表面活性剂与EDTA对紫色土镉的化学行为影响

Elemental composition and geochemical characteristics of iron-manganese nodules from nine main soils in China were studied by chemical and multivariate statistical analyses to better understand the reactions and functions of ironmanganese nodules in soils and sediment. Compared to the corresponding soils, Mn, Ba, Cd, Co and Pb had strong accumulation, Ni had moderate accumulation, while Ca, Cu, Fe, Na, P, Sr and Zn accumulated to a minor degree in the iron-manganese nodules. In contrast, Si, Al, K, Mg and Ti were reduced in the iron-manganese nodules. The contents of Ba, Cd, Co, Cu, Ni, Pb and Zn were positively and significantly correlated with that of MnO2 in the iron-manganese nodules, while the contents of Cr, Cu, Ni, Pb and Zn were positively and significantly correlated with that of Fe2O3 in soils. Based on a principle component analysis, the elements of iron-manganese nodules were divided into four groups： 1） Mn, Ba, Cd, Co, Cu, Li, Ni, Pb and Zn that were associated with Mn oxides, 2） Fe, Cr and P that were associated with Fe oxides, 3） Si, K, and Mg that were included in the elemental composition of phyllosilicate, and 4） Ca, Na, Al and Ti that existed in todorokite, birnessite, lithiophorite and phyllosilicate. It was suggested that accumulation, mineralization and specific adsorption were involved in the formation processes of soil iron-manganese nodules.     

Effectiveness of the Iodide Ligand Along with two Surfactants on Desorbing Heavy Metals from Soils

Used with one of two surfactants (SDS, an anionic surfactant, and Triton X-100, a nonionic surfactant), the ligand, I^? was evaluated as a washing agent for the desorption of Cd from naturally and artificially contaminated soils. Increasing amounts of the ligand, I^?, with a surfactant, specifically removes higher levels of Cd but not Cu, Zn and Pb. After seven washings, the ligand, I^? with the nonionic surfactant, Triton X-100, removed 65 and 90% of the Cd from soils I and II, containing respectively, to 15 and 1275 mg of Cd/kg. The ligand, I^?, and the anionic surfactant, SDS, removed 35 and 70% of the Cd from soils I and II, respectively. Before washing, the carbonate fraction of soil I contained the most Cd (48%) while the exchangeable and carbonate fractions of soil II contained 29 and 33% of the total Cd, respectively. For soil I, SDS with/ without the ligand desorbed Cd mainly from the carbonate and oxide fractions, while only Triton X-100 with ligand I^? could remove Cd from the exchangeable fraction. For soil II, Cd was desorbed from the exchangeable fraction only when either surfactant was used in combination with the ligand. Thus, a surfactant with ligand can extract specific heavy metals from soils and selective sequential extraction is useful in identifying which fraction can be targeted by the surfactant – ligand agent.     

有机溶剂对有机氯农药污染场地土壤淋洗修复效用研究

Problems associated with organochlorine pesticide (OCP)-contaminated sites in China have received wide attention. To solve such problems, innovative ex-situ methods of site remediation are urgently needed. We investigated the feasibility of the extraction method with different organic solvents, ethanol, 1-propanol, and three fractions of petroleum ether, using a soil collected from Wujiang (WJ), China, a region with long-term contamination of dichlorodiphenyltrichloroethanes (DDTs). We evaluated different influential factors, including organic solvent concentration, washing time, mixing speed, solution-to-soil ratio, and washing temperature, on the removal of DDTs from the WJ soil. A set of relatively better parameters were selected for extraction with 100 mL L 1 petroleum ether (60-90℃): washing time of 180 min, mixing speed of 100 r min 1 , solution-to-soil ratio of 10:1, and washing temperature of 50℃. These selected parameters were also applied on three other seriously OCP-polluted soils. Results demonstrated their broad-spectrum effectiveness and excellent OCP extraction performance on the contaminated soils with different characteristics.     

