Qualitative Analysis of Carbohydrates
Solubility Test
Molisch’s Test
Fehling’s Test
Benedict’s Test
Iodine Test
Tollen’s Test
Barfoed’s Test
Sellwanoff’s Test
Bial’s Test
Small amount of samples in test tubes A,B,C & D (Gl.Sc.St.Lc)+ H2O + Shaking + Observation of solubility
2ml Aq. Solution of samples in test tubes A,B & C + 6 drops of Molisch’s reagent + Few drops H2SO4
2ml Fehling’s soln. A+ 2ml of samples + 2ml Fehling’s soln. B + Heat in water bath for few minutes (Glucose reduces red ppts of CuO Ion)
2ml Benedict’s reagent+ Aq. Solution of samples + Mix + Heat in water bath for few minutes (Glucose shows yellow, red, green & orange)
2ml Aq. Solution of sample in test tube + 2 drops of Iodine solution (Applicable to starch & other polysaccharides)
2ml Tollen’s reagent+ 2ml Aq. Solution of samples (Gl.St.Sc) + Mix + Heat in water bath for 10 minutes (Just glucose shows colour change= Ag-ions----Elemental Silver)
1ml Barfoed’s reagent+ 1ml Aq. Solution of samples (Gl.St.Sc) + Mix + Heat in water bath for 5 minutes (Monosaccharaides reduce CuAt & can’t by low power Red.sugars)
1ml of sugars(aldose/Ketose)+ 3.5 ml of 0.5 g of resorcinol per litter+ 10% HCl+ Boil (Ketose dehydrate quickly within 30 seconds)
2-3 drops of Bial’s reagent+ 2ml sample (pentose or any other sugar) + Boil (Furfural are produced due to dehydration of pentose sugars and solution becomes greenish blue)
Biuret’s Test
Ppt Proteins Test
HM Salt Test
Acid Rg. Test
Ninhydrin’s Test
Xanthoprtic Test
Million’s Test
Sakaguchi’s Test
Hopkin-Cole Test
Dead At / SR Test
For SP[2ml protein+5/6 drops CuSo4+1/10 dil.FS+3ml 40% NaOH] = For ISP[3ml acetone+1.5 ml H2O+1 drop dil.NaOH+ protein+ Boil+Cool+0.5 ml 40% NaOH+2 drops FS A]
5 ml of protein solution + Small amount of NH3SO4(1g @ Time) + Agitation (Ppt Agents destabilizes protein = Precipitate formation=Salting Out Method)
3ml of protein solution + Few drops of HgNO3 (Heavy metals salts disrupts the salt bridges in proteins making their propitiates )
3ml of protein solution + Few drops of Tri-chloro-Acetic Acid + Observation (Acid anions & Protein particles make insoluble salt precipitates ) Test is effective for all proteins
5 drops of 0.1 % Ninhydrin’s solution + 2 ml of each of 3 protein suspension + heat for 10 minutes (Detect Alpha Amino Group)
1ml conc. HNO3 + 3ml proteins+ Heat for 30 seconds+ Cool+ Saturated NaOH till alkaline (Detect presence of pyro sine & tryptophan due to coloured benzene ring formation)
5 drops of fresh Million’s reagent + 3ml protein suspension + Heat for 5 minutes+ Observation (By adding much reagent colour will vanish during heating the solution)
1ml of 10 % NaOH + 1 ml 0.02% Alpha-Naphtol +3 ml protein solution+ 2-4 ml of Br-water after 3 minutes (To stabilize colour change add urea & destroy excess hypo bromine)
2ml of Hopkin’s reagent + 3ml of protein suspension + Mixing+ 1 ml concentrated H2SO4 (Slowly sidewise and don’t stir) + Wait for 1-2 minutes + Observation
5ml of 5% NaOH + Few crystals of Pb[II]Ac + 3ml of protein suspension + Heat for 5-10 minutes + Mix occasionally (Detect presence of Cysteine & some other amino acids)
Solubility Test
Ninhydrin’s Test
Alkli stabilty Test
Xanthoprtic Test
Million’s Test
Hopkin-Cole Test
PbSO4 Test
Ehrlich Test
Sakaguchi’s Test
Nitropurside Test
Pauly Diazo Test
Histidine Test
Felines MS’s Test
Istain Test
Small amount of samples in test tubes A, B & C (Gl.Ty.Cy)+ H2O/Ethanol + Heating + Litmus paper to determine solubility (Repeat with dil. HCl & NaOH)
