IONIȚĂ Ana Corina1, MITITELU Magdalena1*, NICOLESCU Teodor Octavian2, NICOLESCU Florica3, HÎRJĂU Mircea4, NEACȘU Sorinel Marius1, HÎNCU Lucian4, DĂRĂBAN Adriana5
1Clinical Laboratory and Food Hygiene Department, Faculty of Pharmacy, ”Carol Davila” University of Medicine and Pharmacy, 6, Traian Vuia Street, 020956, Bucharest (ROMANIA)
2*Organic Chemistry Department, Faculty of Pharmacy, ”Carol Davila” University of Medicine and Pharmacy, 6, Traian Vuia Street, 020956, Bucharest (ROMANIA)
3Toxicology Department, Faculty of Pharmacy, ”Carol Davila” University of Medicine and Pharmacy, 6, Traian Vuia Street, 020956, Bucharest (ROMANIA)
4Pharmaceutical Technology Department, Faculty of Pharmacy, ”Carol Davila” University of Medicine and Pharmacy, 6, Traian Vuia Street, 020956, Bucharest (ROMANIA)
5Phaculty of Pharmacy, „Vasile Goldis” Western University of Arad (ROMANIA)
*corresponding author: email@example.com
Marine lipids generally and mussel lipids in particular are valuable because of their rich composition in fat-soluble vitamins (like A, D, E) and polyunsaturated fatty acids. The purpose of this paper is to prepare and to characterise some cosmetic creams and ointments with total lipid extract from mussel tissue. The preparations obtained were analysed according to the present standards in order to determine their main characteristics and stability in time.
Keywords: mussel total lipids, dermatocosmetic preparations, polyunsaturated fatty acids
Seafood is a very good source of quality protein, with a high nutritional value and a balanced profile of healthy fats. Mussels are edible, their meat is a delicacy much appreciated by nutritionists. They are grown in aquaculture systems, in special marine plantations. Mussels are filter organisms and therefore the quality of the marine environment strongly influences their chemical composition. Thus, in heavily polluted marine waters the accumulation of pollutants in mussels is very high [1,2].
Mussels are characterized by a delicate pleasant taste, high nutritional value and extremely effective healing properties, useful for improving metabolism, stimulating the blood formation process, increasing overall tone and improving immunity. Mussel meat is rich in glycogen and minerals, contains over 30 different macro and microelements, almost the entire group of vitamins, including provitamin D3 and a number of enzymes that improve digestion [3,4]. The complex biochemical composition of the total lipid extract in mussel meat gives it valuable therapeutic properties. The high content of unsaturated fatty acids, along with fat-soluble vitamins make the extract a valuable ingredient for cosmetic preparations with emollient, nourishing, antioxidant and regenerative properties [5,6]. In addition, it has been found experimentally that the extract also has anti-inflammatory action [7,8,9]. Compared to the protein content, the lipid fraction is found in a much smaller proportion in the meat of mussels, up to 4% of the dry matter, but it must be remembered that it has a particular biological value. Thus, polyunsaturated fatty acids (such as linoleic, arachidonic) represent 30-40% of total fatty acids, such an amount being rare in animal fats. In addition, prostaglandins and their precursors (polyunsaturated fatty acids) have been identified in the lipids of mussel meat, which are particularly important in medical practice, acting as immunomodulators, hypocholesterolemics [10-15].
After isolating and purifying the total lipid extract from the mussel meat, a series of topical preparations for external use were developed: emollient and nourishing cosmetic creams and anti-inflammatory and healing ointments. To obtain the lipid extract, adult specimens of Mytilus galloprovincialis were used, with a length of 5-7 cm harvested in April 2019 from the Romanian Black Sea coast. After washing under running water, the live mussels were kept in an aquarium for 48 hours.
The Stages of the Process for Obtaining Topical Preparations with Total Lipid Extract from Mussels:
- fresh tissue (180g) was treated with 400 ml of chloroform:methanol mixture (1:2 v/v), and total lipids were extracted with Soxhlet;
- the total lipid extract was then purified in a separatory funnel in which a mixture of chloroform: methanol: 0.9% KCl solution (10:10:9 v/v) was introduced; the lower layer was retained and concentrated in vacuo to give the purified lipid extract.
Tables 1 and 2 show the formulas of topical preparations with total lipid extract from mussels. In the formulation of cosmetic preparations, substances were used which, together with the total lipid extract from mussels, contribute to soaking and softening the skin. From the category of fat-soluble emollients we used: cetyl alcohol, wax, cetaceum, lanolin, paraffin oil and olive oil, and from the category of hydrophilic emollients: glycerol and 70% sorbitol solution.
