6 Automated Instruments for the Determination of Total Carbon, Hydrogen, Nitrogen, Sulfur, and Oxygen Keith A. Smith The University of Edinburgh, Edinburgh, Scotland M. Ali Tabatabai Iowa State University, Ames, Iowa, U.S.A. I. INTRODUCTION The use of automated laboratory instruments for the determination of total carbon, C, and nitrogen, N, by dry combustion has become well established in recent years. This has become possible through the development of simple and rapid combustion procedures and by coupling them with modern high- sensitivity gas analysis systems in single integrated instruments. As instruments have become more sophisticated, the capability of simultaneous measurement of sulfur, S, and/or hydrogen, H, along with the C and N determinations, has been added to some commercially available systems. Yet another optional add-on is the capacity to determine oxygen, O, in the same samples. Prior to such developments, the techniques available for dry combustion systems were complicated and time-consuming, and manual wet oxidation methods were generally preferred for analysis of soils, plants, and other environmental materials. However, the balance now lies in the opposite direction. This chapter describes the principles on which automated dry combustion instruments and automated analyzers dedicated to the TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. determination of dissolved organic C are based. It examines the particular features available with a selection of different commercial systems and reviews recent information from the scientific literature concerning the application of these instruments to the analysis of soils, plants, waters, and other environmental materials. This chapter is an updated and extended version of that on dry combustion analyzers in the previous edition (Tabatabai and Bremner, 1991), to which reference should be made for information on evaluations of earlier commercial instruments for soil analysis. II. DRY COMBUSTION SYSTEMS A. Dumas Systems for C, H, N Analysis The various automated dry combustion systems for the analysis of C, H, and N in soils, plant materials, and other agricultural and environmental samples generally have several important features in common. They all employ high-temperature combustion (oxidation) of the sample, the determination of C and H as CO 2 and H 2 O vapor, respectively, and the determination of N as N 2 . The historical developments, the principles involved, and a description of the mode of operation of this type of analytical system can be found in Pella (1990a,b), and further material relating particularly to C analysis is also contained in Nelson and Sommers (1996). The essential components of a CHN analyzer operating on the dry combustion principle are: 1. An automated sample introduction system 2. A high-temperature oxidation zone in which the sample is combusted to CO 2 ,H 2 O vapor, N 2 and NO x , and other gases 3. A carrier gas system, to sweep the products of combustion through the remaining stages of the analyzer 4. A gas purification/reduction train in which the oxides of N produced in the combustion are reduced to N 2 , and unwanted gases (e.g., halogen compounds and S oxides) removed 5. A gas separation stage, e.g., a gas chromatographic system or a series of selective traps for individual gases 6. A single, or multiple, gas detector(s) 7. A signal recording/readout system The major developments between the first appearance of commercial C, N, CN, and CHN analyzers and the present generation of instruments are in 236 Smith and Tabatabai TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. the incorporated computer control and data analysis systems, and in the employment of new types of gas detectors in some products. The essential oxidation, reduction, and gas separation operations carried out within the analyzers are usually much as they were in earlier-generation instruments, as can be seen in the next section. B. Examples of Commercial Instrument Systems The intention of this section is to present the main features and operating