METHODS Flashcards
glucose + boiling water —-> eneidol (enol anion form)
Chemical methods : oxidation-reduction
phosphomolybdic acid —> phosphomolybdenum blue
Folin-Wu
arsenomolybdic acid —> arsenomolybdenum blue
Nelson-Somogyi
neocuproine (7,9- dimethyl-1,10-phenanthroline) —> cuprous-neocuproine complex (yellow-orange)
Campbell and King method
ferricyanide —> ferrocyanide
reverse colorimetric method
glucose + ortho-toluidine —-> N-glycosylamine: Schiff base (green) 630 nm
Ortho-toluidine method by Dubowski
glucose + O2 + H2O — GOD —> gluconic acid + H2O2
Glucose Oxidase method
H2O2 + reduced chromogen — POD—> oxidized chromogen + H2O
Glucose Oxidase method
3-methyl-2-benzathiazolinone hydrazone + N,N-dimethylaniline – -> indamine dye (590-66- nm)
Gochman and Schmitz
p-aminophenazone + phenol —-> purple product (quinoneimine)
Trinder
2,2-azine-di(3-ethyl bezothiazoline-(6)-sulfonic acid —> colored chromogen
Miskieweis
glucose + ATP —hexokinase —> glu-6-phosphate + ADP
Hexokinase method
glu-6-phosphate + NADP —G-6-PD —> NADPH + H + 6-phosphogluconate (340 nm)
Hexokinase method
Glucose + NAD —— GDH —– Gluconolactone + NADH + H+
Glucose Dehydrogenase (GDH)
Hb A1c — acid — 5-HMF
COLORIMETRY
acetoacetic acid + ferric chloride —> red color
Gerhardt
acetoacetic acid + NADH + H —β-HBDH—> NAD + beta-hydroxybutyric acid
enzymatic method
NPN + H2SO4 NH4HSO4
KJELDAHL DIGESTION
NH3 + HgI2.2KI NH2Hg2I2 + KI + NH4I
NESSLERIZATION
NH3 + NaOCl + Phenol Indophenol + NaCl + H2O
BERTHELOT METHOD
NH3 + α-ketoglutarate + NADH + H Glutamate + NAD
Kaplan, Manoukian – Fawaz; Kallet – Cook Reaction
Isoindoline + Naphthylethylenediamine colored compound
Diacetyl Monoxime (Fearon)
Creatinine + alkaline picrate Creatinine picrate (red orange/yellow)
JAFFE REACTION
Creatinine —————-→N-methylhydantoin + NH3
F. Lim
Creatinine ———————————–→Creatine (Creatinine amidohydrolase)
G.A. Moss
Uric Acid + PTA ——- OH——– Allantoin + CO2 + Tungsten blue (710 nm)
Phosphotungstic Acid (PTA)
Uric Acid Allantoin + CO2
Blaunch and Koch
Uric Acid + O2 + 2H2O Allantoin + CO2 + H2O2
Trinder –Uricase method
Uric Acid Allantoin + CO2 + H2O2
Uricase –catalase system
Cuprous ions + neocuproine copper neocuproine complex
Bittner method
Ferrous ions + TPTZ blue colored complex (590 nm)
TPTZ Method by Morin
specimen is alkalinized to convert NH4 ions to NH3
Conway and Cook Diffusion Method
NH3 is trapped in acid medium of diffusion cell
Conway and Cook Diffusion Method
Quantitated by titration or colorimetry
Conway and Cook Diffusion Method
Time consuming with poor accuracy and precision
Conway and Cook Diffusion Method