The Scientific World Journal

Volume 2013 (2013), Article ID 212156, 7 pages

http://dx.doi.org/10.1155/2013/212156

Research Article

Direct Synthesis of ESBO Derivatives-¹⁸O Labelled with Dioxirane

Stefano La Tegola,¹ Cosimo Annese,2,3 Michele Suman,⁴ Immacolata Tommasi,² Caterina Fusco,³ and Lucia D'Accolti2,3

¹Chemist at Agenzia delle Dogane e dei Monopoli, Italy

²Department of Chemistry, Università degli Studi di Bari Aldo Moro, Via Orabona 4, 70126 Bari, Italy

³CNR-Istituto di Chimica dei Composti Organometallici (ICCOM), Sezione di Bari, Via Orabona 4, 70126 Bari, Italy

⁴Barilla G. R. F.lli SpA, R&D Central Research Labs, Via Mantova 166, 43100 Parma, Italy

Received 22 July 2013; Accepted 12 August 2013

Academic Editors: A. Nacci and Y. Tang

Copyright © 2013 Stefano La Tegola et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract.

This work addresses a new approach developed in our laboratory, consisting in the application of isolated dimethyldioxirane (DDO, 1a) labelled with ¹⁸O for synthesis of epoxidized glyceryl linoleate (Gly-LLL, 2). We expect that this work could contribute in improving analytical methods for the determination of epoxidized soybean oil (ESBO) in complex food matrices by adopting an ¹⁸O-labelled-epoxidized triacylglycerol as an internal standard.

1. Introduction

The packaging has an essential rule in the food manufacture: it increases the life of food, while guarantees protection from physical and microbiological contaminations [1].

In order to improve the packaging materials performances, in recent decades the use of different types of additives has been significant development (plasticizers, stabilizers, etc.) [2–4], like vegetable oils, that represent an interesting renewable source for the production of useful chemicals and new materials [5–8]. In particular, the use of epoxidized soybean oil (ESBO; see Figure 1) has increased in recent years as a valid alternative to phthalates, since it shows relatively good compatibility with PVC and low toxicity to humans [9]. In this respect ESBO is listed as an authorized substance according to food packaging materials legislation (European Commission 2002).

Figure 1: Structure of (Gly-LLL-ox 2a): one of chemical species in ESBO (from linoleic in sn-1, sn-2, and sn-3).

However, a known potential is the migration of ESBO from the packaging into the food, especially oily foods and fats. This risk in food contamination was investigated by the European Food Safety Authority (EFSA), which set a specific migration limit for ESBO in baby foods [10]. Several analytical methods have been reported for the analysis of ESBO in PVC gaskets and food, although they are very time-consuming and not so user-friendly, especially due to low concentrations of migrants and the complexity of the food matrix [10, 11].

The aim of this work was to obtain a selective synthesis of exa-epoxy-three linoleate glyceryl-¹⁸O labelled (2a) in one or multiple positions. This target compound could be used as internal standard in quantitative ESBO analysis, in order to simplify and increase the accuracy of our previous LC-MS/MS method [10].

Despite the fact that the synthesis of ESBO [12] of epoxy esters of unsaturated fatty acids [13] could be obtained by different methods yielding conversion and selectivity higher than 90%, the preparation of its ¹⁸O labelled epoxide is quite difficult [14]. A possibility is to prepare the corresponding peroxide compound labelled R¹⁸O¹⁸OR′, generated from hydrogen peroxide-¹⁸O-¹⁸O (Scheme 1) [15].

Scheme 1

The standard procedure now mentioned is very arduous and delicate if one takes into account the fact that hydrogen peroxide-¹⁸O-¹⁸O is synthesized by passing H₂¹⁸O vapour through a tube discharge (V 2000) [16] or using 97% ¹⁸O labelled O₂ gas [17].

