Development and validation of a simple high performance liquid chromatography – evaporative light scattering detector method for direct quantification of native cyclodextrins in a cyclization medium

Rojas, Mayerlenis J.; Castral, Thaís Corrêa; Giordano, R. L. C.; Tardioli, Paulo Waldir

doi:10.1016/j.chroma.2015.07.097

Cited by 7 publications

(6 citation statements)

References 30 publications

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“…The concentrations of α-, β-, and γ-CDs were determined using the chromatographic method described by Rojas et al [62]. Briefly, the CDs were separated through a pyramid NUCLEODUR ® C18 column (150 mm × 4.6 mm, 5 µm, Macherey-Nagel, Düren, Germany), at 30 • C, using acetonitrile (solvent B) and water, containing 1% (v/v) acetic acid (solvent A) at a flow rate of 0.3 mL/min, using a linear gradient, as follows: 0-20 min with 100 to 80% of A and 0 to 20% of B; 20.1-30 min with 100% of A.…”

Section: Chromatographic Methods For Analysis Of Cdsmentioning

confidence: 99%

“…The temperature, enzyme loading, and reaction time for the production of CDs were selected based on previous studies with soluble enzyme [60]. Samples were withdrawn at 0.25, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 h for analysis of the α-, βand γ-CD concentrations by ELSD-HPLC chromatographic method [62]. The total yield of CD production (Y CD ) was calculated following Equation 5:…”

Section: Production Of Cyclodextrinsmentioning

confidence: 99%

“…At the end of each batch (five 3-h cycles), the biocatalyst was separated from the reaction medium by centrifugation and washed with distilled water. The CDs were analyzed by ELSD-HPLC method [62].…”

Section: Thermal and Operational Stabilitymentioning

confidence: 99%

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Preparation of Crosslinked Enzyme Aggregates of a Thermostable Cyclodextrin Glucosyltransferase from Thermoanaerobacter sp. Critical Effect of the Crosslinking Agent

et al. 2019

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Crosslinked enzyme aggregates (CLEAs) of a thermostable cyclodextrin glucosyltransferase (CGTase) from Thermoanaerobacter sp. have been prepared for the production of cyclodextrins (CDs). Different parameters in the precipitation (nature and concentration of precipitant) and crosslinking steps (time of reaction with cross-linker, nature and concentration of the crosslinker) were evaluated on the production of CLEAs of CGTase. Among the seven studied precipitants, acetone with a 75% (v/v) concentration produced the aggregates of CGTase with higher activity, which retained 97% of the initial activity. Concerning the cross-linker (glutaraldehyde, starch–aldehyde, and pectin–aldehyde), starch–aldehyde produced the most active CLEAs. The use of bovine serum albumin as co-feeder decreased the expressed activity. Addition of polyethylenimine at the end of cross-linking step prevented the leakage of the enzyme and the subsequent Schiff’s bases reduction with sodium borohydride permitted to maintain 24% of the initial activity even with the large dextrin as substrate. The optimal conditions for the immobilization process required were defined as 75% (v/v) acetone as precipitation reagent for 1 h at 20 °C, 20 mM starch–aldehyde as crosslinking reagent for 2 h at 20 °C, treatment with 1 mg/mL of polyethylenimine for 5 min, reduction with 1 mg/mL of sodium borohydride. The CLEAs of CGTase were active catalyst (similarly to the free enzyme) in the production of cyclodextrins at 50 °C and pH 6.0 for 6 h reaction, maintaining intact their structures. Besides this, after five cycles of 3 h the total cyclodextrin yield was 80% of the initial value (first batch, with around 45% CD yield).

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Section: Chromatographic Methods For Analysis Of Cdsmentioning

confidence: 99%

Section: Production Of Cyclodextrinsmentioning

confidence: 99%

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Preparation of Crosslinked Enzyme Aggregates of a Thermostable Cyclodextrin Glucosyltransferase from Thermoanaerobacter sp. Critical Effect of the Crosslinking Agent

et al. 2019

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“…The precision of the method was examined using the intraday repeatability and interday repeatability. 13 Each concentration of each type of matrix was conducted in triplicate within one batch of experiment for intraday repeatability. Interday repeatability was determined using measurements of different batches on different days.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Development of a Thiolysis HPLC Method for the Analysis of Procyanidins in Cranberry Products

