm using reagent blank as the reference solution must be
performed 10-30 minutes after the combined reagent is added. This procedure will be done in 5
replicates.
When using high wavelength, the samples’ color and turbidity usually does not interfere
with the result. But for highly colored samples and with great turbidity, a blank is prepared by
addition of all reagent except from ascorbic acid and antimony potassium tatrate and this is
subtracted from the absorbance of each sample.
Individual calibration curves can be created by preparing 6 phosphate standards that
concentration ranges from 0.01-6 mg/L. Plot absorbance vs. phosphate concentration to give a
straight line passing through the origin. Test at least one phosphate standard with each set of
samples.
III. Determination of Turbidity

Water turbidity as an essential measurement to determine the quality of water describes
the level of its cloudiness. It is mostly caused by a massive number of small and colloidal
particles that are suspended in the water. Mining, construction and agriculture are just a few
examples of the cause of turbidity in nearby bodies of water. Historically, the standard method
for determination of turbidity has been based on the Jackson candle turbidimeter, however the
minimum value of turbidity that can be measured in this device is 25 Jackson Turbidity Units
(JTU). Since water samples that are being treated usually fall within the range of 0 to 1 units,
other indirect methods were developed such as the nephelometric method.
The nephelometric method is based on the comparison on degree or intensity of scattered
light under certain conditions with standard reference suspension under with the same condition
also. Formazin polymer is the primary standard reference suspension that will be used.
Apparatus
1. Turbidimeter consisting of a nephelometer that satisfies the following:
 Light source—Tungsten-filament lamp operated at a color temperature between 2200 and
3000°K.
 Distance traversed by incident light and scattered light within the sample tube—Total not
to exceed 10 cm.
 Angle of light acceptance by detector—Centered at 90° to the incident light path and not
to exceed ±30° from 90°. The detector, and filter system if used, shall have a spectral
peak response between 400 and 600 nm.
2. Sample tubes (clear colorless glass)
1. Turbidity-free water
For a method that has a turbidity of as low as 0.02 Nephelometric Turbidity unit (NTU), pass
distilled water through a membrane filter having precision-sized holes of 0.2 µm. Some
commercial demineralized water are nearly free of particles and can be used if their turbidity is
lower than what can be obtained in a laboratory.
2. Stock turbidity suspension:
 Solution I—Dissolve 1.000 g hydrazine sulfate in distilled water and dilute to 100 mL in
a volumetric flask.
 Solution II—Dissolve 10.00 g hexamethylenetetramine, (CH2)6N4, in distilled water and
dilute to 100 mL in a volumetric flask.
 In a 100-mL volumetric flask, mix 5.0 mL Solution I and 5.0 mL Solution II. Let stand
24 h at 25 ± 3°C, dilute to mark, and mix. The turbidity of this suspension is 400 NTU.
3. Standard turbidity suspension
Dilute 10.00 mL stock turbidity suspension to 100 mL with turbidity-free water.
Prepare daily. The turbidity of this suspension is defined as 40 NTU.
4. Dilute turbidity standards
Dilute portions of standard turbidity suspension with turbidity-free water as
required.
Procedure
In calibrating the turbidimeter, the manufacturer’s operating instructions must be
followed. In the absence of a pre calibrated scale, prepare calibration curves for each range of the
instrument. Check accuracy of any supplied calibration scales on a pre-calibrated instrument by
using appropriate standards. Run at least one standard in each instrument range to be used. Make
certain that the turbidimeter gives stable readings in all sensitivity ranges used.
For measurement of turbidites less than 40 NTU, thoroughly shake each sample and wait
until air bubbles disappear and then pour the sample into the turbidimeter tube. The turbidity can
be obtained from the instrument scale or from appropriate calibration curve. For measurement of
turbidites above 40 NTU, dilute samples with turbidity-free water until turbidity of 30-40 NTU is
achieved. The turbidity can be calculated from the original sample from the turbidity of diluted
sample and dilution factor. Calculation of turbidity units is:
𝐴(𝐵 + 𝐶)
𝑁𝑇𝑈 =
𝐶
Where:
A = NTU found in diluted sample
B = volume of dilution water in mL
C = Sample volume taken for dilution in mL
IV. Biological Oxygen Demand (BOD)

