Rosemary and sage as antioxidants - Extraction methods

Rosemary and sage as antioxidants

Introduction

Rosemary is one of the most effective spices, widely used in food processing. It is the only spice commercially available for use as an antioxidant in Europe and the United States. One of the main potential uses is the suppression of warmed over flavour (WOF .

However, because of their prime use as flavouring agents, rosemary extract products are not technically listed as natural preservatives or antioxidants.

Extraction methods

The first use of an extract of rosemary leaves as an antioxidant was reported by Rac and Ostric-Matijasevic in 5. Berner and Jacobson obtained a patent in 3 for production of an antioxidant extract from rosemary using oil as a solvent. Chang et al. reported a process for the extraction of rosemary and sage, followed by vacuum steam distillation in an edible oil or fat to obtain a colourless, odourless natural antioxidant. Bracco et al. described an extraction process using peanut oil, followed by micronization, heat treatment and molecular distillation. Inahata et al. obtained a patent in 6 for production of odourless and safe antioxidants from rosemary by repeated extraction, evaporation, purification and dissolving procedures. More recently another technique, supercritical carbon dioxide extraction, has been used to product extracts of rosemary and sage.

Antioxidant properties

Antioxidant properties of rosemary have been well documented. ⁵ Rosemary was considered both lipid antioxidant and metal helator. ² Rosemary extract was found also to scavenge superoxide radicals. The application of rosemary extracts in food has resulted in a variability in the results depending on the test model being used.

Many different solvents have been used for the extraction of the antioxidative compounds. ⁹ Chang et al. extracted rosemary leaves with hexane, benzene, ethyl ether, chloroform, ethylene dichloride, dioxane and methanol. The extracts %) were tested during oxidation of lard at 60sC in the dark. It was established that the greatest antioxidant activity was located in the methanol extract. The methanol extract was further purified, and the resultant fraction showed an outstanding activity in potato chips fried in sunflower oil and held at 0sC in the dark for 0 days.

Marinova et al., Chen et al., and Pokorny et al. found that the hexane extracts from rosemary were better antioxidants for lard, rapeseed and sunflower oils, than methanol or ethanol extracts. Hexane extract ) caused a fold increase of the oxidation stability of lard determined at 0sC, and the use of 0 % ethanol extract resulted in a 20-fold increase. In bulk rapeseed oil hexane extracts from rosemary and sage were also more efficient than ethylacetate or acetone extracts. It was established that rosemary extracts were more active than sage extracts, and that rapeseed oil was more efficiently stabilized than sunflower oil.

The antioxidative effect of rosemary ethanol extract on butter, ^{, 2} as well as on filleted and minced fish during frozen storage was studied. Rosemary antioxidants were found suitable for deep frying in edible oils, ⁴ especially in the presence of ascorbyl palmitate. Reblova et al. investigated the effect of acetone and ethyl acetate extracts on the changes in rapeseed oil and in an oil containing polysiloxanes during frying of potatoes. The authors established that the rosemary extract inhibited the formation of polar substances, polymers and decomposition of polyunsaturated triacylglycerols, especially in the case of rapeseed oil, and improved the sensory attributes of French fries. Barbut et al. ¹ studied the effectiveness of rosemary oleoresin (RO) in turkey breakfast sausages. The authors found that RO was as effective as the combination of BHA, or butylated hydroxytoluene (BHT), with citric acid in suppressing oxidative rancidity. A standardized RO has many different phenolic components. It is thought that they act in

synergy to provide antioxidant activity. Results from the oxidation of stripped soybean oil exposed to fluorescent light, in the

presence of rosmariquinone (RQ) and RO ⁷ indicated that RO contained compounds, such as chlorophyll, pheophytin and mono- and diglycerides, which under light interfere with the antioxidant components, thus reducing the antioxidant activity. This was confirmed by the highest level of antioxidant activity exhibited by the RQ in comparison to RO.

Lai et al. ⁸ and Murphy et al. ⁹ investigated the antioxidant properties of RO alone or in combination with sodium tripolyphosphate (STPP) in controlling lipid oxidation in restructured chicken nuggets and in precooked roast beef slices during refrigerated and frozen storage. Stoick et al. ⁰ studied the oxidative stability of restructured beef steaks processed with RO, tertiary butylhydroxyquinone (TBHQ), and STPP. They found that the addition of RO gave no benefit over STPP. The RO/STPP combination was equivalent to TBHQ/STPP treatment in preventing oxidation.