表面活性剂与EDTA对雪菜吸收镉的影响

采用表面活性剂与螯合剂处理,强化雪菜吸收土壤镉的盆栽试验表明,影响植物吸收镉的主要因子是表面活性剂类型。阴离子型与非离子型表面活性剂的强化修复效果要优于阳离子型表面活性剂,其中以十二烷基硫酸钠和Tween80为好。表面活性剂与EDTA复合使用,可以降低土壤对镉的吸附(顺序依次为EDTA/DBSSEDTA/TX-100EDTA/CTABEDTADBSSTX-100CTAB),增加土壤对镉的解吸,进而促使土壤镉向植物转移,有利于强化镉污染土壤的植物修复。     

The influence of aminopolycarboxylates on the sorption of copper (II) cations by (Hydro)oxides of iron,Aluminum, and manganese

The influence of some complexing agents of (poly)aminopolycarboxylic acids (diethylenetriaminopentaacetic acid (DTPA), ethylenediaminotetraacetic acid (EDTA), nitrilotriacetic acid (NTA), and iminodiacetic acid (IDA)) on the sorption of Cu²⁺ by crystal and amorphous (hydr)oxides of Fe(III), Al(III), and Mn(IV) that are widespread mineral components of soils was studied. The obtained results are considered in terms of complex-formation in the solution and on the sorbent’s surface. The effect of the complexing agents on the metal sorption (mobilization/immobilization) is determined by (1) the stability, structure, and sorption capability of compexonates formed in the solution; (2) the acidity, and (3) the nature of the sorbent. The desorption effect on Cu²⁺ cations was found to change in the following sequence of complexing agents: EDTA > DTPA ? NTA > IDA. The high-dentate complexing agents (EDTA, DTPA) had the greatest impact on ?u²⁺ cations bound with crystalline (hydr)oxides of Fe, Al, and Mn. The low denticity of the complexing agents (IDA, NTA) and binding of ?u²⁺ with amorphous sorbents leads to the weakening of desorption. The decrease in acidity promoted the mobilization of the metal under the influence of complexing agents; the increase in acidity caused its immobilization. The growth in the mobility of heavy metals bound with soil (hydr)oxides of Fe, Al, and Mn due to the complexing agents entering the surface and ground water is considered a factor of ecological risk.     

Remediation of Organochlorine Pesticide-Contaminated Soils by Surfactant-Enhanced Washing Combined with Activated Carbon Selective Adsorption

Remediation of organochlorine pesticide (OCP)-contaminated soils is urgently required especially in China. The present study investigated the removal of OCPs from two soils by triton X-100 (TX-100)-enhanced washing coupled with powdered activated carbon (PAC) adsorption treatment of the solution. Two contaminated soils, including a silt clay contaminated with chlordene, 1,1-dichloro-2,2-bis(p-chlorophenyl)ethanes (DDDs), and dichlorodiphenyltrichloroethane (DDT) and a sandy loam containing chlordanes and mirex, were selected. Effects of varied operating parameters, including TX-100 dosage, liquid/soil (L/S) ratio, and extraction time, on the contaminant removal were examined. For both soils, OCP removal was clearly enhanced with increasing TX-100 in extraction solution and L/S ratio. Meanwhile, the removal efficacy was also impacted by soil texture and characteristics and contents of the contaminants. Moreover, by using PAC as an absorbent, a significant decrease in the contaminants in the extraction solutions was detected, and TX-100 could be further reused. Our investigation suggests that surfactant-enhanced washing combined with activated carbon selective adsorption would be an alternative to remediate OCP-contaminated soils.     

重金属高污染农田土壤EDTA淋洗条件初探

通过室内振荡淋洗试验研究了乙二胺四乙酸二钠(EDTA)浓度、淋洗时间、液固比、淋洗次数对甘肃省白银市某高污染农田土壤中重金属去除效果的影响,并测定了EDTA淋洗前后土壤中重金属形态的变化。结果表明：淋洗剂浓度和液固比越高、淋洗时间越长、淋洗次数越多,对重金属的去除效果越好。在EDTA浓度为5 mmol/L、液固比为2.5、连续振荡淋洗3次、每次1 h时,对土壤中Cd、Cu、Pb、Zn 4种重金属的总去除率分别为 55.2%、21.9%、19.3% 和20.9%,其中Cd 淋洗效率最高。EDTA对土壤中交换态、碳酸盐结合态和铁锰氧化物结合态重金属的去除效果明显,但不能有效去除有机及硫化物态和残余态土壤重金属。     

EDTA Leaching of Cu Contaminated Soils Using Ozone/UV for Treatment and Reuse of Washing Solution in a Closed Loop

