TRAACS REAGENT RECIPES
GEL Revision Date:
3 August 2004
Index
Ammonia………………………………………………………….……………………2
Chloride……………………………………………………………….……….……….4
Methyl Orange Alkalinity………………………………………….……………….…6
Nitrate / Nitrite……………………………………………………….……………..…8
Nitrogen, Total Kjeldahl / Salicylate Ammonia…………………………..…….…...11
Phosphate…………………………………………………………………………..…14
General Notes:
1. GEL revisions are indicated by italics.
2. Unless otherwise specified, all chemicals should be of ACS grade or equivalent.
3. Use Nanopure deionized water (>M ohm-cm) for all solutions specifying water as
solvent.
4. Chemical and reagent locations:
DC
Dry Chemical Cabinet (GWC 675)
SC
Standards Cabinet (GWC 675)
WIR Walk-in Refrigerator (GWC 675)
AC
Acid Cabinet (GWC 675)
BC
Base Cabinet (GWC 675)
OC
Organic Solvent Cabinet (GWC 675)
RC
Reagent Cabinet (GWC 637)
2
Industrial Method No. 804-86T
AMMONIA IN WATER AND SEAWATER
August 1987
List of Raw Materials
Brij-35, 15% Solution (Polyoxyethylene Lauryl Ether) ((C2H4O)23C12H25O)–CAS 9002-92-0,
(DC 675)
Disodium EDTA (Ethylenedinitrilo)-tetraacetic acid – CAS 6381-92-6, (DC 675)
Phenol (C6H5OH), crystalline – CAS 108-95-2, (WIR 675)
Sodium Hydroxide, (NaOH), low N grade – CAS 1310-73-2, (BC 675)
Sodium Hypochlorite, 5% (NaOCl), Commercial Grade; e.g. Clorox (institutional and
germicidal), (WIR 675)
Sodium Nitroprusside (Sodium nitroferricyanide) [Na42(NO)Fe(CN) 5·2H2O] – CAS 13755-389, (DC 675)
Sulfuric Acid, 95 – 98% (H2SO4) – CAS 7664-93-9, (AC 675)
BUFFER FOR NON ACID PRESERVED SAMPLES
By Volume: In a 1 (0.5) L volumetric flask, dissolve 50.0 (25) g disodium
ethylenediamine tetraacetic acid (Na2EDTA) and 225 (117.5) mL 1 M sodium
hydroxide (Reagent 4) (or 15 mL 15 M NaOH or use 9 g solid NaOH*) in
approximately 700 (350) mL DI water. Dilute to the mark and mix with a magnetic
stirrer until dissolved. Prepare fresh monthly.
* Use the NaOH in base cabinet for nitrogen determination (9 g is for prep of 1 L of
buffer). --CK 03/01/01
* Transfer 100 mL of buffer to beaker and add 0.2 mL 15% Brij-35. Mix before use.
* Use 2 mL Brij-35 15% solution per 1L of buffer-add after transferring to bottle.
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SODIUM PHENOLATE (RC 637)
*Add Phenol last. (Used also for QC8000) Add NaOH to a portion of the H2O and allow to
cool. Use the remainder of H2O to rinse in phenol.
CAUTION: Wear gloves! Phenol causes severe burns and is rapidly absorbed into the body
through the skin.
By Volume: In a 1 (0.5) [0.25] L volumetric flask, dissolve 83 (41.5) [20.75] g
crystalline phenol (C6H5OH) is approximately 600 (300) [150] mL DI water. While
stirring, slowly add 32 (16) [8] g sodium hydroxide (NaOH). Cool, dilute to the
mark and invert to mix. Do not degas this reagent. Prepare fresh every 3 to 5 days.
Discard when reagent turns brown. *Use solid NaOH for N determination in base
cabinet.
3
By Weight: QC 8000-Good Baseline. To a tared 1 (0.5) L container, add 888 (444)
g DI water. Add 83 (41.5) g crystalline phenol (C6H5OH). While stirring, slowly
add 32 (16) g sodium hydroxide (NaOH). Cool and invert to mix thoroughly. Do
not degas this reagent. Prepare fresh every 3 to 5 days. Discard when reagent turns
brown. *Use solid NaOH for N determination in base cabinet.
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SODIUM HYPOCHLORITE, 1% SOLUTION (RC 637)
Sodium Hypochlorite (Clorox bleach)
DI Water, q.s.
