Spectrophotometric determination of acetone using vanillin

Amlathe, Sulbha; Gupta, V. K.

doi:10.1039/an9901501385

Cited by 14 publications

(7 citation statements)

References 4 publications

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“…For the screening, α-methylstyrene was chosen as a model substrate, which is oxidatively cleaved to acetophenone by TM1459 ( Hajnal et al, 2015 ). The detection system is based on the base-catalyzed aldol condensation reaction of vanillin with ketones ( Amlathe and Gupta, 1990 ; Figure 2 ). With the model product acetophenone, a yellow colored complex is formed, while no background reaction occurs with the corresponding alkene substrate.…”

Section: Resultsmentioning

confidence: 99%

Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity

et al. 2016

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Oxidative cleavage of alkenes is a widely employed process allowing oxyfunctionalization to corresponding carbonyl compounds. Recently, a novel biocatalytic oxidative alkene cleavage activity on styrene derivatives was identified in TM1459 from Thermotoga maritima. In this work we engineered the enzyme by site-saturation mutagenesis of active site amino acids to increase its activity and to broaden its substrate scope. A high-throughput assay for the detection of the ketone products was successfully developed. Several variants with up to twofold improved conversion level of styrene derivatives were successfully identified. Especially, changes in or removal of the C-terminus of TM1459 increased the activity most significantly. These best variants also displayed a slightly enlarged substrate scope.

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Section: Resultsmentioning

confidence: 99%

Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity

et al. 2016

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“…Acetone has also been quantified photometrically by its reaction with vanillin, which produces a yellow-orange dye which absorbs at 430 nm [21], and by its reaction with alkaline salicylaldehyde [22].…”

Section: Introductionmentioning

confidence: 99%

Determination of acetone in seawater using derivatization solid-phase microextraction

2007

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Acetone plays an important role in the chemistry of both the atmosphere and the ocean, due to its potential effect on the tropospheric HO(x) (= HO + HO(2)) budget, as well as its environmental and health effects. We discuss the development of a mobile, sensitive, selective, economical and facile method for the determination of acetone in seawater. The method consists of derivatizing acetone to its pentafluorobenzyl oxime using 1,2,3,4,5-pentafluorobenzylhydroxylamine (PFBHA), followed by solid-phase microextraction (SPME) and analysis by gas chromatography/mass spectrometry (GC/MS). A detection limit of 3.0 nM was achieved. The buffering capacity of seawater imposes challenges in using the method's optimum pH (3.7) on seawater samples, requiring calibration standards to be made in buffered salt water and the acidification of seawater samples and standards prior to extraction. We employed the technique for analysis of selected surface seawater samples taken on the Nordic seas during the ARK-XX/1 cruise (R.V. Polarstern). An upper limit of 5.5-9.6 nM was observed for acetone in these waters, the first acetone measurements reported for far North Atlantic and Arctic waters. Simplified schematic of transformations of organic compounds at the atmosphere-ocean interface.

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“…To specifically quantify acetone, we adapted a spectrophotometric method, originally used in 10 mL flasks for acetone determination in clinical and biological samples (Amlathe and Gupta, 1990), to a more sensitive high‐throughput 96‐well plate format. The reported protocol for the vanillin‐based colorimetric analysis of acetone (Amlathe and Gupta, 1990) included sodium bisulfite, which was omitted in our assay, as its influence on the colour development was found to be negligible. To match a 96‐well plate format, the overall volume of the assay was scaled down 50‐fold, to a final 200 μl.…”

Section: Resultsmentioning

confidence: 99%

“…In the early 20 th century, several spectrophotometric methods to determine ketone and aldehyde concentrations were developed before gas or liquid analytic chromatography became available in most laboratories. These methods include the use of 2,4‐dinitrophenylhydrazine (2,4‐DNPH) (Lappin, 1951) and vanillin (Amlathe and Gupta, 1990). Here, we optimized two spectrophotometric methods of ketone and aldehyde determination with further potential for high throughput screening of microbial production strains.…”

Section: Introductionmentioning

confidence: 99%

High‐throughput colorimetric assays optimized for detection of ketones and aldehydes produced by microbial cell factories

Kozaeva

Mol

Nikel

et al. 2022

Microbial Biotechnology

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Summary Randomized strain and pathway engineering are critical to improving microbial cell factory performance, calling for the development of high‐throughput screening and selection systems. To facilitate this effort, we have developed two 96‐well plate format colorimetric assays for reliable quantification of various ketones and aldehydes from culture supernatants, based on either a vanillin‐acetone reaction or the 2,4‐dinitrophenylhydrazine (2,4‐DNPH) reagent. The vanillin‐acetone assay enabled accurate and selective measurement of acetone titers up to 2 g l−1 in a minimal culture medium. The 2,4‐DNPH‐based assay can be used for a wide range of aldehydes and ketones, shown here through the optimization of conditions for 15 different compounds. Both assays were implemented to improve acetone production from different substrates by an engineered Escherichia coli strain. The fast and user‐friendly colorimetric assays proposed here open the potential for iterative rounds of (automated) strain and pathway engineering and screening, facilitating the efforts towards further boosting production titers of industrially relevant ketones and aldehydes.

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Spectrophotometric determination of acetone using vanillin

Cited by 14 publications

References 4 publications

Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity

Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity

Determination of acetone in seawater using derivatization solid-phase microextraction

High‐throughput colorimetric assays optimized for detection of ketones and aldehydes produced by microbial cell factories

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