Antiproliferative terpenoids from almond hulls (Prunus dulcis): identification and structure-activity relationships

Bioassay-guided fractionation of the EtOAc crude extract from Sicilian almond hulls, a waste material from Prunus dulcis crop, allowed identification of 10 constituents, isolated as pure compounds (1-5, 7, and 10) or unseparable mixtures (5 + 6 and 8 + 9). All compounds were subjected to spectroscopic analysis and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide bioassay on MCF-7 human breast cancer cells. In addition to the main components oleanolic (1), ursolic (2), and betulinic (3) acids, the 2-hydroxy analogues alphitolic (4), corosolic (5), and maslinic (6) acids, as well as the related aldehydes, namely, betulinic (7), oleanolic (8), and ursolic (9), were identified. From a more polar fraction, the beta-sitosterol 3-O-glucoside (10) was also identified. A sample of commercially available betulin (11) was also included in bioassays as further support to a structure-activity relationship study. Betulinic acid showed antiproliferative activity toward MCF-7 cells (GI50 = 0.27 microM), higher than the anticancer drug 5-fluorouracil.     

New prenylated benzoic acid and other constituents from almond hulls (Prunus amygdalus Batsch).

One new prenylated benzoic acid derivative, 3-prenyl-4-O-beta-D-glucopyranosyloxy-4-hydroxylbenzoic acid, and three known constituents, catechin, protocatechuic acid, and ursolic acid, have been isolated from the hulls of almond (Prunus amygdalus). Complete assignments of the proton and carbon chemical shifts for the new prenylated benzoic acid derivative were accomplished on the basis of high-resolution 1D and 2D nuclear magnetic resonance data. All of these compounds except ursolic acid are being reported from almond hulls (P. amygdalus) for the first time.     

Separation and HPLC-MS identification of phenolic antioxidants from agricultural residues: almond hulls and grape pomace

Almond hulls and grape pomace are residues abundantly generated by agricultural industries, which could be processed to obtain bioactive products. To this purpose, crude ethanol extracts from both agricultural byproducts were attained and subsequently fractionated in order to obtain an organic/water fraction (FOW). Extracts and fractions were analyzed for antioxidant power and their phenolic components tentatively identified by HPLC-MS. Chromatographic peaks of almond hull extracts showed the occurrence of hydroxybenzoic and cinnamic acid derivatives, with minor presence of flavan-3-ols (ECG, EGCG), whereas the FOW fraction offered the additional presence of epicatechin (EC) and glycosylated flavonols. In the composition for extracts of white and red grape pomace several of these compounds were also detected but basically consisted of glycosylated flavonols (quercetin, kaempferol). As a difference between both grape pomaces, myricetin glycosyde was found in that from the red variety, whereas flavan-3-ols (EC, afzelechin) were only identified in white pomace. When their FOW fractions were analyzed, gallic acid and some hydroxybenzoic acids were additionally detected. Antioxidant activity was assessed by DPPH and TBARS assays. Almond hulls showed inhibition percentages lower than 50% in both assays, while the inhibition percentage ranged from 80% to 90% in pomace extracts. Red grape pomace extract was the most efficient antioxidant, with an EC50 value of 0.91 g/L for TBARS and 0.20 g/L for DPPH. Even appearing as two quite different vegetal matrixes, the composition of phenolics in grape pomace and almond hulls is quite similar, the main difference being the major occurrence of flavonols in grape pomace. This fact could presumably explain the lower antiradical activity of hull extracts.     

Antioxidant constituents of almond [Prunus dulcis (Mill.) D.A. Webb] hulls

Almond hulls (Nonpareil variety) were extracted with methanol and analyzed by reversed phase HPLC with diode array detection. The extract contained 5-O-caffeoylquinic acid (chlorogenic acid), 4-O-caffeoylquinic acid (cryptochlorogenic acid), and 3-O-caffeoylquinic acid (neochlorogenic acid) in the ratio 79.5:14.8:5.7. The chlorogenic acid concentration of almond hulls was 42.52 +/- 4.50 mg/100 g of fresh weight (n = 4; moisture content = 11.39%). Extracts were tested for their ability to inhibit the oxidation of methyl linoleate at 40 degrees C. At an equivalent concentration (10 microg/1 g of methyl linoleate) almond hull extracts had higher antioxidant activity than alpha-tocopherol. At higher concentrations (50 microg/1 g of methyl linoleate) almond hull extracts showed increased antioxidant activity that was similar to chlorogenic acid and morin [2-(2,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-1-benzopyran-4-one] standards (at the same concentrations). These data indicate that almond hulls are a potential source of these dietary antioxidants. The sterols (3beta,22E)-stigmasta-5,22-dien-3-ol (stigmasterol) and (3beta)-stigmast-5-en-3-ol (beta-sitosterol) (18.9 mg and 16.0 mg/100 g of almond hull, respectively) were identified by GC-MS of the silylated almond hull extract.     

