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Communications in Soil Science and Plant Analysis

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Phosphorus Determination after Mehlich 3 Extraction and Anion Exchange Resin in an Agricultural Soil of Northwestern Spain
Jorge Paz-Ferreiro , Eva Vidal Vzquez & Cleide Aparecida de Abreu
a c a b

Centro de Investigaciones Agrarias de Mabegondo (CIAM), La Corua, Spain

b c

Facultad de Ciencias, Universidade da Corua, La Corua, Spain Instituto Agronmico de Campinas, Campinas, Brazil

Available online: 30 Jan 2012

To cite this article: Jorge Paz-Ferreiro, Eva Vidal Vzquez & Cleide Aparecida de Abreu (2012): Phosphorus Determination after Mehlich 3 Extraction and Anion Exchange Resin in an Agricultural Soil of Northwestern Spain, Communications in Soil Science and Plant Analysis, 43:1-2, 102-111 To link to this article: http://dx.doi.org/10.1080/00103624.2012.634688

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Communications in Soil Science and Plant Analysis, 43:102111, 2012 Copyright Taylor & Francis Group, LLC ISSN: 0010-3624 print / 1532-2416 online DOI: 10.1080/00103624.2012.634688

Phosphorus Determination after Mehlich 3 Extraction and Anion Exchange Resin in an Agricultural Soil of Northwestern Spain
JORGE PAZ-FERREIRO,1 EVA VIDAL VZQUEZ,2 AND CLEIDE APARECIDA DE ABREU3
Centro de Investigaciones Agrarias de Mabegondo (CIAM), La Corua, Spain Facultad de Ciencias, Universidade da Corua, La Corua, Spain 3 Instituto Agronmico de Campinas, Campinas, Brazil
2 1

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Differences in soil phosphorus (P) contents measured by various techniques may have implications for agronomic and environmental testing. Reduced-tillage systems combined with surface manure application increase the potential risk of nutrient losses by surface runoff. A eld trial was conducted to evaluate the effect of livestock slurry on nutrient accumulation at the surface layer of an acidic soil rich in organic matter with excess P levels and loamy texture. Two reduced-tillage systems, no tillage (NT) and minimum tillage (MT), and four different fertilizer treatments were assessed. The amounts of P extracted by anion-exchange resin (AER) and by Mehlich 3 (M3) were compared; in addition, differences between two determination methods of P contents extracted by Mehlich 3, namely molybdic acid colorimetric standard procedure (M3-COL) and inductively coupled plasmamass spectroscopy (M3-ICP), were evaluated. Ninety-six soil samples were taken from the 0- to 5-cm surface layer in three successive dates after increasing manure addition. Colorimetric Mehlich 3 P ranged from 49 to 431 mg dm3 . The ranks of mean extractable soil P concentrations were AER < M3-COL < M3-ICP. The linear correlation coefcient between M3-COL and M3-ICP was highly signicant (R2 = 0.89; P < 0.01), but a two-straight-lines model or a quadratic relationship were more adequate for describing the dependence between the two determination procedures after M3 extraction. Relative and absolute differences between M3-COL and M3-ICP showed a tendency to increase as organic carbon content increased. Phosphorus content extracted by AER and M3-COL or M3-ICP reported a signicant but much less predictable relationship with R2 values of 0.27 and 0.21 (n = 96), respectively. The P in the surface layer accumulated more under NT than under MT. Keywords Colorimetric analysis, ICP, ion exchange resin, livestock manure, Mehlich 3, phosphorus

Introduction
Phosphorus (P) is an essential element for crop growth, and it is necessary to maintain protable agriculture. However, in agricultural areas with high cattle manure and/or slurry inputs, soil P has accumulated to levels in excess of crop needs and has the potential to enrich surface runoff. Thus, P export in watershed runoff can trigger the eutrophication
Address correspondence to Jorge Paz-Ferreiro, Centro de Investigaciones Agrarias de Mabegondo (CIAM), Apartado 10, 15080, La Corua, Spain. E-mail: jpaz@udc.es

