a p p l i c at i o n N o t e

Atomic Absorption

Author
Praveen Sarojam, Ph.D.
PerkinElmer
Global Application Center
Mumbai, India

Analysis of Micronutrients Introduction

Soil is used in agriculture, where it serves as the primary
in Soil by Using AA 800 nutrient base for plants. Soil material is a critical
component in the mining and construction industries.
Atomic Absorption Soil serves as a foundation for most construction projects.

Spectrophotometer Soil resources are critical to the environment, as well

as to food and fiber production. Waste management
often has a soil component. Land degradation is a
human-induced or natural process which impairs the
capacity of land to function. Soils are the critical
component in land degradation when it involves acidification, contamination etc. Soil contamination
at low levels is often within soil capacity to treat and assimilate. Many waste treatment processes rely
on this treatment capacity. Exceeding treatment capacity can damage soil biota and limit soil func-
tion. Derelict soils occur where industrial contamination or other development activity damages the
soil to such a degree that the land cannot be used safely or productively. The analysis of soils is an
excellent measure of soil fertility. It is a very inexpensive way of maintaining good plant health and
 maximum crop productivity. The standard soil test A PerkinElmer Multiwave™ 3000 microwave oven was used
provides the status of phosphorous (P), potassium (K), for the microwave-assisted digestion of soil samples. This is
calcium (Ca), magnesium (Mg), pH, cation exchange an industrial-type oven which can be equipped with various
capacity, lime requirement index, and base saturation. accessories to optimize the sample digestion. In this case,
Additional tests are also available for iron (Fe), zinc (Zn), the samples were digested in the Rotor 8XF100 comprising
manganese (Mn), soluble salts, and nitrates. Soil fertility eight 100 mL high pressure vessels made of PTFE-TFM in
fluctuates throughout the growing season each year. The their respective protective ceramic jackets. TFM is chemically
quantity and availability of mineral nutrients are altered by modified PTFE that has enhanced mechanical properties at
the addition of fertilizers, manure, compost, mulch, and lime high temperatures compare to conventional PTFE. This vessel
or sulfur, in addition to leaching. Furthermore, large quantities has a “working” pressure of 60 bar (870 psi) and can operate
of mineral nutrients are removed from soils as a result of at temperatures up to 260 ˚C. A Pressure/Temperature (P/T)
plant growth and development, and the harvesting of crops. Sensor Accessory was also used for this work. The P/T sensor
The analysis of soils will determine the current fertility status. simultaneously measures temperature and pressure for one
It also provides the necessary information needed to main- vessel. All vessels’ temperatures were monitored with the
tain the optimum fertility year after year. IR Temperature Sensor Accessory. This device gives thermal
(over temperature) protection to the reactions in all of the
In the present work we compare the performance of vessels by measuring the temperature remotely at the bottom
Mehlich-I extraction with that of microwave digestion for surface of each vessel during the digestion process.
the determination of several micronutrients in soil samples.

Standards, chemicals and certified reference

Experimental material
The measurements were performed using the PerkinElmer® PerkinElmer NIST® traceable calibration standards in acid
AAnalyst™ 800 atomic absorption spectrophotometer for atomic spectroscopy were used as the stock standards
(PerkinElmer, Inc., Shelton, CT, USA) (See Figure 1) for preparing working standards. All the working standards
equipped with WinLab32™ for AA Version 6.5 software, were prepared daily with ASTM® type I water acidified in
which features all the tools needed to analyze samples, Suprapur® nitric acid (Merck®, Germany) in polypropylene
report and archive data and ensure regulatory compliance. vials (Sarstedt®) on volume-by-volume dilution. Micropipettes
PerkinElmer’s high-efficiency double-beam optical system with disposable tips (Eppendorf®, Germany) were used for
and solid-state detector provide outstanding signal-to-noise pippetting solutions. Certified Reference Standard for trace
ratios and Deuterium background correction eliminates most metals in river sediment (Lot # 0800230) and soil solution
interferences. A PerkinElmer corrosion-resistant nebulizer, (Lot # 0733733) from High Purity Standards were used for
which can be used for solutions containing hydrofluoric validating the developed method. Multi element ICP standard
acid, was used for all flame absorption measurements. for trace metal ions in 5% HNO3 from Spex Certiprep®,
A single slot air-acetylene 10-cm burner head was used (New Jersey, USA) prepared at midpoint of the calibration
for all air-acetylene experiments. curve was used as quality control check standard. The other
acids used (Sulphuric acid and hydrochloric acid) were also
of Suprapur grade. (Merck®, Germany).