In this context, we have shown that the dioxiranes R¹R²CO₂ (1) [18, 19] three-member ring strained peroxides, generated in situ, were able to transfer electrophilic oxygen to a variety of donor substrates (S:), yielding the corresponding oxidation products (SO) and the parent ketone (Figure 2).

Figure 2: Dioxiranes commonly employed in the isolated form (in solution of the parent ketone).

Undoubtedly very attractive applications of these reagents are regio-, stereo-, and enantioselective epoxidations of alkenes. In fact simple [18] or supported ketones [20] and optical active ketones as precursors of chiral dioxiranes in situ [21, 22] could be used in organic synthesis.

Further a possible alternative to obtain labelled epoxides consists of using DDO (1a) ¹⁸O labelled, generated in situ, from acetone and potassium monoperoxysulfate (KHSO₅) K + -O-SO₂-¹⁸O¹⁸OH (caroate) ¹⁸O-labelled—as reported in Scheme 2 [23].

Scheme 2

This procedure requires a careful preparation, since the caroate is also able to give molecular oxygen, with loss of ¹⁸O₂, as background reaction [19].

Until now the setup of appropriate techniques allowed the isolation of a few dioxirane volatile species, such as dimethyldioxirane (CH₃)₂CO₂ (1a) [24] and the powerful methyl(trifluoromethyl)dioxirane (CF₃)(CH₃)CO₂ (1b) [25]. These dioxiranes have been found to be a versatile epoxidizing reagent for alkenes of various kinds, either electron rich or electron poor [26] and allylic alcohols [27].

In continuation of our work on selective polyepoxidations [28, 29] we now report on the epoxidation of glyceryl-trilinoleate (2) using the isolated ¹⁸O DDO (1a). Our preliminary results are described herein.

2. Materials and Methods

The GC analyses were run using a ZB-1 column (30 m × 0.25 μm id); in most cases, 1,1,1,2-tetrachloro-2,2-difluoroethane (Freon A112) was employed as inert internal standard. The ¹H NMR were recorded on a 500 MHz resonances and are referenced to residual isotopic impurity CHCl₃ (7.26 ppm) of solvent CDCl₃ and/or to TMS. The ¹³C NMR spectra (125.76 MHz) are referenced to the middle peak of CDCl₃ solvent (77.0 ppm). MS experiments were performed using a Micromass M@LDITM–LR (Waters MS Technologies, Manchester, UK) TOF mass spectrometer equipped with a nitrogen UV laser (337 nm wavelength).

Commercial acetone and methylene chloride (AnalaR grade) were purified by standard methods, stored over 4 Å molecular sieves. H₂O labelled with ¹⁸O (isotopic purity 95%), sulfuric acid (H₂SO₄), calcium chloride (CaCl₂), magnesium sulfate (MgSO₄), sodium hydroxide pellets (NaOH), sodium thiosulfate (Na₂S₂O₃), methyl p-tolyl-sulfide, disodium salt of ethylenediaminetetraacetic acid (EDTANa₂), and sodium bicarbonate (NaHCO₃), commercial products at high purity degree, were used without further purification.

Curox triple salt 2KHSO₅KHSO₄K₂SO₄ (a gift by Peroxid-Chemie GmbH, Munich, Germany) was our source of potassium peroxymonosulfate (caroate); it was used as received for the synthesis of dioxiranes 1a.