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of surface water bodies (Gartley and Sims 1994; Sims et al. 2000, 2002; Kronvang et al. 2005). The accelerated nutrient enrichment or eutrophication of surface waters has become a signicant environment problem in many developed countries, and agriculture has been identied as a signicant P source (Gartley et al. 2002; Kronvang et al. 2005). This is also the case in the Atlantic temperate humid region of Galicia, northwestern Spain (Sande Fouz 2005). No-tillage (NT) systems are efcient in reducing soil losses caused by rainfall erosion, when compared with conventional tillage. However, these effects are generally less on mineral and organic nutrient losses. In the case of P, Bertol et al. (2007) found greater concentrations in the supercial 0- to 0.025-m soil layer of NT than in conventionally tilled erosion plots. The marked accumulation of P in the supercial soil layer in systems with reduced tillage was responsible for the greater concentration of this element in runoff and sediment from NT plots when compared with conventional tillage. Soil testing is a useful diagnostic tool to assess plant P availability. Most soil tests typically involve extracting the soil with a specic solution for a given amount of time. The Mehlich 3 (M3) universal extractant (Mehlich 1984) is suitable for acidic to neutral soils, which include the majority of soil types used for cropping in our study area, northwestern Spain. Soil tests for M3 soil P use a colorimetric analysis method to measure the P extracted from the soil (Mehlich 1984; Jones 1998; Zbiral and Nemec 2000; Caridad Cancela 2002). However, increasing soil-test interpretations for a new version of the M3 P test have been provided (Eckert and Watson 1996; Jones 1998; Eralshidi and Mays 2003). This new version uses inductively coupled plasma (ICP) techniques to measure the P extracted from the soil. The new method was adopted rapidly because the ICP instrument can be used to measure amounts of other nutrients in the same soil extract. This notwithstanding, the P value with ICP is not always comparable with the colorimetric P value that has been previously used to set up fertilizer P recommendations. Using the ICP method may result in greater P test values (Eckert and Watson 1996; Eliason, Lamb, and Rehm 2001; Nathan et al. 2002; Mallarino 2003; Pittman et al. 2005; Sikora et al. 2005). The reason is that soil extracts for all P tests have the orthophosphate P form and also small amounts of other P forms (such as other inorganic and simple organic forms). The ICP instrument is thought to measure all P forms in the sample (Mallarino 2003). Instead of using extractants for soil P analysis, chemical sinkbased tests that rely on P sorptiondesorption reactions have been proposed. One test is based on ionexchange resins, either resin beads or plastic membranes (Olsen and Sommers 1982; Olsen, Watanabe, and Bowman 1983). Recent research suggests that anion-exchange resin (AER) may be as effective as commonly used routine P tests based on a chemical extraction at predicting crop response to P fertilization. Therefore, it would be a useful tool for both agronomic and environmental soil P testing. Universal extractants and ion-exchange resins are capable of simultaneously determining a multiple number of elements in a single step. The adoption of cheaper soil-testing methods can lead to more precise fertilizer application, thereby reducing environmental impact of excess levels. However, differences in P contents measured by various techniques may have implications for agronomic and environmental testing. The objectives of this research were (1) to compare P concentrations as extracted by M3, determined both by colorimetry and ICP, and obtained by ion-resin exchange and (2) to differentiate between the effects of two reduced-tillage systems on P accumulation at the soil surface layer.

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Material and Methods

The experimental site was located near Pastoriza, Lugo Province, Galicia, northwestern Spain. A eld trial took place from 2004 to 2006 for testing four fertilizer management conditions (mineral fertilizer as a control and livestock manure applied at rates of 30, 60, and 90 Mg ha1 ), which were factorially combined with two reduced-tillage systems: minimum tillage (MT), consisting of chisel plowing at 60 cm deep followed by direct drilling, and no tillage (NT). The experimental design was a split plot with four replications, given 32 individual plots. Before to our trial, the experimental eld was conventionally tilled. The crop rotation was maize and winter cereal. Mineral fertilizer and livestock manure were applied on the soil surface in March or April (spring) or in October or November (winter). Samples were collected at three different dates. Table 1 lists the cumulative mineral or organic liquid manure on the four different treatments before each of the three sampling dates. Soil was sampled at 05 cm deep, because this depth is more relevant to the potential for erosive movement of nutrients dissolved or suspended in runoff. Samples were air dried, crushed, and sieved with a 2-mm sieve. Soil general properties were determined by routine analysis as follows: texture by the pipette method (Guitian and Carballas 1976) and pH (H2 O) and organic-matter content (wet oxidation) according to procedures described in detail in van Raij et al. (2001). On average, sand, silt, and clay contents were 29.1%, 46.7%, and 24.2%, respectively; gravel content was 39.8%. Table 2 summarizes results for organic-matter content and pH during the three successive sampling dates. Mean organicmatter content showed a trend to increase from the rst to the second sampling date, with values of 64.2 and 69.7 g kg1 respectively, but these differences were not signicantly different (P < 0.01). Minimum organic-matter contents steadily increased with values of 42.56 and 59 g kg1 during the rst, second, and third sampling dates, respectively. Values of pH were rather stable along the successive sampling dates with mean values of 5.7, 5.8, and 5.7. Soil P was extracted with the M3 test (Mehlich 1984), as briey described: 5 cm3 of soil and 50 mL of the extractant solution [0.2 mol L1 acetic acid (CH3 COOH), 0.25 mol L1 ammonium nitrate (NH4 NO3 ), 0.015 mol L1 ammonium uoride (NH4 F), 0.013 mol L1 nitric acid (HNO3 ), and 0.001 mol L1 ethylenediaminetetraacetic acid (EDTA), adjusted at pH 2.5] were taken. The suspensions were placed in polyethylene asks covered with plastic stoppers and shaken by horizontal and circular movements at

