Method 8048

PHOSPHORUS, REACTIVE (0.00 to 5.00 mg/L PO43-)

PhosVer 3 Method, Test ’N Tube Procedure For water, wastewater, and seawater
USEPA accepted for reporting wastewater analysis*

1. Enter the stored 2. Press: 82 ENTER 3. Insert the COD/TNT 4. Use a TenSette Pipet
program number for Adapter into the cell to add 5.0 mL of sample
The display will show
reactive phosphorus holder by rotating the to a Reactive
mg/L, PO4 and the
(PO43-), Test ’N Tube. adapter until it drops Phosphorus Test 'N Tube
ZERO icon.
into place. Then push Dilution Vial. Cap
Press: PRGM
Note: For alternate forms down to fully insert it. and mix.
The display will show: (P, P2O5), press the CONC
key. Note: A diffuser band Note: For samples with
PRGM ? covers the light path holes extreme pH, see the
on the adapter to give Interference section.
Note: For most accurate increased performance.The
results, perform a Reagent band should NOT be
Blank Correction using removed.
deionized water (see
Section 1).

* Procedure is equivalent to USEPA Method 365.2 and Standard Method 4500-P E for wastewater.

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PHOSPHORUS, REACTIVE, continued

5. Clean the outside of 6. Place the sample vial 7. Tightly cover the 8. Press: ZERO
the vial with a towel. into the adapter. sample vial with the
The cursor will move to
instrument cap.
Note: Wiping with a damp Push straight down on the right, then the
towel, followed by a dry the top of the vial until it display will show:
one, will remove seats solidly into the
fingerprints or other marks. adapter. 0.00 mg/L PO4
Note: For multiple
Note: Do not move the vial
samples, zero only on the
from side to side as this can
first sample. Read the
cause errors.
remaining samples after
adding the PhosVer 3
Reagent.

9. Using a funnel, add 10. Cap the vial tightly 11. Press: 12. Immediately after
the contents of one and shake for 10-15 the timer beeps, place
TIMER ENTER
PhosVer 3 Phosphate seconds. the sample vial in the
Powder Pillow to the A 2-minute reaction adapter.
Note: The powder will not
vial. completely dissolve. time will begin. Push straight down on
Note: Read samples the top of the vial until it
between 2 and 8 minutes seats solidly into the
after the addition of the adapter.
PhosVer 3 reagent.
Note: Do not move the vial
Note: A blue color will from side to side as this can
develop if phosphate is cause errors.
present.

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PHOSPHORUS, REACTIVE, continued

13. Tightly cover the 14. Press: READ

vial with the instrument
The cursor will move to
cap.
the right, then the result
in mg/L phosphate
(PO43-) will be displayed.
Note: Standard Adjust may
be performed using a
prepared standard (see
Section 1).

Sampling and Storage

Collect samples in plastic or glass bottles that have been acid
cleaned with 1:1 Hydrochloric Acid Solution and rinsed with
deionized water. Do not use commercial detergents containing
phosphate for cleaning glassware used in this test.

Analyze samples immediately after collection for best results. If

prompt analysis is impossible, preserve samples for up to 48
hours by filtering immediately and storing at 4 °C. Warm to room
temperature before analyzing the sample.

Accuracy Check
Note: Clean glassware Standard Additions Method
with 1:1 hydrochloric acid a) Fill three 25-mL graduated mixing cylinders with 25 mL of
solution. Rinse again with sample.
deionized water. Do not
use detergents containing
b) Snap the neck off a Phosphate PourRite Ampule Standard,
phosphates to clean
glassware. 50 mg/L as PO43-.

c) Use the TenSette Pipet to add 0.1 mL, 0.2 mL and 0.3 mL,
respectively, to the three 25-mL aliquots of sample
prepared in step a. Mix well.

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PHOSPHORUS, REACTIVE, continued
d) Analyze each sample as described in the procedure; use
5.0 mL of the prepared standard additions for each test.
The concentration should increase as follows: 0.2 mg/L,
0.4 mg/L, 0.6 mg/L
PO43-, respectively.

e) If these increases do not occur, see Standard Additions in

Section 1 for more information.

Standard Solution Method

To check accuracy, use a 1.0 mg/L Phosphate Standard Solution
listed under Optional Reagents. Or, prepare a 1.0-mg/L PO43-
standard by pipetting 2 mL of solution from a Phosphate Voluette
Ampule Standard for Phosphate, 50 mg/L as PO43-, into an acid-
washed, Class A 100-mL volumetric flask. Dilute to the mark
with deionized water. Substitute this standard for the sample and
perform the procedure as described.
Method Performance
Precision
In a single laboratory, using a standard solution of 5.00 mg/L
PO43- and two lots of reagent with the instrument, a single
operator obtained a standard deviation of ±0.08 mg/L PO43-.
Estimated Detection Limit (EDL)
The EDL for program 82 is 0.07 mg/L PO43-. For more
information on derivation and use of Hach’s estimated detection
limit, see Section 1.