5 drops of 0.2 % Ninhydrin’s solution in acetone + 1 ml of each of amino acid + Boil for 2 minutes + Cool + Observation (Detect Ammonia or p/s ary amines @ 4pH)
1ml Glycine + 1ml Amides in separate test tubes + 1 ml dil. NaOH +Boil + Test with wet litmus paper (A.acids don’t evolve NH3 when boiled with alkali but amide & amines do)
2ml conc. HNO3 + 2ml amino acid + Heat for 2 minutes + Cool+ Saturated NaOH till alkaline (Detect presence of aromatic amino acid containing activated benzene ring)
2 ml Acetic Acid + 1-2 drops of Million’s reagent + Heat for 10 minutes (Detection of Phenolic amino acids & its reagent contain Hg and HNO3)
Few ml of glacial acetic acid + 1-2 drops of amino acids + 1-2 ml H2SO4 (layer form underneath acetic acid & Indole group of Tryptophan react with Glyoxilic acid)
[5ml NaOH+2ml PbAt+ Boil--Dissolve Pb(OH)2 ppts to make Na_Plumbate] + Boil 2ml amino acid sol. with 40% NaOH for 2 minutes + Cool+ Few drops of NaPb
0.5ml amino acid solution+ 2 ml Ehrilch solution+ Observation + Repeat 3 times for Urea, Glycine and Tryptophan (Detection of aromatic amines and organic compounds)
1ml of 10 % NaOH + 2 drops of Alpha-Naphtol +3 ml Arginine solution+ 5-6 drops of Br-water
2ml amino acid solution+ 0.5 ml Nitropurside solution+ 4-5 drops of Br-Solution (Detection of Cysteine by colour change due to reaction of –SH & Nitropurside)
Few drops of unknown solution + Place in ice bucket+ Sulphanic Acid+ Na 2CO3 drop wise (Histidine & Tyrosine can be detected by this analysis)
2ml of unknown solution+ 5% Br in 33% Acetic acid + 25oC for 10 minutes +2 ml Ammonium Carbonate+ Heat for 10 minutes
1ml of unknown solution+ Few drops NaOH + Few drops Glycine and Sodimnitropurside + 40oC for 15 minutes +0.5 ml of 6N HCl+ Water
Drop of unknown solution in filter paper + Dry + Apply Istain + Wait & watch (Detection of Amino acids by colour change)
Solubility Test
Molisch’s Test
Fehling’s Test
Benedict’s Test
Iodine Test
Tollen’s Test
Barfoed’s Test
Sellwanoff’s Test
Bial’s Test
Small amount of samples in test tubes A,B,C & D (Gl.Sc.St.Lc)+ H2O + Shaking + Observation of solubility
2ml Aq. Solution of samples in test tubes A,B & C + 6 drops of Molisch’s reagent + Few drops H2SO4
2ml Fehling’s soln. A+ 2ml of samples + 2ml Fehling’s soln. B + Heat in water bath for few minutes (Glucose reduces red ppts of CuO Ion)
2ml Benedict’s reagent+ Aq. Solution of samples + Mix + Heat in water bath for few minutes (Glucose shows yellow, red, green & orange)
2ml Aq. Solution of sample in test tube + 2 drops of Iodine solution (Applicable to starch & other polysaccharides)
2ml Tollen’s reagent+ 2ml Aq. Solution of samples (Gl.St.Sc) + Mix + Heat in water bath for 10 minutes (Just glucose shows colour change= Ag-ions----Elemental Silver)
1ml Barfoed’s reagent+ 1ml Aq. Solution of samples (Gl.St.Sc) + Mix + Heat in water bath for 5 minutes (Monosaccharaides reduce CuAt & can’t by low power Red.sugars)
1ml of sugars(aldose/Ketose)+ 3.5 ml of 0.5 g of resorcinol per litter+ 10% HCl+ Boil (Ketose dehydrate quickly within 30 seconds)
2-3 drops of Bial’s reagent+ 2ml sample (pentose or any other sugar) + Boil (Furfural are produced due to dehydration of pentose sugars and solution becomes greenish blue)
Biuret’s Test
Ppt Proteins Test
HM Salt Test
Acid Rg. Test
Ninhydrin’s Test
Xanthoprtic Test
Million’s Test
Sakaguchi’s Test
Hopkin-Cole Test
