Special Solutions And Reagents
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********************{ Special Solutions And Reagents }*************************
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ALUMINON(Qualitative test for aluminum): Aluminon is the trade name for the
ammonium salt of aurin tricarboxylic acid.Dissolve 1g of the salt in 1 liter of
distilled water.Shake the solution well to insure thorough mixing.
BANG'S REAGENT(For glucose estimation): Dissolve 100g of K CO ,66g of KCl and
2 3
160g of KHCO in about 700ml of water at 30 degrees Celcius.Add 4.4g of CuSO
3 4
dilute to 1 liter after the CO has evolved.This solution should only be shaken
2
in such a manner as to prevent the entry of air.After 24 hours 300ml are diluted
to 1 liter with saturated KCl solution,shake gently and use after 24 hours;50ml
equivalent to 10mg glucose.
BARFOED'S REAGENT: SEE CUPRIC ACETATE.
BAUDISH'S REAGENT: SEE CUPFERRON.
BENEDICT'S REAGENT(Qualitative reagent for glucose): With the aid of heat
dissolve 173g of sodium citrate and 100g of Na CO in 800ml of water.Filter if
2 2
necessary,and dilute to 850ml.Dissolve 17.3g of CuSO 5H O in 100ml of water.
4 2
Pour the latter solution ,with constant stirring,into the carbonate-citrate
solution and make up to 1 liter.
BENZADINE HYDROCHLORIDE SOLUTION(For sulphate determination):Make a paste of 8g
of (C H (NH ) 2HCl)[Benzidine Hydrochloride] and 20ml of water,add 20ml of
12 5 2 2
HCl(spec.grade 1.12) and dilute to 1 liter with water.Each ml of this solution
is equivalent to 0.00357g of H SO .
2 4
BERTRAND'S REAGENT(Glucose estimation): Consists of the following solutions:
A) Dissolve 200g of rochelle salts and 150g of NaOH in enough water to make 1
liter.
B) Dissolve 40g of CuSO in enough water to make 1 liter of solution.
4
C) Dissolve 50g of Fe (SO ) and 200g of H SO (spec.grade 1.84) in enough
2 4 3 2 4
water to make 1 liter of solution.
D) Dissolve 5g of KMnO in enough water to make 1 liter of solution.
4
BIAL'S REAGENT(For pentose): Dissolve 1g of CH C H (OH) [orcinol] in 500ml of
3 6 3 2
30% HCl to which 30 drops of a 10% solution of FeCl has been added.
2
BOUTRON-BOUDET SOAP SOLUTION:
A) Dissolve 100g of pure castile soap in about 2500ml of 56% ethyl alcohol.
B) Dissolve 0.59g of Ba(NO ) in 1 liter of water.
3 2
Adjust the castile soap solution so that 2.4ml of it will give a permanent
lather with 40ml of solution B).When adjusted,2.4ml of soap solution is
equivalent to 220 parts per million of hardness(as CaCO ) for a 40ml sample.
3
BRUCK'S REAGENT(Protein precipitation): SEE POTASSIUM IODIDE-MERCURIC IODIDE
Clarke's Soap Solution:Estimation of hardness in water.
A) Dissolve 100g of pure powdered castile soap in 1 liter of 80% ethyl alcohol
and allow to stand overnight.
B) Prepare a standard solution of CaCO in HCl(spec.grade 1.19),neutralize with
3
NH OH and make slightly alkaline to litmus,and dilute to 500ml.1ml is equivalent
4
to 1mg of CaCO . Titrate (A) against (B) and dilute (A) with 80% ethyl alcohol
3
until 1ml of the resulting solution is equivalent to 1ml of (B)after making
allowance for the lather factor(the amount of standard soap solution required
to produce a permanent lather in 50ml of distilled water).1ml of the adjusted
solution after subtracting the lather factor is equivalent to 1mg of CaCO .
3
COBALTICYANIDE SOLUTION(Rinnmann's test for Zn): Dissolve 4g of K Co(CN) and 1g
3 6
KClO in 100ml of water.Soak filter paper in solution and dry at 100deg.Celcius
3
Apply drop of zinc solution and burn in an evaporating dish.A green disk is
obtained if zinc is present.
COCHINEAL: Extract 1g of cochineal for 4 days with 20ml of alcohol and 60ml of
distilled water.Filter.
CONGO RED: Dissolve 0.5g of congo red in 90ml of distilled water and 10ml of
alcohol.
CUPFERRON(For iron analysis): Dissolve 6g of the ammonium salt of nitroso-
phenyl-hydroxyl-amine in 100ml of H O.Reagent is good for 1 week only and must
2
be kept in the dark.
CUPRIC ACETATE(For reducing mono-saccharides): Dissolve 66g of cupric acetate
and 10ml of glacial acetic acid in water and dilute to 1 liter.
CUPRIC OXIDE,AMMONIACAL(For dissolving cotton,silk and linen,but not wool):
1): Dissolve 5g of cupric sulphate in 100ml of boiling water and add sodium
hydroxide until precipitation is complete.Wash the precipitate well and
dissolve it in a quantity of ammonium hydroxide.
2): Bubble a slow stream of air through 300ml of strong ammonium hydroxide
containing 50g of fine copper turnings,continue for 1 hour.
CUPRIC SULPHATE IN GLYCERIN-POTASSIUM HYDROXIDE(Reagent for silk): Dissolve 10g
of cupric sulphate in 100ml of water and add 5g of glycerin.Add KOH solution
slowly until a deep blue solution is obtained.
CUPRON(Benzoin Oxime): Dissolve 5g in 100ml of 95% alcohol.
CUPROUS CHLORIDE,ACIDIC(Reagent for carbon dioxide in gas analysis):
1): Cover the bottom of a 2 liter flask with a layer of cupric oxide about 1/2
inch deep,suspend a bunch of copper wire so as to reach from the bottom of the
flask to the top of the solution,and fill the flask with HCl(spec.grade 1.10).
Shake occasionally.When the solution becomes nearly colorless,transfer to
reagent bottles,which should also contain copper wire.The stock bottle may be
refilled with dilute HCl until either the cupric oxide or the wire is used
up.Copper sulphate may be substituted for copper oxide in the above procedure.
