碳氮稳定同位素检测能力的验证——2013年实验室间比对分析结果的汇总

稳定性同位素示踪技术,特别是碳、氮稳定性同位素已广泛地应用于农业化学、地球化学和环境化学。碳、氮同位素比值的质谱检测结果直接关系到示踪试验的可靠性。在目前缺乏富集碳、氮同位素的标准物质的条件下,通过实验室间的比对可以验证各实验室对碳、氮同位素比值检测能力、检测结果的准确性和可比性。我所在2013年组织和实施了一次由全国14个单位的检测实验室共17台仪器参加的农业、生态和环境样品中碳氮百分含量及其稳定性同位素比值的实验室间比对。我所制备了多种被检测的样品,有含碳氮的化学肥料、土壤和植物样品;在稳定性同位素富集度上,有自然丰度的和不同富集程度的同位素样品。除固体样品外,还有3种不同丰度的N2O和CO2气体样品。在进行检测结果的统计和评价时,采用Z比分衡量各实验室检测结果的可信度。本文汇总了2013年实验室间比对的结果。     

土壤宏基因组学研究方法与进展

土壤微生物驱动着土壤中的物质循环和养分转化。在土壤学的研究中,长期将土壤作为一个黑箱系统来对待,对其中的生物组成及其参与的生化过程知之甚少。土壤中绝大部分微生物目前尚难以分离培养,因此基于传统的培养方法对于认识土壤微生物群落组成和功能有其局限性。宏基因组学直接从环境样品中提取全部微生物的DNA,或通过测序探究环境中微生物的群落结构和功能(序列驱动),或构建宏基因组文库,筛选新的基因或生物活性物质(功能驱动),克服了传统培养方法的缺陷,极大地丰富了对土壤微生物多样性及其功能的认知。本文在综述土壤宏基因组学研究基本流程的基础上,重点介绍了日益重要的第二代测序平台在土壤宏基因组学研究中的应用及其产生的海量数据的分析处理方法,并简要探讨了宏基因组学在土壤微生物生态学中的应用。最后,作者建议在国家层面上展开相关土壤宏基因组学研究,调查微生物群落及其变化,为生物资源开发、农业生产和环境保护作出应有的贡献。     

虱螨脲在土壤生态系统中的检测方法及持久性研究

采用模拟土壤生态系统的方法,研究了虱螨脲在土壤中的消解动态,应用高效液相色谱仪测定了虱螨脲在土壤中的残留量。结果表明,样品用甲醇提取,二氯甲烷萃取,用高效液相色谱检测,样品前处理简单有效,方法检出限和灵敏度均能达到要求。虱螨脲在土壤中消解较快：2007年试验半衰期为2.51 d,土壤原始附着量为0.180 mg.kg^-1;2008年试验半衰期为2.47 d,土壤原始附着量为0.209 mg.kg^-1。     

除草剂苄嘧磺隆降解菌Hansschlegelia zhihuaiae S113菌剂的研发

磺酰脲类除草剂苄嘧磺隆残留期较长，造成农田土壤的残留污染问题。以苄嘧磺隆高效降解菌株Hansschlegelia zhihuaiae S113为材料制备降解菌剂，优化液体菌剂的保护剂种类与配比，筛选固体菌剂的最佳载体，初步应用固体菌剂并评价修复效果。研究结果表明：（1）向液体菌剂中添加合适的保护剂（0.30% 柠檬酸钠、0.20% 羧甲基纤维素、0.10% CaCl2）可使活菌数提高37.25%，保存30 d的液体菌剂对50 mg/L苄嘧磺隆的降解率为94.25%。（2）筛选出猪粪有机肥为固体菌剂的最佳载体，保存60 d时固体菌剂活菌数为7.45×107 cfu/g，对土壤中10 mg/kg苄嘧磺隆的降解率为91.22%。（3）固体菌剂的添加可有效减轻土壤中苄嘧磺隆残留对玉米的药害。     