principles of some of the available commercial CHN analyzer systems in a generic way, and to give the reader an overall understanding of the capabilities of this type of analyzer and of its potential application to future analytical requirements. It must be pointed out, however, that instrument models come and go from the market, as new developments occur and as companies merge (Table 1). Within any one product range, modifications are frequently made, and new accessories become available from time to time. Thus any precise details presented on current instrumentation will inevitably become out of date in due course. A potential user should always check with the supplier to confirm the detailed specifications of any particular instrument and to identify new features that may have become available. 1. CE Instruments Systems The CE Instruments range (Table 1) includes the EA 1110 Elemental Analyzer, applicable to C, H, and N analysis, and the NA 2500 N and CN systems, applicable to N only, and to C and N, respectively. These instruments have many common features; they are also direct descendants of well-known models such as the NA 1500 based on similar principles (see Table 1), many of which are still in use and continuing to yield valuable analytical data (see Sec. IV). The typical CE sample introduction system consists of an autosampler containing one or more sample carousels, each of which may hold up to 50 samples. Solid samples (e.g., dried soils or plant materials), ranging in weight from a few mg to 500 mg, are contained in crimped tin capsules. Standards are introduced in the same manner. For normal operation, at the start of the analytical cycle the carousel is rotated to the inlet position, allowing the first sample capsule to drop into the oxidation zone. On the completion of the analysis, which is normally controlled by a computer program, the carousel moves to the next position, the next sample is introduced, and the whole cycle is repeated until the preset number of analyses has been completed. Although the system is primarily intended for Automated Instruments 237 TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. Table 1 Some Current/Recent Automated Dry Combustion CNS Analyzer Systems, Related Earlier Models, Operating Principles, and Applications Manufacturer Address/website Model(s) Related earlier makes/models Operating principle/ detection system(s) Typical applications Antek Instruments Inc. 300 Bammel Westfield Rd, Houston, Texas 77090, USA www.antekhou.com 9000 Series N, S, NS Analyzers Combustion at selected temperatures up to 1100  C. N detection by chemiluminescence, S by fluorescence method N and/or S in soils, waters, feces & urine, foods & beverages, plant materials, pesticides CE Instruments (formerly Carlo Erba, then Fisons, now part of ThermoQuest Corp.) Strada Rivoltana 20090 Rodano, Milan, Italy www.ceinstruments.it EA-1110 Elemental Analyser; NA-2100 N & Protein Analyser; NA-2500 Carlo Erba NA-1400 & NA-1500; Fisons NA-1500; Carlo Erba 1102, 1104, 1106, 1108; CE NA-2000 Tin capsules/flash combustion with O 2 / reduction on hot Cu. O determination by pyrolysis. GC separation. Injector for liquid samples. Detection by TCD or MS C, H, N, (O) or C, H, N, S (O) or N only in soils, sediments, waste waters, liquid fertilizers, plant materials LECO Corp. 3000 Lakeview Ave., St. Joseph, Michigan 49085-2396, USA www.leco.com CN-2000, CHN- 2000, CNS-2000, FP-2000; 144 Series; 400 Series 70-sec C Analyzer; CR-412; CHN-600, -800 & -1000; FP-228N; IR-12; DC-12; CS-46; TN-15; FP-228N; TC-36; UO 14SP; Sulfur Analyzer; SC-32; SC-132 Combustion with accelerator & O 2 / reduction on hot Cu. O determination by adding external pyrolysis furnace. Detection by IR for C, H, S, O; by TCD for N Various combinations of C, N, and/or S in soils, plant tissue, fertilizers TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. Exeter Analytical Inc. 7 Doris Drive, Unit 1, N. Chelmsford, MA 01863, USA www.eail.com CE-440 Combustion in tin or Al cups in pure O 2 / reduction on hot Cu. O by pyrolysis. S determined as SO 2 . Detection by TCD C, H, N (and O/S) in agric. materials, foodstuffs, air and water filters, oil residues, etc. PDZ Europa Ltd. Hill St, Elsworth, Sandbach, Cheshire CW11 3JE, UK www.europa-uk.com ANCA-GSL and ANCA-SL, with 20-20 or GEO-Series mass spectrometer Europa Scientific SL CN Analyzer; Europa ANCA-NT Tin capsules/flash com- bustion with O 2 /reduc- tion on hot Cu. O by pyrolysis. GC separa- tion. Detection by isotope ratio MS, giving total element concn. & stable isotope composition C, H, N, S, (O) in soils, plant material, plus 13 C, 2 H, 15 N, 34 S, 18 O content Perkin-Elmer Analytical Instruments (including former Coleman Instruments) 761 Main Avenue Norwalk, Connecticut 06859-0001, USA www.perkinelmer.com 2400 Series II CHNS/O Analyzer; 2410 Series II Nitrogen Analyzer. Coleman Models 29 & 29A Combustion with O 2 in Pt boat/reduction on hot Cu. Detection by TCD C, H, N, S (O) in soils, sediments, fertilizers, plant materials Skalar Analytical BV PO Box 3237, 4800 DE Breda, The Netherlands; www.skalar.com Primacs SC, Primacs SN Dual oven system: total C combustion with O 2 , catalyst at 950–1100  C; inorg C reaction with acid at 20–150  C; TOC by difference. Detection by nondispersive IR Total org C, total N in sludges, sediments, soils TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. the analysis of solid samples, liquid samples may be introduced in special sealed tin capsules, or by fully automated syringe sampling from small vials, followed by injection into the combustion chamber. The oxidation zone is composed of a quartz tube packed with an oxidizing agent (CrO 2 ), within an induction furnace maintained at 1000 Æ50  C. The sample combustion process is aided by a pulse of O 2 gas, introduced at the same time as the sample. The strongly exothermic combustion of the tin capsule and its contents raises the temperature to ca. 1700–1800  C, ensuring the complete combustion of organic compounds present to CO 2 , water vapor, N 2 , and oxides of N (NO and NO 2 ). The gaseous products are swept in a stream of carrier gas (helium) through the CrO 2 packing, where any pyrolysis products such as hydrocarbons are completely oxidized to CO 2 and H 2 O. Carbon monoxide is commonly oxidized to CO 2 by passage over copper oxide as a catalyst. The gas stream then passes through a purification zone, over reagents such as silver vanadate on silver wool or silver-coated Co 3 O 4 , to remove halogens and sulfur oxides. The gases then pass through a reduction column packed with copper granules at 650  C to remove excess O 2 and reduce NO x to N 2 (the Dumas reaction). In instruments configured for C and N determinations, the water vapor is then removed by absorption in a trap filled with a drying agent, e.g., magnesium perchlorate (Fig. 1). The gas stream then passes through a gas chromatographic (GC) column packed Figure 1 Schematic diagram of the CE-Instruments CN analyzer system. 240 Smith and Tabatabai TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. with a material such as Porapak Q, where N 2 and CO 2 are separated, and then through a thermal conductivity detector (TCD). The output signal of the detector is converted to give a final result in terms of units of C and N in the original sample. In CE Instruments analyzers configured to include the determination of H, as well as the C and N content of the sample, the water vapor is not trapped out but instead passes to the GC column. Here it is separated from the N 2 and CO 2 and measured by the TCD. 2. The Leco Range Another widely used range of instruments is that produced by Leco (Laboratory Equipment Corporation, St. Joseph, MI, USA) (Table 1). Several early applications in soil analysis featured Leco models that are now superseded but may still be in use in some laboratories (see Tabatabai and Bremner, 1991, and Table 1). Second-generation automated elemental analyzers such as the Leco CHN-600 and CHN-800 Series have been used widely in soil and environmental investigations through the 1990s (see Sec. VI). These particular models permit the simultaneous determination