2): Dissolve 340g of CuCl 2H O in 600ml of conc.HCl and reduce the cupric
2 2
chloride by adding 190ml of a saturated solution of stannous chloride or until
the solution is colorless.The stannous chloride is prepared by treating 300g of
metallic tin in a 500ml flask with conc. HCl until no more tin goes into
solution.
3): Add a mixture of 86g of CuO and 17g of finely divided matallic Cu,made by
the reduction of CuO with hydrogen,to a solution of HCl,made by diluting 650ml
of conc. HCl with 325ml of water.After the mixture has been added slowly and
with frequent stirring,a spiral of copper wire is suspended in the bottle,
reaching all the way to the bottom.Shake occasionally,and when the solution
becomes colorless it is ready for use.
CUPROUS CHLORIDE,AMMONIACAL(Reagent for CO in gas analysis):
2
1): The acid solution of cuprous chloride as prepared above is neutralized with
ammonium hydroxide until an ammonia odor persists.An excess of metallic copper
must be kept in the solution.
2): Pour 800ml of acidic cuprous chloride prepared by method (3) into 4 liters
of water.Transfer the precipitate to a 250ml graduated tube.After several
hours,siphon off the liquid above the 50ml mark and refill with 7.5% NH OH
4
solution which may be prepared by diluting 50ml of conc. NH OH with 150ml of
4
water.The solution is well shaken and allowed to stand for several hours.It
should have a faint odor of ammonia.
DICHLOROFLOURECSEIN INDICATOR: Dissolve 1g in 1 liter of 70% alcohol or 1g of
the sodium salt in 1 liter of water.
DIMETHYLGLYOXIME: 0.01N;Dissolve 0.6g of Dimethylglyoxime,(CH CNOH) ,in 500ml
3 2
of 95% ethyl alcohol.This is an especially sensitive test for nickel,a very
definite crimson color being produced.
DIPHENYLAMINE(Reagent for rayon):Dissolve 0.2g in 100ml of conc. sulfuric acid.
DIPHENYLAMINE SULPHONATE(For titration of iron with K Cr O ):
2 2 7
Dissolve 0.32g of the barium salt of diphenylamine sulphonic acid in 100ml of
water,add 0.5g of sodium sulphate and filter off the precipitate of BaSO .
4
DIPHENYLCARBAZIDE: Dissolve 0.2g of diphenylcarbazide in 10ml of glacial acetic
acid and dilute to 100ml with 95% ethyl alcohol.
ESBACH'S REAGENT(Estimation of protein): To a water solution of 10g of picric
acid and 20g of citric acid,add sufficient water to make 1 liter of solution.
ESCHKA'S COMPOUND: 2 parts of calcined (light) magnesia are thoroughly mixed
with 1 part of anhydrous sodium carbonate.
FEHLING'S SOLUTION(Reagent for reducing sugars):
A):Dissolve 34.66g of copper sulphate in water and dilute to 500ml.
B):Dissolve 173g of potassium tartrate and 50g of NaOH in water and dilute when
cold to 500ml.
For use,mix equal volumes of both solutions at time of use.
FERRIC-ALUM INDICATOR: Dissolve 140g of ferric-ammonium sulphate crystals in
400ml of hot water.When cool,filter;and make up to a volume of 500ml with
dilute (6N)nitric acid.
FOLIN'S MIXTURE(For uric acid): To 650ml of water add 500g of (NH ) SO ,5g of
4 2 4
uranium acetate and 6g of glacial acetic acid.Dilute to 1 liter.
FORMALDEHYDE-SULPHURIC ACID(Marquis' reagent for alkalloids):Add 10ml of
formaldehyde solution to 50ml of sulphuric acid.
FUCHSIN(Reagent for linen): Dissolve 1g of fuchsin in 100ml of alcohol.
FUCHSIN-SULPHUROUS ACID(Schiff's reagent for aldehydes):Dissolve .5g of fuchsin
and 9g of sodium-bisulphite in 500ml of water,and add 10ml of HCl.Keep in well
stoppered bottles and protect from light.
GUNZBERG'S REAGENT(Detection of HCl in gastric juice): Prepare as needed a
solution containing 4g of phloroglucinol and 2g of vanillin in 100ml of
absolute ethyl alcohol.
HANUS SOLUTION(For iodine number): Dissolve 13.2g of resublimed iodine in 1
liter of glacial acetic acid which will pass the dichromate test for reducible
matter.Add sufficient bromine to double the halogen content,determined by
titration(3ml is about the proper amount).The iodine may be dissolved by the
aid of heat,but the solution should be cold when the bromine is added.
IODINE,TINCTURE OF: To 50ml of water add 70g of I and 50g of KI.Dilute to 1
2
liter with alcohol.
IODO-POTASSIUM IODIDE:Dissolve 2g of iodine and 6g of KI in 100ml of water.
LITMUS(Indicator):Extract litmus powder 3 times with alcohol.each treatment for
1 hour.Reject the alcohol extract.Treat residue with an equal weight of cold
water and filter;then exhaust with 5 times it's weight in boiling water,cool
and filter.Combine with aqueous extracts.
MAGNESIA MIXTURE(Reagent for phosphates and arsenates): Dissolve 55g of
magnesium chloride and 105g of ammonium chloride in water,barely acidify with
hydrochloric acid,and dilute to 1 liter.The ammonium hydroxide may be omitted
until just previous to use.The reagent,if completely mixed and stored for any
length of time becomes turbid.
MAGNESIUM URANYL ACETATE: Dissolve 100g of UO (C H O ) 2H O in 60ml of glacial
2 2 3 2 2 2
acetic acid and dilute to 500ml.Dissolve 330g of Mg(C H O ) 2H O in 60ml of
2 3 2 2 2
glacial acetic acid and dilute to 200ml.Heat solutions to the boiling point
until clear,pour the magnesium solution into the uranyl solution,cool and
dilute to 21 liter.Let stand overnight and filter if necessary.
MAYER'S REAGENT
(White precipitate with most alkaloids in slightly acid solutions)
Dissolve 1.358g of HgCl in 60ml of water and pour into a solution of 5g of KI
2
in 10ml of pure water.Add sufficient water to make 100ml.