Figure 1. PerkinElmer AAnalyst 800 atomic absorption spectrophotometer.

Figure 2. PerkinElmer Multiwave 3000 microwave digestion system.

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Sample preparation check standard was prepared at the midpoint of calibration
Representative soil samples were collected from three dif- for all elements, from a multi-element ICP grade standard.
ferent locations and were finely ground and then passed The quality control check performed immediately after
through a 20 mesh sieve to obtain very fine particles. 5.0 g calibration and also at the end of analysis was ensured to
of an air-dried, ground and sieved sample was placed in an fall within EPA set limits of 90-110%. The two reference
Erlenmeyer flask and 20 mL of the extracting solution (0.05 N materials from High Purity Standards, river sediment and
HCl + 0.025 N H2SO4) was added to it. Then it was placed in the soil solution were analyzed to validate the method
a magnetic stirrer and the mixture was stirred for 20 minutes. developed. The recoveries from both the solutions were
The resulting solution was filtered through a Whatman® No excellent. Further post-digestion spike recovery studies were
42 filter paper into a 50 mL polypropylene vial and diluted performed for one selected sample for all the nutrients under
to 50 mL with the extracting solution. The analytical reagent study and the results are given in the table. From Table 7
blanks were also prepared and these contained only the one can see the clear difference between the amounts of
acids. The above mentioned procedure is in accordance metal ions recovered by using the two different procedures.
with Mehlich-I extraction. The Mehlich-I extraction gave lower values as expected
which gave indication about the extractable metal ions and
The microwave digestion of soil samples were done in the microwave digestion gave the total metal content in the
accordance with EPA Method 3052. This method is applicable soil samples. The agreement between sample duplicates was
to the microwave assisted acid digestion of siliceous matrices, excellent. If one needs to carry out further characterization
organic matrices and other complex matrices. This method is of soil samples, procedures regarding sample collection and
provided as a rapid multi-element, microwave assisted acid treatment can be modified to suit the recommendations of
digestion prior to analysis protocol so that decisions can be regulatory agencies.
made about the site or material. The goal of this method
is total sample decomposition and, with judicious choice Conclusions
of acid combinations, this is achievable for most matrices.
A method was developed for the sequential determination
Samples of soil were weighed directly into the PTFE-TFM
of essential nutrients in soil by using AAnalyst 800 atomic
digestion vessel liners. Sample weights were approximately
absorption spectrophotometer. Sensitivity and accuracy in
0.1 g. To each sample, 9 mL of concentrated nitric acid
flame AA are directly correlated to the optimization of the
and 3 mL of concentrated hydrofluoric acid were added.
burner position as well as the gas flows. Careful optimization
Some vessels contained only the acids with no sample to
of these parameters, which are different for each element,
act as analytical reagent blanks. The vessels were sealed and
maximizes sensitivity and can significantly reduce or eliminate
placed into the Rotor 8XF100 for the microwave digestion.
interferences. The unique computer-controlled, motorized
After the digestion process, the digestate were transferred
burner system and gas controls allow precise, automated
in to 50 mL polypropylene vials and were made up to 25 mL
adjustments. Since WinLab32 software allows optimized
with ASTM® type I water.
burner position and gas flows to be stored with an element
method, each element in a sequential multi-element run can
Results and discussion be determined using its own optimized parameters. The ability
The Mehlich-I extraction will give an indication about the of PerkinElmer/Anton-Paar Multiwave 3000 microwave
amount of extractable micronutrients in soils. If one needs digestion system to digest the soil samples in accordance
to know the total metal content, complete decomposition of with EPA Method 3052 was demonstrated. The method
samples with a microwave digestion is the preferred choice. detection limits obtained indicates that the reliable analysis
A four point calibration with three standards and one blank of micronutrients in difficult matrices such as soil samples is
was used to calibrate all the micronutrients of interest. The possible with the AAnalyst 800 atomic absorption spectro-
WinLab version 6.5 software has several quality control photometer. The analysis of reference materials and excellent
check options. A minimum correlation coefficient of 0.995 spike recovery results gave further proof to the accuracy of
was assigned in the software for QC check. Quality control the developed method.