Table 1. Composition of cosmetic creams with total lipid extract from mussels
|COMPONENTS||Cream 1 (C1) (g)||Cream 2 (C2) (g)||Cream 3 (C3) (g)|
|LIPOPHILIC PHASE||Cetyl alcohol||7||–||2|
|Lipid extract from mussels||1.5||1||1|
|HYDROPHILIC PHASE||Tween 80||–||–||4|
|Preservative solution||at 100||at 100||at 100|
Table 2. Composition of ointments with total lipid extract from mussels
|COMPONENTS||Ointment 1 (O1) (g)||Ointment 2 (O2) (g)|
|Lipid extract from mussels||2.5||1.5|
Marigold tincture is obtained from marigold flowers (Calendula officinalis) by maceration with dilute alcohol (20%). Marigolds have as active principles triterpene saponosides, carotenoids, volatile oil, bitter substances, mucilages, cholesterol esters of fatty acids, protein substances, calendula, vitamin C. Marigold tincture is a yellow-orange liquid with a characteristic, pleasant smell. Store in colored glass bottles tightly closed. Externally, it has an anti-inflammatory and healing effect by stimulating blood circulation to the tissues .
Camphor is obtained from the crystallizable part of the oil extracted by distillation from the wood and leaves of the species Cinnamomum camphora (Lauraceae). Natural camphor is levorotatory, and synthetic camphor is dextrorotatory or racemic. It comes in the form of a crystalline mass, translucent or white crystalline powder, with a characteristic odor and a hot-burning taste. It volatilizes at room temperature. It has a slightly anesthetic and revulsive action.
Propolis or “bee glue”, in its natural state or in the form of extracts obtained by applying gentle processes, is a particularly interesting natural source of products with complex biological action. Resin, waxes, volatile oils, pollen, carbohydrates, amino acids, vitamins, enzymes, mineral salts and impurities have been reported mainly in the chemical composition of crude propolis. The chemical composition of propolis is neither constant nor reproducible, it varies depending on the geographical area and the plant source visited by bees to collect the raw materials from which it is produced. However, in all the propolis samples studied, the predominant components are flavonoid compounds (flavones, flavonones, flavonols), phenolic acids and their derivatives – compounds that have demonstrated strong biological action in laboratory and clinical experiments [17,18]. In principle, the process for preparing the standardized propolis fluid extract with 10% bioflavanoids expressed in chrysin is based on obtaining an extract solution from the crude propolis, concentrating this solution under low pressure at temperatures below 50°C and standardizing the extract obtained at the concentration of 10% bioflavonoids expressed in chrysin. For the preparation of cosmetic creams and ointments, the process of mixing the two phases (lipophilic and hydrophilic) heated to the same temperature was used [19,20]. Thus, the lipophilic components are melted on a water bath, adding in descending order the melting points and the hydrophilic phase is heated to about 70°C and added in a thin line under continuous stirring over the lipophilic phase. After emulsification and cooling to about 30°C, the tincture of marigold or propolis is added dropwise and the homogenization is continued until complete cooling.
Quality control of preparations
The quality of the creams and ointments made for the selection of the best formulas was followed. The preparations were stored in suitable, tightly closed containers at room temperature (maximum 25 ° C). The following control tests were performed: appearance, color, smell, type of emulsion, pH, tensile capacity, viscosity, drip point, penetration capacity, stability at low and high temperatures.
Determination of appearance: determination of appearance was made in accordance with the provisions of Xth Romanian Pharmacopoeia  of the monograph “Ointment”.
Determination of the type of emulsion: – Dilution method: it is based on the principle that an emulsion is miscible with a liquid similar to the external phase; the test is performed by adding a drop of emulsion in 5mL of water or oil. Shake gently. The emulsion droplet diffuses uniformly in water when the emulsion is O/W type or disperses homogeneously in oil when the emulsion is W/O type.
– Staining method: in the presence of a water-soluble dye, the O/W emulsions are stained evenly. Use 0.25% methylene blue solution. W/O emulsions are uniformly stained in the presence of a lipophilic dye (Sudan III 1%). The observation can be made with the naked eye or under a microscope.
Determination of pH: the pH value of the aqueous phase is determined after extraction of 1 g of sample with 5 g of water (maintained at 60°C for 10 minutes). After cooling, the two phases are separated and the pH is determined potentiometrically.
Determination of tensile capacity: the test was performed by the Ojeda-Arbussa method .
Viscosity determination: viscosity was determined using the Brookfield rotary viscometer according to the working technique presented in FRX.
Determination of the drip point: the drip capacity was determined using the Ubbelohde apparatus according to the working technique presented in Xth Romanian Pharmacopoeia.
Determination of penetration capacity: it was determined using the Labor type penetrometer according to the working technique presented in Xth Romanian Pharmacopoeia.