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Table 1 Cumulative amounts of mineral or organic fertilizers applied during three successive sampling dates Treatment 1 [mineral (P2 O5 ), kg ha1 ] 300 400 500 Treatment 2 (livestock manure, 30 Mg ha1 ) 120 150 180 Treatment 3 (livestock manure, 60 Mg ha1 ) 180 240 300 Treatment 4 (livestock manure, 90 Mg ha1 ) 270 360 450

Sampling date 1 2 3

Phosphorus Detection Methods Table 2 Summary statistics for organic-matter content and pH during three successive sampling dates Organic-matter content (g dm3 ) Sampling date 1 2 3 Mean 64.2 69.7 68.8 Maximum 80 102 85 Minimum 42 56 59 Mean 5.7 5.8 5.7 pH Maximum 6.1 6.2 6.3

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Minimum 5.2 5.3 5.1

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240 oscillations per minute for 5 min. After ltration, the P content was determined both by colorimetry and ICP. Phosphorus analysis by ion-exchange resin was performed as outlined by van Raij et al. (2001): 2.5 cm3 of soil and 25 mL water were taken and the soil suspension was shaken for 15 mins. Then 2.5 cm3 resin were added, and the suspension of soil plus resin was shaken for 16 h. The suspension is sieved with a 0.4-mm sieve. Resin was transferred in polyethylene asks using 50 mL ammonium chloride (NH4 Cl) 0.8 mol L1 solution in hydrochloric acid (HCl) 0.2 mol L1 . Soil P desorbed by the resin and extracted by M3 was determined by the ascorbic acid ammonium molybdate colorimetric method based on the Murphy and Riley (1962) method following procedures described in van Raij et al. (2001). For the ICP P determination, M3 extracts were analyzed with a Jobin Yvon JY 50-P ICP emission spectrophotometer (Jobin Yvon Emission Instruments, Longjumeau, France) (van Raij et al. 2001).

Results and Discussion

Comparison of Phosphorus Amounts Extracted by Soil Tests Averaged over all three sampling data, the amounts of P measured by AER, M3-COL, and M3-ICP were 182.0, 267.5, and 296.8 mg dm3 , respectively. The range of measured P values was from 62 to 364 mg dm3 for resin, from 49 to 431 mg dm3 for M3 colorimetric method, and from 45.6 to 698.8 mg dm3 for M3-ICP. The relationship between ICP and colorimetric P determination in M3 extracts of the 96 soil samples in this study is shown in Figure 1. During the rst sampling date, when M3 extractable P was relatively low, P concentrations determined by ICP were generally less than colorimetric values, except for one out of 32 samples. However, during the second and the third sampling dates, P concentrations determined by ICP were greater than colorimetric values, except for one out of 64 samples. A signicant correlation was found between the two methods of M3 determination (R2 = 0.898) by linear regression. However, the intercepts of the line shown in Figure 1 equals 90.3 mg dm3 , meaning that when the extractable P in the soil is low a single linear model would predict even greater colorimetric values than those experimentally measured. Therefore, models different from the lineal model have been tested. A quadratic relationship exhibits a greater determination coefcient (R2 = 0.939). The tted equation was ICP P = 0.0008 (COL P) 2 + 1.131 (COL P) + 17.086. Also, a twostraight-line model with a break at 300 mg dm3 was superior to a single lineal model

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500 Colorimetric-P (mg.dm3) 400 300 200 100 0 0

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y = 0.596x + 90.3 R2 = 0.898

100

200

300

400

500

600

700

ICP-P (mg.dm3)

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Figure 1. Relationship between P contents measured by extraction with Mehlich 3 and two different determination methods, based on inductively coupled plasma emission spectroscopy (ICP-P) and colorimery (colorimetric P, COL-P) (color gure available online).

