High-intensity ultrasonication impact on the chlorothalonil fungicide and its reduction pathway in spinach juice

Among different novel technologies, sonochemistry is a sustainable emerging technology for food processing, preservation, and pesticide removal. The study aimed to probe the impact of high-intensity ultrasonication on chlorothalonil fungicide degradation, reduction pathway, and bioactive availability of spinach juice. The chlorothalonil fungicide-immersed spinach juice was treated with sonication at 360 W, 480 W, and 600 W, 40 kHz, for 30 and 40 min at 30 ± 1 °C. The highest reduction of chlorothalonil fungicide residues was observed at 40 min sonication at 600 W. HPLC-MS (high-performance liquid chromatography-mass spectroscopy) analysis revealed the degradation pathway of chlorothalonil and the formation of m-phthalonitrile, 3-cyno-2,4,5,6-tetrachlorobenamide, 4-dichloroisophthalonitrile, trichloroisophtalonitrile, 4-hydoxychlorothalonil, and 2,3,4,6-tetrachlorochlorobenzonitrile as degradation products. High-intensity sonication treatments also significantly increased the bioavailability of phenolic, chlorophyll, and anthocyanins and the antioxidant activity of spinach juice. Our results proposed that sonication technology has excellent potential in degrading pesticides through free radical reactions formation and pyrolysis. Considering future perspectives, ultrasonication could be employed industrially to reduce pesticide residues from agricultural products and enhance the quality of spinach juice.     

Effect of Nano-anatase TiO2 on Spectral Characterization of Photosystem Ⅱ Particles from Spinach

The photosystem I (PS Ⅱ) particles were purified by means of nano-anatase TiO2 treatment of spinach and studied by spectroscopy. The results show that the electron transport and the oxygen-evolving rate of PS I are accelerated after it has been treated with nano-anatase TiO2; the UV-Vis absorption spectrum of PS I particles is increased; the red shift of fluorescence emission peak of PS I is 2 nm; the peak intensity is decreased; the PS Ⅱ signal I s of low temperature electron paramagnetic resonanace(EPR) spectrum is intensified under light, and the PS I circular dichroism(CD) spectrum is similar to that of control. It is suggested that nano-anatase TiO2 might bind to the PS I reaction center complex and intensify the function of the PS I electron donor, however, nano-anatase TiO2 treatment does not change the configuration of the PS Ⅱ reaction center complex.     

The acclimative response of the main light-harvesting chlorophyll a/b-protein complex of photosystem II (LHCII) to elevated irradiances at the level of trimeric subunits

The changes in structural organization of the major light-harvesting chlorophyll a/b–protein complex of photosystem II (LHC II) at the level of trimeric subcomplexes were studied in spinach plants grown under low light conditions (50 μmol quanta m⁻² s⁻¹) and then acclimated to elevated irradiances. By monitoring photochemical quenching of fluorescence yield (q_P), photosystem II (PS II) functional status was assessed in leaves of plants acclimated to a range of elevated irradiances. Three separate acclimative irradiances were selected for the experiments, reflecting: limiting light conditions (150 μmol quanta m⁻² s⁻¹), near to the inflexion point on the irradiance curve conditions (300 μmol quanta m⁻² s⁻¹) and an excessive light, causing a moderate stress in the form of down regulation of PS II (450 μmol quanta m⁻² s⁻¹). An immunoblot analysis showed that there was a clear decline in an abundance on chlorophyll basis of Lhcb1-3 apoproteins as an acclimative irradiance increased from 50 to 450 μmol quanta m⁻² s⁻¹, with Lhcb1 decreasing to a lesser extent than Lhcb2 and Lhcb3 (only at excessive irradiance). When analyzed by non-denaturing isoelectric focusing BBY membrane fragments (PSII-enriched, stacked thylakoid membranes) isolated from low light-grown plants were resolved into nine fractions, seven of which (labelled 3–9) were established by us previously [Jackowski and Pielucha, J. Photochem. Photobiol. B: Biol. 64 (2001) 45] to be LHC II subcomplexes representing mixed populations of closely similar trimers, comprising permutations of Lhcb1 and Lhcb2 (subcomplexes 3–7) or Lhcb1-3 (subcomplexes 8 and 9). A heterogeneity with regard to accumulation behaviour of LHC II subcomplexes in response to elevated irradiances was revealed. The subcomplexes 5 and 6 were accumulating at similar level, regardless of the light irradiance experienced. Another group consisting of the subcomplexes 3 and 4 (the most basic ones) showed a progressive increase in relative abundance with increasing an irradiance intensity whereas the subcomplexes 7–9 (the most acidic ones) exhibited a progressive decline in their relative abundance during an acclimation of spinach plants to elevated irradiances thus they may collectively represent an elevated irradiance-responsive subunit of LHCII.     