Determination of stability: it was done by keeping the samples in two temperature conditions (at 2ºC and at 40ºC) as follows: in weighing ampoules provided with a lid, 5g of sample from the prepared preparations are introduced, and the ampoules are kept for 8 hours at the mentioned temperatures (at oven and refrigerator), after which the appearance of the samples is examined. Determination of the water index: weigh 10g of ointment into a calibrated capsule, introduce small amounts of water to the point of incorporation. Leave to stand for 24 hours and remove excess water. Weigh the capsule, and the difference is the amount of water incorporated in 10g of ointment.
The experimental results are shown in Tables 3, 4 and 5 and in Fig. 1 and 2.
Table 3. Characteristics of cosmetic creams with mussel lipid extract
|CHARACTERISTICS||Cream 1 (C1)||Cream 2 (C2)||Cream 3 (C3)|
|Organoleptic examination||Homogeneous appearance, consistent, yellow-brown color, specific smell||Homogeneous appearance, consistent, yellow-brown color, specific smell||Homogeneous appearance, consistent, yellow-brown color, specific smell|
|Drip point||39.5º C||38.5º C||39º C|
|Viscosity||275 cP||268 cP||272 cP|
Table 4. Characteristics of ointments with lipid extract from mussels
|CHARACTERISTICS||Ointment 1 (O1)||Ointment 2 (O2)|
|Organoleptic examination||Homogeneous appearance, consistent, brown color, camphorated specific smell||Homogeneous appearance, consistent, brown color, specific smell|
|Drip point||38.5º C||38º C|
|Viscosity||295 mcP||280 mcP|
From the data presented in Tables 3 and 4 it is found that all preparations are homogeneous and have a pH compatible with that of the skin. In addition, all preparations maintained their homogeneous appearance after being kept for 8 hours at 2ºC and 40ºC, respectively. The values of the penetration capacity are presented in Table 5:
Table 5. Penetration capacity values (mm)
|Cream 1 (C1)||Cream 2 (C2)||Cream 3 (C3)||Ointment 1 (O1)||Ointment 2 (O2)|
|Test after 30 days||19||22||16||24||21|
The values of the stretching capacity of the preparations are presented in the figures below:
Fig. 1. Strength surface (S) values depending on the weights applied to the cosmetic creams (weight of the glass plate 153 g)
Fig. 2. Strength surface (S) values depending on the weights applied to the ointments with mussel lipid extract (weight of glass plate 153 g)
The rheological studies carried out on the prepared preparations showed in general a good consistency for all the preparations, in addition the preparations are not sticky and they spread well on the skin. Among the cosmetic creams, the best penetration and stretching capacity was presented by formula C2. Both ointment formulas showed good and close values for penetration and spreading capacity.
With the total lipid extract isolated and purified from mussel meat, a series of topical preparations for external use were developed: three emollient and nourishing cosmetic creams, an anti-inflammatory ointment and a healing ointment.
The control tests carried out on the prepared preparations showed a homogeneous appearance for all preparations, a good stability and a pH compatible with that of the skin. Rheological studies have also shown good consistency for all preparations.Cosmetic creams have a good stretching capacity and, in addition, when applied to the skin are not sticky or oily, leaving behind a pleasant feeling of softness. Future investigations will highlight the optimal way to capitalize on this extract in various pharmaceutical forms.
- Mititelu, M., Moroşan, E., Neacșu, S. M., Ioniţă, E. I. (2018). Research regarding the pollution degree from romanian Black Sea coast. Farmacia 66(6), pp. 1059-1063
- Mititelu, M., Ghica, M., Ioniţă, A. C., Moroşan, E. (2019). The influence of heavy metals contamination in soil on the composition of some wild edible mushrooms. Farmacia 67(3), pp. 398-404
- Buşuricu, F. (2004). Aplicaţii în industria alimentară a fosfatazei alkaline izolate din Mytilus galloprovincialis, Rev. De med. şi farm. – Orvosi es gyogyszereszeti 0szmle, 50, Supl. II, pp. 247-249
- Buşuricu, F. (2005). The study of enzimatic cinetic alkalkine phosphatases in the presences metalic ions, TMJ, Vol.55, Nr.5, pp. 305-308