Dead At / SR Test
For SP[2ml protein+5/6 drops CuSo4+1/10 dil.FS+3ml 40% NaOH] = For ISP[3ml acetone+1.5 ml H2O+1 drop dil.NaOH+ protein+ Boil+Cool+0.5 ml 40% NaOH+2 drops FS A]
5 ml of protein solution + Small amount of NH3SO4(1g @ Time) + Agitation (Ppt Agents destabilizes protein = Precipitate formation=Salting Out Method)
3ml of protein solution + Few drops of HgNO3 (Heavy metals salts disrupts the salt bridges in proteins making their propitiates )
3ml of protein solution + Few drops of Tri-chloro-Acetic Acid + Observation (Acid anions & Protein particles make insoluble salt precipitates ) Test is effective for all proteins
5 drops of 0.1 % Ninhydrin’s solution + 2 ml of each of 3 protein suspension + heat for 10 minutes (Detect Alpha Amino Group)
1ml conc. HNO3 + 3ml proteins+ Heat for 30 seconds+ Cool+ Saturated NaOH till alkaline (Detect presence of pyro sine & tryptophan due to coloured benzene ring formation)
5 drops of fresh Million’s reagent + 3ml protein suspension + Heat for 5 minutes+ Observation (By adding much reagent colour will vanish during heating the solution)
1ml of 10 % NaOH + 1 ml 0.02% Alpha-Naphtol +3 ml protein solution+ 2-4 ml of Br-water after 3 minutes (To stabilize colour change add urea & destroy excess hypo bromine)
2ml of Hopkin’s reagent + 3ml of protein suspension + Mixing+ 1 ml concentrated H2SO4 (Slowly sidewise and don’t stir) + Wait for 1-2 minutes + Observation
5ml of 5% NaOH + Few crystals of Pb[II]Ac + 3ml of protein suspension + Heat for 5-10 minutes + Mix occasionally (Detect presence of Cysteine & some other amino acids)
Solubility Test
Ninhydrin’s Test
Alkli stabilty Test
Xanthoprtic Test
Million’s Test
Hopkin-Cole Test
PbSO4 Test
Ehrlich Test
Sakaguchi’s Test
Nitropurside Test
Pauly Diazo Test
Histidine Test
Felines MS’s Test
Istain Test
Small amount of samples in test tubes A, B & C (Gl.Ty.Cy)+ H2O/Ethanol + Heating + Litmus paper to determine solubility (Repeat with dil. HCl & NaOH)
5 drops of 0.2 % Ninhydrin’s solution in acetone + 1 ml of each of amino acid + Boil for 2 minutes + Cool + Observation (Detect Ammonia or p/s ary amines @ 4pH)
1ml Glycine + 1ml Amides in separate test tubes + 1 ml dil. NaOH +Boil + Test with wet litmus paper (A.acids don’t evolve NH3 when boiled with alkali but amide & amines do)
2ml conc. HNO3 + 2ml amino acid + Heat for 2 minutes + Cool+ Saturated NaOH till alkaline (Detect presence of aromatic amino acid containing activated benzene ring)
2 ml Acetic Acid + 1-2 drops of Million’s reagent + Heat for 10 minutes (Detection of Phenolic amino acids & its reagent contain Hg and HNO3)
Few ml of glacial acetic acid + 1-2 drops of amino acids + 1-2 ml H2SO4 (layer form underneath acetic acid & Indole group of Tryptophan react with Glyoxilic acid)
[5ml NaOH+2ml PbAt+ Boil--Dissolve Pb(OH)2 ppts to make Na_Plumbate] + Boil 2ml amino acid sol. with 40% NaOH for 2 minutes + Cool+ Few drops of NaPb
0.5ml amino acid solution+ 2 ml Ehrilch solution+ Observation + Repeat 3 times for Urea, Glycine and Tryptophan (Detection of aromatic amines and organic compounds)
1ml of 10 % NaOH + 2 drops of Alpha-Naphtol +3 ml Arginine solution+ 5-6 drops of Br-water
2ml amino acid solution+ 0.5 ml Nitropurside solution+ 4-5 drops of Br-Solution (Detection of Cysteine by colour change due to reaction of –SH & Nitropurside)
Few drops of unknown solution + Place in ice bucket+ Sulphanic Acid+ Na 2CO3 drop wise (Histidine & Tyrosine can be detected by this analysis)