Wada and Fang ¹ observed a strong synergistic effect between rosemary extract

and -tocopherol %) in sardine oil at 30sC and in frozen-crushed fish meat models. The authors suggested that rosemary extract functions as a hydrogen atom donor regenerating the -tocopheroxyl radical to -tocopherol. Synergistic effects were also found between rosemary and sage extracts, and tocopherols or soybean meal hydrolysates in a linoleic acid emulsion.

Basaga et al. reported that rosemary extract and BHT, when added as mixtures of

, 0 and 5 had a synergistic effect on preventing oxidation of soybean oil. A

very pronounced synergistic effect was seen between citric acid and rosemary extract.

Chemical structure

Concurrent with the evaluation of rosemary extracts as antioxidants to inhibit lipid oxidation in food systems, research was also focused on isolation, identification and testing of the active compounds contained in the extracts. In a study of 6 compounds isolated from rosemary Bracco et al. concluded that the antioxidant activity of rosemary extracts is primarily related to two phenolic diterpenes, carnosol and carnosic acid. This conclusion was confirmed by other investigators. Nakatani and Inatani ⁵ identified rosmanol and carnosol and found that both were more effective than -tocopherol, BHT and BHA. The same authors also isolated rosmadial from rosemary.

Several other antioxidative diterpenes such as epirosmanol and isorosmanol, rosmaridiphenol and rosmariquinone have been reported to contribute to the antioxidant activity of rosemary extracts. During the storage and extraction of rosemary carnosic acid is partially converted either into carnosol or into other diterpenes such as rosmanol.

Rosmarinic acid (RA) was reported by Gerhardt and Schro ter to be the second most frequently occurring caffeic acid ester, following chlorogenic acid, and to have antioxidant activity equivalent to that of caffeic acid. The authors detected RA in rosemary, balm, sage, thyme, oregano, marjoram, savory, peppermint, and for the first time in basil.

There are many data in the literature concerning the antioxidative properties of the individual compounds isolated from rosemary. Brieskorn and Domling ³ showed that carnosic acid and carnosol were as effective as BHT and that their effectiveness was concentration dependent. The authors noted that the activity of both compounds was due to the cooperation of their ortho phenolic groups with their isopropyl group.

It was also reported that rosmanol had greater antioxidant activity than carnosol, with carnosic acid being more potent than carnosol. ⁴ In soybean oil carnosic acid was found to be more active than BHT and BHA, but less active than TBHQ. Carnosic acid and carnosol showed the ability to chelate iron and were effective radical scavengers of peroxyl radicals. ⁴ It has been established ⁷ that the molecules of carnosol and the radicals formed from them participate in the reactions of chain initiation and propagation to a much lower degree than is the case with most natural and synthetic antioxidants. Houlihan et al. ⁷ found rosmaridiphenol to be more active than BHA in lard and equivalent to BHT in this test system. They reported also that RQ was superior to BHA and equivalent to BHT in controlling the oxidation of lard. ⁸ RQ has been shown to have good antioxidant activity also in soybean oil. ⁷ Hall et al. proved that RQ acted as a hydrogen-donating antioxidant. Isorosmanol and epirosmanol showed high activity in both lard and linoleic acid; ⁶ in lard they were four times more active than BHA and BHT. Nakamura et al. ⁶ reported that RA exhibited a significantly higher superoxide

scavenging activity than ascorbic acid.

As far as the complex food systems are concerned, it is important to clarify the antioxidative behaviour not only in bulk oil, but also in oil-in-water emulsions, ⁹ as well as in microsomal and liposomal systems. ⁴ Frankel et al. ⁷ reported that in bulk corn oil rosemary extract, carnosic and rosmarinic acids were significantly more active than carnosol. In contrast, in corn oil-in-water emulsion, the rosemary compounds were less active than in bulk oil, and the rosemary extract, carnosic acid and carnosol were more active than rosmarinic acid. The decreased antioxidant activity of the polar hydrophilic rosemary compounds in the emulsion system may be explained by their interfacial partitioning into water, thus becoming less protective than in the bulk oil system.

Carnosol and carnosic acid were powerful inhibitors of lipid peroxidation in microsomal and liposomal systems.