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 Table 1. Experimental Conditions of AAnalyst 800.
Element Cu Fe Mn K Na Mg Zn
Wavelength (nm) 324.8 248.3 279.5 766.5 589 285.2 213.9
Slit (nm) 0.7 0.2 0.2 0.7 0.2 0.7 0.7
Mode AA AA AA AA AA AA AA
Flame Air-Ac Air-Ac Air-Ac Air-Ac Air-Ac Air-Ac Air-Ac
Burner Head 10 cm 10 cm 10 cm 10 cm 10 cm 10 cm 10 cm
Calibration Linear through Linear through Linear through Linear through Linear through Non linear Linear through
zero zero zero zero zero through zero zero
Lamp HCL HCL HCL HCL HCL HCL HCL
Lamp current,
mA 15 30 20 12 8 6 10
Standards
(mg/L) 1.0, 2.5, 5.0 1.0, 2.5, 5.0 0.1, 0.25, 0.5 0.1, 0.25, 0.5 0.1, 0.25, 0.5 0.1, 0.25, 0.5 0.1, 0.25, 0.5
Spiked conc.
(mg/L) 2.5 2.5 0.25 0.25 0.25 0.25 0.25
Read Time
(seconds) 3.0 3.0 3.0 3.0 3.0 3.0 3.0
Replicates 3 3 3 3 3 3 3
Air Flow (L/min) 17.0 17.0 17.0 17.0 17.0 17.0 17.0
Acetylene flow
(L/min) 1.5 1.5 1.5 1.5 1.5 1.5 1.3

Table 2. Results of Soil CRM analysis (Lot # 0733733). Table 4. Results of QC Recoveries.
Metal Certified % Recovery % Recovery Metal QC 1 (%) QC 2 (%) QC 1 (%) QC 2 (%)
Value in Solution in Solution Extraction Extraction MDS MDS
(µg/mL) (in HCl and H2SO4) (in HNO3 ) Cu 107.6 100.6 101.1 99.96
Cu 0.30 103.2 104.1 Fe 107.5 106.3 100.1 101.9
Fe 200.0 95.2 96.6 Mg 108 108 100.5 101.5
Mg 70.0 98.1 98.1 Zn 104.3 104.5 103.5 100.8
Zn 1.0 105.7 100.7 Na 100.9 99.96 100.2 98.8
Na 70.0 105.6 98.4 K 99.2 91.4 98.8 97
K 200.0 95.4 95.2 Mn 101 104.9 99.64 96.6
Mn 0.10 103.5 102.7

Table 3. Results of River Sediment CRM Analysis Table 5. Results of Spike Recoveries.
(Lot # 0800230). Metal Spike Recovery (%) Spike Recovery (%)
Metal Certified % Recovery % Recovery for Extracted Sample for Digested Sample
Value in Solution in Solution Cu 108.5 105.1
(µg/mL) (in HCl and H2SO4) (in HNO3 )
Fe 95.8 Conc. too high for spiking
Cu 1.00 102.5 105.8
Mg 97.7 98.8
Fe 1200 102.8 96.4
Zn 102.8 100.4
Mg 70.0 96.9 96.1
Na 106.6 106.5
Zn 15.0 96.0 98.8
K 92.1 96.9
Na 50.0 101.6 93.8
Mn 101.8 97
K 150.0 100.0 98.9
Mn 8.0 96.1 102.8

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 Table 6. Method Detection Limits (MDLs).
Metal MDL (mg/kg)
Cu 0.08
Fe 0.31
Mg 0.009
Zn 0.013
Na 0.02
K 0.02
Mn 0.04

Table 7. Results of Soil Analysis.

Sample
(mg/Kg) Cu Fe Mg Zn Na K Mn
Extraction MDS Extraction MDS Extraction MDS Extraction MDS Extraction MDS Extraction MDS Extraction MDS

Soil-1 26 257 2124 116305 408 4060 1.1 167 30 1424 379 3833 370 2268
Soil-1 duplicate 27 299 2255 131302 406 3720 1.1 191 37 1722 390 3815 387 2749
Soil-2 23 257 1489 126035 392 2138 1.1 170 35 1162 329 2068 259 2439
Soil-2 duplicate 22 312 1490 135730 383 1802 1.0 198 31 1121 309 1481 249 2640
Soil-3 15 248 418 134645 418 2975 0.3 119 25 562 104 2940 71 1907
Soil-3 duplicate 14 288 374 129465 412 2593 0.3 138 24 587 95 4442 82 2210

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