(Figure 1). We are not aware of any published study that has specically evaluated this break in the linear relationship between ICP P and COL P extracted by M3 solution. In our study, colorimetric P was on average 90% of the P determined by ICP, but as previously stated, values in the lower range for COL P were greater than their corresponding ICP P values. Mallarino (2003) and Pittman et al. (2005) found M3 results to be greater when P was measured with ICP as compared to a colorimetric approach for low ranges of extractable P, where P fertilizer would be recommended, that is, values generally less than 60 mg kg1 . Sikora et al. (2005) found the slopes of correlations between ICP and colorimetric P determination in M3 extracts were much closer to 1 (1.02 and 0.968 for two data sets) as compared to the results of Pittman et al. (2005) and Mallarino (2003). However, when M3-extractable P was low and P fertilizer would be recommended, P concentrations determined by ICP were greater than colorimetric values more frequently than the reverse. Jacoby (2005) evaluated 23 elds where manure had been applied either via a manure spreader, grazing cattle, or both on feed yards and dairies. The range in colorimetric P was from 31 to 472 mg kg1 and ICP P ranged from 40 to 772 mg kg1 . Determination coefcient between M3 P by ICP P and the colorimetric method was R2 = 0.76 with a 47 mg kg1 P intercept. The colorimetric P was 55% of the P determined by ICP. The relative difference (ratio) ICP P / COL P was between 0.62 and 1.6. The absolute differences, ICP P minus COL P, were between 37.8 and 60 mg dm3 . Figure 2 shows the relation between the ratio ICP P / COL P and the absolute differences ICP P minus COL P versus colorimetric P. A positive and signicant relationship (P < 0.01) was found in both cases. In the range of M3-COL P < 80 mg kg1 , Mallarino (2003) found a exponential negative relationship between the ratio ICP P / COL P and colorimetric P; at low P contents, relative differences between both P determination methods could be as high as 3.5. Therefore, soil P content plays a role in the relationship between the two determination methods but does not explain differences between them. Weak positive relationships were found between the ratio ICP P / COL P versus organic carbon (R2 = 0.165) and also between the absolute difference ICP P minus COL

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1.8 1.6 M3-ICP/M3-COL P ratio 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0 0 100 200 300 400 500 y = 0.7181e0.0015x R2 = 0.583

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M3-COL P (mg dm3)

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(M3-ICP) - (M3-COL) P(mg dm3)

80 60 40 20 0
20 40 60

y = 0.1591x 33.991 R2 = 0.614

100

200

300

400

500

M3-COL P (mg dm3)

Figure 2. Relationship between soil P measured by extraction with Mehlich 3 and with determination based on colorimetry and the relative (M3-COL/M3-ICP) or absolute (M3-COL minus M3-ICP) differences from soil P determined by ICP and colorimetry.

P and organic carbon (R2 = 0.141). This result suggests that organic carbon changes along time are responsible for the differences between ICP and colorimetric procedure. A possible explanation for the greater values obtained by colorimetric over ICP in the rst sampling date includes the presence of organic by-products. Such substances may absorb light at the specic wavelength corresponding to the colored molybdic complex, thereby increasing the colorimetric P reading. On the other hand, several authors indicate that the commonly held belief is that ICP would measure greater P concentrations than colorimetric procedures because the high-temperature environment of the plasma would allow the measurement of organic P compounds or other soluble P complexes that would not be measured colorimetrically (Mallarino 2003; Pittman et al. 2005). It is likely that organic P species plays a role in the discrepancy between ICP and colorimetric P analyses when ICP results are greater; other as yet unidentied factors may also play roles. However, currently there is little insight into what may cause colorimetric P analyses to be greater than ICP results for water extracts of organic by-product materials. Phosphorus content extracted by AER when compared to M3-COL or M3-ICP reported a signicant but much less predictable relationship with R2 values of 0.27 and

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Resin-P (mg dm3)

300 200 100 0 0 100 200 300 ICP-P 1st sampling 400 500 600 700 800 (mg dm3) 3rd sampling

2nd sampling

Figure 3. Relationship between resin P and P extracted by Mehlich 3 with determination by ICP (color gure available online).