METHYL ORANGE INDICATOR: Dissolve 1g of methyl orange in 1 liter of water.
Filter if necessary.
METHYL ORANGE(Modified):Dissolve 2g of methyl orange and 2.8g of xylene cyanole
FF in 1 liter of 50% alcohol.
METHYL RED INDICATOR: Dissolve 1g of methyl red in 600ml of alcohol and dilute
with 400ml of water.
METHYL RED(Modified): Dissolve 0.5g of methyl red and 1.25ml of xylene cyanole
FF in 1 liter of 90% alcohol.Or,dissolve 1.25g of methyl red and 0.825g of
methylene blue in 1 liter of 90% alcohol.
MILLON'S REAGENT(For albumin's and phenols): Dissolve 1 part of mercury in 1
part of cold fuming nitric acid.Dilute with twice the volume of water and
decant the clear solution after several hours.
MIXED INDICATOR: Prepared by adding about 1.4g of xylene cyanole FF to 1g of
methyl orange.The dye is seldom pure enough for these proportions to be
satisfactory.Each new lot of dye should be tested by adding addl.amounts of dye
until a test portion gives the proper color change.The acid color of this
indicator is that permaganate;the neutral color is gray;the alkaline color is
green.
NAPTHOL(Molisch's reagent for wool):Dissolve 15g of napthol in 100ml of alcohol
or chloroform.
NESSLER'S REAGENT(For ammonia): Dissolve 50g of KI in the smallest possible
quantity of cold water(50ml).Add a saturated solution of mercuric chloride
(about 22g in 350ml of water)until an excess is indicated by the formation of a
precipitate.Then add 200ml of 5N NaOH and dilute to 1 liter.Let settle and draw
off the clear liquid.
NICKEL OXIDE AMMONIACAL(Reagent for silk): Dissolve 5g of nickel sulphate in
100ml of water,add sodium hydroxide solution until nickel oxide is completely
precipitated.Wash the precipitate well and dissolve in 25ml of conc. ammonium
hydroxide and 25ml of water.
NITROBENZENE-AZO-RESORCINOL(Reagent for magnesium): Dissolve 1g of the dye in
10ml of N NaOH and dilute to 1 liter.
NITRON(Detection of nitrate-radical): Dissolve 10g of nitron in 5ml of glacial
acetic acid and 95ml of water.The solution may be filtered with slight suction
through an alundum crucible and kept in a dark bottle.
NYLANDER'S SOLUTION(Carbohydrates): Dissolve 20g of bismuth subnitrate and 40g
of rochelle salt in 1 liter of 8% NaOH solution.Cool and filter.
OBERMAYER'S REAGENT(For indoxyl in urine): Dissolve 4g of FeCl in 1 liter of
2
HCl(spec.grade 1.19).
OXINE: Dissolve 14g of HC H ON in 30ml of glacial acetic acid.Warm slightly if
9 6
necessary.Dilute to 1 liter.
OXYGEN ABSORBANT: Dissolve 300g of ammonium chloride in 1 liter of water and
add 1 liter of conc. ammonium hydroxide solution.Shake the solution thoroughly.
For use as an oxygen absorbent,a bottle half full of copper turnings is filled
nearly half full with the solution and the gas is passed through.
PAVY'S SOLUTION(Glucose reagent): To 120ml of "Fehling's solution",add 300ml of
NH OH (spec. grade 0.88)and dilute with 1 liter of water.
4
PHENANTHROLINE FERROUS ION INDICATOR: Dissolve 1.485g of phenanthroline
monohydrate in 100ml of 0.0025M ferrous sulphate solution.
PHENOLPHETHALINE: Dissolve 1g of phenolphethaline in 50ml of alcohol and add
50ml of water.
PHENOLSULFONIC ACID(determination of nitrogen as nitrate): Dissolve 25g of
phenol in 150ml of conc. sulphuric acid,add 75ml of fuming sulphuric acid.Stir
well and heat at 100 degrees celcius for 2 hours.
PHOSPHORIC ACID-SULPHURIC ACID MIXTURE: Dilute 150ml of conc. H SO with 100ml
2 4
of H PO (85%) with water to a volume of 1 liter.
3 4
PICRIC ACID(For alkaloids,wool and silk):Dissolve 100ml of picric acid in 100ml
of water.
POTASSIUM ANTIMONATE(Reagent for sodium): Boil 22g of potassium antimonate in 1
liter of water until nearly all the salt has dissolved;cool quickly and add
35ml of 10% potassium hydroxide.Filter after standing overnight.
POTASSIUM-CADMIUM IODIDE(Reagent for alkaloids): Add 2g of CdI to a boiling
2
solution of 4g of KI in 12ml of water, then mix with 12ml of saturated KI
solution.
POTASSIUM HYDROXIDE(For carbon dioxide absorption): Dissolve 300g of KOH in
water and dilute to 1 liter.
POTASSIUM IODIDE-MERCURIC IODIDE (Brucke's reagent for proteins):
Dissolve 50g of KI in 500ml of water, and saturate with mercuric iodide (about
120g).Dilute to 1 liter.
POTASSIUM PYROGALLATE (for oxygen absorption): For mixtures of gases containing
less than 28% oxygen,add 100ml of KOH solution (50g of KOH to 100ml of water)
to 5g of pyrogallol.For mixtures containing more than 28% oxygen,the KOH
solution should contain 120g of KOH to 100ml of water.
PYROGALLOL, ALKALINE:
Create the 2 following solutions:
a) Dissolve 75g of pyrogallic acid in 75ml of water.
b) Dissolve 500g of KOH in 250ml of water. When cool,adjust until sp.
gr. is 1.55.for use, add 270ml of solution (b) to 30ml of solution (a).
ROSOLIC ACID (indicator):Dissolve 1g of rosolic acid in 10ml of alcohol and add
100ml of water.