浅谈土壤常规检测中的几个关键环节

从土壤常规检测角度,结合工作实践,着重强调土壤样品检测方法遵循的原则、取样及样品制备、仪器设备质量保证及过程质量控制等几个关键环节,以确保土壤检测数据的准确性和有效性,促进我国农业的发展。     

固、液态磷源在石灰性土壤中的移动性及其对土壤有效磷含量影响的研究

采用同心圆扩散法对固体和酸性液体磷肥在石灰性土壤中的移动性与有效性进行研究。结果表明,土壤Olsen-P含量随施肥点圈层由内向外层而递减,液肥处理的Olsen-P量在施肥点以外的3个土壤圈层中均高于固肥处理。固体肥料大部分停留在距施肥点0~7.5 mm圈层,占施磷总量的55%~70%;液体肥料虽在0~7.5 mm层的含量低于固体肥料(约占15%~40%),但在远离施肥点的后3个圈层(7.5~43 mm)中均高于固体肥料处理,其中7.5~13.5 mm最为明显(液肥占35%,固肥占20%)。3种施磷量在砂土、壤土及粘土上均表现出一致的变化规律。肥料对土壤pH值的影响仅限制在距施肥点最近的0~7.5 mm范围,该层的pH值显著低于后层。本研究说明在石灰性土壤中液体磷的移动性、有效性明显高于固体磷,但对土壤pH值无显著影响。     

电感耦合等离子体发射光谱法测量土壤中有效锌含量的不确定度评估

对电感耦合等离子体发射光谱法(ICP-OES)测定土壤有效锌含量的不确定度进行评定,分析了整个检测过程产生不确定度的来源,对称样量、浸提液体积、标准系列溶液配制、线性标准曲线拟合、测量重复性等产生的不确定度分量进行计算,量化给出扩展不确定度。待测土壤中有效锌含量最终结果表示:w(Zn)=(1.12±0.10)mg/kg,包含因子k=2,置信概率为95%。测量过程中,标准溶液制备所产生的不确定度最大。因此,在ICP-OES法测定土壤样品有效锌时应足够重视标准溶液制备与曲线拟合过程,以减小测量不确定度。本文研究结果为控制ICP-OES法测定土壤有效锌数据质量提供了理论依据。     

内蒙古草原土壤有机物提取分离及鉴定

土壤中的有机物对于斥水性的研究具有重要的作用,但是目前关于内蒙古草原土壤的研究主要集中在有机碳上,而对土壤中有机物种类的研究很少。选取内蒙古锡林浩特市南部的牧草土壤样品,使用溶剂超声萃取的方法,对土壤中的有机物进行了分析。结果发现,石油醚的粗提物最少,异丙醇/氨水(7∶3)的粗提物最多。粗提物经紫外、红外和GC-MS测定,结果表明用石油醚、异丙醇和甲醇作为萃取剂,得到的有机物主要为直链或环烃化合物,含量占总提取物的95%以上;氯仿及异丙醇/乙酸(7∶3)和异丙醇/氨水(7∶3)提取物除了含有直链或环烃化合物外,还含有酯、酰胺和甾类化合物,且甾类化合物在异丙醇/氨水提取物中含量相对最高。采用异丙醇/氨水对所有土壤样品进行萃取,发现NM1和NM7提取物中甾类化合物的含量较高,占总提取物的25%左右。异丙醇/氨水的粗取物用石油醚:乙酸乙酯(4∶1)作洗脱剂,经柱层析分离主要得到A、B两部分;经GC-MS测定、质谱图库检索表明A为甾酮类物质,B为不含氧的甾烯类或者甾醇类物质。     

