Saponifiable Fraction Research Articles

Extraction and characterization of the saponifiable lipid fraction from microalgae Chlamydomonas sp. cultivated under stress

Microalgae when subjected to nutrient restriction conditions, inducing environmental stress, are prone to total lipid accumulation and changes in the fatty profile. In this context, the green microalgae Chlamydomonas sp. was submitted for 3 days to the absence of nutrients. Thus, the efficacy of the extraction of the crude hexane lipid fraction (CHF), using mechanical stirred associated with ultrasound technique, was evaluated in two different times (2 and 4 h), using scanning electron microscopy and thermogravimetric analysis (TG). Saponifiable fractions, CHF extracts and crude chloroform fraction (CCF) were esterified and characterized by TG, FTIR and GC/MS to identify their fatty profile. Residual biomass (RB) was analyzed by elemental analysis, and the resulting data were employed to estimate the higher heating value (HHV) and crude protein percentage. The micrographs of the biomass surfaces before and after the extraction process showed significant fragmentation only for the time of 4 h. A pronounced mass loss in the typical lipid range was evidenced by TG being consistent with the significant difference of material extracted in the time of 2 h (2.95 ± 0.28)% and 4 h (10.54 ± 0.46)%. The total lipid fraction (CHF and CCF) was approximately 29%. The RB revealed values of HHV and crude protein of 18 MJ kg−1 and 56%, respectively. The TG data for the CCF extract revealed a material consisting mainly of saponifiable fractions, unlike CHF which is composed predominantly of unsaponifiables. Therefore, the TG curve ratified a better conversion rate to CCF (approximately 89%), consistent with the value obtained by GC/MS (about 86%). The fatty profile for the saponifiable fractions of both extracts showed that the major fatty acids are C16:0 and C18:3 (ω-3 and ω-6). The ester profile with elevated concentration of polyunsaturated fatty acids (C18:3) is unfeasible in their application for biodiesel production; nevertheless, the fatty profile of microalgae suggests pharmacological potential for diet or therapy, since some of the main components are reported as bioactive metabolites.

Biological removal of pharmaceutical compounds using white-rot fungi with concomitant FAME production of the residual biomass

The efficiency of two white-rot fungi (WRF), Trametes versicolor and Ganoderma lucidum, to eliminate thirteen pharmaceutical pollutants with concomitant biodiesel production from the accumulating lipid content after treatment, was examined. The removal efficiency was studied using both individual and combined strains. The results of individual and combined strains showed a total removal (100%) of diclofenac (DCF), gemfibrozil (GFZ), ibuprofen (IBP), progesterone (PGT) and ranitidine (RNT). Lower removals were achieved for 4-acetamidoantipyrin (AAA), clofibric acid (ACF), atenolol (ATN), caffeine (CFN), carbamazepine (CZP), hydrochlorothiazide (HCT), sulfamethoxazole (SMX) and sulpiride (SPD), although the combination of both strains enhanced the system’s efficiency, with removals ranging from 15 to 41%. This increase of the removal efficiency when combining both strains was attributed to the interactions developed between them (i.e., competition). Results from enzymatic and cytochrome P450 examination suggested that both extracellular (laccase, MnP, LiP) and intracellular oxidation mechanisms participate in the biological removal of pharmaceuticals. On the other hand, the “green” potential of the fungal sludge generated during the biological removal process was assessed for biodiesel production by means of one-step direct (in-situ) transformation. This process consists of the simultaneous extraction and conversion of lipids contained in the sludge by catalytic esterification/transesterification using a robust acid heterogeneous Zr-SBA-15 catalyst. This catalytic system provided conversions close to 80% of the saponifiable fraction (including free fatty acids and glycerides) in the presence of high amount of impurities. The overall weight FAME yield, based on the initial dried mass, was close to 30% for both strains.

