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Chemistry

Employing Pressurized Hot Water Extraction (PHWE) to Explore Natural Products Chemistry in the Undergraduate Laboratory

Published: November 7th, 2018

DOI:

10.3791/58195

1School of Natural Sciences - Chemistry, University of Tasmania, 2School of Chemistry, The University of Sydney
* These authors contributed equally

Here, we employ a pressurized hot water extraction (PHWE) method, which utilizes an unmodified household espresso machine to introduce undergraduates to natural products chemistry in the laboratory. Two experiments are presented: PHWE of eugenol and acetyleugenol from cloves and PHWE of seselin and (+)-epoxysuberosin from the Australian plant Correa reflexa.

A recently developed pressurized hot water extraction (PHWE) method which utilizes an unmodified household espresso machine to facilitate natural products research has also found applications as an effective teaching tool. Specifically, this technique has been used to introduce second- and third-year undergraduates to aspects of natural products chemistry in the laboratory. In this report, two experiments are presented: the PHWE of eugenol and acetyleugenol from cloves and the PHWE of seselin and (+)-epoxysuberosin from the endemic Australian plant species Correa reflexa. By employing PHWE in these experiments, the crude clove extract, enriched in eugenol and acetyleugenol, was obtained in 4-9% w/w from cloves by second-year undergraduates and seselin and (+)-epoxysuberosin were isolated in yields of up to 1.1% w/w and 0.9% w/w from C. reflexa by third-year students. The former exercise was developed as a replacement for the traditional steam distillation experiment providing an introduction to extraction and separation techniques, while the latter activity featured guided-inquiry teaching methods in an effort to simulate natural products bioprospecting. This primarily derives from the rapid nature of this PHWE technique relative to traditional extraction methods that are often incompatible with the time constraints associated with undergraduate laboratory experiments. This rapid and practical PHWE method can be used to efficiently isolate various classes of organic molecules from a range of plant species. The complementary nature of this technique relative to more traditional methods has also been demonstrated previously.

The isolation and identification of natural products are of fundamental importance to the scientific community and society more generally.1 Bioprospecting, the search for valuable organic molecules found in nature,remains an indispensable process in the discovery of new drug leads and potential therapeutic agents. It is estimated that, from 1981-2014, ~75% of all approved small molecule pharmaceutical drugs were natural products, natural product-derived or natural product-inspired.1 Furthermore, natural products possess enormous structural and chemical diversity. For this reason, they also represent valuable chemical sca....

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NOTE: It is advisable that all procedures are performed in a fume hood. Students must wear appropriate personal protective equipment at all times in the laboratory and the safety data sheets (SDS) associated with each reagent must be consulted before use.

1. PHWE of cloves: isolation of eugenol and acetyleugenol

  1. Extraction of eugenol and acetyleugenol from cloves
    1. Place coarsely ground cloves (12.5 g) in a 250-mL beaker.
    2. Add s.......

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PHWE of cloves. When attempting to perform the liquid-liquid extraction step, students often encountered emulsions (the addition of brine was typically not effective). At this stage, students were instructed to allow the mixture to stand in the separating funnel while they explored the effects of eluent composition on the separation of eugenol and acetyleugenol by TLC. It should be noted that hexane can be substituted with either heptane or dichloromethane in the liquid-liquid extraction .......

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The classical procedure for isolating eugenol from cloves by steam distillation has been part of the intermediate chemistry laboratory program at the University of Sydney for decades but was modernized to employ PHWE methodology in 2016 (Figure 1).9,18 This provided a number benefits. Firstly, utilizing household espresso machines in the laboratory environment immediately fascinated and engaged students by illustrating the applicatio.......

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The authors acknowledge the School of Natural Sciences - Chemistry, University of Tasmania and the School of Chemistry, The University of Sydney for financial support. B.J.D. and J.J. thank the Australian Government for Research Training Program Scholarships.

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Name Company Catalog Number Comments
espresso machines Breville/Sunbeam Breville espresso machine model 800ES / Sunbeam EM3820 Café Espresso II
rotary evporators Buchi and Heidolph
cloves (plant material) Dijon Food Pty Ltd Cloves must be ground in a food processor for students.
Correa reflexa (plant material) sample obtained in Tasmania Sample collected from mature shrubs in the Thomas Crawford Reserve at the University of Tasmania
sand Ajax 1199
ethanol Redoc Chemicals E95 F3
hexanes Ajax 251
magnesium sulfate Ajax 1548
diethyl ether Merck 1009215000
silica on aluminium TLC plates Merck 1055540001
eugenol Merck 1069620100
eugenyl acetate Aldrich W246905
acetone Redox Chemicals Aceton13
cyclohexane ChemSupply CA019
silica gel 60 Trajan 5134312 40 - 63um (230-400mesh)
Congo red paper ChemSupply IS070-100S
32% hydrochloric acid Ajax 256

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