Determination of total and pancreatic α-amylase in human serum with 2-chloro-4-nitrophenyl-α-d-maltotrioside as substrate

Gella, F. Javier; Gubern, Gemma; Vidal, Ricardo; Canalías, Francesca

doi:10.1016/s0009-8981(96)06481-9

Cited by 74 publications

(41 citation statements)

References 11 publications

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“…The in vitro a-amylase inhibition activity of extracts and fractions from J. phoenicea leave was determined based on the spectrophotometric assay using acarbose as the reference compound (Gella et al 1997). The studied sample was dissolved in DMSO to give concentrations of 50, 100, and 200 mg/mL.…”

Section: A-amylase Inhibition Assay By Cnpg3 Methodsmentioning

confidence: 99%

LC-MS–MS and GC-MS analyses of biologically active extracts and fractions from Tunisian Juniperus phoenice leaves

Keskes

Belhadj

Jlail

et al. 2016

Pharmaceutical Biology

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Context: Despite some studies related to Juniperus phoenicea L. (Cupressaceae), phytochemical and biological investigations of this plant remain unexplored. Objective: This work is the first report dealing with the identification and characterization of volatile components and flavonoids in hexane and methanol extracts from J. phoenicea leaves Materials and methods: Antioxidant activity of hexane, and methanol extracts from J. phoenicea leaves were determined by DPPH-radical scavenging assay. a-Amylase inhibitory activity was evaluated by enzyme inhibition using in vitro assay (each extract was dissolved in DMSO to give concentrations of 50, 100 and 200 mg/mL). The chemical composition of fractions (Fr1-Fr3) from methanol extract was determined by high-performance liquid chromatography coupled with mass spectroscopy (HPLC-MS) analysis. Results and discussion: The hexane extract was analyzed by GC-MS technique which allowed the identification of 32 compounds. The main constituents were a-humulene (16.9%), pentadecane (10.2%) and a-cubebene (9.7%). Fraction Fr 2 exhibited a strong DPPH radical-scavenging activity (IC 50 ¼ 20.1 lg/mL) compared to that of BHT as well as the highest a-amylase inhibitory activity (IC 50 ¼ 28.4 lg/mL). Three flavonoids were identified in these fractions using HPLC-MS analysis: Quercetin 3-O-glucoside, isoscutellarein 7-O-pentoside and quercetin 3-O-pentoside. In addition, the more active fraction (Fr 2) was purified with semi-preparative HPLC affording one pure compound (amentoflavone) using 1 H NMR analysis. This compound exhibited powerful DPPH radical-scavenging (IC 50 ¼ 14.1 lg/mL) and a-amylase inhibition (IC 50 ¼ 20.4 lg/mL) effects. Conclusion: This study provides scientific support to some medicinal uses of J. phoenicea found in North Africa. ARTICLE HISTORY

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Section: A-amylase Inhibition Assay By Cnpg3 Methodsmentioning

confidence: 99%

LC-MS–MS and GC-MS analyses of biologically active extracts and fractions from Tunisian Juniperus phoenice leaves

Keskes

Belhadj

Jlail

et al. 2016

Pharmaceutical Biology

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“…By the classical initial rate method, no chromogenic substrates for serum lipase assay are satisfactory due to their low solubility (Tsao et al, 2007;Yamada and Fujita, 2007); chromogenic maltotrioside substrates for direct assay of serum α-amylase are also unsatisfactory due to the cost (Winn-Deen et al, 1988;Gella et al, 1997). Taken together, when concentrations of chromogenic substrates are limited by their solubility, cost, substrate inhibition/activation, and/or measurable ranges on instruments, the integration strategy is an effective and general alternative to measure serum enzymes with expanded linear ranges and practical efficiency, and the optimizations of reaction duration and regular intervals to monitor reaction curves can be general guidelines to validate the integration strategy at low concentrations of chromogenic substrates.…”

Section: Further Discussionmentioning

confidence: 99%

Kinetic analysis of γ-glutamyltransferase reaction process for measuring activity via an integration strategy at low concentrations of γ-glutamyl p-nitroaniline