Isolation and structural elucidation of secondary metabolites obtained from the soft coral Sinularia candidula present in the Egyptian Red Sea

The present study described the isolation and structural elucidation of five compounds (1 – 5) obtained from the Red Sea soft coral Sinularia candidula. Sinularia is a soft coral belonging to the phylum Cnidaria, class Alcyonaria, and family Alcyoniidae. Its taxonomic identification is largely based on the examination of the spicules, and a fairly reliable taxonomic key is available to guide species identification [1]. The presently investigated species yielded two sterols namely, Cholesterol (1), 24-methylene cholesterol (2), two oxygenated compounds namely chimyl alcohol (3), heptadecanoic acid (4), along with the ceramide (R)-2'-hydroxy-N-[(2S,3S,4R)-1,3,4-trihydroxypentacosan-2-yl] nonadecanamide (5). Their structures were elucidated by means of 1D and 2D NMR spectroscopic techniques. Cytotoxicity testing for Sinularia candidula extracts of different polarities was performed against HELA, HCT, MCF-7 and PC3 cell Lines [2], where the hexane-ethyl acetate (1:1) fraction revealed the highest percentage of inhibition by 73.77% and 78.92% against MCF-7 and PC3 cell lines, respectively, while the ethyl acetate fraction showed the highest percentage of inhibition by 67.29% against HCT cell line. This study also includes the investigation of lipoidal matter of the Red sea soft coral Sinularia candidula, where the hydrocarbons as well as the fatty acid contents were identified in both the unsaponifiable and saponifiable fractions of the total extract using GC/MS.

Phytochemical Analysis and Anti-cancer Investigation of Boswellia serrata Bioactive Constituents In Vitro.

Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography- mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate β-boswellic acid and identification of the pure compound was done using UV, mass spectra, 1H NMR and 13C NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and β-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1- hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with IC50 values equal 1.58 and 5.82 μg/mL at 48 h, respectively which were comparable to doxorubicin with an IC50 equal 4.68 μg/mL at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with IC50 values equal 0.12 and 6.59 μg/mL at 48 h, respectively which were comparable to 5-fluorouracil with an IC50 equal 3.43 μg/ mL at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.

Municipal sewage sludge to biodiesel by simultaneous extraction and conversion of lipids

Two different approaches have been investigated for the production of biodiesel from glycerides and free fatty acids (FFAs) extracted from sewage sludge. The first one is a two-step process consisting of organic solvent extraction followed by acid-catalyzed esterification/transesterification of the isolated oil fraction. The second one is a one-step direct transformation consisting of the simultaneous extraction and conversion of the lipid fraction contained in the sewage sludge. In both alternatives, a heterogeneous acid Zr-SBA-15 catalyst has been used. In the two-step extraction–reaction process, conversion close to 90% of the saponifiable fraction (including FFAs and glycerides) were achieved. Remarkably, the catalyst provided such high conversion in the presence of high amounts of unsaponifiable matter. Furthermore, the catalyst kept its activity in successive catalytic runs in presence of this low-quality lipid fraction. In the one-step direct conversion of the dried sludge, the overall weight FAME yield, based on the initial mass of dried sewage sludge, was around 15.5wt% for primary and 10.0wt% for secondary sludge. In contrast, this FAME yield was lower than 6wt% for two-step process when processing primary sludge (being negligible for the secondary sludge). Finally, the results of this work proof the high potential of Zr-SBA-15 as catalyst for the production of biodiesel from a low quality oleaginous feedstock such as municipal sewage sludge.