2.1. Preparation of ¹⁸O-DDO (1a)

Dioxiranes in solution of the parent ketones (i.e., acetone for 1a) could be obtained by following a procedure already reported in detail [24, 30] starting with buffered (pH 7, NaHCO₃) aqueous potassium peroxomonosulfate KHSO₅ and the ketone. A 500 mL four-necked round-bottomed vessel is equipped with a mechanical stirrer, a splash-guard adapter connected to an air-cooled straight condenser, a solids addition funnel, a gas inlet tube extending into the reaction mixture, and a thermometer. The air condenser is connected laterally to the top entry of an efficient jacketed spiral condenser cooled at −70°C; the bottom exit of this condenser is fitted to a two-way fraction-collector adapter carrying two 50 mL receiving flasks and is also cooled at −70°C. In succession to the adapter, two cold traps are placed, the first is cooled at −70°C and the second at liquid nitrogen temperature. The main vessel is charged with 35 mL of bidistilled water, the mixture of ¹⁸O acetone (30 mL) and H₂¹⁸O (6 mL) Na₂EDTA₂ (0.1 g), and NaHCO₃ (22 g). Mechanical stirring is initiated, and the solid potassium peroxomonosulfate (0.189 mol, 66 g of corresponding to 2.86 mmol/g of Caroat triple salt iodometry) is quickly added during 1-2 minutes to the reaction vessel, while passing a gentle stream of Ar (or N₂) gas through the mixture. Shortly after the addition is initiated, the end two-way adaptor is switched to insert the main collection flask and during 10 min ca. 25 mL of weak-yellow solution of 1a in acetone is collected. This solution had a 0.083 M concentration in dioxirane (iodometry).

2.2. General Procedure for Oxidation of Methyl-p-tolyl Sulfide (3) with Dioxirane ¹⁸O-DDO (1a)

A solution of methyl-p-tolyl-sulfide (3) (9.1 mg, 0.066 mmol) in 5 mL of CH₂Cl₂ dry was prepared. To 0.3 mL of this standard solution kept at 0°C 0.01 mL of ¹⁸O-DDO (1a) solution 0.083 M in acetone (2 equivalents) was added. The reaction progress was monitored by GC analysis of microaliquots periodically withdrawn from the reaction flask. In each experiment, the values were measured at various reaction times; data from two or more independent runs were averaged. Upon reaction completion, the reaction mixture was characterized by a combination of spectroscopic techniques (¹H, ¹³C NMR, IR, and MS) collecting data in agreement with the literature. The mass spectrum of the resulting labelled 3a revealed two peaks at (ri) 154 (M⁺, 100) and 156 (M⁺ + 2, 13.9), while for the same unlabelled 3a     154 (M⁺, 100) and 156 (M⁺ + 2, 5.9). The percent of ¹⁸O was estimated from the .

2.3. Oxidation of Glyceryl-trilinoleate with ¹⁸O-DDO (1a)

To a solution of Gly-LLL (2) (0.213 g 0.24 mmoli), in 20 mL of CH₂Cl₂ dry, kept at 20°C, a solution ¹⁸O-DDO (1a) 0.083 M in ¹⁸O acetone of 21 mL (7.2 equiv.) was added. The reaction was monitored using TLC (silica gel hexane/ethyl ether 50 : 50). The precious labelled acetone was recovered by careful distillation of the crude reaction (18 mL, 20% labelled). Upon completion, pure ¹⁸O-LLLox  (2a): was simply obtained by evaporation of the volatile solvents in vacuum and full characterized with MALDI-Tof MS ¹H e {¹H}  ¹³C-NMR.

¹H-NMR (CDCl₃, 500 MHz): 0.86 (m, 9H, CH₃), 1.25–1.80 (m, 66H, CH₂), 2.28 (s, 6H, CH₂ in al C=O), 2.90–3.08 (m, 12H, CH epox), 4.11–4.22 (m, 4H, CH₂ glycerol), 5.22 (m, 1H, CH glycerol); {¹H}  ¹³C-NMR (CDCl₃, 125 MHz): 13.89, 22.47, 24.67, 24.70, 26.04, 26.14, 26.35, 26.47, 26.82, 27.11, 27.70, 27.73, 27.78, 27.80, 28.85, 28.89, 29.08, 29.21, 31.56, 33.87, 34.03, 54.01, 54.23, 54.25, 56.56, 56.58, 56.64, 56.85, 56.91, 61.99, 68.80, 172.6, 173.10; MALDI-TOF (2577 V pulse voltage) (ir) = 997.65 (100, C₅₇H₉₈O₁₂Na, M), 999.65, (64.7 M + 2).