Gao

Cunningham²,

Liu³

et al. 2018

J. Agric. Food Chem.

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The objective of this study was to develop a thiolysis HPLC method to quantify total procyanidins, the ratio of A-type linkages, and A-type procyanidin equivalents in cranberry products. Cysteamine was utilized as a low-odor substitute of toluene-α-thiol for thiolysis depolymerization. A reaction temperature of 70 °C and reaction time of 20 min, in 0.3 M of HCl, were determined to be optimum depolymerization conditions. Thiolytic products of cranberry procyanidins were separated by RP-HPLC and identified using high-resolution mass spectrometry. Standards curves of good linearity were obtained on thiolyzed procyanidin dimer A2 and B2 external standards. The detection and quantification limits, recovery, and precision of this method were validated. The new method was applied to quantitate total procyanidins, average degree of polymerization, ratio of A-type linkages, and A-type procyanidin equivalents in cranberry products. Results showed that the method was suitable for quantitative and qualitative analysis of procyanidins in cranberry products.

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“…The μPMC workflow used ELSD quantitation to determine isolated amounts of compound (μg) similar to methods used by Rojas. 43 All desired fractions were reformatted in DMSO at 30, 10, and 4 mM concentrations for the tPMC, aPMC, and μPMC workflows, respectively. A 5 μL aliquot was taken and diluted on a Tecan system (Tecan Group Ltd., Switzerland) to 0.5 mg/mL for quality control (QC) to assess compound purity via UPLC, with the remaining majority being sent to biological assays, solution storage, or dry storage, as indicated by the synthetic chemist.…”

mentioning

confidence: 99%

An Automated Purification Workflow Coupled with Material-Sparing High-Throughput ¹H NMR for Parallel Medicinal Chemistry

Bellenger,

Koos,

Avery

et al. 2024

ACS Med. Chem. Lett.

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In medicinal chemistry, purification and characterization of organic compounds is an ever-growing challenge, with an increasing number of compounds being synthesized at a decreased scale of preparation. In response to this trend, we developed a parallel medicinal chemistry (PMC)-tailored platform, coupling automated purification to mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR) on a range of synthetic scales (∼3.0−75.0 μmol). Here, the generation and acquisition of 1.7 mm NMR samples is fully integrated into a high-throughput automated workflow, processing 36 000 compounds yearly. Utilizing dead volume, which is inaccessible in conventional liquid handling, NMR samples are generated on as little as 10 μg without consuming material prioritized for biological assays. As miniaturized PMC synthesis becomes the industry standard, we can now obtain quality NMR spectra from limited material. Paired with automated structure verification, this platform has the potential to allow NMR to become as important for high-throughput analysis as ultrahigh performance liquid chromatography (UPLC)-MS.

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Development and validation of a simple high performance liquid chromatography – evaporative light scattering detector method for direct quantification of native cyclodextrins in a cyclization medium

Cited by 7 publications

References 30 publications

Preparation of Crosslinked Enzyme Aggregates of a Thermostable Cyclodextrin Glucosyltransferase from Thermoanaerobacter sp. Critical Effect of the Crosslinking Agent

Preparation of Crosslinked Enzyme Aggregates of a Thermostable Cyclodextrin Glucosyltransferase from Thermoanaerobacter sp. Critical Effect of the Crosslinking Agent

Development of a Thiolysis HPLC Method for the Analysis of Procyanidins in Cranberry Products

An Automated Purification Workflow Coupled with Material-Sparing High-Throughput ¹H NMR for Parallel Medicinal Chemistry

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Development and validation of a simple high performance liquid chromatography – evaporative light scattering detector method for direct quantification of native cyclodextrins in a cyclization medium

Cited by 7 publications

References 30 publications

Preparation of Crosslinked Enzyme Aggregates of a Thermostable Cyclodextrin Glucosyltransferase from Thermoanaerobacter sp. Critical Effect of the Crosslinking Agent

Preparation of Crosslinked Enzyme Aggregates of a Thermostable Cyclodextrin Glucosyltransferase from Thermoanaerobacter sp. Critical Effect of the Crosslinking Agent

Development of a Thiolysis HPLC Method for the Analysis of Procyanidins in Cranberry Products

An Automated Purification Workflow Coupled with Material-Sparing High-Throughput 1H NMR for Parallel Medicinal Chemistry

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About

An Automated Purification Workflow Coupled with Material-Sparing High-Throughput ¹H NMR for Parallel Medicinal Chemistry