Enhanced microbial oil production by activated sludge microorganisms from sugarcane bagasse hydrolyzate

The use of sugarcane bagasse hydrolyzate as a carbon source for enhanced oil production by activated sludge microbial cultures was investigated. Cultivation of raw activated sludge inoculum using pure xylose as carbon source was necessary prior to bagasse hydrolyzate feeding for microbial acclimation to this pentose sugar, the major component of the hydrolyzate. Lipid contents from 40 to 47% (dry cell weight) were achieved under high C:N ratio following bagasse hydrolyzate feeding; however nutrient supplementation was found to be necessary in order to maintain viable cell biomass levels (>10 g/L) to achieve a high lipid titer (7.62 g/L). Hence, a process involving sequential batch feeding of hydrolyzate with and without nutrients was proposed and simulated using the Logistic and Luedeking–Piret models. Analysis of the product lipids showed up to 50% saponifiable fractions and dominance of C16 and C18 fatty acids, demonstrating their suitability as biofuel feedstock.

Insecticidal activity of chicory (Cichorium intybus L.) extracts against two dipterous insect-disease vectors: Mosquito and housefly

The plant chicory, Cichorium intybus L. (Asteraceae), was subjected to a bioassay-guided fractionation scheme, starting with successive extraction of the whole plant powder with petroleum ether, chloroform, ethyl acetate and methanol. Toxicity screening against larvae and adults of the mosquito (Anopheles pharoensis) and the housefly (Musca domestica), revealed high potency for petroleum ether and chloroform extracts, with LC50 of 15.3mgkg−1 and 0.023mg/cm2 against larvae and adults of mosquitoes, respectively. The LC50 for housefly larvae equaled 65.8mgkg−1 and the LD50 for adults equaled 0.112μg/insect. Saponification of petroleum ether extract resulted in saponifiable and unsaponifiable fractions, the latter was highly toxic than the former. Their activity was referred to the presence of fatty acid methyl esters in the saponifiable fraction, and sterols and hydrocarbons in the unsaponifiable fraction. Successive TLC fractionation to the chloroformic extract resulted in isolation and identification of two biologically active compounds, e.g., lactucopicrin-15-oxalate (compound I) and chicoralexin (compound II); both showed high toxicity towards the two tested insects. Compound II was more toxic than compound I to the mosquito larvae, while the opposite was obtained for the housefly larvae. The two compounds possessed equitoxic values against the adult stages of both insects. It was concluded that fractionation of the chicory plant (C. intybus) was in favor of toxicity increase towards the two insect pests used in the present study. Moreover, the obtained results provide new data on the insecticidal efficacy of this native plant against pests of medical importance such as mosquitoes and housefly.

Effects of Topical and Dietary Use of Shea Butter on Animals

Shea butter is the fat extracted from the nut of Africa Shea tree (Vitellaria paradoxa). It is used in cosmetic formulations and as a substitute for Cocoa butter in chocolate industries. It is edible and used cooking fat in Africa. The saponifiable fraction of Shea butter is composed primarily of stearic and oleic acids with lesser amounts of palmitic, linoleic and arachidic acids while the unsaponifiable fraction of Shea butter is composed of bioactive substances that are responsible for Shea butter’s medicinal properties. Shea butter is a solid at room temperature and melts at body temperature. It is therefore useful for skin care as it has sun screening properties and acts as an emollient and skin moisturizer. Topical use of Shea butter has also demonstrated anti-aging and anti-inflammatory properties. Dietary intake of Shea butter has hypocholesterolemic effect and reduces serum and organ protein concentrations.

Effects of Oral and Topical Use of the Oil from the Nut of Vitellaria paradoxa

Shea butter is the fat extracted from the nut of Africa Shea tree (Vitellaria paradoxa). It is used in cosmetic formulations and as a substitute for Cocoa butter in chocolate industries. It is edible and used cooking fat in Africa. The saponifiable fraction of Shea butter is composed primarily of stearic and oleic acids with lesser amounts of palmitic, linoleic and arachidic acids while the unsaponifiable fraction of Shea butter is composed of bioactive substances that are responsible for Shea butter’s medicinal properties. Shea butter is a solid at room temperature and melts at body temperature. It is therefore useful for skin care as it has sun screening properties and acts as an emollient and skin moisturizer. Topical use of Shea butter has also demonstrated anti-aging and anti-inflammatory properties. Dietary intake of Shea butter has hypocholesterolemic effect and reduces serum and organ protein concentrations.