S AND O REAGENT (Suitsu and Okuma's test for Mg): Dissolve 0.5g of the dye
(o-p-dihydroxy-monazo-p-nitrobenzene) in 100ml of 0.25N NaOH
SCHEIBLER'S REAGENT: SEE PHOSPHOTUNGSTIC ACID
SCHIFF'S REAGENT: SEE FUCHSIN-SULFUROUS ACID
SCHWEITZER'S REAGENT: SEE CUPRIC OXIDE, AMMONIACAL
SOAP SOLUTION:(Reagent for hardness in water).Dissolve 100g of dry castile soap
in 1 liter of 80% alcohol (5 parts alcohol to 1 part water).Allow to stand
several days and dilute with 70% to 80% alcohol until 6.4ml produces a
permanent lather with 20ml of standard calcium solution.The latter solution is
made by dissolving 0.2g of CaCO in a small amount of dilute HCL,evaporating to
3
dryness and making up to 1 liter.
SODIUM BISMUTHATE (oxidation of manganese): Heat 20 parts of NaOH nearly to
redness in an iron or nickel crucible and add slowly 10 parts of basic bismuth
nitrate which has been previously dried.Add two parts of sodium peroxide and
pour the brownish-yellow fused mass on an iron plate to cool.When cold,break up
in a mortar,extract with water,and collect on an asbestos filter.
SODIUM HYDROXIDE (for CO absorption): Dissolve 330g of NaOH in water and
2
dilute to 1 liter.
SODIUM NITROPRUSSIDE (reagent for hydrogen sulfide and wool): Use a freshly
prepared solution of 1g of sodium nitroprusside in 10ml of water.
SODIUM OXALATE: According to Sorensen (primary standard). Dissolve 30g of the
commercial salt in 1 liter of water,make slightly alkaline with sodium
hydroxide,and let stand until perfectly clear.Filter and evaporate the filtrate
to 100ml.Cool and filter.Pulverize the residue and wash it several times with
small volumes of water.The procedure is repeated until the mother liquor is
free from sulfate and is neutral to phenolphethaline.
SODIUM PLUMBITE (reagent for wool): Dissolve 5g of sodium hydroxide in 100ml of
water.Add 5g of litharge and boil until dissolved.
SODIUM POLYSULFIDE: Dissolve 480g of Na 8.9H O in 500ml of water,add 40g of
2
NaOH and 18g of sulfur.Stir thoroughly and dilute to 1 liter with water.
SONNENSCHEIN'S REAGENT: SEE PHOSPHOMOLYBDIC ACID
STARCH SOLUTION:
1. Make a paste with 2g of soluble starch and 0.01g of HgI with a small
2
amount of water.Add the mixture slowly to 1 liter of boiling water and boil for
a few minutes.Keep in a glass stoppered bottle.If other than soluble starch is
used,the solution will not clear on boiling;it should be allowed to stand and
the clear liquid decanted.
2. A solution of starch which keeps indefinitely is made as follows:Mix 500ml
of saturated NaCl solution (filtered),80ml of glacial acetic acid,20ml of water
and 3g of starch.Bring slowly to a boil and boil for two minutes.
3. Make a paste with 1g of soluble starch and 5mg of HgI using as little
2
cold water as possible.Then pour about 200ml of boiling water on the paste and
stir immediately.This will give a clear solution if the paste is prepared
correctly and the water actually boiling.Cool and add 4g of KI.Starch solution
decomposes on standing due to bacterial action,but this solution will keep a
long time if stored under a layer of Toluene.
STOKE'S REAGENT: Dissolve 30g of FeSO and 20g of tartaric acid in water and
4
dilute to 1 liter.Just before using,add concentrated NH OH until the
4
precipitate first formed is redissolved.
STOKES REAGENT:Dissolve 30g of FeSO and 20g of tartaric acid in water and
4
dilute to 1 liter.Just before using,add concentrated NH OH until the
4
precipitate first formed is redissolved.
SULFANILLIC ACID: (Reagent for nitrates)Dissolve 0.5g of sulfanillic acid in a
mixture of 15ml glacial acetic acid and 135ml of recently boiled water.
SULFOMOLYBDIC ACID: (Reagent for alkaloids and glucosides)Dissolve 10g of
molybdic acid or sodium molybdate in 100ml of conc.H SO .
2 4
TANNIC ACID: (Reagent for albumin,alkaloids and gelatin)Dissolve 10g of tannic
acid in 10ml of alcohol and dilute with water to 100ml.
o-TOLIDINE SOLUTION: (Residual chlorine in water analysis)Prepare 1 liter of
dilute HCl(100ml - HCl/sp.gr. 1.19) in sufficient water to make 1 liter.
Dissolve 1g of o-tolidine in 100ml of dilute HCl and dilute to 1 liter with
dilute HCl solution.
TURMERIC PAPER: Impregnate white,unsized paper with the tincture and dry.
TURMERIC TINCTURE: Digest ground turmeric root with several quantities of water
which are discarded.Dry the residue and digest it several days with 6 times it
weight of alcohol,filter.
UFFELMANN'S REAGENT: To a 2% solution of pure phenol in water,add a water
solution of FeCl until the phenol solution becomes violet in color.
2
WAGNERS SOLUTION: Dissolve 25g of citric acid and 1g of salicylic acid in water
and dilute to 1 liter.Use 50ml of the reagent.
WIJ'S IODINE MONOCHLORIDE SOLUTION:Dissolve 13g of resublimed iodine in 1 liter
of glacial acetic acid which will pass the dichromate test for reductible
matter.Set aside 25ml of this solution.Pass into the remainder of the solution
dry chlorine gas until the characteristic color of free iodine has been
discharged.Now add the iodine solution which was reserved,until all free
chlorine has been destroyed.A slight excess of iodine does little or no harm,
but an excess of chlorine must be avoided.Preserve in well stoppered amber
colored bottles.Avoid usage of solutions older than 30 days.
WIJ'S SPECIAL SOLUTION: To 200ml of glacial acetic acid that will pass the
dichromate test for reductible matter,add 12g of dichloroamine T(paratoluene-
sulphonedichloroamide) and add 16.6g of dry KI(in small quantities until all of
the KI has dissolved).Make up to 1 liter with the same quality of acetic acid
used above and preserve in dark colored bottles.
ZIMMERMAN-REINHARDT SOLUTION: Dissolve 70g of MnSO x 4H O in 500ml of water,add
4 2
125ml of conc. H SO and 125ml of 85% H PO and dilute to 1 liter.