of total C, H, and N in solid or liquid organic materials. The CHN-600 Analyzer, shown schematically in Fig. 2, is a macrosample instrument capable of analyzing samples ranging in weight from 100 to 200 mg and is thus well suited to soil and plant samples, whereas the CHN-800 Analyzer is a microsample instrument capable of analyzing samples ranging in weight from 3 to 15 mg and is aimed more at the analysis of organic compounds. Figure 2 Schematic diagram of the Leco CHN-600 analyzer system. Automated Instruments 241 TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. In total N analysis by these instruments, a weighed sample is encapsulated in tin or copper and dropped into a reusable ceramic crucible centered in the primary hot zone of a U-shaped combustion tube located in a resistance furnace, and the sample is burned in O 2 at 950  C. The potential combustion products are CO 2 , water vapor, oxides of N or N 2 , and oxides of S. Oxides of S are removed with a reagent in the secondary hot zone to prevent the formation of H 2 SO 4 . The secondary hot zone also ensures the complete combustion of all volatile gases. The remaining products of combustion (CO 2 ,H 2 O, O 2 ,N 2 , and NO X ) are collected and mixed thoroughly in a glass tube under a sliding PVC piston. The CO 2 and H 2 O levels are constantly monitored during combustion by two independent selective IR detectors, and when they drop to predetermined levels, combustion is terminated. At this stage, an aliquot of the combustion products is removed automatically and carried by H 2 gas through a reagent train containing hot Cu for the reduction of NO X to N 2 , ascarite for the removal of CO 2 , and anhydrone for the removal of H 2 O. The N 2 thus obtained is then collected and measured by a thermal conductivity detector. The measurements are weight-compensated and displayed digitally as percent C, H, and N. Total analysis time for all three elements is 4–5 min with the CHN-600 Analyzer and less than 2.5 min with the CHN-800 Analyzer. The successors to the CHN-600 have been the CHN-1000, and most recently the 2000 Series (Table 1). These latter instruments are micro- computer-based, use nondispersive infrared detection systems, and are designed to measure the C, H, and N content in a wide variety of organic compounds and environmental materials. The CN-2000/CNS-2000 models can accommodate samples weighing up to 2 g, and these are introduced in sample boats into a horizontal combustion system. This makes it possible to remove the sample ash, along with the boat, after each analysis, thus reducing the problem of ash accumulation. However, the standard operating procedures for CN, or CNS, analysis in soils and plant materials recommend sample weights of 0.15–0.8 g, depending on the sample. For soils, the furnace temperature is set at 1350–1450  C, while for CN analysis of plant material, 1050  C is sufficient. Use of the lower temperature extends the life of the combustion tube. The procedure for noncarbonate C (total organic C) in soils involves manual treatment of the sample with HCl in nickel-lined combustion boats, until all reaction with carbonate has ceased, followed by evaporation on a hotplate, before introduction into the analyzer. The FP-2000 variant can determine N in 1–2 g soil samples in reusable ceramic crucibles, in plant samples up to 4 g, and in 0.25 g samples of fertilizer materials. 242 Smith and Tabatabai TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. 