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PHOSPHORUS, REACTIVE, continued
Interferences
The following may interfere when present in concentrations
exceeding these listed below:

Substance Interference Level and Treatment

Aluminum 200 mg/L

Arsenate Interferes at any level
Chromium 100 mg/L
Copper 10 mg/L
Iron 100 mg/L
Nickel 300 mg/L
Silica 50 mg/L
Silicate 10 mg/L
Sulfide 6 mg/L. Sulfide interference may be removed by
oxidation with Bromine Water as follows:
1. Measure 25 mL of sample into a 50-mL beaker.
2. Swirling constantly, add Bromine Water drop-
wise until a permanent yellow color develops.
3. Swirling constantly, add Phenol Solution
dropwise until the yellow color just disappears.
Proceed with step 1.
Turbidity (large amounts) May cause inconsistent results because the acid
present in the powder pillows may dissolve some of
the suspended particles and because of variable
desorption of orthophosphate from the particles.
Zinc 80 mg/L
Highly buffered samples May exceed the buffering capacity of the reagents
or extreme sample pH and require sample pretreatment; see pH
Interferences (Section 1).

The PhosVer 3 Phosphate Reagent Powder Pillows should be

stored in a cool, dry environment.

Sample Disposal Information

Final samples will contain molybdenum. In addition, final
samples will have a pH less than 2 and are considered corrosive
(D002) by the Federal RCRA.

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PHOSPHORUS, REACTIVE, continued
Summary of Method
Orthophosphate reacts with molybdate in an acid medium to
produce a phosphomolybdate complex. Ascorbic acid then
reduces the complex, giving an intense molybdenum blue color.

REQUIRED REAGENTS
Reactive Phosphorus Test ’N Tube Reagent Set.................................50 tests ............. 27425-45
Includes: (1) 21060-46, (50) Orthophosphate Dilution Vials*
Quantity Required
Description Per Test Unit Cat. No.
PhosVer 3 Phosphate Reagent Powder Pillows ..................1 ..............50/pkg ............. 21060-46
50 Orthophosphate Test ‘N Tube Dilution Vials ................1 ..............50/pkg.................................. *

REQUIRED APPARATUS
COD/TNT Adapter .............................................................1 ..................each ............. 48464-00
Funnel, micro ......................................................................1 ..................each ............. 25843-35
Pipet, TenSette, 1 to 10 mL.................................................1 ..................each ............. 19700-10
Pipet Tips, for 19700-10 TenSette Pipet .............................1 ..............50/pkg ............. 21997-96
Test Tube Rack ............................................................... 1-3 ..................each ............. 18641-00

OPTIONAL REAGENTS
Bromine Water, 30 g/L......................................................................... 29 mL ............... 2211-20
Drinking Water Standard, Inorganic, F–, NO3–N, PO43–, SO42–.... .............500mL.......... 28330-49
Hydrochloric Acid Standard Solution, 6.0 N (1:1) ............................ 500 mL ................. 884-49
Phenol Solution, 30 g/L ....................................................................... 29 mL ............... 2112-20
Phosphate Standard Solution, 1 mg/L as PO43- .................................. 500 mL ............... 2569-49
Phosphate Standard Solution, Voluette ampule,
50 mg/L as PO43-, 10 mL ...............................................................16/pkg ................. 171-10
Phosphate Standard Solution, PourRite ampule,
50 mg/L as PO43-, 2 mL .................................................................20/pkg .............. 171-20H
Wastewater Effluent Standard, Inorganic
(NH3–N, NO3–N, PO4, COD, SO4, TOC)......................................500 mL.......... 28332-49
Water, deionized........................................................................................ 4 L ................. 272-56

* These items are not sold separately.

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PHOSPHORUS, REACTIVE, continued
OPTIONAL APPARATUS
Ampule Breaker, Pour Rite (2-mL ampule)............................................ each..............24846-00
Ampule Breaker Kit ................................................................................ each..............21968-00
Aspirator, vacuum ................................................................................... each................2131-00
Cylinder, graduated, mixing, 25 mL (3 required) ................................... each..............20886-40
Dispenser, Repipet Jr., 2 mL ................................................................... each..............22307-01
Filter Holder, 47 mm, 300 mL, graduated............................................... each..............13529-00
Filter, membrane, 47 mm, 0.45 microns ........................................... 100/pkg..............13530-00
Flask, filtering, 500 mL........................................................................... each..................546-49
Flask, volumetric, Class A, 100 mL........................................................ each..............14574-42
pH Indicator Paper, 1 to 11 pH units ............................................. 5 rolls/pkg..................391-33
pH Meter, sension™1, portable with electrode ...................................... each..............51700-10
Pipet, TenSette, 0.1 to 1.0 mL ................................................................. each..............19700-01
Pipet Tips, for 19700-01...................................................................... 50 pkg..............21856-96
Pipet Tips, for 19700-01 ...................................................... .......... 1000/pkg..............21856-28
Pipet Filler, Safety Bulb .......................................................................... each..............14651-00
Pipet, volumetric, Class A, 5.00 mL ....................................................... each..............14515-37
Pipet, volumetric, Class A, 2.00 mL ....................................................... each..............14515-36

For Technical Assistance, Price and Ordering

In the U.S.A. call 800-227-4224
Outside the U.S.A.—Contact the Hach office or distributor serving you.

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