Identification and quantification of flavonol glycosides in almond seedcoats using MALDI-TOF MS

Interest in the molecular composition of almonds is growing, due to their popularity in a wide variety of food formulations. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a powerful new technique that can be used to rapidly identify and quantify possible bioactive compounds in these popular tree nuts. Four flavonol glycosides were identified in almond seedcoats for the first time: isorhamnetin rutinoside, isorhamnetin glucoside, kaempferol rutinoside, and kaempferol glucoside. A MALDI-TOF MS methodology was developed using rutin (quercetin-3-rutinoside) as an internal standard to quantitatively determine each of the four flavonol glycosides. Results of MALDI-TOF MS analysis were verified by high performance liquid chromatography.     

Strain of Fusarium oxysporum isolated from almond hulls produces styrene and 7-methyl-1,3,5-cyclooctatriene as the principal volatile components

An isolated strain of Fusarium oxysporum from the hulls of Prunus dulcis (sweet almond) was found to produce relatively large quantities of the hydrocarbons styrene and two isomers of 7-methyl-1,3,5- cyclooctatriene (MCOT). Production of styrene and MCOT was reproduced on a small scale using potato dextrose agar as a growth medium and scaled up using 1 L of inoculated potato dextrose broth. The compounds were trapped as volatile organic compounds (VOCs) onto solid-phase microextraction (SPME) for small scale and Tenax for large scale and then isolated using standard high-performance liquid chromatography (HPLC) methods. Styrene was authenticated by a comparison to the retention times, fragmentation patterns, and calculated retention indices of a commercially available sample. The identity of MCOT was verified by a short chemical synthesis and a comparison of spectroscopic data to the isolated sample. A biosynthetic scheme of styrene is proposed on the basis of a (13)C-labeling study. This is the first report of MCOT isolated as a natural product.     

Antioxidant polyphenols in almond and its coproducts

Antioxidant efficacy of defatted almond whole seed, brown skin, and green shell cover extracts was evaluated by monitoring inhibition of human low-density lipoprotein (LDL) oxidation, inhibition of DNA scission, and metal ion chelation activities. The total phenolic contents of ethanolic extracts of brown skin and green shell cover of almond were 10 and 9 times higher than that of the whole seed, respectively. Brown skin extract at 50 ppm effectively inhibited copper-induced oxidation of human LDL cholesterol compared to whole seed and green shell cover extracts, which reached the same level of efficacy at 200 ppm. Green shell cover extract at 50 ppm level completely arrested peroxyl radical-induced DNA scission, whereas 100 ppm of brown skin and whole seed extracts was required for similar efficiencies. All three almond extracts exhibited excellent metal ion chelation efficacies. High-performance liquid chromatographic (HPLC) analysis revealed the presence of quercetin, isorhamnetin, quercitrin, kaempferol 3-O-rutinoside, isorhamnetin 3-O-glucoside, and morin as the major flavonoids in all extracts.     

杏仁油微胶囊制备工艺的优化

杏仁油中不饱和脂肪酸含量较高,为防止杏仁油的氧化,扩大杏仁油的用途,以麦芽糊精和牛乳分离蛋白为壁材,大豆卵磷脂为乳化剂,采用喷雾干燥法对超临界CO<,2>萃取的杏仁油进行了微胶囊化研究,采用响应面分析法研究了微胶囊的最佳配方,正交法确定了喷雾干燥的最佳工艺参数.结果表明:杏仁油微胶囊包埋最佳配方为杏仁油质量分数25%、牛乳分离蛋白质量分数25%、大豆卵磷脂质量分数2%和麦芽糊精质量分数48%,在20 MPa压力下均质处理;喷雾干燥适宜的工艺参数为进风温度180℃,出风温度80℃,进料温度50℃.在上述优化条件下可制得高质量高包埋率的杏仁油微胶囊产品,包埋率达94.7%.     