Ozone and UV irradiation were used for oxidative decomposition of EDTA-Cu complexes in washing solution obtained during multi-step leaching of Cu (344,1?±?36.5 mg kg^?1) contaminated vineyard soil with EDTA as a chelant. The released Cu was absorbed from the washing solution on a commercial mixture of metal absorbing minerals, and the treated washing solution then reused for removal of soil residual Cu-EDTA complexes in a closed-loop process. Six consecutive leaching steps (6?×?2.5 mmol kg^?1 of EDTA) removed 38.8 % of Cu from soils, and reduced Cu soil mobility, determined using the toxicity characteristic leaching test (TCLP), by 28.5%. The final washing solution obtained after soil remediation was colourless, with a pH close to neutral (7.5?±?0.2) and with low concentrations of Cu and EDTA (0.51?±?0.22 mg L^?1 and 0.083 mM, respectively). The proposed remediation method has therefore potential not just to recycle and save process water, but also not to produce toxic wastewaters. Soil treatment did not substantially alter the soil properties determined by pedological analysis, and had relatively little impact on soil hydraulic conductivity and soil water sorption capacity.     

淋洗法修复铬渣污染场地实验研究

铬渣污染场地已经列为我国土壤污染重点治理对象,急需修复铬渣污染土壤的关键技术。本文通过室内模拟实验,采用振荡淋洗的方法研究了水、EDTA、EDDS、柠檬酸、草酸、盐酸、磷酸、环糊精和十二烷基苯磺酸钠等淋洗剂对铬渣污染场地Cr的淋洗效果,探讨了淋洗剂浓度、淋洗时间、土水比、淋洗次数等对淋洗效果的影响,并研究了不同淋洗剂复合对Cr的淋洗效果以及不同浓度柠檬酸淋洗前后重金属形态的变化。结果表明：水、EDTA、EDDS、环糊精和十二烷基苯磺酸钠对Cr的去除率较低,柠檬酸在浓度为0.5mol·L-1、土水比1：20、反应时间为24h、淋洗次数为2次的条件下可以达到最佳淋洗效果,Cr的去除率可达到82.8%;SDBS、EDTA与柠檬酸单独组合顺序或者混合淋洗,都没有增加柠檬酸对Cr的去除率。重金属形态分析表明,柠檬酸淋洗有效地改变了Cr的形态从而达到较高的修复效果。当柠檬酸浓度小于0.25mol·L-1时,对铬的去除主要以C（rⅥ）为主;当柠檬酸浓度大于0.25mol·L-1时,对铬的去除主要以C（rⅢ）为主。本文的研究结果为异位淋洗修复铬渣污染土壤提供理论依据。     

Assessing the Efficiency of Humic Substances as Washing Agents for Oil–Contaminated Soils and Peats under Model Experimental Conditions

Two humic preparations of different origins have been compared as washing agents for oil-contaminated soils and peat under model experimental conditions using a sample from the plow horizon of soddypodzolic soil artificially contaminated with oil or diesel fuel and a sample of high-moor peat contaminated with crude oil because of a spill occurred 15 years ago. Soil and peat were washed by shaking with solutions of the humic preparations Gumat Sakhalinskii and Lignogumat in a 1: 10 (m/v) ratio. Control samples were washed with distilled water. Washing with a synthetic surfactant (sodium dodecyl sulfate) was also added to the experimental design. After washing, soil and peat samples were air-dried and used for the determination of the total content of petroleum hydrocarbons; the characterization of their hydrocarbon composition; and the assessment of hydrophobicity from the contact angle and the efficiency of colonization by oil-destructing microorganisms Rhodococcus sp. and Candida sp., which are components of the preparation Bioros recommended for oil contaminations. It has been shown that the extraction efficiency of petroleum hydrocarbons by humic preparations did not differ from the extraction efficiency by water and was less than that by sodium dodecyl sulfate in all cases. No appreciable changes in the contact angles of soil and peat have been observed at the use of water and humic preparations as washing agents, while the contact angle decreased to less than 90o after washing with sodium dodecyl sulfate, which indicated the hydrophobicity of the surface of substrate particles. It has been found that humic preparations favor the colonization of soil and peat by oil-destructing microorganisms Rhodococcus sp. and Candida sp. Based on the obtained results, humic preparations have been recommended for further study as preparations favoring the ability of oil-destructing microorganisms to colonize oil-contaminated substrates.     