Promotion of nano-anatase TiO2 on the spectral responses and photochemical activities of D1/D2/Cyt b559 complex of spinach

Previous researches approved that photocatalysis activity of nano-TiO₂ could obviously increase photosynthetic effects of spinach, but the mechanism of improving light energy transfer and conversion is still unclear. In the present we investigated effects of nano-anatase TiO₂ on the spectral responses and photochemical activities of D1/D2/Cyt b559 complex of spinach. Several effects of nano-anatase TiO₂ were observed: (1) UV–vis spectrum was blue shifted in both Soret and Q bands, and the absorption intensity was obviously increased; (2) resonance Raman spectrum showed four main peaks, which are ascribed to carotene, and the Raman peak intensity was as 6.98 times as that of the control; (3) the fluorescence emission peak was blue shifted and the intensity was decreased by 23.59%; (4) the DCPIP photoreduction activity showed 129.24% enhancement; (5) the oxygen-evolving rate of PS II was elevated by 51.89%. Taken together, the studies of the experiments showed that nano-anatase TiO₂ had bound to D1/D2/Cyt b559 complex, promoted the spectral responses, leading to the improvement of primary electron separation, electron transfer and light energy conversion of D1/D2/Cyt b559 complex.     

Chemical environment heterogeneity around the two chlorophyll a′ molecules in photosystem I

Treatment of whole and fractionated plant tissues with hydrophilic and hydrophobic solvent mixtures of varied volume ratio liberates two chlorophyll (Chl) a′ molecules from the photosystem (PS)I core when the solvent hydrophilicity exceeds a critical level, whereas only one molecule is extracted in hydrophobic media. The PSI core proteins, PSI-A and PSI-B, which form a heterodimer, appear to bind one Chl a′ molecule each, in local environments significantly different regarding their hydrophilicity or hydrophobicity.     

低强度微波电磁场作用下菠菜和烟草光合细胞叶绿素荧光动力学过程及光合色素系统的差异

在300MHz低强度微波电磁场作用下,菠菜和烟草叶片光合细胞叶绿素荧光动力学过程和光合色素呈现不同的变化.菠菜的荧光动力学参量F0和FVI/FV减小,FV/F0、ΔFV/T和FV/Fm升高;烟草的荧光动力学参量F0升高,FVI/FV没有明显变化,FV/F0、ΔFV/T和FV/Fm降低.在微波作用下,菠菜和烟草叶片的光合色素系统的变化也有差异,菠菜和烟草的叶绿素含量均降低,但菠菜的类胡萝卜素含量明显升高.这些结果表明,低强度微波电磁场使烟草叶片光合机构受到抑制,光合色素系统受到破坏,因而光合作用下降;菠菜则通过光合细胞的光合机构中PSⅡ活性中心异质性的转变和光合色素中类胡萝卜素含量的升高来适应微波辐射的环境,使光合作用维持在较高水平.     

Use of graphitic carbon black and primary secondary amine for determination of 17 organophosphorus pesticide residues in spinach

Graphitized carbon black (GCB) and primary secondary amine (PSA) as dispersive-SPE sorbents were applied to optimize the method for the determination of 17 organophosphorus pesticides in spinach which contained so many pigments using GC with flame photometric detector (FPD). The sample was extracted with ACN, and an aliquot of the extract was concentrated to near dryness. Ethyl acetate or acetone was chosen as the dissolving solvent. Subsequently, dispersive-SPE was used for cleanup, and the type and quantity of sorbents (GCB, PSA and activated carbon) were tested in the experiments. The best results were when acetone was used to dissolve and 30 mg each of GCB and PSA for cleanup. In this condition, recoveries of pesticides analyzed were between 52-117% with RSD below 10%, and LOQ ranged from 10 to 20 microg/kg. This method was simple, effective and efficient, and can protect the GC system to some extent.     

Impact of high-intensity thermosonication treatment on spinach juice: Bioactive compounds,rheological, microbial,and enzymatic activities

To study the impacts of thermosonication (TS), the spinach juice treated with TS (200 W, 400 W, and 600 W, 30 kHz, at 60 ± 1 °C for 20 mint) were investigated for bioactive compounds, antioxidant activities, color properties, particle size, rheological behavior, suspension stability, enzymatic and microbial loads. As a result, TS processing significantly improved the bioactive compounds (total flavonols, total flavonoids, total phenolic, carotenoids, chlorophyll, and anthocyanins), antioxidant activities (DPPH and FRAP assay) in spinach juice. Also, TS treatments had higher b*, L*, hue angle (h⁰), and chroma (C) values, while minimum a* value as compared to untreated and pasteurized samples. TS processing significantly reduced the particle size, improved the suspension stability and rheological properties (shear stress, apparent viscosity, and shear rate) of spinach juice as compared to the untreated and pasteurized sample. TS plays a synergistic part in microbial reduction and gained maximum microbial safety. Moreover, TS treatments inactivated the polyphenol oxidase and peroxidase from 0.97 and 0.034 Abs min⁻¹ (untreated) to 0.31 and 0.018 Abs min⁻¹, respectively. The spinach juice sample treated at a high intensity (600 W, 30 kHz, at 60 ± 1 °C for 20 mint, TS3) exhibited complete inactivation of microbial loads (<1 log CFU/ml), the highest reduction in enzymatic activities, better suspension stability, color properties, and highest bioactive compounds. Collectively, the verdicts proposed that TS processing could be a worthwhile option to pasteurize the spinach juice to enhance the overall quality.     