Detection and stability of the major almond allergen in foods

Almond major protein (AMP or amandin), the primary storage protein in almonds, is the major allergen recognized by almond-allergic patients. A rabbit antibody-based inhibition ELISA assay for detecting and quantifying AMP in commercial foods has been developed, and this assay, in conjunction with Western blotting analyses, has been applied to the investigation of the antigenic stability of AMP to harsh food-processing conditions. The ELISA assay detects purified AMP at levels as low as 87 +/-16 ng/mL and can detect almond at between 5 and 37 ppm in the tested foods. The assay was used to quantify AMP in aqueous extracts of various foods that were defatted and spiked with known amounts of purified AMP or almond flour. In addition, AMP was quantified in commercially prepared and processed almond-containing foods. Neither blanching, roasting, nor autoclaving of almonds markedly decreased the detectability of AMP in subsequent aqueous extracts of almonds. Western blots using both rabbit antisera and sera from human almond-allergic patients confirm a general stability of the various peptides that comprise this complex molecule and show that the rabbit antibody-based assay recognizes substantially the same set of peptides as does the IgE in sera from almond-allergic patients.     

Evaluation of extracts from Gevuina avellana hulls as antioxidants

The antioxidant activity of the extracts from Gevuina avellana hulls was evaluated and compared with that of BHT (butylated hydroxytoluene) and BHA (butylated hydroxyanisole), using the beta-carotene bleaching assay, the accelerated oxidation of crude soybean oil, and the 2,2-diphenyl-beta-picrylhydrazyl (DPPH) radical scavenging method. Solvents of different polarity were used to obtain the extracts. Both the extraction yield and the antioxidant activity were strongly dependent on the solvent. The ethanol and diethyl ether soluble fractions were the most active with the beta-carotene assay. Ethanol and methanol extracts were the most active in hydrogen radical scavenging activity. Water and methanol inhibited more efficiently the oxidation of soybean oil at 70 and 80 degrees C, respectively. As a general trend, increased antioxidant activity was observed for increased extract concentration. Except the acetone extracts, all were stable after 6 months storage at 4 degrees C. The ethanol solubles from G. avellana hulls present antioxidant activity similar to that of synthetic antioxidants and to other reported residual agroindustrial materials.     

Discriminating significance of the free amino acid profile in almond seeds

Known statistical techniques have been applied to the free amino acid composition of 107 samples from 10 different almond cultivars (Marcona, Desmayo-Largueta, Guara, Tuono, Ferragnes, Masbovera, Non Pareil, Titan, Texas, and Primorskyi) cultivated in seven different locations and growing conditions. It is concluded that free amino acid composition can constitute a basis for classifying and typifying these cultivars into five groups: (1) Marcona and Texas, (2) Ferragnes and Masbovera (and probably Primorskyi), (3) Tuono and Guara, (4) Non Pareil (and probably Titan), and (5) an isolated cultivar (Desmayo Largueta). As a result, an easy decision tree is proposed to discriminate the cultivar of an almond flour, as used in confectionery, if it consists of a single cultivar.     

Enhancement of frost tolerance of almond flowers using potassium

Abstract

Using mineral nutrients is recommended as a plan to alleviate or avoid spring frost damage to blossoms. The purpose of this study was to investigate the effects of potassium application on reducing or preventing spring frost damage of almond (Prunus dulcis Mill.) flowers. Treatments were four levels of K₂SO₄ (0, 1, 2, and 4%). The nutrition solutions were sprayed on trees in two times (bud swell stage and green tip stage) in 2018–2019. The spurs containing flower buds were collected during the full blooming phase. The spurs segments were exposed to three different freeze test temperatures (0, ?2, and ?4?°C), and some relevant physiochemical changes were evaluated. The application of K₂SO₄, especially 4% K₂SO₄, decreased freezing injury, electrolyte leakage percentage, hydrogen peroxide (H₂O₂), and malondialdehyde (MDA) concentrations of flowers after freezing tests compared with control. The nutrient treatments, especially 4% K₂SO₄, increased significantly the contents of soluble proteins, soluble carbohydrates, total phenolic compounds, and proline of flowers. The results of the present study could have implications for the use of potassium in freezing tolerance improvement of other temperate fruit trees grown in low temperatures regions.     