冻融-化学淋洗法协同修复重金属Cd和Pb污染黏性土

针对质地黏重、低渗透性黏性土的淋洗效率低下,该文提出冻融协同化学淋洗的修复方案,并以某冶炼厂受Cd、Pb污染场地黏性土为研究对象,选用乙二胺四乙酸二钠(ethylene diaminetetraacetic acid disodium salt,EDTA)为淋洗剂,进行了冻融-淋洗土柱的实证试验。结果表明,土体的反复冻融(冻胀-吸水、融沉-排水)破坏土体颗粒原有结构,有助于淋洗液与污染物充分接触,淋洗效果明显,经7次冻融后,Cd、Pb去除率分别达到77.24%、37.78%。采用改进的BCR(European Communities Bureau of Reference)连续提取法分析了土柱中Cd、Pb的赋存特征,经7次冻融后,土壤中弱酸提取态、可还原态、残渣态结合的Cd质量分数较淋洗前分别降低了41.46%、63.02%、26.33%,而土壤中可还原态和残渣态结合的Pb质量分数分别降低了32.32%、67.36%。冻融协同化学淋洗修复技术的淋洗剂用量远小于传统淋洗法,为今后利用寒区冻融交替现象,大规模对季冻区重金属污染土壤的异位修复提供了新的思路。     

重金属污染土壤清洗导致几种营养元素流失的研究

通过HCl、柠檬酸、EDTA这3种提取剂对Pb、Cd、Cu、Zn污染土壤的漫提试验，研究了土壤清洗洗脱重金属的效果，并探讨了土壤清洗导致K、Ca、Mg、Fe、Mn这几种营养元素的流失和有效性的改变。试验结果表明，土壤清洗能有效地洗脱土壤中的Ph、Cd、Cu、Zn，伺时也造成了土壤中K、Ca、Mg、Fe、Mn的流失，降低了其中部分营养元素的有效性。EDTA对重金属的洗脱能力大于HCl和柠檬酸。HCl漫提造成的K、Ca、Mg的流失量大于柠檬酸和EDTA，柠檬酸浸提造成的Fe流失量大于HCl和EDTA，EDTA漫提造成的Mn流失量大于HCl和柠檬酸。HCl和柠檬酸浸提降低了土壤K、Ca、Mg的有效性EDTA浸提提高了土壤K的有效性。降低了土壤Ca的有效性。在高浓度时（50mmol／L），EDTA显著地降低了土壤Mg的有效性。HCl漫提导致土壤K的有效性下降比柠檬酸严重，高浓度的EDTA浸提导致土壤Ca的有效性下降比HCl和柠檬酸严重。HCl、柠檬酸、EDTA浸提都能提高土壤中Fe的有效性．且EDTA的效应大于其它2种萃取剂；在低浓度时，3种提取剂都能提高土壤中Mn的有效性，且以柠檬酸的作用最为明显，当提取剂浓度增加时，Mn的有效性有所回落。     

表面活性剂对老化土壤中PAHs的生物有效性的影响

采用聚2,6-二苯基对苯醚(poly(2,6-diphenyl-p-phenylene oxide),Tenax-TA)提取和蚯蚓(Eisenia Fetida)富集实验研究曲拉通-100(聚乙二醇辛基苯基醚,Triton X-100)、吐温-80(聚氧乙烯脱水山梨醇油酸酯,Tween-80)和羟丙基-β环糊精(HPCD)表面活性剂对老化农田土壤中多环芳烃(PAHs)可提取组分以及生物有效性的影响。结果显示,表面活性剂可以显著增加PAHs的可提取性和在蚯蚓体内的蓄积浓度;和对照相比,增加可提取性42.3%~269%,蚯蚓体内的蓄积量增加10%~340%。研究表明在使用表面活性剂进行土壤修复时老化PAHs类污染物可能重新释放到环境中并带来环境风险。     

Extraction kinetics of copper,zinc, iron,and manganese from contaminated sediment using Disodium Ethylenediaminetetraacetate

One technique for cleansing heavy metal contaminated soils is to wash the excavated soil with an extraction solution of a chelating agent. The rate of extraction is an important parameter when considering the length of time needed for soil clean-up and the amount and concentration of wash solution required. The extraction kinetics of copper, zinc, iron and manganese from a contaminated sediment of the Clark Fork River in western Montana, U.S.A., with Disodiun Ethylenediaminetetraacetate (Na₂EDTA) as the extraction agent, were investigated. The results showed the extraction process consisted of rapid extraction in the first minutes followed by much slower extraction for the remainder of the experiment. The rate of extraction, particularly in the rapid phase, demonstrated clear pH dependence: the lower the pH, the faster the extraction rate. In the EDTA concentration range of 0.01 M to 0.05 M, the effect of the EDTA concentration on the extraction rate was not important compared with that of the solution pH. Extraction kinetics for different size particles were similar, although in the first few minutes, EDTA extracted more metals from clay and silt than sand. The two reaction, diffusion, and two-constant kinetic models were compared to experimental results. The two reaction model did not fit any of the data well, and only iron extraction could be described with a simple diffusion model. In general the extraction rates can be well described by the two-constant model, C=A t ^B, up to 600 minutes and under different conditions such as solution pH, EDTA concentration, and different sediment particle size.     