Cuvelier et al. ⁰ found no correlation between the antioxidative effectiveness of the rosemary extracts from different pilot-plant or commercial sources and their composition in 0 specific phenols, a finding which illustrates the complex influence of the various factors on lipid oxidation stability.

Sage: antioxidant properties

Salvia officinalis L, commonly known as sage (Dalmatian sage), is used in foods for flavouring and seasoning. It was found that, along with rosemary, it had the best antioxidant activity among the numerous herbs, spices and teas tested. ¹ Its extracts are also well known as efficient antioxidants.

Since methanol and ethanol were found to be the most suitable solvents for extraction of antioxidants from the plant materials, a number of publications have dealt with further purification of the alcohol extracts. Vacuum steam distillation⁴ or molecular distillation are recommended for use on production scale.

Since rosemary and sage belong to the Labiatae family, it is not surprising to find the same antioxidants in both plants: carnosol, ⁴ carnosic acid, ⁷ rosmanol, rosmadial, ⁰ rosmarinic acid. ⁶ Various methyl and ethyl esters of carnosol, rosmanol, and carnosic acid can be found in sage, as well as in other Labiatae plant extracts; in most cases the compounds are believed to be artefacts from the extraction procedures.

The main antioxidative effect of sage was reported to relate to the presence of carnosic acid, carnosol and rosmarinic acid.

References

1 VALENZUELA A B and NIETO S K, Synthetic and natural antioxidants: food quality protectors , Grasas y Aceitas, 6 7

2 RAC M and OSTRIC B, Les proprietes antioxigenes du romarin, Rev Franc Corps

Gras, 5 2

BERNER D L and JACOBSON G A, Spice antioxidant principle and process for the extraction thereof , US Patent, 3 3 2

4 CHANG S S, OSTIC-MATIJASAVIC B, HSIEH O A L and HUANG C L, Natural antioxidants from rosemary and sage, J Food Sci, 7 2

5 BRACCO U, LO LIGER J and VIRET J-L, Production and use of natural antioxidants, J Amer Oil Chem Soc, 1 8

6 INAHATA K, NAKASAKI T, MATSUMORA S and NAKAHARA T, Odorless and safe antioxidants derived from rosemary and their preparation , Jpn Kokai Tokkyo Koho JP, 9 6 8 7 7

7 GERARD G, QUIRIN K-W and SCHWARZ E CO -extracts from rosemary and sage

Food Market Technol, 5 )

8 LOREZ-SEBASTIAN S, RAMOS E, IBANEZ E, BUENO J M, BALLESTER L, TABERA J and REGLERO G, Dearomatization of antioxidant rosemary extracts by treatment with supercritical carbon dioxide, J Agric Food Chem, 9 8 6

9 HUISMAN M, MADSEN H L, SKIBSTED L H and BERTELSEN G, The combined effect of rosemary (Rosmarinus officinalis L) and modified atmosphere packaging as

protection against warmed over flavour in cooked minced meat, Z Lebensmittel

Untersuch Forsch, 4 8

0 WU J W, LEE M-H, HO C-T and CHANG S S Elucidation of the chemical structures of natural antioxidants isolated from rosemary, J Amer Oil Chem Soc, 2 9

1 BARBUT S, JOSEPHSON D B and MAURER J, Antioxidant properties of rosemary oleoresin in turkey sausage, J Food Sci, 5 0 1

2 NOZAKI K, Antioxidant activity of rosemary, New Food Ind (Japan), 9 1

3 FANG X and WADA S, Enhancing the antioxidant effect of alpha-tocopherol with rosemary in inhibiting catalysed oxidation caused by Fe ⁺ and hemoprotein, Food Research Intern, 3 6 1.

4 ARUOMA O I, SPENCER J P E, ROSSI R, AESCHBACH R, KHAN A, MAHMOOD N, MUNOZ A, MURCIA A, BUTLER J and HALLIWELL B, An evaluation of antioxidant and antiviral action of extracts of rosemary and Provencal herbs, Food Chem Toxicol, 6

5 BASAGA H, TEKKAYA C and ACKIKEL F, Antioxidative and free radical scavenging properties of rosemary extract, Food Sci Technol (London), 97 0

6 PAZOLA Z, KORCZAK J and GOGOLEWSKI M, Studies on the antioxidative properties of spices from the Labiatae family. II. Attempt at identification of antioxidative components of rosemary and sage, Roczn Acad Roln Pozn, 9 0 CCXVIII