20 mL
100 mL
Preparation: Dilute 20 mL of sodium hypchlorite, 5% solution to 100 mL with DI water
and mix thoroughly. Prepare the 1% working solution fresh daily. Keep the stock 5%
solution refrigerated.
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SODIUM NITROPRUSSIDE, 0.05% (RC 637)
Sodium Nitroprusside
DI Water, q.s.
0.5 g
1000 mL
Preparation: Dissolve 0.5 g of sodium nitroprusside in one liter of DI water and mix
thoroughly. Store in a light resistant container. Prepare every 1 – 2 weeks.
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SAMPLER (BASELINE) SOLUTION (Use only with seawater samples. This reagent is
not recommended for fresh water).
Isopropanol
DI Water, q.s.
100 mL
1000 mL
Preparation: Dilute 100 mL of isopropanol to one liter with DI water and mix thoroughly.
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4
Industrial Method No. 783-86T
CHLORIDE
LIST OF RAW MATERIALS
Brij – 35, 15% Solution (Polyoxyethylene Lauryl Ether) ((C2H4O)23C12H25O)–CAS 9002-92-0,
(DC 675)
Ferric Nitrate [Fe(NO3) 3·9H2O] – CAS 7782-61-8, (DC 675)
Mercuric Thiocyanate (Hg(SCN)2 – CAS 592-85-8, (DC 675)
Methanol (CH3OH) – CAS 67-56-1, (OC 675)
Nitric Acid (HNO3) – CAS 7697-37-2, (AC 675)
STOCK MERCURIC THIOCYANATE SOLUTION (Used also for QC8000) (RC 637)
For ½
Mercuric thiocyanate
4.17 g
2.085 g
Methanol, q.s.
1000 mL
500 mL
Dissolve 4.17 g of mercuric thiocyanate in about 800 mL of methanol. Dilute to one liter
with methanol, mix thoroughly and filter.
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STOCK FERRIC NITRATE SOLUTION (Used also for Lachat QC8000) (RC 637)
Ferric Nitrate
Nitric Acid
DI Water, q.s.
202 g
25 mL
1000 mL
101 g
12.5 mL
500 mL
Dissolve 202 g of ferric nitrate in about 600 mL of water. While mixing, slowly add 25 mL
of nitric acid. Dilute to one liter with DI water and mix thoroughly. Filter through paper
and store in a well sealed amber reagent container.
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5
CHLORIDE COLOR REAGENT (Used also for Lachat QC8000) (RC 637)
Stock Mercuric Thiocyanate Solution
Stock Ferric Nitrate Solution
DI water, q.s.
87 mL
87 mL
500 mL
To about 150 mL of DI water, add 87 mL of stock mercuric thiocyanate, 87 mL of stock
ferric nitrate solution and mix. Dilute to 500 mL with DI water and mix thoroughly.
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DILUENT WATER
On day of analysis, add 4 drops 15% Brij-35 to 50 mL of water.
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6
Industrial Method No. 788-86T
METHYL ORANGE ALKALINITY
August 1987
LIST OF RAW MATERIALS
Brij-35 (Polyoxyethylene Lauryl Ether) ((C2H4O)23C12H25O)–CAS 9002-92-0, (DC 675)
Hydrochloric Acid, conc. (HCl) – CAS 7647-01-0, (AC 675)
Methyl Orange (C14H14N3NaO3S) – CAS #, (DC 675)
Potassium Acid Phthalate (Potassium Hydrogen Phthalate) (C8H5KO4) –CAS 877-24-7, (DC 675)
BUFFER, pH 3.1
Potassium Acid Phthalate
0.1N Hydrochloric Acid
DI Water, q.s.
10.2 g
175 mL (Approx.)
1000 mL
Dissolve 10.2 g of potassium acid phthalate in 700 mL of DI water. Add 0.1 N
hydrochloric acid (8.3 mL hydrochloric acid – sp.gr. 1.19/L DI water) to bring to pH 3.1.
Dilute one liter with DI water. Store in a glass container.
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METHYL ORANGE (RC 637)
Methyl Orange
DI Water, q.s.
144 mg
1000 mL
Dissolve 144 mg of methyl orange in 800 mL of DI water and dilute to one liter. Filter this
solution through glass wool if necessary. Store in an amber glass container.
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WORKING BUFFER
Buffer, pH 3.1
DI Water, q.s.