2ml of unknown solution+ 5% Br in 33% Acetic acid + 25oC for 10 minutes +2 ml Ammonium Carbonate+ Heat for 10 minutes
1ml of unknown solution+ Few drops NaOH + Few drops Glycine and Sodimnitropurside + 40 oC for 15 minutes +0.5 ml of 6N HCl+ Water
Drop of unknown solution in filter paper + Dry + Apply Istain + Wait & watch (Detection of Amino acids by colour change)
Solubility Test
Molisch’s Test
Fehling’s Test
Benedict’s Test
Iodine Test
Tollen’s Test
Barfoed’s Test
Sellwanoff’s Test
sBial’s Test
Small amount of samples in test tubes A,B,C & D (Gl.Sc.St.Lc)+ H2O + Shaking + Observation of solubility
2ml Aq. Solution of samples in test tubes A,B & C + 6 drops of Molisch’s reagent + Few drops H2SO4
2ml Fehling’s soln. A+ 2ml of samples + 2ml Fehling’s soln. B + Heat in water bath for few minutes (Glucose reduces red ppts of CuO Ion)
2ml Benedict’s reagent+ Aq. Solution of samples + Mix + Heat in water bath for few minutes (Glucose shows yellow, red, green & orange)
2ml Aq. Solution of sample in test tube + 2 drops of Iodine solution (Applicable to starch & other polysaccharides)
2ml Tollen’s reagent+ 2ml Aq. Solution of samples (Gl.St.Sc) + Mix + Heat in water bath for 10 minutes (Just glucose shows colour change= Ag-ions----Elemental Silver)
1ml Barfoed’s reagent+ 1ml Aq. Solution of samples (Gl.St.Sc) + Mix + Heat in water bath for 5 minutes (Monosaccharaides reduce CuAt & can’t by low power Red.sugars)
1ml of sugars(aldose/Ketose)+ 3.5 ml of 0.5 g of resorcinol per litter+ 10% HCl+ Boil (Ketose dehydrate quickly within 30 seconds)
2-3 drops of Bial’s reagent+ 2ml sample (pentose or any other sugar) + Boil (Furfural are produced due to dehydration of pentose sugars and solution becomes greenish blue)
Biuret’s Test
Ppt Proteins Test
HM Salt Test
Acid Rg. Test
Ninhydrin’s Test
Xanthoprtic Test
Million’s Test
Sakaguchi’s Test
Hopkin-Cole Test
Dead At / SR Test
For SP[2ml protein+5/6 drops CuSo4+1/10 dil.FS+3ml 40% NaOH] = For ISP[3ml acetone+1.5 ml H2O+1 drop dil.NaOH+ protein+ Boil+Cool+0.5 ml 40% NaOH+2 drops FSA]
5 ml of protein solution + Small amount of NH3SO4(1g @ Time) + Agitation (Ppt Agents destabilizes protein = Precipitate formation=Salting Out Method)
3ml of protein solution + Few drops of HgNO3 (Heavy metals salts disrupts the salt bridges in proteins making their propitiates )
3ml of protein solution + Few drops of Tri-chloro-Acetic Acid + Observation (Acid anions & Protein particles make insoluble salt precipitates ) Test is effective for all proteins
5 drops of 0.1 % Ninhydrin’s solution + 2 ml of each of 3 protein suspension + heat for 10 minutes (Detect Alpha Amino Group)
1ml conc. HNO3 + 3ml proteins+ Heat for 30 seconds+ Cool+ Saturated NaOH till alkaline (Detect presence of pyro sine & tryptophan due to coloured benzene ring formation)
5 drops of fresh Million’s reagent + 3ml protein suspension + Heat for 5 minutes+ Observation (By adding much reagent colour will vanish during heating the solution)
1ml of 10 % NaOH + 1 ml 0.02% Alpha-Naphtol +3 ml protein solution+ 2-4 ml of Br-water after 3 minutes (To stabilize colour change add urea & destroy excess hypo bromine)
2ml of Hopkin’s reagent + 3ml of protein suspension + Mixing+ 1 ml concentrated H2SO4 (Slowly sidewise and don’t stir) + Wait for 1-2 minutes + Observation
5ml of 5% NaOH + Few crystals of Pb[II]Ac + 3ml of protein suspension + Heat for 5-10 minutes + Mix occasionally (Detect presence of Cysteine & some other amino acids)