********************{ Special Solutions And Reagents }*************************
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ALUMINON(Qualitative test for aluminum): Aluminon is the trade name for the
ammonium salt of aurin tricarboxylic acid.Dissolve 1g of the salt in 1 liter of
distilled water.Shake the solution well to insure thorough mixing.
BANG'S REAGENT(For glucose estimation): Dissolve 100g of K CO ,66g of KCl and
2 3
160g of KHCO in about 700ml of water at 30 degrees Celcius.Add 4.4g of CuSO
3 4
dilute to 1 liter after the CO has evolved.This solution should only be shaken
2
in such a manner as to prevent the entry of air.After 24 hours 300ml are diluted
to 1 liter with saturated KCl solution,shake gently and use after 24 hours;50ml
equivalent to 10mg glucose.
BARFOED'S REAGENT: SEE CUPRIC ACETATE.
BAUDISH'S REAGENT: SEE CUPFERRON.
BENEDICT'S REAGENT(Qualitative reagent for glucose): With the aid of heat
dissolve 173g of sodium citrate and 100g of Na CO in 800ml of water.Filter if
2 2
necessary,and dilute to 850ml.Dissolve 17.3g of CuSO 5H O in 100ml of water.
4 2
Pour the latter solution ,with constant stirring,into the carbonate-citrate
solution and make up to 1 liter.
BENZADINE HYDROCHLORIDE SOLUTION(For sulphate determination):Make a paste of 8g
of (C H (NH ) 2HCl)[Benzidine Hydrochloride] and 20ml of water,add 20ml of
12 5 2 2
HCl(spec.grade 1.12) and dilute to 1 liter with water.Each ml of this solution
is equivalent to 0.00357g of H SO .
2 4
BERTRAND'S REAGENT(Glucose estimation): Consists of the following solutions:
A) Dissolve 200g of rochelle salts and 150g of NaOH in enough water to make 1
liter.
B) Dissolve 40g of CuSO in enough water to make 1 liter of solution.
4
C) Dissolve 50g of Fe (SO ) and 200g of H SO (spec.grade 1.84) in enough
2 4 3 2 4
water to make 1 liter of solution.
D) Dissolve 5g of KMnO in enough water to make 1 liter of solution.
4
BIAL'S REAGENT(For pentose): Dissolve 1g of CH C H (OH) [orcinol] in 500ml of
3 6 3 2
30% HCl to which 30 drops of a 10% solution of FeCl has been added.
2
BOUTRON-BOUDET SOAP SOLUTION:
A) Dissolve 100g of pure castile soap in about 2500ml of 56% ethyl alcohol.
B) Dissolve 0.59g of Ba(NO ) in 1 liter of water.
3 2
Adjust the castile soap solution so that 2.4ml of it will give a permanent
lather with 40ml of solution B).When adjusted,2.4ml of soap solution is
equivalent to 220 parts per million of hardness(as CaCO ) for a 40ml sample.
3
BRUCK'S REAGENT(Protein precipitation): SEE POTASSIUM IODIDE-MERCURIC IODIDE
Clarke's Soap Solution:Estimation of hardness in water.
A) Dissolve 100g of pure powdered castile soap in 1 liter of 80% ethyl alcohol
and allow to stand overnight.
B) Prepare a standard solution of CaCO in HCl(spec.grade 1.19),neutralize with
3
NH OH and make slightly alkaline to litmus,and dilute to 500ml.1ml is equivalent
4
to 1mg of CaCO . Titrate (A) against (B) and dilute (A) with 80% ethyl alcohol
3
until 1ml of the resulting solution is equivalent to 1ml of (B)after making
allowance for the lather factor(the amount of standard soap solution required
to produce a permanent lather in 50ml of distilled water).1ml of the adjusted
solution after subtracting the lather factor is equivalent to 1mg of CaCO .
3
COBALTICYANIDE SOLUTION(Rinnmann's test for Zn): Dissolve 4g of K Co(CN) and 1g
3 6
KClO in 100ml of water.Soak filter paper in solution and dry at 100deg.Celcius
3
Apply drop of zinc solution and burn in an evaporating dish.A green disk is
obtained if zinc is present.
COCHINEAL: Extract 1g of cochineal for 4 days with 20ml of alcohol and 60ml of
distilled water.Filter.
CONGO RED: Dissolve 0.5g of congo red in 90ml of distilled water and 10ml of
alcohol.
CUPFERRON(For iron analysis): Dissolve 6g of the ammonium salt of nitroso-
phenyl-hydroxyl-amine in 100ml of H O.Reagent is good for 1 week only and must
2
be kept in the dark.
CUPRIC ACETATE(For reducing mono-saccharides): Dissolve 66g of cupric acetate
and 10ml of glacial acetic acid in water and dilute to 1 liter.
CUPRIC OXIDE,AMMONIACAL(For dissolving cotton,silk and linen,but not wool):
1): Dissolve 5g of cupric sulphate in 100ml of boiling water and add sodium
hydroxide until precipitation is complete.Wash the precipitate well and
dissolve it in a quantity of ammonium hydroxide.
2): Bubble a slow stream of air through 300ml of strong ammonium hydroxide
containing 50g of fine copper turnings,continue for 1 hour.
CUPRIC SULPHATE IN GLYCERIN-POTASSIUM HYDROXIDE(Reagent for silk): Dissolve 10g
of cupric sulphate in 100ml of water and add 5g of glycerin.Add KOH solution
slowly until a deep blue solution is obtained.
CUPRON(Benzoin Oxime): Dissolve 5g in 100ml of 95% alcohol.
CUPROUS CHLORIDE,ACIDIC(Reagent for carbon dioxide in gas analysis):
1): Cover the bottom of a 2 liter flask with a layer of cupric oxide about 1/2
inch deep,suspend a bunch of copper wire so as to reach from the bottom of the
flask to the top of the solution,and fill the flask with HCl(spec.grade 1.10).
Shake occasionally.When the solution becomes nearly colorless,transfer to
reagent bottles,which should also contain copper wire.The stock bottle may be
refilled with dilute HCl until either the cupric oxide or the wire is used
up.Copper sulphate may be substituted for copper oxide in the above procedure.