Liu

Yang

et al. 2011

J. Zhejiang Univ. Sci. B

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At 0.12 mmol/L γ-glutamyl p-nitroaniline (GGPNA), an improved integrated method was developed for kinetic analysis of γ-glutamyltransferase (GGT) reaction process and the integration with the classical initial rate method to measure serum GGT. For the improved integrated method, an integrated rate equation, which used the predictor variable of reaction time and considered inhibitions by both GGPNA and products, was nonlinearly fit to GGT reaction processes. For the integration strategy, classical initial rates were estimated when GGPNA consumption percentages were below 50%; otherwise, maximal reaction rates of GGT were estimated by the improved integrated method and converted into initial rates according to the differential rate equation at 0.11 mmol/L GGPNA. The integration strategy was validated using optimized GGT kinetic parameters and 10-s intervals to record reaction curves within 8.0 min. By the integration strategy, there was a linear response from 0.9 to 32.0 U/L GGT, coefficients of variation were below 3.5% for GGT from 8.0 to 32.0 U/L (n=5), and GGT activities in clinical sera responded linearly to their classical initial rates at 2.00 mmol/L GGPNA with an expected slope. Therefore, the integration strategy was successful in measuring GGT at 0.12 mmol/L GGPNA.

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“…Harvested samples were centrifuged at 4000 rpm during 10 minutes and supernatant were decanted to measure alpha-Amylase concentration that same day. Alpha-amylase activity was determined through kinetic enzymatic method which uses CNP-G3 (2-chloro-4-nitrophenyl-alpha-maltotriosid) (Gella et al, 1997) as substrate. Briefly, on the recommendation of fabricant, 1000 µL of reagent was mixed with 25 µL of serum or saliva; saliva sample were diluted.…”

Section: Data Collection and Processingmentioning

confidence: 99%

Effects of Daily Alcohol Intake on Serum and Salivary Alpha-Amylase Activity Associated With Two Local Alcoholic Drinks of Benin

Gomina

Sina²,

Gandekon³

et al. 2017

JBLS

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This study was initiated to assess the effect of daily amount of alcohol intake on serum and salivary alpha-amylase activity of regular adult consumers of tchoukoutou and sodabi, two local alcoholic drinks made in Benin. It was a descriptive, cross-sectional and analytical study carried out from 1 st of April to 31 st of August, 2012. The study population consisted of 50 subjects as regular consumers of tchoukoutou (titrated with 3% of alcohol), 50 regular consumers of sodabi (titrated with 40% of alcohol) and 50 non-consumers of alcohol. Alpha-amylase activity in saliva and serum were measured in each subject. There was no significant difference in the activity of serum and salivary alpha-amylase between consumers of low quantity of tchoukoutou and sodabi (˂ 30 g/ day) on one hand (P = 0.24 and 0.99 respectively), and between moderate consumers (30-79 g/ day) on the other (P = 0.31 and 0.48 respectively). Daily amount of alcohol intake had a positive effect on the serum alpha-amylase activity when taking into consideration the entire group at the threshold of 1%, in tchoukoutou consumers (r = +0.887), and in women sodabi consumers at the threshold of 5% (r = +0.928). Therefore, serum alpha-amylase activity is positively associated with the consumption of these two local alcoholic drinks made in Benin.

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Determination of total and pancreatic α-amylase in human serum with 2-chloro-4-nitrophenyl-α-d-maltotrioside as substrate

Cited by 74 publications

References 11 publications

LC-MS–MS and GC-MS analyses of biologically active extracts and fractions from Tunisian Juniperus phoenice leaves

LC-MS–MS and GC-MS analyses of biologically active extracts and fractions from Tunisian Juniperus phoenice leaves

Kinetic analysis of γ-glutamyltransferase reaction process for measuring activity via an integration strategy at low concentrations of γ-glutamyl p-nitroaniline

Effects of Daily Alcohol Intake on Serum and Salivary Alpha-Amylase Activity Associated With Two Local Alcoholic Drinks of Benin

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