Solubility Test
Ninhydrin’s Test
Alkli stabilty Test
Xanthoprtic Test
Million’s Test
Hopkin-Cole Test
PbSO4 Test
Ehrlich Test
Sakaguchi’s Test
Nitropurside Test
Pauly Diazo Test
Histidine Test
Felines MS’s Test
Istain Test
Small amount of samples in test tubes A, B & C (Gl.Ty.Cy)+ H2O/Ethanol + Heating + Litmus paper to determine solubility (Repeat with dil. HCl & NaOH)
5 drops of 0.2 % Ninhydrin’s solution in acetone + 1 ml of each of amino acid + Boil for 2 minutes + Cool + Observation (Detect Ammonia or p/s ary amines @ 4pH)
1ml Glycine + 1ml Amides in separate test tubes + 1 ml dil. NaOH +Boil + Test with wet litmus paper (A.acids don’t evolve NH3 when boiled with alkali but amide & amines do)
2ml conc. HNO3 + 2ml amino acid + Heat for 2 minutes + Cool+ Saturated NaOH till alkaline (Detect presence of aromatic amino acid containing activated benzene ring)
2 ml Acetic Acid + 1-2 drops of Million’s reagent + Heat for 10 minutes (Detection of Phenolic amino acids & its reagent contain Hg and HNO 3)
Few ml of glacial acetic acid + 1-2 drops of amino acids + 1-2 ml H2SO4 (layer form underneath acetic acid & Indole group of Tryptophan react with Glyoxilic acid)
[5ml NaOH+2ml PbAt+ Boil--Dissolve Pb(OH)2 ppts to make Na_Plumbate] + Boil 2ml amino acid sol. with 40% NaOH for 2 minutes + Cool+ Few drops of NaPb
0.5ml amino acid solution+ 2 ml Ehrilch solution+ Observation + Repeat 3 times for Urea, Glycine and Tryptophan (Detection of aromatic amines and organic compounds)
1ml of 10 % NaOH + 2 drops of Alpha-Naphtol +3 ml Arginine solution+ 5-6 drops of Br-water
2ml amino acid solution+ 0.5 ml Nitropurside solution+ 4-5 drops of Br-Solution (Detection of Cysteine by colour change due to reaction of –SH & Nitropurside)
Few drops of unknown solution + Place in ice bucket+ Sulphanic Acid+ Na 2CO3 drop wise (Histidine & Tyrosine can be detected by this analysis)
2ml of unknown solution+ 5% Br in 33% Acetic acid + 25oC for 10 minutes +2 ml Ammonium Carbonate+ Heat for 10 minutes
1ml of unknown solution+ Few drops NaOH + Few drops Glycine and Sodimnitropurside + 40 oC for 15 minutes +0.5 ml of 6N HCl+ Water
Drop of unknown solution in filter paper + Dry + Apply Istain + Wait & watch (Detection of Amino acids by colour change)
Reducing Sugars are soluble & VV
Red & Violet rings at junction due to Gl.St.Sc
Red Ppt=Red.sugar & N.red.sugar=No Change
YGOR=Red.sugar & N.red.sugar=No Change
Blue-black Colored Starch/ Polysaccharides
Silver mirror is formed at bottom of test tube
Brick red CuOx formed=Monosaccharaides
Red colour within 30 seconds (Gl. on heating)
Green-blue colour indicates pentose & VV
Qualitative Analysis of Proteins
Colour change due to proteins of 2/+ peptides
Ppts forms & exposure of hydrophobic region
White precipitations occurs due to reactions
Protein ppts are formed due to salt formation
Blue-purple complex colour is formed
Colour change will show presence of proteins
Colour change will show existence of proteins
Strong red colour proves Arginine existence
Violet colour will present +ve test of proteins
Black & grey ppts will present positive test
Qualitative Analysis of Amino Acids
Red or pink LP will show solubility & VV
Blue coloured substance is formed
NH3 is not absorbed on LP and that is A.acids
Yellow colour product confirms Aromatic AA
Positive test is identified by brick red colour
Purple colour detects presence of Tryptophan