Brij-35
140 mL
1000 mL
1.0 mL
Dilute 140 mL of buffer to one liter with DI water. Add 1.0 mL Brij-35 per liter. Store in
a glass container. Make fresh the day of analysis.
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7
Industrial Method No. 818-87T
NITRATE/NITRITE IN WATER AND SEAWATER
February 1987
LIST OF RAW MATERIALS
Copper (II) Sulfate (CuSO4·5H2O) – CAS 7758-99-8, (DC 675)
Hydrochloric Acid, 36.5 – 38.0% (HCl) – CAS 7647-01-0, (AC 675)
Imidizole (NHCH:NCH:CH), Highest Purity – CAS 288-32-4, (DC 675)
N-(1-Naphthyl)ethylenediamine Dihydrochloride (C10H2HNCH2CH2NH2·2HCl)–CAS 1465-25-4,
(DC 675)
Nitric Acid, 69.0 – 71.0% (HNO3) – CAS 7697-37-2, (AC 675)
2-Propanol or Isopropanol [(CH3) 2CHOH] – CAS 67-63-0, (OC 675)
Sulfanilamide (4-NH2C6H4SO2NH2) – CAS 63-74-1, (DC 675)
Triton X-100 (Technicon No. T21-0188-06), (SC 675)
50% TRITON X-100 SOLUTION (RC 637)
Triton X-100
Isopropanol
50 mL
50 mL
Preparation: To 50 mL of isopropanol contained in a 100 mL glass-stoppered graduated
cylinder, add 50 mL of Triton X-100. Stopper the cylinder and invert repeatedly until
thoroughly mixed. Transfer to a dropper bottle.
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STOCK IMIDIZOLE BUFFER, 0.1M (RC 637)
Imidizole
Hydrochloric Acid
DI Water, q.s.
6.81 g
As required
1000 mL
Preparation: Dissolve 6.81 g of imidizole in about 900 mL of DI water. Adjust pH to 7.5 ±
0.05 with hydrochloric acid. Dilute to one liter with DI water and mix thoroughly.
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STOCK COPPER SULFATE SOLUTION, ‘A’, 0.01M (RC 637)
(Used to prepare both Stock Copper Sulfate, 0.001M and Copper Activation Solution.)
Copper Sulfate
DI Water, q.s.
2.5 g
1000 mL
Preparation: Dissolve 2.5 g of copper sulfate in about 600 mL of DI water. Dilute to one
liter with DI water and mix thoroughly.
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STOCK COPPER SULFATE SOLUTION ‘B’, 0.001M (RC 637)
Stock Copper Sulfate ‘A’, 0.001M
DI Water, q.s.
10.0 mL
100 mL
Preparation: Dilute 10.0 mL of copper sulfate, 0.01M to 100 mL with DI water and mix
thoroughly.
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WORKING IMIDIZOLE, 0.05M
Stock Imidizole Buffer, 0.1M
Stock Copper Sulfate ‘B’, 0.001M
DI Water, q.s.
50% Triton X-100
50.0 mL
0.5 mL
100 mL
0.1 mL
Preparation: Add 0.5 mL of stock copper sulfate ‘B’, 0.001M to 50 mL of stock
imidizole buffer, 0.1. Dilute to 100 mL with DI water, add 0.1 mL (~3 drops) 50% Triton
X-100 and mix thoroughly. Prepare fresh daily in dedicated bottle.
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WORKING SAN SOLUTION (RC 637)
Sulfanilamide
Hydrochloric Acid
DI Water, q.s.
50% Triton X-100
5.0 g
50 mL
1000 mL
0.5 mL
Preparation: Add 5.0 g of sulfanilamide to a one liter volumetric flask containing 600 mL
of DI water and 50 mL of hydrochloric acid. Swirl to dissolve and dilute to one liter with
DI water. Add 0.5 mL 50% Triton X-100 (~15 drops) and mix thoroughly. Transfer to a
light resistant container.
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WORKING NED SOLUTION (RC 637)
N-(1-naphtyl)ethylenediamine dihydrochloride
DI Water, q.s.
50% Triton X-100
0.5 g
1000 mL
0.5 mL
Preparation: Dissolve 0.5 g of N-(1-naphthyl)ethylenediamine dihydrochloride in about
600 mL of DI water. Dilute to one liter with DI water, add 0.5 mL 50% Triton X-100 (~15
drops) and mix thoroughly. Transfer to a light resistant container.