2): Dissolve 340g of CuCl 2H O in 600ml of conc.HCl and reduce the cupric
2 2
chloride by adding 190ml of a saturated solution of stannous chloride or until
the solution is colorless.The stannous chloride is prepared by treating 300g of
metallic tin in a 500ml flask with conc. HCl until no more tin goes into
solution.
3): Add a mixture of 86g of CuO and 17g of finely divided matallic Cu,made by
the reduction of CuO with hydrogen,to a solution of HCl,made by diluting 650ml
of conc. HCl with 325ml of water.After the mixture has been added slowly and
with frequent stirring,a spiral of copper wire is suspended in the bottle,
reaching all the way to the bottom.Shake occasionally,and when the solution
becomes colorless it is ready for use.
CUPROUS CHLORIDE,AMMONIACAL(Reagent for CO in gas analysis):
2
1): The acid solution of cuprous chloride as prepared above is neutralized with
ammonium hydroxide until an ammonia odor persists.An excess of metallic copper
must be kept in the solution.
2): Pour 800ml of acidic cuprous chloride prepared by method (3) into 4 liters
of water.Transfer the precipitate to a 250ml graduated tube.After several
hours,siphon off the liquid above the 50ml mark and refill with 7.5% NH OH
4
solution which may be prepared by diluting 50ml of conc. NH OH with 150ml of
4
water.The solution is well shaken and allowed to stand for several hours.It
should have a faint odor of ammonia.
DICHLOROFLOURECSEIN INDICATOR: Dissolve 1g in 1 liter of 70% alcohol or 1g of
the sodium salt in 1 liter of water.
DIMETHYLGLYOXIME: 0.01N;Dissolve 0.6g of Dimethylglyoxime,(CH CNOH) ,in 500ml
3 2
of 95% ethyl alcohol.This is an especially sensitive test for nickel,a very
definite crimson color being produced.
DIPHENYLAMINE(Reagent for rayon):Dissolve 0.2g in 100ml of conc. sulfuric acid.
DIPHENYLAMINE SULPHONATE(For titration of iron with K Cr O ):
2 2 7
Dissolve 0.32g of the barium salt of diphenylamine sulphonic acid in 100ml of
water,add 0.5g of sodium sulphate and filter off the precipitate of BaSO .
4
DIPHENYLCARBAZIDE: Dissolve 0.2g of diphenylcarbazide in 10ml of glacial acetic
acid and dilute to 100ml with 95% ethyl alcohol.
ESBACH'S REAGENT(Estimation of protein): To a water solution of 10g of picric
acid and 20g of citric acid,add sufficient water to make 1 liter of solution.
ESCHKA'S COMPOUND: 2 parts of calcined (light) magnesia are thoroughly mixed
with 1 part of anhydrous sodium carbonate.
FEHLING'S SOLUTION(Reagent for reducing sugars):
A):Dissolve 34.66g of copper sulphate in water and dilute to 500ml.
B):Dissolve 173g of potassium tartrate and 50g of NaOH in water and dilute when
cold to 500ml.
For use,mix equal volumes of both solutions at time of use.
FERRIC-ALUM INDICATOR: Dissolve 140g of ferric-ammonium sulphate crystals in
400ml of hot water.When cool,filter;and make up to a volume of 500ml with
dilute (6N)nitric acid.
FOLIN'S MIXTURE(For uric acid): To 650ml of water add 500g of (NH ) SO ,5g of
4 2 4
uranium acetate and 6g of glacial acetic acid.Dilute to 1 liter.
FORMALDEHYDE-SULPHURIC ACID(Marquis' reagent for alkalloids):Add 10ml of
formaldehyde solution to 50ml of sulphuric acid.
FUCHSIN(Reagent for linen): Dissolve 1g of fuchsin in 100ml of alcohol.
FUCHSIN-SULPHUROUS ACID(Schiff's reagent for aldehydes):Dissolve .5g of fuchsin
and 9g of sodium-bisulphite in 500ml of water,and add 10ml of HCl.Keep in well
stoppered bottles and protect from light.
GUNZBERG'S REAGENT(Detection of HCl in gastric juice): Prepare as needed a
solution containing 4g of phloroglucinol and 2g of vanillin in 100ml of
absolute ethyl alcohol.
HANUS SOLUTION(For iodine number): Dissolve 13.2g of resublimed iodine in 1
liter of glacial acetic acid which will pass the dichromate test for reducible
matter.Add sufficient bromine to double the halogen content,determined by
titration(3ml is about the proper amount).The iodine may be dissolved by the
aid of heat,but the solution should be cold when the bromine is added.
IODINE,TINCTURE OF: To 50ml of water add 70g of I and 50g of KI.Dilute to 1
2
liter with alcohol.
IODO-POTASSIUM IODIDE:Dissolve 2g of iodine and 6g of KI in 100ml of water.
LITMUS(Indicator):Extract litmus powder 3 times with alcohol.each treatment for
1 hour.Reject the alcohol extract.Treat residue with an equal weight of cold
water and filter;then exhaust with 5 times it's weight in boiling water,cool
and filter.Combine with aqueous extracts.
MAGNESIA MIXTURE(Reagent for phosphates and arsenates): Dissolve 55g of
magnesium chloride and 105g of ammonium chloride in water,barely acidify with
hydrochloric acid,and dilute to 1 liter.The ammonium hydroxide may be omitted
until just previous to use.The reagent,if completely mixed and stored for any
length of time becomes turbid.
MAGNESIUM URANYL ACETATE: Dissolve 100g of UO (C H O ) 2H O in 60ml of glacial
2 2 3 2 2 2
acetic acid and dilute to 500ml.Dissolve 330g of Mg(C H O ) 2H O in 60ml of
2 3 2 2 2
glacial acetic acid and dilute to 200ml.Heat solutions to the boiling point
until clear,pour the magnesium solution into the uranyl solution,cool and
dilute to 21 liter.Let stand overnight and filter if necessary.
MAYER'S REAGENT
(White precipitate with most alkaloids in slightly acid solutions)
Dissolve 1.358g of HgCl in 60ml of water and pour into a solution of 5g of KI
2
in 10ml of pure water.Add sufficient water to make 100ml.
