Development of quantitative enzymatic method and its validation for the assay of glucose in human serum

Spectrophotometric determination of glucose in human serum samples using para‐phenylenediamine and alpha‐naphthol as a chromogenic reagent

“…The K m value for PPD/AN coupling proposed assay was witnessed to be 526.31 µM, the K m value shows the affection between the enzyme active site with the substrate. Whereas by the reported SSA/Fe (III) system 33 the values were K m = 1075 µM and 2,5‐DMA system, 8 K m = 1192 µM, lower K m value of PPD/AN method advocates that the proposed system two times more catalytic interaction between the substrate and co‐substrate (PPD/AN) with enzyme active site of GOD than the SSA/Fe (III) and 2,5‐DMA system.…”

“…The glucose concentration in the serum sample is practiced with the range of 0.02–2.775 mM and the recovery range of glucose by the proffered method achieved 97.78%–104.35% with 100.75% mean recovery. The proposed enzymatic method shows excellent recovery and the need for a 10 µl serum sample shows that the proposed method is more sensitive than the reported systems 6,8,33 using a simple spectrophotometer. Limit of detection (LOD) and limit of quantification (LOQ) for glucose were found to be 2.175 and 8.56 µmol L –1 , so the proposed system is not significantly affected by the common interferants present in serum samples.…”

“…The proposed enzymatic system showed better recovery (97.78%-104.35%) and the need for 10 µl serum sample as compared to 20, 30, and 40 µl serum samples used by some of the reported methods, recovery studies reveal that the proposed method is more sensitive than the reported systems. 6,8,33 The GOD-catalyzed spectrophotometric determination of glucose by PPD with AN, the linearity of glucose lies in the range of 57.81-1850.2 and 14.45-925.1 µmol L -1 by kinetic and one-time detection method. The established method can analyze a larger number of samples at a time using microtiter plates.…”

“…Determination of blood glucose levels using enzymes is generally practiced in the clinical analysis due to its high reliability, simplicity, and consistency related to any other analytical methods. [6][7][8] Numerous analytical techniques have been reported for monitoring the glucose level in serum samples such as HPLC, 9,10 cyclic voltammetry, 11,12 radiometer biosensor, 13,14 microtiter plate reader, 15,16 flow injection analysis, 17,18 spectrofluorimetric, 19,20 polarographic, 21,22 chemiluminescence, 23,24 Raman scattering, 25,26 chromatography, 27,28 and Colorimetry. 29,30 Every method has its unique merits like the use of <20 µl serum sample and constraints including the need for a sophisticated instrument, operative complexity, high cost to implement, less versatility, involving multiple steps, and the need for immobilized enzymes.…”

A Review on the Impact of the Plant Bioactive Compound &lt;i&gt;β&lt;/i&gt;-Glucans as Phytochemicals for Boosting Human and Animals’ Immune Response—&lt;i&gt;β&lt;/i&gt;-Glucans as Immunostimulants

“…First, samples prior β-Glucans assay is treated with alkaline such as potassium hydroxide (KOH) for the gelatinization (swelling) of polysaccharide structure of β-Glucans. This alkaline treatment is assisting the acid or the enzymes to penetrate and breakdown the polysaccharide of β-Glucans into D-Glucose units [33]. Released D-Glucose from both assay methods is measured by a calorimetric method.…”