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2N HYDROCHLORIC ACID (RC 637)
Hydrochloric Acid
DI Water, q.s.
50% Triton X-100
165 mL
1000 mL
0.5 mL
Preparation: Cautiously, while swirling, slowly add 165 mL of hydrochloric acid to about
600 mL of DI water. Cool to room temperature and dilute to one liter with DI water. Add
0.5 mL of 50% Triton X-100 and mix thoroughly.
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10
2N NITRIC ACID
Nitric Acid
DI Water, q.s.
12.5 mL
100 mL
Preparation: Cautiously, while swirling, slowly add 12.5 mL of nitric acid to about 60 mL
of DI water. Cool to room temperature, dilute to 100 mL with DI water and mix
thoroughly.
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SYSTEM WASH SOLUTION
Add 0.5 mL 50% Triton X-100 to one liter of DI water and mix thoroughly.
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COPPER ACTIVATING SOLUTION (RC 637)
Stock Copper Sulfate ‘A’, 0.01M
50 mL
DI Water, q.s.
100 mL
50% Triton X-100
0.1 mL
Preparation: Dilute 50 mL of stock copper sulfate ‘A’, 0.01M to 100 mL of DI water, add
0.1 mL 50% Triton X-100 (~3 drops) and mix thoroughly.
NOTES:
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11
Industrial Method No. 786-86T
NITROGEN, TOTAL KJELDAHL / SALICYLATE AMMONIA
August 1987
LIST OF RAW MATERIALS
Brij-35, 30% Solution (Polyoxyethylene Lauryl Ether) ((C2H4O)23C12H25O)–CAS 9002-92-0,
(DC 675)
Sodium Hydroxide Solution, 50% w/w (NaOH)
Sodium Hypochlorite Solution, 5.25%
Sodium Nitroprusside (Sodium nitroferricyanide) [Na2Fe(CN)5NO·2H2O] – CAS 13755-38-9,
(DC 675)
Sodium Phosphate, dibasic crystal (Na2HPO4·7H2O) – CAS #, ( )
- or Sodium Phosphate, dibasic anhydrous (Na2HPO4) – CAS #, ( )
Sodium Potassium Tartrate (NaKC4O6·4H2O) – CAS #, ( )
Sodium Salicylate (Na2C7H5O3) – CAS #, ( )
Sulfuric Acid, conc. (H2SO4) – CAS 7664-93-9, (AC 675)
SAMPLER IV WASH RECEPTACLE SOLUTION (Not for KCl/dilution soil extracts)
Sulfuric Acid, conc.
DI Water, q.s.
80 mL
2000 mL
Add 80 mL of concentrated sulfuric acid to about 1800 of DI water. Dilute to two liters
with water.
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STOCK SODIUM HYDROXIDE SOLUTION, 20% (Pre-made in BC 675))
Sodium Hydroxide Solution, 50% w/w
DI Water, q.s.
40 g
1000 mL
To 600 mL of DI water, add 400 g of sodium hydroxide solution, 50% w/w. Cool to room
temperature and dilute to one liter with DI water.
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12
STOCK SODIUM POTASSIUM TARTRATE SOLUTION, 20% (WIR 675)
Sodium Potassium Tartrate
DI Water, q.s.
200 g
1000 mL
Dissolve 200 g of sodium potassium tartrate in about 600 mL of DI water. Dilute to one
liter with DI water and mix thoroughly.
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STOCK BUFFER SOLUTION, 0.5 M
(Keep at room temperature)
Sodium Phosphate, dibasic crystal
- or Sodium Phosphate, dibasic anhydrous
Sodium Hydroxide Solution, 50% w/w
DI Water, q.s.
134 g
70 g
40 g
1000 mL
Dissolve 134 g of sodium phosphate, dibasic crystals (or 70 g sodium phosphate, dibasic
anhydrous) in about 800 mL of DI water. Add 40 g of sodium hydroxide solution, 50%
w/w, dilute to one liter with DI water and mix thoroughly.
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WORKING BUFFER SOLUTION* (Make fresh each run)
Stock Buffer Solution, 0.5 M
Stock Sodium Potassium Tartrate Soln, 20%
Stock Sodium Hydroxide Solution, 20%
DI Water, q.s.