磁力搅拌法改进土壤阳离子交换量测定的研究

针对传统土壤阳离子交换量测定方法中的玻璃棒搅拌所存在的分析效率低的问题,开展了运用磁力搅拌替代玻璃棒搅拌的探索性研究。通过平行性实验、条件实验、实验室间对比和不同黏粒含量土壤的CEC测定结果检验,分析了磁力搅拌法的精准度、可操作性、稳定性和适用性。结果表明,磁力搅拌法测定土壤CEC的准确度、精密度均不低于甚至高于玻璃棒搅拌方法,土壤CEC测定值与土壤标准物质认定值吻合。当搅拌时间在1 min、搅拌速度850 r·min–1以上时,延长搅拌时间和提高搅拌速度对分析结果无影响。不同实验室间测定结果相关性好、没有显著性差异。相比传统的玻璃搅拌法,磁力搅拌法具有操作简单、高效,测定结果稳定性好、精准度高等优点,非常适用于大批量土壤CEC的测定。     

二苯醚类除草剂降解菌Bacillus sp. Za微生物制剂的研发与初步应用

以二苯醚类除草剂高效降解菌株Bacillus sp. Za为材料制备微生物制剂，优化液体制剂保护剂的物质配比，筛选固体制剂的最适材料，对固体制剂进行初步应用并评价其降解效果。研究结果表明：（1）液体制剂保护剂（0.20%柠檬酸钠、0.20%羧甲基纤维素、0.30% KCl）可使活菌数提高35.71%，保存30 d的液体制剂对50 mg/L乳氟禾草灵的降解率为83.50%。（2）筛选得到猪粪有机肥作为固体制剂的最适材料，保存60 d时固体制剂活菌数为8.26×108 cfu/g，对土壤中10 mg/kg乳氟禾草灵的降解率为85.52%。（3）添加固体制剂可有效缓解乳氟禾草灵残留对玉米所产生的药害。     

Eine Methode zur quantitativen Bestimmung von Pentachlorphenol in Umweltproben

A method for the quantitative determination of pentachlorophenol in environmental samples An analytical method is presented for the rapid extraction, purification and quantitative detection by gaschromatography of pentachlorophenol in environmental samples (plant, animal and soil samples). The detection sensivity of this method gets up to 5 ppb or 10 pg absolute.     

A sensitive method for detection of sulfamethazine and N4-acetylsulfamethazine residues in environmental samples using solid phase immunoextraction coupled with MALDI-TOF MS

Sulfamethazine (SMT) and its major metabolite, N(4)-acetylsulfamethazine (NA-SMT), were each recovered from spiked water (0.1 ppb) and 10% (w/v) aqueous suspensions of soil (1 ppb) or composted manure (1 ppb), by using a three-stage solid phase immunoextraction (SPIE) system, followed by detection with matrix-assisted laser/desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Sulfonamide recovery rates are reported for separate stages of the SPIE system and for trace-level sulfonamide SPIE extraction from the environmental samples. SPIE MALDI-TOF MS is a rapid and definitive technique with potentially better efficiency relative to other established trace-level sulfonamide analytical methods. SPIE MALDI-TOF MS required 1.5 h per batch (8-24 samples/batch) for sample enrichment, 5 min per batch for probe preparation, and 5 min per sample to acquire and process the spectrum. This is the first time MALDI-TOF MS has been reported as a potential means of detecting trace-level drug residues in complex environmental samples.     

Multiresidue analysis of pesticides in soil by supercritical fluid extraction/gas chromatography with electron-capture detection and confirmation by gas chromatography-mass spectrometry

The applicability of supercritical fluid extraction (SFE) in pesticide multiresidue analysis (organohalogen, organonitrogen, organophosphorus, and pyrethroid) in soil samples was investigated. Fortification experiments were conducted to test the conventional extraction (solid-liquid) and to optimize the extraction procedure in SFE by varying the CO2 modifier, temperature, extraction time, and pressure. The best efficiency was achieved at 400 bar using methanol as modifier at 60 degrees C. For the SFE method, C-18 cartridges were used for the cleanup. The analytical screening was performed by gas chromatography equipped with electron-capture detection (ECD). Recoveries for the majority of pesticides from spiked samples of soil at different residence times were 1, 20, and 40 days at the fortification level of 0.04-0.10 mg/kg ranging from 70 to 97% for both methods. The detection limits found were <0.01 mg/kg for ECD, and the confirmation of pesticide identity was performed by gas chromatography-mass spectrometry in a selected-ion monitoring mode. Multiresidue methods were applied in real soil samples, and the results of the methods developed were compared.     