METHYL ORANGE INDICATOR: Dissolve 1g of methyl orange in 1 liter of water.
Filter if necessary.
METHYL ORANGE(Modified):Dissolve 2g of methyl orange and 2.8g of xylene cyanole
FF in 1 liter of 50% alcohol.
METHYL RED INDICATOR: Dissolve 1g of methyl red in 600ml of alcohol and dilute
with 400ml of water.
METHYL RED(Modified): Dissolve 0.5g of methyl red and 1.25ml of xylene cyanole
FF in 1 liter of 90% alcohol.Or,dissolve 1.25g of methyl red and 0.825g of
methylene blue in 1 liter of 90% alcohol.
MILLON'S REAGENT(For albumin's and phenols): Dissolve 1 part of mercury in 1
part of cold fuming nitric acid.Dilute with twice the volume of water and
decant the clear solution after several hours.
MIXED INDICATOR: Prepared by adding about 1.4g of xylene cyanole FF to 1g of
methyl orange.The dye is seldom pure enough for these proportions to be
satisfactory.Each new lot of dye should be tested by adding addl.amounts of dye
until a test portion gives the proper color change.The acid color of this
indicator is that permaganate;the neutral color is gray;the alkaline color is
green.
NAPTHOL(Molisch's reagent for wool):Dissolve 15g of napthol in 100ml of alcohol
or chloroform.
NESSLER'S REAGENT(For ammonia): Dissolve 50g of KI in the smallest possible
quantity of cold water(50ml).Add a saturated solution of mercuric chloride
(about 22g in 350ml of water)until an excess is indicated by the formation of a
precipitate.Then add 200ml of 5N NaOH and dilute to 1 liter.Let settle and draw
off the clear liquid.
NICKEL OXIDE AMMONIACAL(Reagent for silk): Dissolve 5g of nickel sulphate in
100ml of water,add sodium hydroxide solution until nickel oxide is completely
precipitated.Wash the precipitate well and dissolve in 25ml of conc. ammonium
hydroxide and 25ml of water.
NITROBENZENE-AZO-RESORCINOL(Reagent for magnesium): Dissolve 1g of the dye in
10ml of N NaOH and dilute to 1 liter.
NITRON(Detection of nitrate-radical): Dissolve 10g of nitron in 5ml of glacial
acetic acid and 95ml of water.The solution may be filtered with slight suction
through an alundum crucible and kept in a dark bottle.
NYLANDER'S SOLUTION(Carbohydrates): Dissolve 20g of bismuth subnitrate and 40g
of rochelle salt in 1 liter of 8% NaOH solution.Cool and filter.
OBERMAYER'S REAGENT(For indoxyl in urine): Dissolve 4g of FeCl in 1 liter of
2
HCl(spec.grade 1.19).
OXINE: Dissolve 14g of HC H ON in 30ml of glacial acetic acid.Warm slightly if
9 6
necessary.Dilute to 1 liter.
OXYGEN ABSORBANT: Dissolve 300g of ammonium chloride in 1 liter of water and
add 1 liter of conc. ammonium hydroxide solution.Shake the solution thoroughly.
For use as an oxygen absorbent,a bottle half full of copper turnings is filled
nearly half full with the solution and the gas is passed through.
PAVY'S SOLUTION(Glucose reagent): To 120ml of "Fehling's solution",add 300ml of
NH OH (spec. grade 0.88)and dilute with 1 liter of water.
4
PHENANTHROLINE FERROUS ION INDICATOR: Dissolve 1.485g of phenanthroline
monohydrate in 100ml of 0.0025M ferrous sulphate solution.
PHENOLPHETHALINE: Dissolve 1g of phenolphethaline in 50ml of alcohol and add
50ml of water.
PHENOLSULFONIC ACID(determination of nitrogen as nitrate): Dissolve 25g of
phenol in 150ml of conc. sulphuric acid,add 75ml of fuming sulphuric acid.Stir
well and heat at 100 degrees celcius for 2 hours.
PHOSPHORIC ACID-SULPHURIC ACID MIXTURE: Dilute 150ml of conc. H SO with 100ml
2 4
of H PO (85%) with water to a volume of 1 liter.
3 4
PICRIC ACID(For alkaloids,wool and silk):Dissolve 100ml of picric acid in 100ml
of water.
POTASSIUM ANTIMONATE(Reagent for sodium): Boil 22g of potassium antimonate in 1
liter of water until nearly all the salt has dissolved;cool quickly and add
35ml of 10% potassium hydroxide.Filter after standing overnight.
POTASSIUM-CADMIUM IODIDE(Reagent for alkaloids): Add 2g of CdI to a boiling
2
solution of 4g of KI in 12ml of water, then mix with 12ml of saturated KI
solution.
POTASSIUM HYDROXIDE(For carbon dioxide absorption): Dissolve 300g of KOH in
water and dilute to 1 liter.
POTASSIUM IODIDE-MERCURIC IODIDE (Brucke's reagent for proteins):
Dissolve 50g of KI in 500ml of water, and saturate with mercuric iodide (about
120g).Dilute to 1 liter.
POTASSIUM PYROGALLATE (for oxygen absorption): For mixtures of gases containing
less than 28% oxygen,add 100ml of KOH solution (50g of KOH to 100ml of water)
to 5g of pyrogallol.For mixtures containing more than 28% oxygen,the KOH
solution should contain 120g of KOH to 100ml of water.
PYROGALLOL, ALKALINE:
Create the 2 following solutions:
a) Dissolve 75g of pyrogallic acid in 75ml of water.
b) Dissolve 500g of KOH in 250ml of water. When cool,adjust until sp.
gr. is 1.55.for use, add 270ml of solution (b) to 30ml of solution (a).
ROSOLIC ACID (indicator):Dissolve 1g of rosolic acid in 10ml of alcohol and add
100ml of water.