7 MARINOVA E, YANISHLIEVA N and GANEVA I, Antioxidative effect of Bulgarian rosemary and inhibiting activity of its carnosol , Oxidation Communications,

4

8 CHEN Q, SHI H and HO C-T, Effects of rosemary extracts and major constituents on lipid oxidation and soybean lipoxygenase activity, J Amer Oil Chem Soc, 2

9 POKORNY J, NGUYEN H T T and KORCZAK J, Antioxidant activities of rosemary and sage extracts in sunflower oil, Nahrung, 7 1

0 POKORNY J, REBLOVA Z, TROIAKOVA L, NGUYEN H T T, KORCZAK J and JANITZ W

Antioxidant activities of spices and herbs in rapeseed oil, Proceedings of the World Conference on Oil Seed and Edible Oils Processing, . October , Istanbul, Turkey, Eds Koseoglu S S, Rhee K C and Wilson R F, Champaign, Illinois, Vol. II,

, pp.

1 ZEGARSKA Z, AMAROWICZ R, KARMAC M and RAFALOWSKI R, Antioxidative effect of rosemary ethanolic extract on butter, Milchwissenschaft, 6 1

2 ZEGARSKA Z, RAFALOWSKI R, AMAROWICZ R, KARMAC M and SHAHIDI F, Stabilization of butter with deodorized rosemary extract, Z Lebensmittel Untersuch Forsch,

6

3 VARELTZIS K, KOUFIDIS D, GAVRIILIDOU E, PAPAVEREGOU E and VASILIADOU S

Effectiveness of a natural rosemary (Rosemarinus officinalis) extract on the stability of filleted and minced fish during frozen storage , Z Lebensmittel Untersuch Forsch,

7 5

4 GORDON M H and KOURIMSKA L, The effect of antioxidants on changes in oil during heating and deep frying, J Sci Food Agric, 5 8

5 GORDON M H and KOURIMSKA L, Effect of antioxidants on losses of tocopherols during deep-fat frying, Food Chemistry, 5 2

6 REBLOVA Z, KUDRNOVA J, TROJAKOVA L and POKORNY J, Effect of rosemary extracts on the stabilization of frying oil during deep fat frying, J Food Lipids, 9 6

7 HAL III C, CUPPETT S, WHEELER D and FU X, Effects of bleached and unbleached

rosemary oleoresin and rosemariquinone on light-sensitized oxidation of soybean oil, J Amer Oil Chem Soc, 4 1

8 LAI S-H, GRAY J I, SMITH D M, BOOREN A M, CRACKEL R L and BUCKLEY D J Effect of oleoresin rosemary, tertiary butylhydroquinone, and sodium tripolyphosphate on the development of oxidative rancidity in restructed chicken nuggets, J Food Sci,

9 MURPHY A, KERRY J E, BUCKLEY D J and GRAY J I, The antioxidative properties of rosemary oleoresin and inhibition of off-flavors in precooked roast beef slices , J Sci Food Agric, 8 7

0 STOICK S M, GRAY J I, BOOREN A M and BUCKLEY D J, Oxidative stability of restructed beef steaks processed with oleoresin rosemary, tertiary butylhydroquinone and sodium tripolyphosphate, J Food Sci, 1 6

1 WADA S and FANG X, The synergistic antioxidant effect of rosemary extract and - tocopherol in sardine oil model system and frozen-crushed fish meat, J Food Process Preserv, 2 6

2 KORCZAK J, JANITZ W and NOGALA-KALUCKA M Synergism of natural antioxidants in preserving of lipids, Proceedings of the 7^th Annual Meeting of Polish Academy of Science, Szczecin, Poland, 6, 8 June, pp.

3 CHIPAULT J R, MIZUNO G R, HAWKINS J M and LUNDBERG W O, The antioxidant properties of natural spices, Food Research, 2 7

4 ARUOMA O I, HALLIWELL B, AESCHBACH R and LO LIGER J, Antioxidant and pro- oxidant properties of active rosemary constituents: carnosol and carnosic acid, Xenobiotica, 2 2 7

5 NAKATANI N and INATANI R, Structure of rosmanol a new antioxidant from rosemary

(Rosmarinus officinalis L), Agric Biol Chem, 81 5

6 NAKATANI N and INATANI R, Two antioxidative diterpenes from rosemary

(Rosmarinus officinalis L) and a revised structure for rosmanol, Agric Biol Chem,

4 8

7 HOULIHAN C M, HO C-T and CHANG S S, Elucidation of the chemical structure of a novel antioxidant, rosmaridiphenol, isolated from rosemary, Amer Oil Chem Soc,

4 1

38 HOULIHAN C M, HO C-T and CHANG S S, The structure of rosmariquinone a new antioxidant isolated from Rosmarinus officinalis L, Amer Oil Chem Soc, 1985 62 968.