Hydraulic Core Extraction: Cutting Device for Soil–Root Studies

Abstract

A critical objective of belowground research is to collect and process representative soil samples. Mechanical devices have been developed to quickly take soil cores in the field; however, techniques to rapidly process large‐diameter soil cores are lacking. Our objective was to design and construct a soil extraction–cutting system that could effectively reduce processing time. Soil cores were extracted from large diameter steel core tubes using a custom hydraulic cylinder device that vertically pushes the soil core to a desired depth increment before cutting in a horizontal direction with another hydraulically driven device. As many as eight large cores per hour could be processed with this system. This system has been effectively used in processing soil samples from both agricultural and forestry sites to meet desired experimental goals.     

用于ISE硝态氮检测的土壤原位前处理装置设计与试验

前处理自动化程度是影响土壤养分检测效率和可行性的关键环节之一。该文设计开发了1种适用于电极法硝态氮原位测定的前处理装置,融合土壤水分校准算法,装置具备水分测定、称重、注液浸提、高速旋转离心等功能,性能测定结果表明:装置两自由度机械平台纵向控制精度达到0.1 cm,旋转定位控制精度为1?,注液精度可达0.2 m L,位移量及注液量具有较高重复性;较常规手动操作,利用该装置进行土样前处理耗时明显降低,自动化及时效性明显提高,人工劳动强度下降;利用装置前处理土样,电极法测定结果与标准光学检测结果接近,两者回归方程调整决定系数为0.87,平均绝对偏差为20.97 mg/kg,均方根误差为22.34 mg/kg,较悬液直测精度明显提高。装置浸提环节仍需进一步优化,研究结果可为集成式土壤养分原位检测装置研发提供参考。     

Improved HPLC-MS/MS method for determination of isoxaflutole (balance) and its metabolites in soils and forage plants

A robust multi-residue procedure is needed for the analysis of the pro-herbicide isoxaflutole and its degradates in soil and plant materials at environmentally relevant (<1 microg kg-1) levels. An analytical method using turbo-spray and heat-nebulizer high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed for the analysis of isoxaflutole (IXF) and its two metabolites, diketonitrile (DKN) and the benzoic acid metabolite (BA), at sub-microgram per kilogram levels in soil and plant samples. The average recoveries of the three compounds in spiked soil and plant samples ranged from 84 to 110% and 94 to 105%, respectively. The limits of quantification were validated at 0.06 microg kg-1 for soil and 0.3 microg kg-1 for plant samples. The limits of detection (LOD) for soil analysis were 0.01, 0.002, and 0.01 microg kg-1 for IXF, DKN, and BA, respectively. Corresponding LOD for the plant analysis method were 0.05, 0.01, and 0.05 microg kg-1. The developed method was validated using forage grass and soil samples collected from a field lysimeter study in which IXF was applied to each of four forage treatments. Forage plants and soils were sampled for analyses 25 days after IXF application to the soil. In soils, IXF was not detected in any treatment, and DKN was the predominant metabolite found. In forage plants, the concentrations of DKN and BA were 10-100-fold higher than that in soil samples, but IXF was not detected in any forage plants. The much higher proportion of BA to DKN in plant tissues (23-53%), as compared to soils (0-5%), suggested that these forages were capable of detoxifying DKN. The developed methods provided LODs at sub-microgram per kilogram levels to determine the fate of IXF and its metabolites in soils and forage plants, and they also represent considerable improvements in extraction recovery rates and detection sensitivity as compared to previous analytical methods for these compounds.     