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0.21 (n = 96), respectively. The regression equations were as follows: AER P = 0.362 M3COL+ 84.97 and AER P = 0.204 M3-ICP + 121.4. Figure 3 shows the relationship between AER P versus P extracted by M3-ICP for the three successive sampling dates in this study. The relationship between both variables is stronger when considering each of the sampling date. The respective regression equations and coefcients of determination are as follows: AER P = 0.791 M3-ICP + 17.08 (R2 = 0.626), AER P = 0.320 M3-ICP + 110.9 (R2 = 0.265), and AER P = 0.419 M3ICP + 0.064 for the rst, second, and third sampling dates, respectively. From the slope and intercept of the regression lines, it is apparent that the ratio AER P / ICP P decreases as the fertilizer addition and thus the soil surface layer P content increases. Effect of Tillage and Management Systems Table 3 lists mean P concentrations as extracted by resin and determined by the colorimetric method (AER P) and as extracted by M3 solution and determined by ICP and colorimetry. The P levels were in excess of levels required for maximum crop yield. Manure applications to these soils should be restricted because of risk of loss of P to surface waters. Mineral fertilizer in the control treatment and liquid manure applications in the three remaining treatments signicantly increased soil-test P levels, and these effects were detectable with all the analytical methods. However, the differences in the amounts of P between the second and the third sampling dates were not detected by the resin extraction method. The widest differences between mean P contents during the successive sampling dates were those extracted with M3 and determined by ICP. Differences between liquid organic fertilizer treatments were not signicantly different along the successive sampling dates. Because of considerable spatial variability in the experimental eld, there was considerable scatter between individual plots, which masked clear evidence of P differences between fertilizer treatments. Phosphorus contents extracted by M3 and determined both by ICP and colorimetry from the 0- to 5-cm surface layer were signicantly greater (P < 0.01) in the no-tillage than in the minimum-tillage treatments for all the different mineral or organic manure doses and during all three study sampling dates. In general, AER P concentrations were also greater in NT than in MT treatments, but there were some exceptions. Thus, no tillage

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Table 3 Mean soil P contents resulting from three different methods for two management systems and four fertilizer treatments (mg dm3 ) Minimum tillage Treatment First date 1 2 3 4 Mean Second date 1 2 3 4 Mean Third date 1 2 3 4 Mean Resin 153.3 125.0 147.0 158.0 145.8 239.0 153.0 194.5 201.8 197.1 149.5 134.5 182.0 170.8 159.2 M3-COL 203.5 172.5 200.5 203.3 195.0 219.8 238.8 261.5 244.0 241.0 318.3 318.3 285.5 328.0 312.5 M3-ICP 165.4 160.2 170.7 186.7 170.7 243.0 254.2 280.3 258.1 258.9 384.5 350.7 434.6 406.5 394.1 Resin 158.8 192.0 200.5 216.8 192.0 204.5 211.5 203.8 178.5 199.6 172.8 194.8 226.8 198.8 198.3 No tillage M3-COL 213.8 240.8 240.3 253.3 237.1 260.0 284.3 256.8 243.0 261.0 334.5 326.7 334.5 368.5 341.1 M3-ICP 178.6 222.1 209.8 240.6 212.8 286.5 332.5 268.5 260.6 287.0 415.4 474.7 464.6 474.7 457.3

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was most efcient in accumulation of mineral fertilizer and manure at the uppermost soil surface layer, whereas minimum tillage facilitated incorporation into deeper soil layers than no tillage.

Conclusions
This study conrms that M3 soil tests are an effective means for evaluating the P status of agricultural soils with excess levels and high inputs of organic or mineral fertilizers. Mehlich 3 P determined by ICP was greater than colorimetric P at high values of extractable P, whereas the reverse was true at low values of extractable P. Both determination methods were capable of evaluating excessive soil surface layer P due to mineral and liquid organic fertilization. The absolute and the relative differences between the M3-ICP and M3-COL tests were correlated with the soil P level in the M3-COL P range from 49 to 431 mg dm3 . Phosphorus contents extracted by M3 and determined both by ICP and colorimetry from the 0- to 5-cm surface layer were signicantly greater (P < 0.01) in the no-tillage than in the minimum-tillage treatments.

Acknowledgments
This study was supported by Spanish Education and Science Ministry (MEC) under Project AGL2003-09284-C02-01.

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