9 WENKERT E, FUCHS A and MCCHESNEY J D, Chemical artefacts from the family

Labiatae , J Organ Chem, 5 0

0 SCHWARTZ K and TERNES W, Antioxidative constituents of Rosmarinus officinalis and Salvia officinalis II Isolation of carnosic acid and formation of other phenolic diterpenes , Z Lebensmittel Untersuch Forsch, 2 5

1 HALL III C A and CUPPETT S L, Structure-activities relationship of natural antioxidants, in: Antioxidant Methodology: in vivo and in vitro Concepts, Eds. Auroma O I and Cuppett S L, Champaign, Illinois, AOCS, , pp

2 GERHARDT U and SCHRO TER A, Rosmarinic acid an antioxidant occurring naturally in herbs, Fleischwirtschaft, 3 3

3 BRIESKORN C H and DOMLING H-J Carnosolsaure, der wichtige antioxidativ wirksame Inhaltsstoff des Rosmarin- und Salbeiblattes, Z. Lebensmittel Untersuch Forsch, 9 1

4 RICHHEIMER S L, BERNERT M W, KING G A, KENT M C and BAILEY D T, Antioxidant activity of lipid soluble phenolic diterpenes from rosemary, J Amer Oil Chem Soc,

6 3

5 HALL III C A, CUPPERT S L and DUSSAULT P, Hydrogen-donating mechanism of rosemariquinone, an antioxidant found in rosemary, J Amer Oil Chem Soc, 8

6 NAKAMURA Y, OHTO Y, MURAKAMI A and OHIGASHI H, Superoxide scavenging activity of rosmarinic acid from Perilla frutescens Britton Var. acuta f. viridis , J Agric Food Chem, 8 6

7 FRANKEL E N, HUANG S-W, AESCHBACH R and PRIOR E, Antioxidant activity of rosemary extract and its constituents, carnosic acid, carnosol, and rosmarinic acid, in bulk oil-in-water emulsion, J Agric Food Chem, 6 4

8 HOPIA A I, HUANG S-W, SCHWARZ K, GERMAN J B and FRANKEL E N, Effect of different lipid systems on antioxidant activity of rosemary constituents carnosol and carnosic acid, J Agric Food Chem, 6 4

9 HUANG S-W, FRANKEL E N, SCHWARZ K, AEASCHBACH R and GERMAN J B, Antioxidant activity of carnosic acid and methyl carnosate in bulk oils and oil-in-water emulsions, J Agric Food Chem, 6 4

0 CUVELIER M-E, RICHARD H and BERSET C, Antioxidative activity and phenolic composition of pilot-plant and commercial extracts of sage and rosemary, J Amer Oil Chem Soc, 6 3

1 CHIPAULT J R, MIZUNO G R, HAWKINS J M and LUNDBERG W O, Antioxidant properties of spices in oil-in-water emulsion, Food Research, 9 5 0

2 DJARMATI Z, JANKOV R M, SCHWIRTLICH E, DJULINAC B and DJORDJEVIC A, High antioxidant activity of extracts obtained from sage by supercritical CO₂ extraction, J Amer Oil Chem Soc, 91 8

3 ABDALLA A E and ROOZEN J P, Effect of plant extracts on the oxidative stability of sunflower oil and emulsion, Food Chemistry, 9 4

4 BRIESKORN C, FUCHS A, BREDENBERG J, MCCHESNEY J and WENKERT E, The structure of carnosol, J Organ Chem, 4 9

5 LINDE H Ein neues Diterpen aus Salvia officinalis L und eine Notiz zur Konstitution von Pikrosalvin, Helv Chim Acta, 4 6

6 CUVELIER M-E, BERSET C and RICHARD H, Separation of major antioxidants in sage by high performance liquid chromatography, Sci Aliments, 4 4

7 CUVELIER M-E, BERSET C and RICHARD H, Antioxidant constituents in sage (Salvia officinalis , J Agric Food Chem, 4 2 9