Biological oxygen demand is the most important assessment used in determination of
water quality. It’s a measurement unit that stands for the needed amount of oxygen by aerobic
biological organisms to break down organic matter. The higher the level of water pollution is
also the higher organic matter present and so more oxygen is needed for oxidation, hence the
higher the biological oxygen demand level. Moderately polluted rivers may have a BOD value in
the range of 2 to 8 mg/L. Rivers may be considered severely polluted when BOD values exceed
8 mg/L (Grover and Wats, 2013). Biological oxygen demand test is an indirect measurement of
organic matter; it measures the change in dissolved oxygen caused by microorganisms as they
degrade. In this method, a 5-Day BOD Test will be used. The dissolved oxygen will be measured
before and after incubation.
Apparatus
1. 300-mL Incubation bottles
2. Air incubator or water bath that is thermostatically controlled at 20 ±1 oC. The sample
must not be directed to light to prevent photosynthetic production of dissolved oxygen.
3. Oxygen-sensitive optical probe with appropriate meter.
1. Phosphate buffer solution
Dissolve 8.5 g monopotassium phosphate, 21.75 g dipotassium phosphate, 33.4 g
disodium phosphate heptahydrate, and 1.7 g ammonium chloride in about 500 mL
reagent-grade water and dilute to 1 L. The pH should be around 7.2
2. Magnesium sulphate solution
Dissolve 22.5 g magnesium sulfate heptahydrate in reagent-grade water and dilute
to 1 L.
3. Calcium chloride solution
Dissolve 27.5 g Calcium chloride in reagent grade water and dilute to 1 L.
4. Ferric Chloride
Dissolve 0.25 g ferric chloride hexahydrate in reagent-grade water and dilute to 1
L.
5. For the neutralization of caustic and acidic waste samples, 28 mL of concentrated sulfuric
acid is diluted to 1 L for basic wastes and 40 g sodium hydroxide is dissolved in distilled water
and diluted to 1 L for neutralization of acidic wastes.
6. Sodium sulfite solution
Dissolve 1.575 g sodium sulfite in 1000 mL reagent-grade water. This solution is
unstable and must be freshly prepared before usage.
7. Nitrification inhibitor:
 2-chloro-6-(trichloromethyl) pyridine—Use pure TCMP or commercial preparations*.
 Allylthiourea (ATU) solution—Dissolve 2.0 g allylthiourea (C4H8N2S) in about 500 mL
water and dilute to 1 L. Storeat 4°C. The solution is stable for not more than 2 weeks.
8. Glucose-glutamic acid solution