Phytochemical and Bioactivity Investigations of <i>Macfadyena unguis-cati</i> L. (<i>Bignoniaceae</i>)

GC/MS of the volatile components of the aerial part of Macfadyena unguis-cati L, Fam. Bignoniaceae revealed the presence of 74 compounds, 52 (75.97%) of them were identified. The major compound was n-decane (12.21%) followed by phytol (12.19%). The saponifiable fraction of the petroleum ether extract contained 21 fatty acids identified as methyl esters. In the unsaponifiable fraction, 37 compounds (representing 93.26%) were identified. β-amyrin, squalene, β-sitosterol and 3α,5-cyclo-ergosta-7,22-dien-6-one were identified in the USM. Determination of LD50 of different extracts showed that total ethanol extract was the safest (4.9 g/kg) followed by petroleum ether extract, (4.5 g/kg) and ethyl acetate extract having the least LD50 (3.1 g/kg). The total ethanol extract was found to be the most potent as antipyretic, followed by ethyl acetate extract. The ethanol extract, as well as the coumarin containing fraction exhibited significant analgesic activity. Key words: Macfadyena unguis-cati L., Bignoniaceae, fatty acids, analgesic activity, antipyretic activity, LD 50 .

EXTRACTION OF LIPID FROM ABALONE (HALIOTIS DISCUS HANNAI INO) GONAD BY SUPERCRITICAL CARBON DIOXIDE AND ENZYME-ASSISTED ORGANIC SOLVENT METHODS

The objective of this study was to establish methods to extract lipids from abalone (Haliotis discus hannai Ino) gonad. Results showed that enzyme‐assisted organic solvent extraction allowed recovering 60.00 ± 2.16% of abalone gonad lipid from the samples digested with neutral protease. In addition, a lipid yield of 82.56 ± 2.77% was achieved by supercritical carbon dioxide (SC‐CO2) extraction. Thin‐layer chromatography‐flame ionization detection analysis indicated that lipid extracted by soxhlet method from H. discus hannai Ino gonad contained 89.05 ± 0.97% of triglycerides, 7.34 ± 0.80% of phospholipids, 1.46 ± 0.08% of cholesterol and 2.15 ± 0.09% of free fatty acids. The lipid extracted was divided into the unsaponifiable fraction (sterol) and the saponifiable fraction, and analyzed by gas chromatography mass spectrometry. Results demonstrated that the compositions of sterol and fatty acid were different for samples extracted by different methods, indicating that the extraction method could influence the composition of the lipids. PRACTICAL APPLICATIONS: Shellfish by‐products, such as viscera, contain high levels of nutrient material, such as protein, fat and polysaccharides, and have certain utility value. So far, there is a lack of utilization methods for the recovery of those substances. In our lab, an enzymatic hydrolysis process has been established to recover polysaccharides and protein (peptide) simultaneously from shellfish by‐product (viscera). However, the enzymatic hydrolysis process did not involve the recovery of another important nutrient content, lipid. In the present study, the lipid was extracted from H. discus hannai Ino gonad by enzyme‐assisted organic solvent method and SC‐CO2 method. The two methods could be combined into the established enzymatic hydrolysis process to accomplish the comprehensive utilization of shellfish viscera.