3. Results and Discussion

Considering remarkable success of methods of epoxidation using dioxirane [19, 24, 25], we decide to modify opportunely the technique which allowed for the first time the isolation of ¹⁸O-DDO (1a). In view of the fact that we need to start from a significant amount of acetone and that ¹⁸O-acetone is commercially available but particularly expensive [31], we chose as source of ¹⁸O the H₂¹⁸O 95%, using it in an exchange reaction with acetone, acid-catalysed (H₂SO₄ 0.02 N) (1) [16]:

(1)

The exchange reaction was monitored by GC-MS, quantifying the two species of acetone, labelled or not, by the signal intensity of the corresponding molecular ion ( acetone,   ¹⁸O acetone). Finally we had a labelling for acetone equal to 69%. Dioxirane (1a) could be obtained by the standard procedure (Scheme 3). The only change is the amount of water, because we used the exchange mixture, without further purification.

Scheme 3

Also in this case, it is of considerable importance that the dioxirane synthesis reaction must be perfectly buffered with NaHCO₃, in order to minimize the loss of ¹⁸O-acetone labelling due to water exchange process (acid and base catalyzed). Anyway, the rate of formation of dioxirane is faster than the exchange reaction with water, thanks to the high nucleophilicity of peroxide ((2) alpha effect) [32]:

(2)

Before proceeding with the polyepoxidation of the triglyceride, in order to assess the amount of labelled of ¹⁸O-DDO (1a), it appears necessary to explore its efficacy in the labelled reaction, as both ¹⁶O and ¹⁸O oxygen atoms of the dioxirane can be transferred to substrate [23].

We chose as a target reaction the oxidation of methyl-p-tolyl-sulfide (3), a transformation well known for its efficiency [24]. The level of labelling on the corresponding sulfoxide (3a) was determined by GC-MS analysis, considering the increase of the ratio of the signal intensity M + 2/M (where M is the molecular ion), compared to the same ratio obtained by the GC-MS spectrum of a solution of methyl-p-tolyl sulfoxide standard unlabelled. In this case we obtained a labelled sulfoxide (3a) equal to 8% (3):

(3)

Using the data regarding the oxidation of 3 it is possible to carry out a preliminary theoretical assessment of isotopic enrichment on M + 2 signals relative to our target compound, the ¹⁸O-LLLox (2a), for which there are six successive processes of oxygen insertion. We used the Bayes formula for the determination of probability to give the percent of labelled compound: [33] where is the probability of the elemental event of epoxidation with ¹⁸O, is the probability of the opposite elemental event, , and is the number of ¹⁸O constituent the compound, product of the reaction, from 0 to 6. Based on these calculations we expected an increase of isotope peak M + 2 of our product equal to 32%, and so the theoretical possibility of obtaining a yield of 32% as product (2a) containing a single Atom ¹⁸O. The preliminary oxidation of triglyceride 2 was carried out with unlabelled DDO (1a) in order to optimize the reaction conditions and in particular to determine the best DDO (1a)/substrate ratio required for a complete substrate oxidation; afterwards, the reaction was performed using the 1a labelled.

The easy oxidation procedure involved the stepwise addition (20 min) of standardized dioxirane 1a solution to the glyceryl-three linoleate (2) dissolved in CH₂Cl₂ (20 mL) and kept at 20°C. A moderate excess of dioxirane ¹⁸O-DDO (1a) was applied in order to achieve complete substrate conversion as monitored by TLC (Scheme 4).

Scheme 4

The GC-MS analysis of the reaction mixture showed that a significant residual of labelled acetone is still present. The precious acetone ¹⁸O-enriched was recovered by a careful distillation of the crude reaction.