- Burdge, G. C., Wootton, S. A. (2002). Conversion of α-linolenic acid to eicosapentaenoic, docosapentaenoic and docosahexaenoic acids in young women. Br J Nutr 88(4), pp. 411-420
- Ameur, A., Enroth, S., Johansson, A., et al. (2012). Genetic adaptation of fatty-acid metabolism: a human-specific haplotype increasing the biosynthesis of long-chain omega-3 and omega-6 fatty acids. Am J Hum Genet 90(5), pp. 809-820
- Flock, M. R., Harris, W. S., Kris-Etherton, P. M. (2013). Long-chain omega-3 fatty acids: time to establish a dietary reference intake. Nutr Rev 71(10), pp. 692-707
- Stillwell, W., Wassall, S. R. (2003). Docosahexaenoic acid: membrane properties of a unique fatty acid. Chem Phys Lipids 126(1), pp. 1-27
- Mititelu, M., Nicolescu, T. O., Moroșan, E., Pop, A. L., Nicolescu, F. (2020). Study of Hypolipemiant Activity of Some Mussel Extracts. Proceedings of 6 th International Conference on Interdisciplinary Management of Diabetes Mellitus and its Complications INTERDIAB (Bucharest, Romania, 5-7 March 2020), Filodiritto Editore – Proceedings, pp.338-343, ISBN 978-88-85813-79-3
- Ioniță, A. C., Mititelu, M., Nicolescu, T. O., Nicolescu, F., Moroșan, E., Ozon, E. A., Ioniță, E. I. (2020). Research on the Hypolipidemic Action of Almond Oil and Almond Seeds. Proceedings of 6 th International Conference on Interdisciplinary Management of Diabetes Mellitus and its Complications INTERDIAB (Bucharest, Romania, 5-7 March 2020), Filodiritto Editore – Proceedings, pp. 326-331
- Ioniță, A. C., Ghica, M., Moroșan, E., Nicolescu, F., Mititelu, M. (2019). In vitro effects of some synthesized aminoacetanilides N’- substituted on human leukocytes separated from peripheral blood. Farmacia 67(4), pp. 684-690
- Ioniță, A. C., Nicolescu, T. O., Ioniţă, E. I., Nicolescu, F., Mititelu, M. (2020). Chemical-Toxicological Research on the Evaluation of Serum Malondialdehyde Levels in Patients with Diabetic Arteriopathy. Proceedings of 6 th International Conference on Interdisciplinary Management of Diabetes Mellitus and its Complications INTERDIAB (Bucharest, Romania, 5-7 March 2020), Filodiritto Editore – Proceedings, pp. 320-325
- Mititelu, M., Nicolescu, T. O., Udeanu, D. I., Nicolescu, F. (2020). Research on the Influence of Some Sweeteners on Body Weight and Blood Glucose in Laboratory Mice. Proceedings of 6 th International Conference on Interdisciplinary Management of Diabetes Mellitus and its Complications INTERDIAB (Bucharest, Romania, 5-7 March 2020), Filodiritto Editore – Proceedings, pp. 332-337
- Ghica, M., Băncescu, I., Udeanu, D. I., Tăerel, A., Arsene, A. L., Anuța, V., Velescu, B. Șt., Gerghiceanu, F., Mititelu, M., Ioniță, A. C. (2020). Computational Approach for Small Data in Animal Models with Induced Metabolic Disorders. Proceedings of 6 th International Conference on Interdisciplinary Management of Diabetes Mellitus and its Complications INTERDIAB (Bucharest, Romania, 5-7 March 2020), Filodiritto Editore – Proceedings, pp. 98-106
- Mihele, D., Pop, A., Dãrmãnescu, D., Cocu, F. (2008). Comparative studies regarding the influence of the esters and the ethanol amides of the fatty acids from extra-virgin Oleum Olivae on the experimental ulcers and on the gastric activity on rat Jul 2008 in Planta Medica, Vol. 74, pp. 995 – 1228
- Safdar, W., Majeed, H., Naveed, I., Kayani, W.K., Ahmed, H., Hussain, S., et al. (2010). Pharmacognostical study of the medicinal plant Calendula officinalis L. (family Compositae) Int J Cell Mol Biol.;1:108–16
- Mititelu, M., Moroşan, E., Iosif, M., Ioniţă, E. I. (2018). Analisys of quality of different types of honey from various sources. Proceedings of The Romanian National Congress of Pharmacy – 17th Edition, “21st Century Pharmacy – Between Intelligent Specialization and Social Responsibility”, Filodiritto Editore – Proceedings, pp. 84-87
- Mihai, S., Dumitrescu, D. E., Răducanu, M. A., Stoicescu, I., Badea, V. (2019). Phytochemical Profile and Total Antioxidant Capacity of Sempervivum Ruthenicum Koch Hydroethanolic Extract. Revista de Chimie, 70 (1), pp. 23-26
- Stănescu, V. (1983). Tehnica farmaceutica, Ed.Medicala, Bucuresti, pp.55-67
- Gafițanu, E., Popovici, I., Braha, S. (1996). Tehnologie farmaceutica, Iasi, vol.3, pp. 40-44
- XXX. Farmacopeea Romana X. (1993) Ed. Medicala, Bucuresti, pp. 951-952