Porcine Pancreas: A Superior Source of Cholesterol Esterase for Total Serum Cholesterol Assay by the Enzymatic Kinetic Method

Srisawasdi, Pornpen; Prasertsincharoen, Noppadol; Kroll, Martin H.

doi:10.1002/jcla.21538

Cited by 2 publications

(3 citation statements)

References 20 publications

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“…Specifically, the steady-state kinetic parameters of the reactions were determined by varying the concentrations of (a) ABTS or TMB in the presence of H 2 O 2 and (b) H 2 O 2 in the presence of ABTS or TMB. The concentrations of ABTS (green color) and TMB (blue color) were subsequently calculated with the molar absorption coefficients of 36,000 and 39,000 M –1 cm –1 , respectively. − The results (Michaelis–Menten and Lineweaver–Burk plots) are shown in Figure S8A–D.…”

Section: Resultsmentioning

confidence: 99%

“…Second, the concentrations of enzymes ChOx and GOx in the ranges of 5000–12,000 U L –1 and 3000–12,000 U L –1 , respectively, were optimized. The concentrations of V-porous Co 3 O 4 , ChE, cholesterol standard, and glucose standard were fixed at 75 mg L –1 , 7000 U L –1 (excess reagent for converting the cholesteryl ester level in human serum), 7 and 10 mM, respectively. As shown in Figure S10B,C, the colorimetric responses were found to increase with increasing concentrations of the enzymes and reached a plateau after the concentrations of ChOx and GOx were increased up to 10,000 U L –1 .…”

Section: Resultsmentioning

confidence: 99%

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Vanadium-Doped Porous Cobalt Oxide for Its Superior Peroxidase-like Activity in Simultaneous Total Cholesterol and Glucose Determination in Whole Blood Based on a Simple Two-Dimensional Paper-Based Analytical Device

et al. 2022

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Vanadium-doped porous Co3O4 (V-porous Co3O4) was synthesized via a simple soft-templating method and used as a superior peroxidase mimic for the simultaneous colorimetric determination of glucose and total cholesterol (TC) in whole blood samples on a two-dimensional microfluidic paper-based analytical device (2D-μPAD). The large surface area and the presence of two metals in V-porous Co3O4 contributed to its excellent catalytic activity toward 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 3,3′,5,5′- tetramethylbenzidine (TMB) with Michaelis–Menten constants (K M) of 0.1301 and 0.0141 mM, respectively. The 2D-μPAD was fabricated using simple wax screen-printing and cutting techniques. The colorimetric reactions of both glucose and TC on 2D-μPAD were simultaneously performed by adding a single drop of a whole blood sample on the sample zone made of the LF1 membrane. After the enzymatic reactions, the generated hydrogen peroxide (H2O2) was oxidized by V-porous Co3O4 to produce hydroxy radicals (•OH), inducing ABTS and TMB to generate colored products. The generated H2O2 was proportional to the intensities of the green and blue products of the glucose and TC systems, respectively. The developed 2D-μPAD required a short analysis time (∼5 min) with small volumes of samples (15 μL of whole blood) whereby no sample preparation was needed. Owing to several advantages including simplicity, low cost, long-term stability, and simultaneous readout, the novel V-porous Co3O4 coupled with 2D-μPAD proved to be promising for practical uses as a pioneering portable device for the determination of glucose, TC, and other important biomarkers without the need of technical supports.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Vanadium-Doped Porous Cobalt Oxide for Its Superior Peroxidase-like Activity in Simultaneous Total Cholesterol and Glucose Determination in Whole Blood Based on a Simple Two-Dimensional Paper-Based Analytical Device

et al. 2022

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“…Approximately 30% of cholesterol in serum is present in 'free' form (typically, 40 mg/dl -85 mg/dl) while the remainder is assimilated into cholesteryl esters with long-chain fatty acids [23]. In order to measure the total cholesterol in serum, a triple enzyme assay was used comprising two cholesterol reactions and an optical product step via the activity of peroxidase.…”

Section: B Enzyme Assay Methodsmentioning

confidence: 99%

A Colorimetric CMOS-Based Platform for Rapid Total Serum Cholesterol Quantification

et al. 2017

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Elevated cholesterol levels are associated with a greater risk of developing cardiovascular disease and other illnesses, making it a prime candidate for detection on a disposable biosensor for rapid point of care diagnostics. One of the methods to quantify cholesterol levels in human blood serum uses an optically mediated enzyme assay and a bench top spectrophotometer. The bulkiness and power hungry nature of the equipment limits its usage to laboratories. Here, we present a new disposable sensing platform that is based on a complementary metal oxide semiconductor process for total cholesterol quantification in pure blood serum. The platform that we implemented comprises readily mass-manufacturable components that exploit the colorimetric changes of cholesterol oxidase and cholesterol esterase reactions. We have shown that our quantification results are comparable to that obtained by a bench top spectrophotometer. Using the implemented device, we have measured cholesterol concentration in human blood serum as low as 29 µM with a limit of detection at 13 µM, which is approximately 400 times lower than average physiological range, implying that our device also has the potential to be used for applications that require greater sensitivity.

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Porcine Pancreas: A Superior Source of Cholesterol Esterase for Total Serum Cholesterol Assay by the Enzymatic Kinetic Method

Abstract: Porcine pancreas cholesterol esterase is superior to that obtained from P. fluorescens, C. cylindracea, and bovine pancreas for total serum cholesterol determination by the kinetic method because of its lower cost, better accuracy and precision, less interference, and longer stability.

Cited by 2 publications

References 20 publications

Vanadium-Doped Porous Cobalt Oxide for Its Superior Peroxidase-like Activity in Simultaneous Total Cholesterol and Glucose Determination in Whole Blood Based on a Simple Two-Dimensional Paper-Based Analytical Device

Vanadium-Doped Porous Cobalt Oxide for Its Superior Peroxidase-like Activity in Simultaneous Total Cholesterol and Glucose Determination in Whole Blood Based on a Simple Two-Dimensional Paper-Based Analytical Device

A Colorimetric CMOS-Based Platform for Rapid Total Serum Cholesterol Quantification

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