The polyepoxide 2a (oil) was fully characterized by ¹H- and ¹³C-NMR, yielding spectra in complete agreement with the given structure, and the ¹H NMR spectrum of 2a shows inter alias the multiple at 2.90–3.08 (m, 12H) due to the C-H resonances of the epoxy ring.

In addition, the MALDI-TOF spectrum confirm the structure of the compound (noted as adducts Na + signal at ) and also determine the isotopic enrichment of the M + 2 signal (). We found that it is equal to 42%, and this is in agreement with the previously calculated according to the theory of probability (Figure 3).

Figure 3: MALDI-TOF spectrum of of exa-epoxy-three linoleate glyceryl-¹⁸O labelled (2a).

Data that are representative of the enrichment attainable in the direct dioxirane oxidation of different substrates are collected in Table 1. These show that, using the procedure reported herein, labelled polyepoxide 2a can be synthesized up to 42% labelled yield.

Table 1: Mass spectrometry data.

Products
M (m/z)
Unlabelled
Labelled
18O %a

M + 2
M + 2

Acetoneb
100 (58)
1.01
69.93
69%

Methyl-p-Tolil sulfoxide (3a)b
100 (154)
5.91
13.88
8%

18O-LLLox (2a)c
100 (998)
23.23
64.71
42%

Acetone after DDO (1a) synthesis
100 (58)
1.01
21.02
20%

The percent of ¹⁸O was estimated form the [((M + 2)/M)labelled – ((M + 2)/M)unlabelled]}.

bThe percent of ¹⁸O was determined using the GC/Ms analysis.

cFor the ¹⁸O-LLLox was used the MALDI-TOF analysis.

In summary, by the easily prepared ¹⁸O dimethyldioxirane (1a), natural products as glyceryl-three linoleate can be cleanly converted to the corresponding labelled epoxide in mild conditions. The high efficiency and selectivity of these oxidations provide highly pure products, which can be used directly in subsequent reaction steps, avoiding costly and time-consuming product purification procedures. Besides the oxidant precursor, the expensive acetone ¹⁸O labelled can be recovered and reused for dioxirane regeneration, thus increasing the atom economy of the process.

In conclusion, we believe that these results contribute to reinforce the notion that the application of dioxiranes efficiently provides access to useful new application not only in organic synthesis but also for the setup of new analytical methods.

Conflict of Interests

The authors declare that there is no conflict of interests regarding the publication of this paper. The authors declare not having a direct financial relation that might lead to a conflict of interests for any of the authors.

Authors’ Contribution

Caterina Fusco and Lucia D’Accolti contributed equally to this work.

Acknowledgments

Partial support of this work is by the Ministry of University and Scientific and Research of Italy (COFIN National Project) and Barilla G. R. F.lli SpA, R&D Central Research Labs, Parma, Italy, is gratefully acknowledged. Thanks are due to Dr. D. Calvano (University of Bari, Italy) for performing the MALDI-TOF analyses.

References

A. L. Brody and K. S. Marsh, The Wiley Encyclopedia of Packaging Technology, John Wiley & Sons, New York, NY, USA, 2nd edition, 1997.

W. Soroka, Fundamemals of Packaging Technology, Institute of Packaging Professionals, Herndon, Wash, USA, 2nd edition, 1999.

A. E. Gerbase, C. L. Petzhold, and A. P. O. Costa, “Dynamic mechanical and thermal behavior of epoxy resins based on soybean oil,” Journal of the American Oil Chemists' Society, vol. 79, no. 8, pp. 797–802, 2002.

B. Çakmakli, B. Hazer, I. Ö. Tekin, and F. B. Cömert, “Synthesis and characterization of polymeric soybean oil-g-methyl methacrylate (and n-butyl methacrylate) graft copolymers: biocompatibility and bacterial adhesion,” Biomacromolecules, vol. 6, no. 3, pp. 1750–1758, 2005.