3. Other Systems The Perkin-Elmer Corp. have developed a model CHN-2400 analyzer that simultaneously measures C, H, and N using the principle employed in the traditional Pregl and Dumas procedures (Nelson and Sommers, 1996). A sample contained in a platinum boat is oxidized with O 2 at about 1000 o Cin a combustion tube in the absence of carrier gas (He) flow. After combustion, He flow is initiated, and the CO 2 ,H 2 O, and N 2 gases produced are passed over CuO to convert CO to CO 2 and silver mesh (silver vanadate on silver wool) to remove S and halogen gases. The gases then pass into a tube packed with copper granules between end plugs of silver wool and maintained at 650 o C to reduce the N oxides to N 2 . The gases are brought to constant pressure and volume in a gas-mixing chamber and then allowed to expand into the analyzer portion of the instrument. The analyzer consists of three thermal conductivity detectors (TC) connected in series and separated by two traps. The sequence of the TC detectors and traps enabling quantification of H, C, and N is as follows (Nelson and Sommers, 1996): (1) TC detector 1 (output equals total gas composition), (2) Mg(ClO 4 ) 2 traps to remove H 2 O, (3) TC detector 2 (decrease in output from detector 1 is proportional to H content), (4) soda asbestos plus Mg(ClO 4 ) 2 trap to remove CO 2 , (5) TC detector 3 (decrease in output from detector 2 is proportional to C content), and (6) the remaining gases in the sample are N 2 . The RoboPrep-CN Analyzer marketed by Europa Scientific Ltd., Crewe, England (now PDZ Europa) and its successor models, the ANCA- GSL and GL, are essentially CN analyzers of the CE (Carlo Erba) type. Their special feature is that they can be linked via a capillary interface to a mass spectrometer for the 15 N analysis of the N 2 produced by Dumas combustion, and 13 C analysis of the CO 2 (Fig. 3). These instruments also give total C and N contents of the samples and can be used routinely for such measurements even where isotopic data are not required. The ANCA- GSL system can be fitted with either a 66- or a 130-position autosampler, with alternative versions available for small or large samples. C. Modifications for S and O Analysis 1. Sulfur Some automated analyzers are available with either a CHN or a CHNS capability. In the latter versions, generally the SO 2 formed by reduction of the SO 3 in the gas stream leaving the oxidation furnace is not trapped out but instead is quantitatively determined, along with the gaseous forms of the other target elements: H 2 O vapor, CO 2 , and N 2 . In the CE Instruments Automated Instruments 243 TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. EA-1110 instrument, for example, the SO 2 is separated from these other gases chromatographically and determined by the TCD. Alternatively, analysis of trace sulfur contents down to a few mg kg À1 in the original sample can be performed with this particular analyzer by connecting it to an optional electron capture detector (ECD), which has a much greater sensitivity than the TCD to SO 2 . There seems no reason in principle why a similar adaptation could not be made with various other analyzer systems. In the Leco family of instruments a different approach is used. The sample is combusted in a stream of oxygen, and the SO 2 produced, like CO 2 and H 2 O, is determined by IR absorption. For example, in the Leco model SC-132 automated total S analyzer (an instrument dedicated to S analysis), the sample is mixed with combustion accelerators in a ceramic boat and combusted in a resistance furnace at 1370 o CinanO 2 atmosphere. The SO 2 thus produced is passed through an infrared (IR) cell which is used as both a reference and a measurement chamber. It detects total S, as SO 2 , Figure 3 Schematic diagram of an automatic CN analyzer interfaced with an isotope ratio mass spectrometer. 244 Smith and Tabatabai TM Copyright n 2004 by Marcel Dekker, Inc. All Rights Reserved. [...]... 0.8 568 0.9493 0.8587 2 .67 2 .67 2. 56 2.57 2.55 2 .68 0.175 0.175 0. 160 0. 169 0. 167 0.1 76 0.033 0.034 0.030 0.033 0.032 0.035 0.2358 0.2214 0.2183 0.2341 0.2330 0.2 267 39.27 39.37 39.21 38.91 39. 16 38. 96 3 .67 3 .69 3.7 3 .67 3.71 3 .68 0.300 0.289 0.289 0.289 0.2 96 0.2 86 "  Std dev CV (%) 2 .62 0. 06 2.18 0.170 0.0 06 3.35 0.033 0.002 4.79 "  Std dev CV (%) 39.15 0. 16 0.42 3 .69 0.01 0.40 0.292 0.005 1 .67 Source:... Table 6 Effect of Sample Mesh Size on Total Carbon Analysis of Soils by Leco 70-Second Carbon Analyzer Max particle size Soil Lindley Sharpsburg Grundy Glencoe Mesh mm Range Mean SD 10 40 100 10 40 100 300 10 40 100 300 10 40 100 2000 420 150 2000 420 150 50 2000 420 150 50 2000 420 150 1. 