Dynamic Analysis of Phorbol Esters in the Manufacturing Process of Fatty Acid Methyl Esters from <Emphasis Type="Italic">Jatropha curcas</Emphasis> Seed Oil

Jatropha curcas seed oil, which is unsuitable as an edible oil but has received attention as a novel vegetable fat and oil resource, contains tumor-promoting phorbol esters. Currently, six types of derivatives of 12-deoxy-16-hydroxyphorbol (DHPEs) in J. curcas oil have been identified, and their toxicological safety for humans is being discussed. However, it is reported that most DHPEs disappear during the transesterification process. We investigated the dynamics of phorbol esters in the manufacturing process of fatty acid methyl esters from J. curcas seed oil. With the assumption that the precursor ion was the fragment ion (m/z = 311) from the frame unit of phorbol esters and their derivatives, we developed an LC–MS method for detecting the product ion (m/z = 165), which was obtained by cleavage of the fragment ion. The derivatives generated from the structural changes of the phorbol esters existed in fractions of glycerine–water in the manufacturing process; however, phorbol esters and their derivatives were not detected in the fatty acid methyl esters that were produced via a high-vacuum distillation process. Investigation into the dynamics of phorbol esters confirmed that the contents of phorbol esters, including DHPEs, in the fatty acid methyl esters were under detection limits.     

Jatropha curcas,a promising crop for the generation of biodiesel and value‐added coproducts

The review highlights the specific features of the Jatropha curcas plant and its potential for the production of biofuel, protein concentrates as livestock feed and value‐added products that could enhance the economic viability of Jatropha seed oil‐based biodiesel production. The roles of the plant in carbon capture, enhancing socio‐economic conditions, food production in the tropical regions, and influencing micro‐climate, vegetation and soil quality are discussed. The paper also gives a comparative account of the toxic and non‐toxic genotypes of J. curcas from the point of view of their physical and chemical properties and their potential for biodiesel and livestock feed production. Future areas of research are also presented.     

Removal and Degradation of Phorbol Esters during Pre-treatment and Transesterification of <Emphasis Type="Italic">Jatropha curcas</Emphasis> Oil

Phorbol esters present in Jatropha curcas oil are toxic when consumed and are co-carcinogens. These could be a potential constraint in the widespread acceptance of Jatropha oil as a source of biodiesel. Phorbol esters were quantified in the fractions obtained at different stages of oil pre-treatment and biodiesel production. During degumming some phorbol esters were removed in the acid gums and wash water. This implies that the use of these acid gums in animal feed is not possible and care should be taken when disposing the wash water into the environment. Silica treatment did not decrease the phorbol esters, while stripping/deodorization at 260 °C at 3 mbar pressure with 1% steam injection completely degraded phorbol esters. Phorbol esters were not detected in stripped oil, fatty acid distillate, transesterified oil (biodiesel) and glycerine. The presence of possibly toxic phorbol ester degradation products in these fractions could not be ruled out.     

Investigating Some Physicochemical Properties and Fatty Acid Composition of Native Black Mulberry (<Emphasis Type="Italic">Morus nigra</Emphasis> L.) Seed Oil

The physicochemical properties of seed and seed oil obtained from the native black mulberry (Morus nigra L.) were investigated in 2008 and 2009. The results showed that the seed consisted of 27.5–33% crude oil, 20.2–22.5% crude protein, 3.5–6% ash, 42.4–46.6% carbohydrate and 112.2–152.0 mg total phenolics/100 g. Twenty different fatty acids were determined, with the percentages varying from 0.02% myristic acid (C14:0) to 78.7% linoleic acid (C18:2). According to the GC analysis of fatty acid methyl esters, linoleic acid (C18:2), followed by palmitic acid (C16:0), oleic acid (C18:1) and stearic acid (C18:0) were the major fatty acids, which together comprised approximately 97% of the total identified fatty acids. High C18:2 content (average 73.7%) proved that the black mulberry seed oil is a good source of the essential fatty acid, linoleic acid. Linolenic acid (C18:3) was also found in a relatively lower amount (0.3–0.5%). The α-tocopherol content was found to be between 0.17 and 0.20 mg in 100 g seed oil. The main sterols in the mulberry seed oil were β-sitosterol, Δ5-avenasterol, Δ5, 23-stigmastadienol, clerosterol, sitosterol and Δ5, 24-stigmastadienol. The present study stated that the native black mulberry seed oil can be used as a nutritional dietary substance and has great usage potential.     

TAG Profiles of <Emphasis Type="Italic">Jatropha curcas</Emphasis> L. Seed Oil by Easy Ambient Sonic-Spray Ionization Mass Spectrometry

Easy ambient sonic-spray ionization mass spectrometry (EASI–MS) was used to follow the maturation of Jatropha curcas L. seeds via the monitoring of the triacylglycerides (TAG) profile of the oil. Results show that TAG composition is significantly modified during seed development but remains nearly unchanged during storage. The EASI–MS oil analysis performed herein is simple, requires just a tiny droplet of the oil and is performed without any pre-separation or chemical manipulation. The oil from Jatropha gossypifolia L. was also evaluated, and a very different TAG profile was obtained.     

Quantification of Phorbol Esters in Jatropha curcas by HPLC‐UV and HPLC‐ToF‐MS with Standard Addition Method

Established analytic methods for the quantification of phorbol esters (PE), which are some toxic components in Jatropha curcas L., include HPLC with UV‐detection with the commercially available phorbol myristate acetate (PMA) as internal standard or HPLC coupled with MS detection with an external calibration, mostly also with PMA. The differences in the fatty acid side chains and connection to the base structure of PMA compared to PE leads to different UV absorption and MS ionization effects and cause problems for exact quantitative measurements. In this paper, a method is presented which combines both detection types and shows differences between both results. For this purpose, an extraction routine is performed on a PE‐containing seed oil to get a PE standard in high purity, which was used for a standard addition method on two real J. curcas oil samples, derived from Ghana and Mexico. Furthermore, a detection window of ±10 ppm for the high accurate ToF‐MS detection is set to eliminate isobaric interferences from co‐eluting material. Method evaluation of inter‐ and intra‐day variance as well as the recovery rate are performed and determined. With this method a limit of detection of 62 ng mL^?1 (UV) and 11 ng mL^?1 (MS) can be achieved. Practical Applications: Due to the good biological and technical properties of Jatropha curcas L., its seed oil seems perfect for the application as biodiesel feedstock. The toxicity on the other hand could cause problems when converting side products from the oil production to products of higher value. With the here described method an accurate and precise analysis procedure for the quantification of the toxic compounds namely, phorbol esters, could be applied for toxicity studies or routine checks in industry which is converting plant material from J. curcas, so that no toxic material is used for example as animal feed. In this paper, an exact and robust analysis method is described for the quantification of phorbol esters (PE) in Jatropha curcas L. seed oil. This method procedure includes the extraction of PE in methanol, chromatographic separation on a reverse phase C18 HPLC column and the quantification by standard addition method. For the standard addition method a highly pure PE standard is used, which is extracted and purified by semi preparative HPLC right before the measurements. The used detector for identification and quantification is UV set at 280 nm and ESI‐ToF‐MS with a ±10 ppm mass difference of the deprotonated and formate adduct pseudo molecular ion of PE.

     

Quality of Biodiesel Prepared from Phorbol Ester Extracted <Emphasis Type="Italic">Jatropha curcas</Emphasis> Oil

Jatropha curcas seeds are rich in oil (28–32%), which can be converted to high quality biodiesel. The oil is non-edible due to the presence of toxic compounds, namely, phorbol esters (PEs). PEs have a number of agricultural/medicinal/pharmaceutical applications and hence their recovery generates a value added co-product towards the biodiesel production chain. This study aims to assess the effects of PE extraction on quality of both the residual oil and the biodiesel production from it. Two Approaches (1, use of an Ultra-turrax; and 2, use of a magnetic stirrer) were used with an effective treatment time of 2 and 5 min, resulting in 80 and 78% extraction of PEs, respectively. The phosphorus content was reduced by 70.2 and 75.8%, free fatty acids by 55.3 and 55.6%, and the fatty acid composition did not change in the residual oils. The peroxide value increased from 2.69 (untreated oil) to 3.01 and 3.49 mequiv O₂/kg in the residual oils following Approach 1 and Approach 2, respectively. The biodiesel prepared from both residual oils met European (EN 14214:2008) and American biodiesel standard (ASTM D6751-09) specifications. Oxidative stability indices for both the biodiesels were well within the permitted limit. It is concluded that PEs could be isolated in active forms for various applications by either of the two methods with a high yield and the residual oil can be processed to produce high quality biodiesel.     

Toxic Compound, Anti-Nutritional Factors and Functional Properties of Protein Isolated from Detoxified Jatropha curcas Seed Cake

Jatropha curcas is a multipurpose tree, which has potential as an alternative source for biodiesel. All of its parts can also be used for human food, animal feed, fertilizer, fuel and traditional medicine. J. curcas seed cake is a low-value by-product obtained from biodiesel production. The seed cake, however, has a high amount of protein, with the presence of a main toxic compound: phorbol esters as well as anti-nutritional factors: trypsin inhibitors, phytic acid, lectin and saponin. The objective of this work was to detoxify J. curcas seed cake and study the toxin, anti-nutritional factors and also functional properties of the protein isolated from the detoxified seed cake. The yield of protein isolate was approximately 70.9%. The protein isolate was obtained without a detectable level of phorbol esters. The solubility of the protein isolate was maximal at pH 12.0 and minimal at pH 4.0. The water and oil binding capacities of the protein isolate were 1.76 g water/g protein and 1.07 mL oil/g protein, respectively. The foam capacity and stability, including emulsion activity and stability of protein isolate, had higher values in a range of basic pHs, while foam and emulsion stabilities decreased with increasing time. The results suggest that the detoxified J. curcas seed cake has potential to be exploited as a novel source of functional protein for food applications.     

Toxic Compound,Anti-Nutritional Factors and Functional Properties of Protein Isolated from Detoxified Jatropha curcas Seed Cake

Jatropha curcas is a multipurpose tree, which has potential as an alternative source for biodiesel. All of its parts can also be used for human food, animal feed, fertilizer, fuel and traditional medicine. J. curcas seed cake is a low-value by-product obtained from biodiesel production. The seed cake, however, has a high amount of protein, with the presence of a main toxic compound: phorbol esters as well as anti-nutritional factors: trypsin inhibitors, phytic acid, lectin and saponin. The objective of this work was to detoxify J. curcas seed cake and study the toxin, anti-nutritional factors and also functional properties of the protein isolated from the detoxified seed cake. The yield of protein isolate was approximately 70.9%. The protein isolate was obtained without a detectable level of phorbol esters. The solubility of the protein isolate was maximal at pH 12.0 and minimal at pH 4.0. The water and oil binding capacities of the protein isolate were 1.76 g water/g protein and 1.07 mL oil/g protein, respectively. The foam capacity and stability, including emulsion activity and stability of protein isolate, had higher values in a range of basic pHs, while foam and emulsion stabilities decreased with increasing time. The results suggest that the detoxified J. curcas seed cake has potential to be exploited as a novel source of functional protein for food applications.     

Euphorbia lagascae Spreng., an abundant source of epoxyoleic acid; Seed extraction and oil composition

Euphorbia lagascae Sprengel seed contains an unique epoxy-bearing oil (58–62%, 12,13-epoxyoleic acid) in high quantity (42–50%) with possible industrial potential. Oil from domestically grown seed was equal in quality and quantity to that obtained from Spain. It was readily and completely extracted at room temperatures without lipolysis using low solvent (petroleum naphtha) to seed ratios. The extracted crude oil was about equal in quality to that obtained by cold-pressing. Analytical data on the seed oil are compared with those previously reported for the oil ofVernonia anthelmintica (L.) Willd. (ironweed) seed. E. Utiliz. Res. Dev. Div.,ARS, USDA.     

Hybrid <Emphasis Type="Italic">Hibiscus</Emphasis> seed oil compositions

The seeds of cultivated Hibiscus spp. were extracted with supercritical carbon dioxide, and the resulting extracts were analyzed to identify the major TG components as the corresponding FAME. The seed oils were composed predominantly of oleic and linoleic FA (69.6–83.4%) with lesser amounts of palmitic acid (14.8–27.0%). Minor amounts of C14, C18, and C20 saturated FA were also detected. The oil content of the seeds was determined to be between 11.8 and 22.1 wt% for hybrid varieties and between 8.9 and 29.5 wt% for the native species from which the hybrid varieties were developed. The protein content of the defatted seed meal averaged 20% for the hybrid varieties. The composition of the extracted hibiscus seed oils suggests potential edible applications.     

Development of solid base catalyst X/Y/MgO/γ-Al2O3 for optimization of preparation of biodiesel from Jatropha curcas L.seed oil

The preparation and regeneration conditions of the identified catalyst X/Y/MgO/γ-Al₂O₃ with high catalytic activity were studied and optimized. The biodiesel was prepared by transesterification of Jatropha curcas seed oil produced in Guizhou with methanol at its reflux temoerature in the presence of X/Y/MgO/γ-Al₂O₃. The pilot plant tests were carried out in a 100 L reaction vessel. Both average yield and fatty acid methyl esters (FAME) content reached more than 96.50% under the optimum reaction conditions of the pilot plant tests designed withan oil/methanol molar ratio of 1: 10, catalyst concentration of 1.00%, and reaction time of 3 h at reflux temperature. In addition, analysis shows that the quality of biodiesel meets the standard EN 14214. __________ Translated from Modern Chemical Industry, 2007, 27(Suppl. 2): 452–455 [译自: 现代化工]     

Fatty acid variation in seed oil amongOcimum species

Content, fatty acid composition, and glyceride profile of oil from seeds of seven basil (Ocimum sp.) chemotypes were determined. The species studied includedO. basilicum, O. canum, O. gratissimum, andO. sanctum. The oil content ranged from 18 to 26%, with triglycerides comprising between 94 and 98% of extracted neutral lipids. The major acylated fatty acids were linolenic (43.8–64.8%), linoleic (17.8–31.3%), oleic (8.5–13.3%), and palmitic acid (6.1–11.0%). Linolenic acid was similar among the fourO. basilicum chemotypes (57–62%), highest inO. canum (65%), and lowest inO. sanctum (44%). Basil seed oil appears suitable as an edible oil or can be used for industrial purposes, and could be processed in the same way as linseed oil. Preliminary calculations estimate that a hectare of basil could produce from 300 to 400 kg of seed oil.     

Effects of Geographical Origin on the Conjugated Linolenic Acid of <Emphasis Type="Italic">Trichosanthes kirilowii</Emphasis> Maxim Seed Oil

Trichosanthes kirilowii Maxim (T. kirilowii) seeds from four geographical locations (Changxing, Quzhou, Yuexi, Dongzhi) contained 26.15–49.41% oil and 28.68–37.90% protein. The seed oil was distinguished by the conjugated linolenic acids, punicic acid (PA) and α-eleostearic acid (α-ESA). The main fatty acids in T. kirilowii seed oils were ranked in the following order: punicic acid (33.09–39.15%), linolenic acid (33.77–38.66%), oleic acid (15.15–24.88%), palmitic acid (2.36–4.86%). PA was the main isomer of CLNA (33.09–39.15%). No significant differences were found either in PA content or in α-ESA content of T. kirilowii seed from these geographical locations. Little difference was observed in the quantitative composition of the lipid contents of seeds from different geographical locations. The α-tocopherol content of T. kirilowii seed ranged from 6.34 to 31.74 mg/100 g, with the highest levels in Changxing seeds. The present results showed that T. kirilowii seeds were especially rich in PA, and their contents were not influenced by the geographical locations. Variation in some proximate compositions by geographical locations may be caused by ecological conditions, temperature, climate condition, technical and cultural conditions.     

Antifungal Activity of Leaf Essential Oil and Extracts of <Emphasis Type="Italic">Metasequoia glyptostroboides</Emphasis> Miki ex Hu

Plant diseases constitute an emerging threat to global food security. Many of the currently available antifungal agents for agriculture are highly toxic and nonbiodegradable and cause extensive environmental pollution. Moreover, an increasing number of phytopathogens are developing resistance to them. Therefore, the aim of this study was to assess the antifungal efficacy of the leaf essential oil and the leaf extracts of Metasequoia glyptostroboides against Fusarium oxysporum, Fusarium solani, Phytophthora capsici, Colletotrichum capsici, Sclerotinia sclerotiorum, Botrytis cinerea and Rhizoctonia solani. The oil (1,000 μg/disc) and the extracts (1,500 μg/disc) revealed a remarkable antifungal effect against the tested plant pathogenic fungi with a radial growth inhibition percentage of 41.3–66.3% and 13.4–54.4%, respectively along with their respective MIC values ranging from 62.5 to 1,000 μg/ml and 500–4,000 μg/ml. The oil had a strong detrimental effect on spore germination of all the tested plant pathogens along with the concentration as well as time-dependent kinetic inhibition of Botrytis cinerea. Also, the oil exhibited a potent in vivo antifungal effect against Phytophthora capsici on greenhouse grown pepper plants. The results of this study indicate that the oil and extracts of M. glyptostroboides leaves could become natural alternatives to synthetic fungicides to control certain important plant fungal diseases.     

The Phorbol Ester Fraction from Jatropha curcas Seed Oil: Potential and Limits for Crop Protection against Insect Pests

The physic nut shrub, Jatropha curcas (Euphorbiaceae), has been considered as a “miracle tree”, particularly as a source of alternate fuel. Various extracts of the plant have been reported to have insecticidal/acaricidal or molluscicidal/anthelminthic activities on vectors of medical or veterinary interest or on agricultural or non-agricultural pests. Among those extracts, the phorbol ester fraction from seed oil has been reported as a promising candidate for use as a plant-derived protectant of a variety of crops, from a range of pre-harvest and post-harvest insect pests. However, such extracts have not been widely used, despite the “boom” in the development of the crop in the tropics during recent years, and societal concerns about overuse of systemic chemical pesticides. There are many potential explanations to such a lack of use of Jatropha insecticidal extracts. On the one hand, the application of extracts potentially harmful to human health on stored food grain, might not be relevant. The problem of decomposition of phorbol esters and other compounds toxic to crop pests in the field needing further evaluation before such extracts can be widely used, may also be a partial explanation. High variability of phorbol ester content and hence of insecticidal activity among physic nut cultivars/ecotypes may be another. Phytotoxicity to crops may be further limitation. Apparent obstacles to a wider application of such extracts are the costs and problems involved with registration and legal approval. On the other hand, more studies should be conducted on molluscicidal activity on slugs and land snails which are major pests of crops, particularly in conservation agriculture systems. Further evaluation of toxicity to natural enemies of insect pests and studies on other beneficial insects such as pollinators are also needed.     

