Determination of lignans in flaxseed using liquid chromatography with time-of-flight mass spectrometry

A new method of flaxseed-derived lignan determination was developed using HPLC with high-resolution time-of-flight MS (TOF-MS), optimized, and compared to two existing methods (HPLC/MS/MS and GC/MS). The limits of detection (LODs) for HPLC/TOF-MS (0.002–0.043 pg) were comparable with those of the optimized and improved HPLC/MS/MS (0.001–0.015 pg), whereas the LODs for the optimized GC/MS were higher (0.02–3.0 pg, yet lower than reported before). Using the newly developed detection and separation methods, several key flaxseed sample preparation parameters (including extraction, hydrolysis, and sample purification) were evaluated resulting in the development of efficient protocol for lignan quantification from flaxseed hulls and embryos. The results confirmed the importance of quantification of both aglycones and unhydrolyzed glucosides in order to obtain the total lignan estimates.     

不同产地亚麻籽矿物质元素及脂肪酸组成的主成分分析

亚麻籽是一种具有食用与药用价值的生物质,富含多种营养物质与矿物质元素。我国是亚麻籽的主要产地之一,为探讨不同产地亚麻籽中矿物质元素与脂肪酸组成的主要特征成分,采用电感耦合等离子体质谱法(ICP-MS)分析了10个产地亚麻籽中的17种矿物质元素,利用气相色谱-质谱联用法(GC-MS)分析了于相同条件下提取的10个产地亚麻籽油脂中的脂肪酸组成。采用主成分分析法对10个产地亚麻籽中的矿物质元素及脂肪酸组成进行了分析,探讨了不同产地亚麻籽矿物质元素及脂肪酸组成的差异,分析了矿物质元素及脂肪酸的主要特征成分。结果表明,K,Sr,Mg,Ni,Co,Cr,Cd,Se,Zn和Cu是亚麻籽的主要特征矿物质元素,C16∶0,C18∶0,C18∶2,C18∶3,C20∶0,C20∶1是亚麻籽油脂的主要特征脂肪酸。ICP-MS和GC-MS结合主成分分析,可有效揭示不同产地亚麻籽中矿物质元素与脂肪酸组成的特征与差异,进而为亚麻籽的合理、有效利用提供重要的理论依据。     

High-Throughput Structural Elucidation of Lignans in Flaxseed by High-Performance Liquid Chromatography Coupled with Electrospray Ionization Mass Spectrometry

Methods based on high-performance liquid chromatography coupled with mass spectrometry and quadrupole time-of-flight mass spectrometry were developed for the rapid identification of lignans in flaxseed. The lignans were extracted from dried flaxseed extract power by ultrasonic extraction. The fragment ions were further investigated for structural elucidation and the MS/MS spectra provided an abundance of structural information for the identification. A total of eleven lignans were investigated and three were new compounds. This research demonstrates the rapid characterization of bioactive constituents in plant extracts while avoiding tedious and difficult separation and purification steps.     

Large-Scale Purification of Water-Soluble Polysaccharides from Flaxseed Mucilage, and Isolation of a New Anionic Polymer

Summary Viscous flaxseed mucilage has previously been described as a mixture of two polysaccharides (acidic and neutral). In this study, we have shown that a combination of ion exchange and size exclusion chromatography enables purification of three distinct polysaccharides. Molecular weight (Mw) analysis, performed by size-exclusion chromatography on line with a multi-angle laser light-scattering detector, revealed that the main polysaccharide (75%) was a neutral polymer with a Mw of approximately 1.2 × 10⁶ g mol^–1. The two others were acidic polysaccharides denoted AF₁ (3.75%, 6.5 × 10⁵ g mol^–1) and AF₂ (21.25%, 1.7 × 10⁴ g mol^–1).Presented at: International Symposium on Separation and Characterization of Natural and Synthetic Macromolecules, Amsterdam, The Netherlands, February 5–7, 2003     

Heat-induced changes to lipid molecular structure in Vimy flaxseed: spectral intensity and molecular clustering

Autoclaving was used to manipulate nutrient utilization and availability. The objectives of this study were to characterize any changes of the functional groups mainly associated with lipid structure in flaxseed (Linum usitatissimum, cv. Vimy), that occurred on a molecular level during the treatment process using infrared Fourier transform molecular spectroscopy. The parameters included lipid CH(3) asymmetric (ca. 2959 cm(-1)), CH(2) asymmetric (ca. 2928 cm(-1)), CH(3) symmetric (ca. 2871 cm(-1)) and CH(2) symmetric (ca. 2954 cm(-1)) functional groups, lipid carbonyl CO ester group (ca. 1745 cm(-1)), lipid unsaturation group (CH attached to CC) (ca. 3010 cm(-1)) as well as their ratios. Hierarchical cluster analysis (CLA) and principal components analysis (PCA) were conducted to identify molecular spectral differences. Flaxseed samples were kept raw for the control or autoclaved in batches at 120°C for 20, 40 or 60 min for treatments 1, 2 and 3, respectively. Molecular spectral analysis of lipid functional group ratios showed a significant decrease (P<0.05) in the CH(2) asymmetric to CH(3) asymmetric stretching band peak intensity ratios for the flaxseed. There were linear and quadratic effects (P<0.05) of the treatment time from 0, 20, 40 and 60 min on the ratios of the CH(2) asymmetric to CH(3) asymmetric stretching vibration intensity. Autoclaving had no significant effect (P>0.05) on lipid carbonyl CO ester group and lipid unsaturation group (CH attached to CC) (with average spectral peak area intensities of 138.3 and 68.8 IR intensity units, respectively). Multivariate molecular spectral analyses, CLA and PCA, were unable to make distinctions between the different treatment original spectra at the CH(3) and CH(2) asymmetric and symmetric region (ca. 2988-2790 cm(-1)). The results indicated that autoclaving had an impact to the mid-infrared molecular spectrum of flaxseed to identify heat-induced changes in lipid conformation. A future study is needed to quantify the relationship between lipid molecular structure changes and functionality/availability.     