S AND O REAGENT (Suitsu and Okuma's test for Mg): Dissolve 0.5g of the dye
(o-p-dihydroxy-monazo-p-nitrobenzene) in 100ml of 0.25N NaOH
SCHEIBLER'S REAGENT: SEE PHOSPHOTUNGSTIC ACID
SCHIFF'S REAGENT: SEE FUCHSIN-SULFUROUS ACID
SCHWEITZER'S REAGENT: SEE CUPRIC OXIDE, AMMONIACAL
SOAP SOLUTION:(Reagent for hardness in water).Dissolve 100g of dry castile soap
in 1 liter of 80% alcohol (5 parts alcohol to 1 part water).Allow to stand
several days and dilute with 70% to 80% alcohol until 6.4ml produces a
permanent lather with 20ml of standard calcium solution.The latter solution is
made by dissolving 0.2g of CaCO in a small amount of dilute HCL,evaporating to
3
dryness and making up to 1 liter.
SODIUM BISMUTHATE (oxidation of manganese): Heat 20 parts of NaOH nearly to
redness in an iron or nickel crucible and add slowly 10 parts of basic bismuth
nitrate which has been previously dried.Add two parts of sodium peroxide and
pour the brownish-yellow fused mass on an iron plate to cool.When cold,break up
in a mortar,extract with water,and collect on an asbestos filter.
SODIUM HYDROXIDE (for CO absorption): Dissolve 330g of NaOH in water and
2
dilute to 1 liter.
SODIUM NITROPRUSSIDE (reagent for hydrogen sulfide and wool): Use a freshly
prepared solution of 1g of sodium nitroprusside in 10ml of water.
SODIUM OXALATE: According to Sorensen (primary standard). Dissolve 30g of the
commercial salt in 1 liter of water,make slightly alkaline with sodium
hydroxide,and let stand until perfectly clear.Filter and evaporate the filtrate
to 100ml.Cool and filter.Pulverize the residue and wash it several times with
small volumes of water.The procedure is repeated until the mother liquor is
free from sulfate and is neutral to phenolphethaline.
SODIUM PLUMBITE (reagent for wool): Dissolve 5g of sodium hydroxide in 100ml of
water.Add 5g of litharge and boil until dissolved.
SODIUM POLYSULFIDE: Dissolve 480g of Na 8.9H O in 500ml of water,add 40g of
2
NaOH and 18g of sulfur.Stir thoroughly and dilute to 1 liter with water.
SONNENSCHEIN'S REAGENT: SEE PHOSPHOMOLYBDIC ACID
STARCH SOLUTION:
1. Make a paste with 2g of soluble starch and 0.01g of HgI with a small
2
amount of water.Add the mixture slowly to 1 liter of boiling water and boil for
a few minutes.Keep in a glass stoppered bottle.If other than soluble starch is
used,the solution will not clear on boiling;it should be allowed to stand and
the clear liquid decanted.
2. A solution of starch which keeps indefinitely is made as follows:Mix 500ml
of saturated NaCl solution (filtered),80ml of glacial acetic acid,20ml of water
and 3g of starch.Bring slowly to a boil and boil for two minutes.
3. Make a paste with 1g of soluble starch and 5mg of HgI using as little
2
cold water as possible.Then pour about 200ml of boiling water on the paste and
stir immediately.This will give a clear solution if the paste is prepared
correctly and the water actually boiling.Cool and add 4g of KI.Starch solution
decomposes on standing due to bacterial action,but this solution will keep a
long time if stored under a layer of Toluene.
STOKE'S REAGENT: Dissolve 30g of FeSO and 20g of tartaric acid in water and
4
dilute to 1 liter.Just before using,add concentrated NH OH until the
4
precipitate first formed is redissolved.
STOKES REAGENT:Dissolve 30g of FeSO and 20g of tartaric acid in water and
4
dilute to 1 liter.Just before using,add concentrated NH OH until the
4
precipitate first formed is redissolved.
SULFANILLIC ACID: (Reagent for nitrates)Dissolve 0.5g of sulfanillic acid in a
mixture of 15ml glacial acetic acid and 135ml of recently boiled water.
SULFOMOLYBDIC ACID: (Reagent for alkaloids and glucosides)Dissolve 10g of
molybdic acid or sodium molybdate in 100ml of conc.H SO .
2 4
TANNIC ACID: (Reagent for albumin,alkaloids and gelatin)Dissolve 10g of tannic
acid in 10ml of alcohol and dilute with water to 100ml.
o-TOLIDINE SOLUTION: (Residual chlorine in water analysis)Prepare 1 liter of
dilute HCl(100ml - HCl/sp.gr. 1.19) in sufficient water to make 1 liter.
Dissolve 1g of o-tolidine in 100ml of dilute HCl and dilute to 1 liter with
dilute HCl solution.
TURMERIC PAPER: Impregnate white,unsized paper with the tincture and dry.
TURMERIC TINCTURE: Digest ground turmeric root with several quantities of water
which are discarded.Dry the residue and digest it several days with 6 times it
weight of alcohol,filter.
UFFELMANN'S REAGENT: To a 2% solution of pure phenol in water,add a water
solution of FeCl until the phenol solution becomes violet in color.
2
WAGNERS SOLUTION: Dissolve 25g of citric acid and 1g of salicylic acid in water
and dilute to 1 liter.Use 50ml of the reagent.
WIJ'S IODINE MONOCHLORIDE SOLUTION:Dissolve 13g of resublimed iodine in 1 liter
of glacial acetic acid which will pass the dichromate test for reductible
matter.Set aside 25ml of this solution.Pass into the remainder of the solution
dry chlorine gas until the characteristic color of free iodine has been
discharged.Now add the iodine solution which was reserved,until all free
chlorine has been destroyed.A slight excess of iodine does little or no harm,
but an excess of chlorine must be avoided.Preserve in well stoppered amber
colored bottles.Avoid usage of solutions older than 30 days.
WIJ'S SPECIAL SOLUTION: To 200ml of glacial acetic acid that will pass the
dichromate test for reductible matter,add 12g of dichloroamine T(paratoluene-
sulphonedichloroamide) and add 16.6g of dry KI(in small quantities until all of
the KI has dissolved).Make up to 1 liter with the same quality of acetic acid
used above and preserve in dark colored bottles.
ZIMMERMAN-REINHARDT SOLUTION: Dissolve 70g of MnSO x 4H O in 500ml of water,add
4 2
125ml of conc. H SO and 125ml of 85% H PO and dilute to 1 liter.
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