Bioguided isolation of pentacyclic triterpenes from the leaves of Alstonia scholaris (Linn.) R. Br. growing in Egypt

Ethanolic and aqueous extracts of the leaves and flowers of Alstonia scholaris were evaluated for their antioxidant activity by investigating their effect on blood glutathione levels in alloxan-induced diabetic rats. The ethanolic extract of the leaves was the most active; therefore, its cytotoxicity against HepG2 cells was also tested. Promising GI50 values of 1.96, 4.34 and 4.65 µg mL–1 were observed for the extract, its chloroform and ethyl acetate fractions, respectively. The chloroform active subfraction I (GI50 = 2.97 µg mL–1) yielded betulin (1), betulinic acid (2) and ursolic acid (3) upon purification. Compounds 1–3 were identified using spectroscopic techniques and by comparison with reported data. GLC of unsaponifiable and saponifiable fractions of the hexane extract revealed β-sitosterol (7.37%) and n-tetracosane (54.4%) to be the major sterol and hydrocarbon components, respectively. Linoleic acid (48.89%) was the predominant fatty acid.

Phytochemical and in vitro screening of some Ficus and Morus spp. for hypolipidaemic and antioxidant activities and in vivo assessment of Ficus mysorensis (Roth)

Phytochemical screening of air-dried leaves and fruit juice of certain Ficus and Morus spp. have been studied. In an in vitro study, the ethanol and hexane extracts of the investigated plants were evaluated against hyperlipidaemia by estimating the rate limiting enzyme of cholesterol biothenysis; β-hydroxy-β-methylglutaryl coenzyme A reductase (HMG-CoA reductase). The antioxidant activity was evaluated by reduction of DPPH− free radical. Extra phytochemical screening of Ficus extracts was undertaken, which recorded potent hypolipidaemic and antioxidant activities. The more pronounced extract, Ficus mysorensis (hexane extract), was evaluated in vivo by estimation of the lipid profile and certain antioxidant parameters in hypercholesterolemic rats. The hexane fraction was chromatographed and six isolated compounds were identified. Furthermore, its saponifiable fraction was identified by a MS/MS technique. In conclusion, F. mysorensis recorded hypolipidaemic and antioxidant effects. Detailed studies of the isolated compounds must be undertaken for an evaluation against hypercholesterolemia and free radical elevation.

Hypolipidaemic and antioxidant activities of Ficus microcarpa (L.) in hypercholesterolemic rats

Saponifiable and unsaponifiable fractions of Ficus microcarpa leaves hexane extract have been phytochemically studied and evaluated for its hypolipidaemic and antioxidant effects in hypercholesterolemic rats. The effect of the extract on the lipid profile was assessed by measuring the levels of total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides, phospho and total lipids. Lipid peroxides, glutathione and superoxide dismutase were measured as antioxidants. The work was extended to evaluate liver function indices as well as the histopathological picture of the liver after treatment. Treatment with leaves extract (500 mg kg−1 body weight) 5 times/week for 9 weeks at the same time of cholesterol administration (30 mg/0.3 mL 0.7% tween/animal) recorded an improvement of lipid profile, antioxidants, liver function enzymes and the liver histopathological picture. The lipoidal matters of the unsaponifiable fraction of the hexane extract by GC/MS led to the identification of 22 compounds, while saponifiable fraction by (MS/MS) technique led to identification of 13 unsaturated and saturated fatty acid methyl ester derivatives. It can be concluded that the hexane extract of F. microcarpa L has been proved to have hypolipidaemic and antioxidant effects in hypercholesterolemic rats through its role in counteracting LDL oxidation, enhancement of HDL synthesis and inhibition of lipid peroxidation.

Original article: Extraction of lipid from scallop (Patinopecten yessoensis) viscera by enzyme‐assisted solvent and supercritical carbon dioxide methods

SummaryIn the present study, lipid was extracted from scallop (Patinopecten yessoensis) viscera by using the enzyme‐assisted solvent method and the supercritical carbon dioxide (SC‐CO2) method. Soxhlet extraction with ethyl ether produced a yield of 23.7 ± 0.6 g of lipid 100 g−1 of dry matter. Enzyme‐assisted solvent extraction allowed recovering 60.6 ± 1.5% of P. yessoensis viscera lipid from the samples treated with papain, whereas a lipid recovery rate of 78.3 ± 0.6% was achieved by SC‐CO2 extraction. The lipid extracted was divided into the unsaponifiable fraction (sterol) and the saponifiable fraction (fatty acid) and analysed by gas chromatography mass spectrometry. Results indicated that the fatty acid composition and sterol composition for lipids extracted by different methods were slightly different. Eicosapentaenoic acid and docosahexaenoic acid were dominant polyunsaturated fatty acids accounting for 35–40% of the total fatty acid.