Study on model plant based functional beverage emulsion (non-dairy) using ultrasound – A physicochemical and functional characterization

This study reports the development of non-dairy functional beverage emulsion employing ultrasound (US) of 20 kHz at 130 W and 195 W at processing times of 2 to 8 min using chickpea milk extract and bioactive, flaxseed oil (4%). The pre-emulsion was formed with high shear homogenizer followed by main sonication process. The sonicated emulsions were stored at 4 ± 2 °C till 14 days and characterized for physicochemical and functional properties. A comparative study was carried out using conventional high shear homogenizer (UT) at 10,000 RPM for 5 min. Upon optimization, 130 W − 8 min, 195 W − 6 min and 195 W – 8 min sono-emulsions showed creaming stability of 100%; with particle sizes as 1.12, 0.97 and 0.78 µm; and zetapotential values as − 40.4 mV, −37.52 and −36.91 mV, respectively. The improvement in protein solubility by 86% proved the emulsifying capability of chickpea proteins, which had partially denatured upon physical effects of acoustic cavitation producing stable and finer emulsion droplets. The reduced sedimentation values of sonicated chickpea extract in comparison to UT showed improvement in physical stability of plant-based milk. Oxidative stability is observed for 130 W − 8 min sonicated emulsions with no change in conjugated dienes, indicating the absence of process generated free radicals. The US process did not have any effect on reduction of stachyose content. But extracted chickpea milk had lower amount of stachyose in comparison to raw chickpeas, reducing the flatulence problem, mainly due to adaptation of high temperature pressure cooking process.     

Functional properties of ultrasonically generated flaxseed oil-dairy emulsions

This study reports on the functional properties of 7% flaxseed oil/milk emulsion obtained by sonication (OM) using 20 kHz ultrasound (US) at 176 W for 1–8 min in two different delivery formulae, viz., ready-to-drink (RTD) and lactic acid gel. The RTD emulsions showed no change in viscosity after sonication for up to 8 min followed by storage up to a minimum of 9 days at 4 ± 2 °C. Similarly, the oxidative stability of the RTD emulsion was studied by measuring the conjugated diene hydroperoxides (CD). The CD was unaffected after 8 min of ultrasonic processing. The safety aspect of US processing was evaluated by measuring the formation of CD at different power levels. The functional properties of OM gels were evaluated by small and large scale deformation studies. The sonication process improved the gelation characteristics, viz., decreased gelation time, increased elastic nature, decreased syneresis and increased gel strength. The presence of finer sono-emulsified oil globules, stabilized by partially denatured whey proteins, contributed to the improvements in the gel structure in comparison to sonicated and unsonicated pasteurized homogenized skim milk (PHSM) gels. A sono-emulsification process of 5 min followed by gelation for about 11 min can produce gels of highest textural attibutes.     

Characterization and complete separation of major cyclolinopeptides in flaxseed oil by reversed‐phase chromatography

Organoleptic properties of flaxseed oil deteriorate during storage due to methionine oxidation in its major cyclolinopeptides. Cyclolinopeptide E was previously identified as being responsible for the manifestation of bitter taste with flaxseed oil ageing. We developed a chromatographic procedure to monitor the oxidation of major cyclic peptides in flaxseed oil. We also used liquid chromatography with mass spectrometry and high‐efficiency core–shell reversed‐phase sorbents to study the separation of cyclolinopeptides in detail. The Kinetex^TM family of stationary phases (C₈, C₁₈, phenyl‐hexyl) was tested, along with the standard porous Luna^TM C₁₈(2) media. We found that only the phenyl‐hexyl stationary phase allows for complete resolution of major cyclolinopeptides, thus permitting direct UV monitoring of degree of conversion for cyclolinopeptide B into C and L into E. We also report, for the first time, a significant effect of peak splitting for some methionine S‐oxide (Mso) containing cyclolinopeptides, which most likely appear due to diastereomerization. This results in poor separation efficiency for cyclolinopeptides F, G, and E, and gives baseline resolution of diastereomeric pairs for cyclolinopeptides I and P. Thus, a single oxidation of cyclolinopeptide N yields three distinct chromatographic peaks corresponding to cyclolinopeptide T (cyclo‐MsoLMPFFWV, reported for the first time) and pair of cyclolinopeptide I (cyclo‐MLMsoPFFWV) diastereomers.     

ICP-MS两种模式下测定亚麻籽中微量元素及其不确定度评定

探讨了微波消解结合电感耦合等离子体质谱(ICP-MS)标准(STD)及碰撞池(KED)两种模式测定亚麻籽中微量元素铜、铁、锌、锰方法的有效性。分析比较了两种模式的测定效果,在此基础上选择各元素的有效测定模式,并对四种微量元素的分析结果进行不确定度评价。根据不确定度评判标准JJFl059—1999《测量不确定度评定与表示》中的规定,逐一分析了影响不确定度的主要因素,并对各不确定度进行了计算。结果表明ICP-MS用于测定亚麻籽中微量元素结果令人满意,同时为ICP-MS测定这四种元素的不确定度评价提供了一定的参考价值。