添加螯合剂诱导－栽培红叶菾菜（Beta vulgaris var. cicla L.）修复铅和镉污染土壤效果的研究

以雄安新区安新县重金属污染农田土壤为供试土壤,以Cd超积累植物红叶菾菜（Beta vulgaris var. cicla L.）为供试植物,设置不同浓度EDTA和柠檬酸（0,2.5,5,7.5,10 mmol kg?1）处理进行盆栽试验,探究螯合诱导－红叶菾菜修复Cd、Pb污染土壤的可行性。结果表明:（1）与对照相比,添加EDTA螯合剂使红叶菾菜生长及生物量均受到抑制,一定浓度柠檬酸处理能显著促进植物生长,5 mmol kg?1柠檬酸处理对植物株高、茎粗及生物量与对照相比的上升比例分别为4.52%、44.07%和50%;（2）添加EDTA螯合剂后土壤中Cd、Pb有效态含量相比对照分别提高了108.61% ~ 235.39%、67.98% ~ 224.16%,柠檬酸处理后土壤Cd、Pb有效态含量最大提高了180.07%、186.01%,EDTA对土壤重金属的活化效率显著高于柠檬酸;（3）通过对红叶菾菜地上部Cd、Pb含量及富集系数比较发现,EDTA更能促进红叶菾菜对Pb的吸收,柠檬酸更能促进红叶菾菜对Cd的吸收;（4）螯合剂处理后土壤中铵态氮、有效磷、速效钾含量显著增加。就本文试验条件、供试材料而言,螯合诱导－红叶菾菜修复铅镉复合污染土壤是可行的。     

Distribution of Manganese(Ⅱ) Chemical Forms on Soybean Roots and Manganese(Ⅱ) Toxicity

Distribution of chemical forms of manganese(Ⅱ)(Mn(Ⅱ))on plant roots may affect Mn(Ⅱ)absorption by plants and toxicity of Mn(Ⅱ)to plants at its high level.The chemical forms of Mn(Ⅱ)on soybean roots were investigated to determine the main factors that affect their distribution and relationship with Mn(Ⅱ)plant toxicity.Fresh soybean roots were reacted with Mn(Ⅱ)in solutions,and Mn(Ⅱ)adsorbed on the roots was differentiated into exchangeable,complexed,and precipitated forms through sequential extraction with KNO_3,EDTA,and HCl.The exchangeable Mn(Ⅱ)content on the roots was the highest,followed by the complexed and precipitated Mn(Ⅱ)contents.Mn(Ⅱ)toxicity to the roots was greater at pH 5.5 than at pH 4.2 due to the larger amount of exchangeable Mn(Ⅱ)at higher pH.The cations Al~(3+),La~(3+),Ca~(2+),Mg~(2+),and NH_4~+competed with Mn(Ⅱ)for cation exchange sites on the root surfaces and thus reduced exchangeable Mn(Ⅱ)on the roots,in the order Al~(3+),La~(3+)Ca~(2+),Mg~(2+)NH_4~+.Al~(3+) and La~(3+) at 100μmol L~(-1) decreased exchangeable Mn(Ⅱ)by 80%and 79%,respectively,and Ca~(2+) and Mg2+at 1 mmol L~(-1) decreased exchangeable Mn(Ⅱ)by 51%and 73%,respectively.Organic anions oxalate,citrate,and malate reduced free Mn(Ⅱ)concentration in solution through formation of complexes with Mn(Ⅱ),efficiently decreasing exchangeable Mn(Ⅱ)on the roots;the decreases in exchangeable Mn(Ⅱ)on the roots were 30.9%,19.7%,and 10.9%,respectively,which was consistent with the complexing ability of these organic anions with Mn(Ⅱ).Thus,exchangeable Mn(Ⅱ)was the dominant form of Mn(Ⅱ)on the roots and responsible for Mn(Ⅱ)toxicity to plants.The coexisting cations and organic anions reduced the exchangeable Mn(Ⅱ)content,and thus they could alleviate Mn(Ⅱ)toxicity to plants on acid soils.     