T. Tsujimoto, H. Uyama, and S. Kobayashi, “Synthesis and curing behaviors of cross-linkable polynaphthols from renewable resources: preparation of artificial Urushi,” Macromolecules, vol. 37, no. 5, pp. 1777–1782, 2004.

A. K. Chakraborti, S. Rudrawar, and A. Kondaskara, “Lithium bromide, an inexpensive and efficient catalyst for opening of epoxide rings by amines at room temperature under solvent-free condition,” European Journal of Organic Chemistry, no. 17, pp. 3597–3600, 2004.

G. Lligadas, J. C. Ronda, M. Galià, U. Biermann, and J. O. Metzger, “Synthesis and characterization of polyurethanes from epoxidized methyl oleate based polyether polyols as renewable resources,” Journal of Polymer Science A, vol. 44, no. 1, pp. 634–645, 2006.

U. Biermann, W. Friedt, S. Lang, et al., “New syntheses with oils and fats as renewable raw materials for the chemical industry,” Angewandte Chemie International Edition, vol. 39, no. 13, pp. 2206–2224, 2000.

European Food Safety Authority (EFSA), “Opinion of the Scientific Panel on Food Additives, Flavourings, Processing Aid and Materials in Contact with Food (AFC) on a request from the Commission related to the use of epoxidized soybean oil in food contact materials adopted on 26 May 2004,” EFSA Journal, vol. 64, article 1, 2004, http://www.efsa.europa.eu/

M. Suman, S. La Tegola, D. Catellani, and U. Bersellini, “Liquid chromatography-electrospray ionization-tandem mass spectrometry method for the determination of epoxidized soybean oil in food products,” Journal of Agricultural and Food Chemistry, vol. 53, no. 26, pp. 9879–9884, 2005.

C. Bueno-Ferrer, A. Jiménez, and M. C. Garrigós, “Migration analysis of epoxidized soybean oil and other plasticizers in commercial lids for food packaging by gas chromatography-mass spectrometry,” Food Additives and Contaminants A, vol. 27, no. 10, pp. 1469–1477, 2010.

A. E. Gerbase, J. R. Gregório, M. Martinelli, M. C. Brasil, and A. N. F. Mendes, “Epoxidation of soybean oil by the methyltrioxorhenium-CH₂Cl₂/H₂O₂ catalytic biphasic system,” Journal of the American Oil Chemists' Society, vol. 79, no. 2, pp. 179–181, 2002.

B.-J. Kollbe Ahn, S. Kraft, and X. S. Sun, “Chemical pathways of epoxidized and hydroxylated fatty acid methyl esters and triglycerides with phosphoric acid,” Journal of Materials Chemistry, vol. 21, no. 26, pp. 9498–9505, 2011.

B. Borhan, S. Nazarian, E. M. Stocking, B. D. Hammock, and M. J. Kurth, “An efficient preparation of ¹⁸O-labeled epoxides,” Journal of Organic Chemistry, vol. 59, no. 15, pp. 4316–4318, 1994.

S. Jankowski and R. Kaminski, “Preparation of ¹⁸O-labelled m-chloroperbenzoic acid,” Journal of Labelled Compounds & Radiopharmaceuticals, vol. 36, no. 4, pp. 373–376, 1995.

M. Camporeale, T. Fiorani, L. Troisi, W. Adam, R. Curci, and J. O. Edwards, “On the mechanism of the Baeyer-Villiger oxidation of ketones by bis(trimethylsilyl) peroxomonosulfate. Intermediacy of dioxiranes,” Journal of Organic Chemistry, vol. 55, no. 1, pp. 93–98, 1990.

A. W. Vermilyea, T. C. Dixon, and B. M. Voelker, “Use of H₂¹⁸O₂ to measure absolute rates of dark H₂O₂ production in freshwater systems,” Environmental Science and Technology, vol. 44, no. 8, pp. 3066–3072, 2010.