Physico-Chemical Characteristics of Citrus Seeds and Seed Oils from Pakistan

The physico-chemical characteristics of the seeds and seed oils of four citrus species, Mitha (Citrus limetta), Grapefruit (Citrus paradisi), Mussami (Citrus sinensis), and Kinnow (Citrus reticulata) were investigated. The hexane-extracted oil content of citrus seeds ranged from 27.0 to 36.5%. The protein, fiber and ash contents were found to be 3.9–9.6%, 5.0–8.5%, and 4.6–5.6%, respectively. The extracted oils exhibited an iodine value of 99.9–110.0; refractive index (40 °C), 1.4639–1.4670; density (24 °C), 0.920–0.941 mg/mL; saponification value, 180.9–198.9; unsaponifiable matter, 0.3–0.5%; acid value (mg KOH/g of oil), 0.5–2.2 and color (1-in. cell) 1.4–3.0R + 15.0–30.0Y. The oils revealed a good oxidative stability as indicated by the determinations of specific extinctions at 232 and 270 nm (2.3–4.4 and 0.6–0.9, respectively), p-anisidine value (2.2–3.2) and peroxide value (1.6–2.4 mequiv/kg of oil). The citrus seed oils mainly consisted of linoleic acid (36.1–39.8%). Other prominent fatty acids were palmitic acid (25.8–32.2%), oleic acid (21.9–24.1%), linolenic acid (3.4–4.4%), and stearic acid (2.8–4.4%). The contents of tocopherols (α, γ, and δ) in the oil were 26.4–557.8, 27.7–84.1, and 9.1–20.0 mg/kg, respectively. The results of the present study demonstrated that the seeds of citrus species investigated are a potential source of valuable oil which might be utilized for edible and other industrial applications.     

Fatty Acid Composition and Oil Yield in Fruits of Five <Emphasis Type="Italic">Arecaceae</Emphasis> Species Grown in Cuba

The present study targeted the whole-fruit oil yield and fatty acid composition from five of the most abundant Arecaceae species grown in Cuba. The oil yields (% dry weight), determined by the Soxhlet extraction technique with hexane, were 25.5, 5.3, 6.9, 5.4, and 6.4% for Roystonea regia, Colpothrinax wrightii, Sabal maritima, Sabal palmetto and Thrinax radiata, respectively. The free fatty acid (FFA) content varied from 2.7 to 6.8%. Fatty acid (FA) profiles of the oils indicated that lauric acid (13.7–44.4%), myristic acid (9.4–22.4%) and palmitic acid (9.2–17.1%) as major saturated FA; whereas oleic acid (9.6–42.7%) and linoleic acid (9.3–17.0%) as major unsaturated FA. R. regia fruit seemed the most promising among Arecaceae grown in Cuba because of its high oil yield and low oil FFA content.     

Search for new industrial oils. XIII. Oils from 102 species of cruciferae

Seed from additional species of Cruciferae have been analyzed for crude protein, oil and fatty acids in the oil. Oils were like those reported earlier from other crucifers, except forCardamine impatiens which is unique among known seed oils because it contains some 25% dihydroxy acids. Erucic acid is present (0.3–55%) in about three-fourths of the 102 samples. Eicosenoic acid is a major constituent (32–53%) in four species and monohydroxy acids (45–72%) in another four. Linolenic acid occurs (2–66%) in oil of all species. Presented at the AOCS meeting in Chicago, Ill., October 11–14, 1964. A laboratory of the No. Utiliz. Res. and Dev. Div., ARS, USDA. ARS, USDA.     

Characterization of the Composition of <Emphasis Type="Italic">Caesalpinia bonducella</Emphasis> Seed Grown in Temperate Regions of Pakistan

Caesalpinia bonducella is an oilseed that is indigenous to Pakistan. The hexane-extracted oil content from the seed kernel was 17.3 ± 1.0% DM (dry matter). The proximate analysis of C. bonducella seed estimated protein, fiber and ash contents to be 20.8 ± 1.4, 5.3 ± 1.0 and 4.6 ± 0.8%, respectively. Trace metals were determined comparable to commonly consumed legume seeds. α-Tocopherol was the predominant tocopherol ranging from 345.10 to 460.21 mg/kg of oil, followed by γ- and δ-tocopherol. The major sterols were β-sitosterol, stigmasterol, campesterol, Δ5-avenasterol, Δ7-stigmastenol and Δ7 avenasterol. The kernel oil was found to contain a high level of linoleic acid (72.7 ± 1.0%) followed by oleic, stearic and palmitic acids. The high percentage of linoleic acid revealed that this oil is a potential source for the manufacture of cosmetics, paints, varnishes, soaps, liquid soaps and other products including biodiesel. These investigations suggest that C. bonducella oil is potentially an important dietary source of essential fatty acids and protein which could be employed for edible and commercial applications in various industries of Pakistan.