Desorption of Cadmium(II) from Artificially Contaminated Sediments

The desorption kinetics of cadmium from natural sediments was investigated. Three sediment samples from Maryland water bodies were artificially contaminated with Cd(II) and subsequently exposed to a wide range of chemical conditions in batch reactors. Dissolved Cd(II) was monitored. The highest desorption rates and extents were observed at low pH and high concentrations of CaCl₂ and EDTA (a strong Cd chelator), with reduced rates at higher pH. Nearly all of the desorption occurred within the first 30 minutes of exposure. Adsorption of Cd(II) was greatest, and desorption slowest on the sediments with the lowest sand content. A simple desorption model was developed assuming proton competition with cadmium. This model was fit to experimental data at different pH using a non-linear, least-squares analysis to obtain rate constants.     

Phytoremediation of Mixed Soil Contaminants

Tests were conducted to study the influence of non-ionic surfactants Triton X-100 and Tween 80 on the removal of mixed contaminants from a sandy soil using phytoremediation. Cd(II) and Pb(II) were used to form the inorganic contaminant, while used engine oil was selected to form the organic contaminant. The Indian mustard (Brassica juncea) plant was the plant chosen for phytoremediation of the sandy soil that contained the mixed contaminant. Thirty days after the plants were grown in the greenhouse, surfactants were applied to test pots in which the soil had been spiked with 50 mg kg⁻¹ of CdCl₂, 500 mg kg⁻¹ of PbCl₂ and 500 mg kg⁻¹ of used engine oil. Two control tests were conducted in this study. Planted and unplanted control tests were conducted using soil without surfactants. Following these tests, the tests were completed using the plants and surfactants at different concentrations. Test results showed that Triton X-100 and Tween 80 at concentrations higher than their critical micellar concentration enhanced Cd(II) and Pb(II) accumulation in the plant roots. Further, test data showed that translocation of contaminants to plant shoots occurred for Cd(II) but not for Pb(II). At the same concentrations, Tween 80 was more effective than Triton X-100 in facilitating rhizodegradation of used engine oil. This study demonstrates that simultaneous phytoremediation of Pb(II), Cd(II) and oil can be enhanced by using non-ionic surfactant Tween 80. Leaching test results indicated that the enhanced phytoremediation could remove the mixed contaminants safely from the point of view of limiting groundwater contamination.     

EDTA Extraction of Heavy Metals from Different Soil Fractions and Synthetic Soils

A laboratory-prepared contaminated soil was partitioned into four fractions, namely carbonate, Fe/Mn oxides, organic matter and clay mineral, according to the form in which the heavy metal bound with soil constituents. Individual contaminated soil fractions and synthetic soils were prepared for the study of soil extraction using ethylenediaminetetraacetic acid (EDTA). The effect of contact time and EDTA concentration were evaluated for both individual soil fractions and synthetic soils. The extraction reached equilibrium rapidly, after about 30 min. A 0.01 M EDTA solution was less effective than a 0.05 M or a 0.10 M EDTA. EDTA was proved to be effective for metal removal from the four individual soil fractions and synthetic soils. In general, approximately 90% of metals were removed from synthetic soils by 0.10 M EDTA. EDTA extraction of Pb from a contaminated carbonate fraction was thought to be affected by the formation of lead carbonates. A simple equation based on the sum of the released heavy metal from the individual components is used to check if there are interactions among the different soil components when mixed. The estimated values agreed well with the experimentally measured results only for the 0.10 M EDTA system.     

Pre-lysis washing improves DNA extraction from a forest soil

A pre-lysis buffer washing procedure was introduced to DNA extraction from a forest soil with high organic matter and iron oxide contents. Sodium phosphate of 0.1 M (pH 7.5) was used as a buffer to wash soil samples when subsequent lysis buffer was phosphate, and 20 mM EDTA (pH 7.5) was used when subsequent lysis buffer included EDTA. Initial experiments were not successful because the DNA extracts could not be amplified by polymerase chain reaction (PCR). The consideration of introducing a pre-lysis washing procedure was based on the idea that the washing should promote soil dispersion and homogeneity, decrease DNA adsorption by soil components (e.g. iron oxides), and remove covalent cations and those easily-dissolving organic compounds from the soil samples. Results revealed that humic substance content decreased by 31%, but DNA yield increased by 24% in the DNA extracts of the pre-lysis washing procedures, compared to the non-washing procedures. DNA extracted by the pre-washing procedure needed less purification for subsequent 18S and 16S rDNA PCR amplifications. It was recommended that the pre-lysis buffer washing should be used for DNA extraction from those difficult environmental samples, such as the forest soil with high contents of organic matter and iron oxides.