Évaluation de la biodiversité de quelques cucurbitacées d’Afrique subsaharienne à partir des données chromatographiques (CGC et CLHP)

Two triacylglycerol quantitative analysis methods, needing none standard, using liquid chromatography coupled with light scattering detection, as well as fatty acid methyl esters quantitative analysis using capillary gas chromatography were led. In application, seed oil saponifiable fraction composition of eight groups of cucurbitacee's seeds from equatorial and west Africa was determinated Their correct classification and their differentiation using respectively factorial discriminent and principal component analysis were investigated. It is demonstrated that the best method for charac- terizing an oil is the new original triacylglycerol quantitative analysis reported in this article. The interest of the two other methods is also reported.

Évaluation de la biodiversité de quelques cucurbitacées d’Afrique subsaharienne à partir des données chromatographiques (CGC et CLHP)

Article recu le 15 juin 2007 accepte le 25 mai 2008 Abstract: Two triacylglycerol quantitative analysis methods, needing none standard, using liquid chromatography coupled with light scattering detection, as well as fatty acid methyl esters quantitative analysis using capillary gas chromatography were led. In application, seed oil saponifiable fraction composition of eight groups of cucurbitacee’s seeds from equatorial and west Africa was determinated Their correct classification and their differentiation using respectively factorial discriminent and principal component analysis were investigated. It is demonstrated that the best method for characterizing an oil is the new original triacylglycerol quantitative analysis reported in this article. The interest of the two other methods is also reported.

Identification of Organic Compounds in Fictile Unguentaria from Two Sicilian Necropolis of Greek Age (5th Century, b.C.) by GC‐MS Analysis

A study to obtain more knowledge on funeral set in Greek age, (5th Century, b.c.) was carried out on thirteen ancient unguentaria, small vessels used as containers of balms or ointments, founded in two different Sicilian necropolis: Adranon and Hymera. Every find was subjected to three extractions by increasing polarity solvents. All crude extracts, unsaponifiables and methyl esters of saponifiable fraction were analysed by GC-MS. Analysis showed difference between two groups of finds: the unguentaria from Adranon show abundant traces of lipids used in balm making, while those from Hymera resulted empty and buried for ritual purpose. Even if in the two towns, flourished in the same period, adopted probably different funerary uses.

Phytochemical and bioactivity investigations of Macfadyena unguis-cati L. (Bignoniaceae)

GC/MS analysis of the volatile components of the aerial parts of Macfadyena unguis-cati L. (Bignoniaceae) revealed the presence of 74 compounds, 52 of them (representing 75.97%) were identified [1]. The major compound is n-decane (21.21%) followed by phytol (12.19%). The saponifiable fraction of the petroleum ether extract showed 21 fatty acids identified as methyl esters. 37 compounds were identified in the unsaponifiable fraction; representing 93.26%. β-amyrin, squalene, β-sitosterol and 3α', 5-cyclo-ergosta-7,22-dien-6-one were identified in the USM.

NATURAL HAIR RECIPE

Biological evaluation of natural preparations used for treatment of hair loss revealed that the saponifiable fraction was the active fraction. The main constituents of this fraction were palmitic, stearic saturated fatty acids and oleic and linoleic unsaturated fatty acids. Certain modifications were done to increase the activity by addition of Cantharidin or Pilocarpine to the most active subfraction of saponifiable matter. The superior effect on treating hair loss was shown with the formulation containing cantharidin than that which contain pilocarpine