Brij-35, 30% Solution
200 mL
250 mL
200 mL
1000 mL
2 mL
For ½
100 mL
125 mL
100 mL
500 mL
1 mL
For ¼
50 mL
62.5 mL
50 mL
250 mL
0.5 mL
Combine the reagents in the stated order: add 250 mL of stock sodium potassium tartrate
solution, 20% to 200 mL of stock buffer solution, 0.5 M while swirling. Slowly, while
swirling, add 200 mL of sodium hydroxide solution, 20%. Dilute to one liter with DI
water, add 2 mL of Brij-35, 30% solution (~60 drops) and mix thoroughly.
* For NH4-N add 10 mL KCl (150 g KCl + 5 mL 50% conc. 6 N HCl in 1 L nanopure).
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13
SODIUM SALICYLATE/SODIUM NITROPRUSSIDE SOLUTION (Last in first out)
Sodium Salicylate
Sodium Nitroprusside
DI Water, q.s.
175 g
0.35 g
1000 mL
Dissolve 175 g of sodium salicylate and 0.35 g sodium nitroprusside in about 800 mL of
DI water. Filter through fast filter paper into a 1000 mL volumetric flask, dilute to volume
with DI water and mix thoroughly. Store in a light resistant container. Make up fresh
monthly.
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SODIUM HYPOCHLORITE SOLUTION
Sodium Hypochlorite Solution, 5.25% (if 3.25% use 10.8 mL)
DI Water, q.s.
7 mL
100 mL
Dilute 7 mL of sodium hypochlorite solution to 100 mL with DI water and mix
thoroughly. Prepare fresh daily. [Any commercial bleach solution (e.g. Clorox) containing
5.25% available chlorine is satisfactory.]
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14
Method #J-004-88 B (Multitest MT3A)
PHOSPHATE IN SEAWATER AND FRESH WATER
LIST OF RAW MATERIALS
Antimony potassium tartrate, K(SbO)C4H4O6·1/2H2O – CAS #, (DC 675)
Ascorbic acid, C6H8O6 – CAS 50-81-7, (DC 675)
Sodium chloride, NaCl – CAS 7647-14-5, (DC 675)
Sodium hydrogen carbonate, NaHCO3 – CAS #, (DC 675)
Sodium hydroxide, NaOH – CAS 1310-73-2, (BC 675)
Sodium laureth sulfate (Sodium Dodecyl Sulfate or Sodium Laurel Sulfate) (C12H25OSO3Na)
– CAS 151-21-3, (DC 675)
Sodium molybdate, Na2MoO4·2H2O - CAS 7631-95-0, (DC 675)
Sulfuric acid, H2SO4 – CAS 7664-93-9, (AC 675)
REAGENT MAKE-UP
Vacuum filter reagents through a filter with pore size 0.5 μm or less for best results.
SAMPLER WASH SOLUTION (Seawater only)
Sodium Chloride
Sodium hydrogen carbonate
DI water, q.s.
35 g
0.5 g
1000 mL
Dissolve 35 g of sodium chloride and 0.5 g of sodium hydrogen carbonate in about 900
mL of DI water. Dilute to one liter with DI water and mix thoroughly.
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DILUENT
Sulfuric acid*
Sodium laureth sulfate
DI Water, q.s.
1 mL
0.075 g
100 mL
For 5x
5 mL
0.38 g
495 mL
*Important! Use good, fresh acid.
Slowly add 1 mL of sulfuric acid to 60 mL of DI water and mix. Cool to room temperature.
Dilute to 100 mL with DI water and add 0.075 of sodium laureth sulfate. Mix thoroughly.
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15
STOCK SODIUM MOLYBDATE (RC 637)
Sodium molybdate
Antimony potassium tartrate
Sulfuric acid
DI water, q.s.
8.0 g
0.17 g
50 mL
1000 mL
Dissolve 8.0 g sodium molybdate and 0.17 g of antimony potassium tartrate in about 800
mL of DI water. Add 50 mL of sulfuric acid. Dilute to one liter and mix thoroughly. The
reagent must not show any clearly visible blue color.
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COLOR REAGENT
Stock sodium molybdate
Ascorbic acid
Sodium laureth sulfate
100 mL
0.8 g
0.3 g
For ½
50 mL
0.4 g
0.15 g
Dissolve 0.8 g of ascorbic acid and 0.3 g of sodium laureth sulfate in 100 mL of Stock
sodium molybdate. Prepare fresh daily.
*Stir on highest speed (cover top with parafilm) until all SLS dissolves (~ 10 minutes).
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16