Simultaneous determination of ethidimuron, methabenzthiazuron, and their two major degradation products in soil

An analytical method has been developed for the quantification of two herbicides (ethidimuron and methabenzthiazuron) and their two main soil derivatives. This method involves fluidized-bed extraction (FBE) prior to cleanup and analysis by reverse-phase liquid chromatography with UV detection at 282 nm. FBE conditions were established to provide efficient extraction without degradation of the four analytes. (14)C-labeled compounds were used for the optimization of extraction and purification steps and for the determination of related efficiencies. Extraction was optimal using a fexIKA extractor operating at 110 degrees C for three cycles (total time = 95 min) with 75 g of soil and 150 mL of a 60:40 v/v acetone/water mixture. Extracts were further purified on a 500 mg silica SPE cartridge. Separation was performed on a C18 Purosphere column (250 mm x 4 mm i.d.), at 0.8 mL min(-1) and 30 degrees C with an elution gradient made up of phosphoric acid aqueous solution (pH 2.2) and acetonitrile. Calibration curves were found to be linear in the 0.5-50 mg L(-1) concentration range. Besides freshly spiked soil samples, method validation included the analysis of samples with aged residues. Recovery values, determined from spiked samples, were close to 100%. Limits of detection ranged between 2 and 3 microg kg(-1) of dry soil and limits of quantification between 8 and 10 microg kg(-1) of dry soil. An attempt to improve these performances by using fluorescence detection following postcolumn derivatization by orthophthalaldehyde-mercaptoethanol reagent was unsuccessful.     

Rapid analysis of inositol phosphates

Fast and simple analytical methods for the determination of inositol bis- to hexakisphosphates or only inositol hexakisphosphate in foods and feces are presented. The methods are both faster and simpler with regard to analytical detection and sample pretreatment as compared to previously reported methods. The samples are pretreated using extraction and centrifugal ultrafiltration and analyzed using high-performance ion chromatography (HPIC) with gradient or isocratic elution. The analytes are detected using ultraviolet detection after postcolumn reaction. The methods are efficient, highly selective, and appropriate for analyzing inositol phosphates in food and feces samples. The between- and within-day variances were generally below 8 and 5% (relative standard deviation), respectively, for the presented HPIC method with gradient elution.     

基于频域法的便携式无线土壤水分测量装置设计与试验

针对农田土壤水分测量的实际需要,研制了一种便携式无线土壤水分测量装置。该装置结构一体化设计采用"T"型结构,将土壤水分传感器和信息采集与发送单元融合,可在0~300 mm的不同深度下测量土壤水分,并采用蓝牙传输技术,将测量数据实时发送给Android手机,手机可通过App软件对数据进行分析处理,实现了农田数据的大容量存储和智能化处理。在实验室环境下,使用砂土和壤土2种土样对测量装置进行了标定试验,土壤容积含水率与传感器输出电压服从二次曲线关系,决定系数均达到0.99以上;将测量装置与波兰Easy Test TDR土壤测试仪进行对比试验,二者测量结果呈线性相关关系,决定系数为0.987。试验结果表明该装置可准确测量土壤水分含量。     

Sample extraction method combining micellar extraction-SPME and HPLC for the determination of organochlorine pesticides in agricultural soils

A method for the determination of organochlorine pesticides in soil samples combining microwave assisted micellar extraction (MAME) with solid-phase microextraction (SPME) and high-performance liquid chromatography-UV has been developed. A mixture of two nonionic surfactants (polyoxyethylene 10 lauryl ether and polyoxyethylene 10 stearyl ether) was used for the extraction of pesticides from agricultural soils, and different types of SPME fibers were compared. The different parameters which affect extraction efficiency in the SPME procedure were optimized such as extraction time and temperature. The method developed involves extraction and preconcentration for the target analytes in soil samples. The analytical parameters were also studied and good recoveries obtained, RSD being lower than 10% and detection limits ranging between 36 and 164 ng g(-1) for the pesticides studied. The proposed method was successfully applied to the determination of some organochlorine pesticides in several kinds of agricultural soil samples with different characteristics.