Brown colour shows presence of Sulphides
Colour change for the amines shows result
Arginine is detected by the change in colour
Colour absolutely changes if Cysteine present
Red colour appears and detect their presence
Histidine presence detected by blue colour
Red colour indicates presence of Methionine
Blue colour spot on FP indicates amino acids
Qualitative Analysis of Carbohydrates
Reducing Sugars are soluble & VV
Red & Violet rings at junction due to Gl.St.Sc
Red Ppt=Red.sugar & N.red.sugar=No Change
YGOR=Red.sugar & N.red.sugar=No Change
Blue-black Colored Starch/ Polysaccharides
Silver mirror is formed at bottom of test tube
Brick red CuOx formed=Monosaccharaides
Red colour within 30 seconds (Gl. on heating)
Green-blue colour indicates pentose & VV
Qualitative Analysis of Proteins
Colour change due to proteins of 2/+ peptides
Ppts forms & exposure of hydrophobic region
White precipitations occurs due to reactions
Protein ppts are formed due to salt formation
Blue-purple complex colour is formed
Colour change will show presence of proteins
Colour change will show existence of proteins
Strong red colour proves Arginine existence
Violet colour will present +ve test of proteins
Black & grey ppts will present positive test
Qualitative Analysis of Amino Acids
Red or pink LP will show solubility & VV
Blue coloured substance is formed
NH3 is not absorbed on LP and that is A.acids
Yellow colour product confirms Aromatic AA
Positive test is identified by brick red colour
Purple colour detects presence of Tryptophan
Brown colour shows presence of Sulphides
Colour change for the amines shows result
Arginine is detected by the change in colour
Colour absolutely changes if Cysteine present
Red colour appears and detect their presence
Histidine presence detected by blue colour
Red colour indicates presence of Methionine
Blue colour spot on FP indicates amino acids
Qualitative Analysis of Carbohydrates
Reducing Sugars are soluble & VV
Red & Violet rings at junction due to Gl.St.Sc
Red Ppt=Red.sugar & N.red.sugar=No Change
YGOR=Red.sugar & N.red.sugar=No Change
Blue-black Colored Starch/ Polysaccharides
Silver mirror is formed at bottom of test tube
Brick red CuOx formed=Monosaccharaides
Red colour within 30 seconds (Gl. on heating)
Green-blue colour indicates pentose & VV
Qualitative Analysis of Proteins
Colour change due to proteins of 2/+ peptides
Ppts forms & exposure of hydrophobic region
White precipitations occurs due to reactions
Protein ppts are formed due to salt formation
Blue-purple complex colour is formed
Colour change will show presence of proteins
Colour change will show existence of proteins
Strong red colour proves Arginine existence
Violet colour will present +ve test of proteins
Black & grey ppts will present positive test
Qualitative Analysis of Amino Acids
Red or pink LP will show solubility & VV
Blue coloured substance is formed
NH3 is not absorbed on LP and that is A.acids
Yellow colour product confirms Aromatic AA
Positive test is identified by brick red colour
Purple colour detects presence of Tryptophan
Brown colour shows presence of Sulphides
Colour change for the amines shows result
Arginine is detected by the change in colour
Colour absolutely changes if Cysteine present
Red colour appears and detect their presence
Histidine presence detected by blue colour
Red colour indicates presence of Methionine
Blue colour spot on FP indicates amino acids