Chemical Variability of Rosmarinus officinalis Essential Oil According to the Geographical Origin

Introduction

Rosmarinus officinalis L is a medicinal and aromatic plant native to the Mediterranean basin. It is cultivated all over the world for medicinal, pharmaceutical, cosmetic and food purposes. Rosemary contains several active components among which; flavonoids [1], diterpenes such as carnosolic acid [2], tannins [3], rosmarinic acid [4,5]. These compounds are responsible for a wide variety of activities; including anti-cancer activity [2], antioxidants for food preservation [6], anti-inflammatory and analgesic effect [4-7].

Rosemary is also a source of an essential oil whose majority compounds are generally 1.8 cineole, camphor, camphene and α-pinene [8-10]. Several studies have focused on the biological activities of rosemary essential oil (REO). Thus, the REO has a great antioxidant effect [11], anticancer effect [12] and antileishmanial activity [13], in addition to the antimicrobial effects including antibacterial and antifungal activity [10,14,15]. Other studies have shown ovicidal and repellent effect of ROE [16] and also larvicidal properties [17].

In Morocco, R. officinalis L is intensively exploited in the form of dried leaves and essential oils. In 2014, Morocco exported nearly 8,000 tonnes of dry matter rosemary more to the production of essential oils [18]. However, the majority of the essential oils of Moroccan rosemary are intensively extracted from spontaneous plants [19-21].

This work focuses on the qualitative and quantitative analysis of the essential oils of wild rosemary from several regions of Morocco in comparison with the rosemary cultivated in the region of Fez in the aim to determine the influence of the origin and the climate of the growth zone rosemary on the yield and the chemical composition of the essential oils.

The purpose of this analysis is to encourage the production of essential oils from cultivated rosemary under suitable conditions in order to increase the quantitative yield of essential oils and to contribute to the conservation of the species for sustainable development in the regions threatened.

Materials and Methods

Regions of study and plant materials

For cultivated rosemary, the study area is an open space Fez which is part of northern Morocco Saïss basin characterized by the semi-arid bioclimatic stage with temperate winter.

For wild rosemary, the samples were collected from different Moroccan regions. Geographic and climatic characteristics of each region are:

1. Tafoughalt (Berkane) (34° 48′ 20″ N, 2° 24′ 41″ W); this region belongs to the Rif mountain range and it has as climax Ras Foughal (1532 m).

The region is dominated by a semi-arid Mediterranean climate except on the massif of Beni Snassen where a subhumid climate with a cold and wet winter is altered.

2. Taferdouste - Skoura M’daz (Boulemane) (33°28'25.8"N 4°39'05.8"W); it is part of the mountainous area of the middle atlas which is located in the center north of Morocco. This region is a part of the high hills of Tafrdouste with altitudes ranging from 700 m to 1400 m; it belongs to the semi-arid to sub-humid bioclimatic stage.

3. Talzemt (Bouiblane); (33°36'51.3"N 4°11'29.7"W); it belongs to the subhumid meso-mediterranean bioclimatic stage with an average temperature of 3 to 20°C and an average rainfall of 450 to 800 mm/year.

4. Ait Ben Akki (Errachidia) (32°05'15.5"N 4°43'58.1"W); it is located in the High Atlas with an altitude of 1515 m. The bioclimatic stage is arid to semi-arid with a definite moderation between summer and winter.

Figure 1 shows the geographical location of harvest areas of wild and cultivated rosemary.

1- Topographical extract from the region of Berkane - Tafoughalt in the 1/50000.

2- Topographical extract from the region of Boulemane - Talzemt in the 1/100000.

3- Topographical extract from the region of Boulmane - Skoura and Taferdouste in the 1/50000.

4- Topographical extract from the region of Errachidia - Aït Ben Akki) in the 1/50000.

Sampling and extraction of essential oils

Leaves, flowers and flowering tops of cultivated or wild rosemary were freshly harvested and dried in the shade at room temperature (25°C) for 3 days. The drying time of the plant has been studied previously in order to get maximum yield in essential oils.

Essential oils extraction was carried out by hydrodistillation in a Clevenger-type apparatus according to the method recommended by the European Pharmacopoeia [22].

All parameters such as quantity of plant, water volume and also time of distillation have been previously optimized. The extraction was performed for 2 h by boiling the samples of 100 g of rosemary in 500 ml of distilled water at a constant temperature (100°C) in a heating mantle. The distillation was carried out in a system consisting of a flask of tow liter surmounted by a column of 60 cm length connected to a condenser. After training with water vapor, a mixture of essential oil with the water was condensed and then recovered, after cooling, the water and the essential oil are separated by difference in density. The essential oil was stored at 4°C in the dark before their use. The yield of essential oil was determined relative to the dry matter.

Physico-chemical characteristics

Two parameters were determined; the refractive index and the density. The refractive index was measured using an Erma 1537 type refractometer at a temperature of 25°C. The density was determined using a Sartorius TP scale.

Gas chromatography-mass spectrometry analysis

The essential oils of R. officinalis samples were analyzed using gas chromatography coupled to mass spectrometry (GC-MS). The essential oil was analyzed using an Agilent-Technologies 6890 N Network GC system equipped with a flame ionization detector and HP-5MS capillary column (30 m × 0.25 mm, film thickness of 0.25 μm; Agilent-Technologies, Little Falls, CA, USA). The carrier gas was helium with a flow rate of the 1.0 ml/min. The essential oils samples were diluted 1: 100 in n-hexane, and 0.1 μl were injected into the GC systems. The injector and detector temperatures were set at 250°C and 280°C, respectively. The column temperature was programmed from 35°C to 250°C at a rate of 5°C/min, with the lower and upper temperatures being held for 3 and 10 min, respectively. All quantifications were carried out using a built-in data-handling programme provided by the manufacturer of the gas chromatograph. The composition was reported as a relative percentage of the total peak area. The constituents of the volatile oils were also identified by comparing their GC retention indices. A mixture of aliphatic hydrocarbons (C8-C24) in hexane (Sigma-Aldrich, St. Louis, USA) was injected as under the above-mentioned temperature programme to calculate the retention indices using the generalized equation of Van den Dool and Kratz [23].

Conclusion

To conclude, the chemical composition of the essential oil of the cultivated rosemary is very rich in major components and it can compete in quantity as well as in quality and consequently in price the essential oils of the wild rosemary. This wealth can be improved by taking into account the extrinsic conditions that seem to influence the quantity and quality of REO. This comparative study is a good approach from an economic point of view; it makes it possible to better target the desired rosemary according to the region from which it comes in order to extract the major component sought in the REO according to its percentage. Our work can serve as a basis for developing the regional economy to generate financial income for rural and urban populations encouraging them to cultivate rosemary.

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