R. Curci, M. Fiorentino, L. Troisi, J. O. Edwards, and R. H. Pater, “Epoxidation of alkenes by dioxirane intermediates generated in the reaction of potassium caroate with ketones,” Journal of Organic Chemistry, vol. 45, no. 23, pp. 4758–4760, 1980.

S. E. Denmark, D. C. Forbes, D. S. Hays, J. S. DePue, and R. G. Wilde, “Catalytic epoxidation of alkenes with oxone,” Journal of Organic Chemistry, vol. 60, no. 5, pp. 1391–1407, 1995.

L. D’Accolti, C. Annese, A. De Riccardis et al., “Dioxirane-mediated heterogeneous epoxidations with potassium caroate. A solid catalyst bearing anchored ketone moieties,” European Journal of Organic Chemistry, vol. 24, pp. 4616–4621, 2012.

M. Frohn and Y. Shi, “Chiral ketone-catalyzed asymmetric epoxidation of olefins,” Synthesis, no. 14, pp. 1979–2000, 2000.

D. Yang, “Ketone-catalyzed asymmetric epoxidation reactions,” Accounts of Chemical Research, vol. 37, no. 8, pp. 497–505, 2004.

D. Yang, M.-K. Wong, Y.-C. Yip, et al., “Design and synthesis of chiral ketones for catalytic asymmetric epoxidation of unfunctionalized olefins,” Journal of the American Chemical Society, vol. 120, no. 24, pp. 5943–5952, 1998.

R. W. Murray, R. Jeyaraman, and M. K. Pillay, “Chemistry of dioxiranes. 6. Electronic effects in the oxidation of sulfides and sulfoxides by dimethyldioxirane,” Journal of Organic Chemistry, vol. 52, no. 5, pp. 746–748, 1987.

R. Curci, L. D'Accolti, and C. Fusco, “A novel approach to the efficient oxygenation of hydrocarbons under mild conditions. Superior oxo transfer selectivity using dioxiranes,” Accounts of Chemical Research, vol. 39, no. 1, pp. 1–9, 2006.

C. Annese, L. D'Accolti, A. Dinoi, C. Fusco, R. Gandolfi, and R. Curci, “Concerning the reactivity of dioxiranes. Observations from experiments and theory,” Journal of the American Chemical Society, vol. 130, no. 4, pp. 1197–1204, 2008.

L. D'Accolti, M. Fiorentino, C. Fusco, A. M. Rosa, and R. Curci, “Chemo- and diastereoselectivities in the oxidation of cyclopentenols with dimethyldioxirane and methyl(trifluoromethyl)dioxirane,” Tetrahedron Letters, vol. 40, no. 45, pp. 8023–8027, 1999.

L. D'Accolti, C. Annese, and C. Fusco, “Direct regio- and stereoselective synthesis of squalene 2,3;22,23-dioxide using dioxiranes,” Tetrahedron Letters, vol. 46, no. 49, pp. 8459–8462, 2005.

C. G. Zambonin, C. D. Calvano, L. D'Accolti, and F. Palmisano, “Laser desorption/ionization time-of-flight mass spectrometry of squalene in oil samples,” Rapid Communications in Mass Spectrometry, vol. 20, no. 2, pp. 325–327, 2006.

L. D'Accolti, C. Fusco, M. R. Rella, and R. Curci, “Concerning synthesis of ring-A fluorinated anthracyclines. The dioxirane shunt,” Synthetic Communications, vol. 33, no. 17, pp. 3009–3016, 2003.

“The current cost of ¹⁸O-acetone 90% is  2.693,50/gr (Aldrich), water H₂¹⁸O  694, Medical Isotopes, Inc., NH, USA”.

N. J. Fina and J. O. Edwards, “Alpha effect. Review,” International Journal of Chemical Kinetics, vol. 5, no. 1, pp. 1–26, 1973.

M. Levin, “A misuse of Bayes's theorem,” Informal Logic, vol. 19, no. 1, pp. 63–66, 1999.