Dry reagent-grade glucose and reagent-grade glutamic acid at 103°C for 1 h. Add
150 mg glucose and 150 mg glutamic acid to distilled water and dilute to 1 L. Prepare
fresh immediately before use unless solution is maintained in a sterile condition. Store all
glucose-glutamic acid mixtures at 4°C or lower. Commercial preparations may be used
but concentrations may vary
9. Ammonium chloride solution:
Dissolve 1.15 g NH4Cl in about 500 mL distilled water, adjust pH to 7.2 with
NaOH solution, and dilute to 1 L. Solution contains 0.3 mg N/mL.
Procedure
The samples that will undergo BOD test must undergo first with pretreatment if it does
not meet the requirements needed in the test. A pH range between 6.5 and 7.5 for every sample
is required. It is urgent to therefore test its pH level, if the pH of the sample does not satisfy the
required value, it can be adjusted by adding 1N sulfuric acid or 1 N sodium hydroxide depending
on the pH value that will be obtained. For 300 mL samples it is recommended that the maximum
volume of acid or base that must be added to samples should not exceed 1.5 mL. Samples that
have residual chlorine must undergo dechlorination as chlorine kills the microorganism present
in samples that lead to less precision of the results. Samples supersaturated with dissolved
oxygen must be shaken or aerated with filtered compressed air to reduce the dissolved oxygen
because these types of samples have high concentration of nitrifying organisms which can lead to
bias in BOD results. Lastly, samples that have extreme conditions such as high temperature,
extreme pH values, and untreated industrial waste may not contain enough microorganisms to
oxidize the biodegradable matter in the samples and should undergo seeding prior to BOD test.
Estimated BOD5 (mg/L) Suggested Sample Volumes (mL)
< 5 200, 250, 300
< 10 100, 150, 200
10 - 30 25, 50, 100
30 - 60 15, 25, 50
60 - 90 10, 15, 25
90 - 150 5, 10, 15
150 - 300 3, 5, 10
300 - 700 1, 3, 5 ***
700 - 1500 0.5, 1, 3 ***
1500 - 2500 0.25, 0.5, 1 ***
Table 2. Suggested Sample Volumes for Estimated BODs
When preparing replicate samples for quality control purposes, prepare the replicate at
exactly the same dilutions as the original sample. Using either volumetric pipette for samples <
50 mL or graduated cylinder for larger sample, measure the desired amount of well-mixed
sample into well cleaned 300 mL bottles. The dilution water can be prepared before use,days, or
weeks ahead of time except when using phosphate buffer as it is essential for stimulating
contaminating growths (it must be added and prepared on the day of usage). Distilled water must
be equilibrated with the incubator with at least 24 hours at 20 oC before use.
Each BOD bottle must be filled by gradual addition of dilution water. When the volume
of samples that will be used exceeds 150 mL. Dilution water must be added to two bottles to be
incubated as blanks. Each bottle must be labeled accordingly.
The dissolved oxygen meter must be calibrated and checked each day of use. The
barometric pressure is also recorded each day. The two dilution water blanks’ dissolved oxygen
are determined together with all samples bottles. Data must be recorded. The samples and
dilution water blanks will be placed in a 20 ± 1 oC incubator for 5 days. The seals must be filled
with distilled water and must be capped to reduce evaporation of samples. After 5 days the
dissolved oxygen can be determined. In general, BOD5 values less than 2.0 mg DO/L should be
reported on DMRs as non-detects ( i.e., < LOD). Using the data recorded for un-seeded samples:
BOD mg/l = (Initial DO - DO5) x Dilution Factor
Dilution Factor = Bottle Volume (300 ml) /Sample Volume
V. Statistical Treatment
Quantitative data on water quality parameters will be compared using Single Factor
ANOVA at 95% level of confidence to determine a significant difference between the water
quality of the Pansipit River before and after the Taal Volcano eruption in various parameters.
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Republic of the Philippines

BATANGAS STATE UNIVERSITY

WATER QUALITY ASSESSMENT OF PANSIPIT RIVER AFTER THE TAAL

SIGNIFICANCE OF THE STUDY

• Sample collection equipment:

- If necessary, equipment for collection of samples from a safe distance

- Container for field measurements

- Container for collecting sample

- Sample bottles for dispatch to laboratory

• Field parameter meters or test kits (e.g. alkalinity/acidity test kit)

• Powder-less nitrile gloves

• Esky or portable refrigerator

• Personal Protective Equipment, first aid and communication

Field Measured Parameters and Laboratory Analyzed Parameters

Minimum Amount of Sample

Procedures for preparation of containers

Chain of Custody Procedures

Table 1. Summary of the Sample Collections

Phosphates G(A) 100 g Cool, ≤ 6oC 48 h 48 h

BOD P, G, FP 1000 g,c Cool, ≤ 6oC 6h 48 h

Laboratory Analyzed parameters

to develop. The precipitated copper can then be eliminated through washing of

9. Stock nitrate solution B

II. Determination of Phosphates

III. Determination of Turbidity

IV. Biological Oxygen Demand (BOD)

8. Glucose-glutamic acid solution

Estimated BOD5 (mg/L) Suggested Sample Volumes (mL)

< 5 200, 250, 300

< 10 100, 150, 200

10 - 30 25, 50, 100

300 - 700 1, 3, 5 ***

700 - 1500 0.5, 1, 3 ***

1500 - 2500 0.25, 0.5, 1 ***

Table 2. Suggested Sample Volumes for Estimated BODs

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