Development of a procedure for the sequential extraction of substances binding trace elements in plant biomass

This work investigates how the amounts of some important substances in a plant, and their behaviour inside the plant, depend on the levels of stress placed on the plant. To this end, model plant spinach (Spinacia oleracea L.) was cultivated on soil treated with sewage sludge. The sewage sludge contained various trace elements (As, Cd, Cu, Zn), and the uptake of these trace elements placed the plant under stress. Following this, a sequential extraction procedure was employed to determine the levels and distributions of trace elements within the most important groups of compounds present in the spinach plants. Since the usual five-step sequential extraction procedure provides only general information on the distributions of elements within individual groups of organic compounds, due to the wide range of organic compounds within the individual fractions, this scheme was extended and improved through the addition of two solvent extraction steps—a butanol step (between the ethyl acetate and methanol solvent steps) and an H₂O step (after the methanol+H₂O solvent step). The distributions and levels of the trace elements within the main groups of compounds in spinach biomass was investigated using this new seven step sequential extraction (water free solvents: petroleum ether (A) ethyl acetate (B) butanol (C) methanol (D) water solvents: methanol+H₂O (1+1; v/v) (E) H₂O (F) methanol+H₂O+HCl (49.3+49.3+1.4; v/v/v) (G)). The isolated fractions were characterized using IR spectroscopy and the trace element contents were determined in the individual fractions. Lipophilic compounds with low contents of Cd, Cu and Zn were separated in the first two fractions (A, B). Compounds with higher As contents (11.5–12.8% of total content) were also extracted in the second fraction, B. These two fractions formed the smallest portion of the isolated fractions. Low molecular compounds from secondary metabolism and polar lipids were separated in the third (C) and fourth (D) fractions, and high molecular compounds (mainly polypeptides and proteins) separated in the fifth and sixth fractions (E, F). The addition of the H₂O solvent step was particularly useful for separating compounds that have a significant impact on trace element bounds. The methanol fraction was dominant for all treatments, and a significant decrease in the spinach biomass separated in this fraction was observed when the soil was treated with sewage sludge. Most of the As (35.5–38.8% of total content), Cu (45.0–51.6%) and Zn (39.8–47.2%) was also determined in this fraction. The G fraction (obtained after acid hydrolysis) contained polar compounds. Most of the Cd was also found in this fraction, as was a significant amount of Zn. Non-extractable residues formed the last fraction (polysaccharides, proteins).     

基于银溶胶表面增强拉曼光谱对菠菜毒死蜱农药的快速检测

利用表面增强拉曼光谱(SERS)技术,建立了一种用于菠菜中毒死蜱农药残留的非破坏、快速检测方法。以碱性环境下盐酸羟胺还原法制备的银溶胶作为表面增强剂滴涂于菠菜样品表面后,采用实验室自行搭建的拉曼系统直接采集样品的拉曼信息,该方法无需对样品进行前处理,可以实现菠菜中毒死蜱含量的实时在线定量分析。采集24片不同毒死蜱含量的菠菜样品拉曼光谱,每个样品采集20个点。拉曼信号采集后,用气相色谱法对24个菠菜样品中毒死蜱含量进行检测。为了消除光谱噪音以及荧光背景对分析建模的影响,分别采用Savitzky-Golay平滑和有效峰线性拟合法对原始拉曼光谱进行预处理。该表面增强拉曼方法具有较好的重复性,实验中对50个相同毒死蜱含量,但不同状态的菠菜进行光谱采集,其相对标准偏差为13.4%,说明该方法具有一定的普适性。光谱预处理后,选取615.5~626.4 cm-1波段为感兴趣区域,建立0.05~37.4 mg·kg-1浓度范围内毒死蜱含量的多元线性预测模型,结果表明感兴趣区域的拉曼信号和毒死蜱浓度呈良好的线性关系,其校正集和验证集相关系数R_C和R_P分别为0.961和0.954。该方法的最低检出含量为0.05 mg·kg-1,低于国家标准规定的农药残留最大限量。该方法简单快速,无需样品前处理,可以实现果蔬的农药残留快速、定量检测。