Cytotoxic and antitumor activity of momilactone B from rice hulls

The cytotoxic and antitumor activity of methanolic extract of rice hulls (MERH) were evaluated by the MTT-dye reduction assay against human colon cancer cells and the colonic aberrant crypt foci (ACF) assay in 1,2-dimethylhydrazine (DMH)-injected F344 male rats, respectively. MERH was found to be highly cytotoxic, with IC50 values of 0.5 microg/mL in vitro. Forty weeks of MERH supplementation (50 mg/kg of body weight/day) reduced colonic pre-neoplastic ACF formation by 35% (p < 0.01). An active compound, momilactone B, was isolated from MERH by silica gel chromatography, Sephadex LH-20 chromatography, and HPLC. The cytotoxic activity of momilactone B was evaluated by the MTT-dye reduction, lactate dehydrogenase (LDH), and colony-forming ability assays in human colon cancer HT-29 and SW620 cells. The results indicated that momilactone B from rice hulls might be a new candidate for chemotherapeutic agent against human colon cancer.     

Antioxidant activity of various fractions of non-tannin phenolics of canola hulls

Cyclone canola hulls were extracted with 70% (v/v) acetone. The dried crude extract was dissolved in ethanol and fractionated on a Sephadex LH-20 column using 95% (v/v) ethanol as the mobile phase. Five major fractions were isolated according to the UV absorption. All fractions exhibited marked antioxidant activity in a beta-carotene-linoleate model system. Fractions I and II showed the best preventive effect against the bleaching of beta-carotene. The scavenging effect of fractions I, III, and V, at 1 mg, on alpha, alpha-diphenyl-beta-picrylhydrazyl (DPPH) radical was 67.4%, 80.7%, and 63.3%, respectively. Fractions II and IV showed weak DPPH scavenging effects. The reducing power of phenolics present in fractions IV and V was greater than that of fractions I-III, and the observed data correlated well (r(2) = 0.937; P = 0.007) with the total content of phenolics present in each fraction.     

Effect of far-infrared radiation on the antioxidant activity of rice hulls

After far-infrared (FIR) radiation onto rice hull, a methanolic extract was prepared for the determination of antioxidant ability. After 30 min of FIR treatment, the radical scavenging activity and total phenol contents of rice hull extracts increased from 47.74 to 79.63% and from 0.12 to 0.19 mM, respectively, compared to control. Inhibition of lipid peroxidation in extracts was also increased from 41.07 to 47.96%. According to the GC-MS analysis, more phenolic compounds (p-coumaric acid, 3-vinyl-1-oxybenzene, p-hydroxybenzaldehyde, vanillin, p-hydroxybenzoic acid, and 4,7-dihydroxyvanillic acid) were detected in FIR-irradiated rice hull extract. These results indicated that FIR radiation onto rice hull could liberate and activate covalently bound phenolic compounds that have antioxidant activities.     

超声波辅助提取黑豆皮色素工艺优化

为了优化超声波辅助提取黑豆皮色素的工艺条件,在单因素试验基础上,采用响应曲面法对影响黑豆皮色素提取效果的3个主要影响因素即超声功率、溶液pH值和液固比进行了优化,建立并分析了各因子与黑豆皮色素提取液吸光值关系的数学模型。结果表明：原料用量0.5 g,在超声功率为80 W、溶液pH值1.5、液固比30 mL/g时,黑豆皮色素提取效果较好,在此条件下色素的提取率达到95.6%。     

超临界CO2流体萃取杏仁油工艺研究

该研究以单因素试验和正交试验相结合的方法对苦杏仁脂肪油的超临界CO2萃取工艺进行了研究.确定了超临界CO2萃取杏仁油的最佳工艺为:萃取压力35 MPa、萃取温度50 ℃、CO2流量24 L/h、粒径60目、萃取时间2 h.各因素影响杏仁的得率的顺序为:粒径＞时间＞萃取压力＞CO2流量＞萃取温度.最佳工艺验证试验的杏仁油的得率为52.98%.本研究的结果为下一步综合、无毒、高效地开发利用苦杏仁奠定了基础.