16 1.41 1.31–1.38 1.35–1.38 2.30–2.45 2.28–2.34 2.28–2.32 2. 26 2.29 2 .60 –2 .69 2 .63 –2.75 2 .62 –2 .69 2 .60 –2 .68 5 .65 6. 05... 2 .60 –2 .68 5 .65 6. 05 5 .68 –5. 86 5.77–5.84 1.29 1.33 1. 36 2. 36 2.31 2.29 2.28 2 .63 2 .68 2 .66 2 .64 5.81 5.78 5.80 0.11 0.03 0.01 0. 06 0.02 0.02 0.01 0.04 0.04 0.03 0.03 0.15 0.07 0.03 a Five analyses SD, standard deviation Source: Tabatabai and Bremner (1970) TM Copyright n 2004 by Marcel Dekker, Inc All Rights Reserved Total C content (%)a 254 Smith and Tabatabai soil mineral N (nitrate, nitrite and nonfixed ammonium)... All Rights Reserved  268 Smith and Tabatabai Table 9 Comparison of a Shimadzu TOC-500 (HTC-type) Analyzer with a Dohrmann DC-80 (Wet Oxidation Type) Analyzer for the Measurement of DOC in Amazonian Freshwater Samples DOC by Shimadzu DOC-500 (mg LÀ1 Æ SD) Sample source Obidos Rio Ica Rio Negro Salmon Pond Portage Bay KPA standarda 2.39 Æ 0. 06 2 .62 Æ 0.07 8. 06 Æ 0.31 2.73 Æ 0. 16 2 .67 Æ 0.04 10.11 Æ 0.20... furnace and a wet-oxidation method indicated that the CHN -6 0 0 gave the most precise results and allowed an operator to perform 90 to 100 analyses in 8 h (Sheldrick, 19 86) Although work by McGeehan and Naylor (1988) showed that the CHN -6 0 0 gave organic C values that were greater for four samples out of five than those obtained by the wet oxidation method of Walkley and Black (1934), this is not surprising and. .. 20 soil samples determined with the Leco CHN -6 0 0 analyzer, versus total N content by Kjeldahl method (From Yeomans and Bremner, 1991.) determination of total C, total N, and 15N in soils and plant materials They showed that this system gave results comparable to those obtained with conventional manual methods Coefficients of variation for C were 1.03–1.41% for plant materials, and 1. 46 1 .62 % for soils;... total C and S in soils, and reasonably acceptable values for total N, which were slightly lower than, but proportional to, those by Kjeldahl analysis Table 5 and Fig 6 show manufacturer’s data for total N analysis of soils, TM Copyright n 2004 by Marcel Dekker, Inc All Rights Reserved Automated Instruments 251 Figure 6 Total N content of soil, plant materials, foods and feedstuffs determined by CE NA-2100... (various); 0.05–50,000 ppm C TOC-4100, -5 000A & 5050A; TOCN-4100; TN-4100 High-temp combustion at 68 0 C Detection of C by NDIR, of N by chemiluminescence Optional nonautomated solid sample module TOC, or C & N, in waters; on-line analysis of aqueous waste streams, cooling waters, river and lake water Formacs HT, Formacs TN, Formacs LT High-temp combustion (Model HT); low-temp UV/persulfate digestion... TOC and/ or total N in drinking, ground, surface, sea waters SGE International Pty Ltd Shimadzu Corp and Shimadzu Scientific Instruments Skalar Analytical BV TM Copyright n 2004 by Marcel Dekker, Inc All Rights Reserved Smith and Tabatabai Manufacturer PO Box 4295 76, Cincinnati, Ohio 4524 2-9 5 76, USA www.tekmar.com www.dohrmann.com Apollo 9000 Tekmar-Dohrmann Co PO Box 4295 76 Cincinnati, Ohio 4524 2-9 5 76, ... CHN -6 0 0 analyzer and the CR-12 carbon analyzer, was satisfactory for routine analysis of soils for total organic C content Their comparison of C measurements on 20 soils made with these two instruments, with those obtained by the Allison (1 960 ) wet oxidation method, is shown in Fig 4 The sample combustion in the CHN -6 0 0 was carried out at 950 C, a much lower temperature than that used in the CR-12 . 144 Series; 400 Series 70-sec C Analyzer; CR-412; CHN -6 0 0, -8 00 & -1 000; FP-228N; IR-12; DC-12; CS- 46; TN-15; FP-228N; TC- 36; UO 14SP; Sulfur Analyzer; SC-32; SC-132 Combustion with accelerator. 2.55 0. 167 0.032 0.2330 39. 16 3.71 0.2 96 0.8587 2 .68 0.1 76 0.035 0.2 267 38. 96 3 .68 0.2 86 "  2 .62 0.170 0.033 "  39.15 3 .69 0.292 Std. dev. 0. 06 0.0 06 0.002 Std. dev. 0. 16 0.01 0.005 CV. 2 .68 0.04 100 150 2 .62 –2 .69 2 .66 0.03 300 50 2 .60 –2 .68 2 .64 0.03 Glencoe 10 2000 5 .65 6. 05 5.81 0.15 40 420 5 .68 –5. 86 5.78 0.07 100 150 5.77–5.84 5.80 0.03 a Five analyses. SD, standard deviation. Source: