a p p l i c at i o n N o t e Atomic Absorption Author: Randy L. Hergenreder PerkinElmer, Inc. Shelton, CT 06484 USA Trace Metals in Waters by GFAAS, in Accordance with U.S. EPA and Health Canada Requirements Introduction Several trace elements are recognized as toxic or carcinogenic and are regulated in drinking water by various environmental agencies worldwide. The U.S. Environmental Protection Agency’s (EPA) Safe Drinking Water Act includes maximum levels permitted in drinking water for the elements arsenic (As), cadmium (Cd), lead (Pb), selenium (Se) and thallium (Tl). The World Health Organization (WHO) and Health Canada also have limits on these elements in drinking water (Table 1). Water contamination sources can range from naturally occurring deposits exposed from erosion, to agriculture and industrial discharges. There can also be direct contamination from: Pb used in plumbing fixtures, Cd found in galvanized pipes, and electronics manufacturing discharges for Tl. Precise and accurate measurements at the regulated levels are an important factor for assuring safe drinking water. U.S. EPA Method 200.91 is the method cited by EPA, Health Canada, and the WHO for the use of graphite furnace atomic absorption spectroscopy (GFAAS). In evaluating a GFAAS system for determination of these elements, it must provide good sensitivity, low noise, limited drift, and accuracy in matrices with high salt content (hard water) that might be found in drinking waters. In this work, the PinAAcle™ 900T, with a unique optical system, is evaluated for the use of EPA Method 200.9 for As, Cd, Pb, Se, and Tl in drinking waters. Table 1. Drinking water limits for regulatory agencies (µg/L). Organization As Cd Pb Se Tl U.S. EPA 10 5 15 50 2 Health Canada 10 5 10 10 – WHO 10 3 10 10 – Experimental Instrumentation A PerkinElmer® PinAAcle 900T flame and longitudinal Zeeman atomic absorption spectrometer was used for all measurements. Samples were automatically pipetted into standard transversely heated graphite atomizer (THGA) tubes (Part No. B0504033) using an AS 900 autosampler. PerkinElmer Lumina™ single-element hollow cathode lamps (HCLs) were used as the light source for lead (Part No. N3050157), cadmium (Part No. N3050115), and thallium (Part No. N3050183); while single-element electrodeless discharge lamps (EDLs) were used for arsenic (Part No. N3050605) and selenium (Part No. N3050672). Figure 2. AS 900 autosampler depositing a droplet of water in the THGA tube as seen using the TubeView furnace camera. Table 2. Instrument settings used for U.S. EPA method 200.9 on the PinAAcle 900T. Wave- Lamp Read Read Element length Slit Lamp Current Delay Time (nm) (nm) Type (mA) (s) (s) As 193.7 0.7 EDL 380 1.0 2.5 Cd 228.8 0.7 HCL 4 0.5 2.5 Pb 283.3 0.7 HCL 10 0.5 3.5 Se 196.0 2.0 EDL 280 1.0 3.0 Tl 276.8 0.7 HCL 6 0.5 3.0 *Baseline Offset Correction (BOC) was 5 seconds for all analytes. Figure 1. PinAAcle 900T atomic absorption spectrometer with AS 900 furnace autosampler. The PinAAcle 900T instrument settings are listed in Table 2 and the furnace program is listed in Table 3. The pyrolysis and atomization temperatures were optimized using a spiked drinking water sample. The Method Development feature of the software (WinLab32™ for AA) automated these experiments. The TubeView™ furnace camera (Figure 2) on the PinAAcle 900T was used to adjust the pipette tip to the most appropriate depth and to watch for potential matrix buildup in the tube. The camera was also used during method development to verify the drying steps, ensuring that no sample boiling or spattering occurred. The furnace programs for individual elements are listed in Tables 3-7. Argon was the normal gas type. The special gas type was 95% Ar / 5% H2 and was used in steps 1-3. Step 4 removed the H2 gas from the tube before atomization and the read step was Step 5 for all elements. All elements used 0.005 mg Pd (Part No. B0190635) + 0.003 mg Mg(NO3)2 (Part No. B0190634) as a matrix modifier. All samples and standards used an injection volume of 20 µL sample plus 5 µL of matrix modifier. Each analyte was run in duplicate (2 replicates). Table 3. Furnace parameters for arsenic (As). Step Temp (˚C) Ramp Time Hold Time Internal Flow 1 110 1 30 250 Special 2 130 15 30 250 Special 3 900 10 20 250 Special 4 900 1 15 250 Normal 5* 2300 0 5 0 Normal 6 2450 1 3 250 Normal *Read step 2 Gas Type Results Table 4. Furnace parameters for cadmium (Cd). Step Temp (˚C) Ramp Time Hold Time Internal Flow Gas Type 1 110 1 30 250 Special 2 130 15 30 250 Special 3 700 10 20 250 Special 4 700 1 15 250 Normal 5* 1700 0 5 0 Normal 6 2450 1 3 250 Normal *Read step Table 5. Furnace parameters for lead (Pb). Step Temp (˚C) Ramp Time Hold Time Internal Flow Gas Type 1 110 1 30 250 Special 2 130 15 30 250 Special 3 850 10 20 250 Special 4 850 1 15 0 Normal 5* 2000 0 5 0 Normal 6 2450 1 3 250 Normal *Read step Table 6. Furnace parameters for selenium (Se). Step Temp (˚C) Ramp Time Hold Time Internal Flow Gas Type 1 110 1 30 250 Special 2 130 15 30 250 Special 3 800 10 30 250 Special 4 800 1 15 250 Normal 5* 2150 0 4 0 Normal 6 2450 1 3 250 Normal *Read step Table 7. Furnace parameters for thallium (Tl). Step Temp (˚C) Ramp Time Hold Time Internal Flow Gas Type 1 110 1 30 250 Special 2 130 15 30 250 Special 3 500 10 20 250 Special 4 500 1 15 250 Normal 5* 1600 0 5 0 Normal 6 2450 1 3 250 Normal Calibration Statistics All elements showed calibration correlation coefficients better than or equal to an r2 value of 0.999. The method detection limit (MDL) was determined by running seven separate samples of a 0.5 µg/L standard for As, Pb, Se and Tl and a 0.17 µg/L standard for Cd (Table 8). The resulting standard deviation was then multiplied by the Student’s t-value (ρ = 0.01). The instrument detection limit (IDL) was calculated by analyzing 10 replicates of a blank and multiplying the standard deviation by 3. Linear ranges are the highest standard that was within 5% of its true value. Table 8. Linear ranges, MDL and IDLs using the PinAAcle 900T and U.S. EPA Method 200.9. IDL IDL MDL Element (µg/L) (pg) (µg/L) Linear Range (µg/L) As 0.30 6.0 0.17 100 Cd 0.056 1.1 0.027 2.50 Pb 0.33 6.6 0.16 100 Se 0.28 5.6 0.30 200 Tl 0.37 7.4 0.18 100 The sensitivity, as indicated by the characteristic mass, was better than the expected value for As, Pb and Tl, while the Se and Cd values were slightly elevated. This could be due to the difference in modifier and lamp type from those detailed in the Atomic Absorption Spectroscopy Analytical Methods “cookbook” guide. Practical Quantification Limits In the Arsenic Rule,2 a standard of 3 µg/L must be recovered within ±30%. This is performed at the practical quantification limit (PQL) for As. Other elements have similar limits3 and these are listed in Table 9, along with the values determined in this study. A lower standard was used here to demonstrate the ease of this test using the PinAAcle 900T. The low standard was read as five separate samples and the average value with percent relative deviation is given. *Read step 3 Table 9. Results of the PQL study in accordance with U.S. EPA standards. Element PQL Level (µg/L) Acceptance Limit (± in %) Standard Concentration Used (µg/L) Average Found (µg/L) %RSD As 3 30 1 0.876 4.45 88.0 Cd 2 20 1 1.03 1.33 103 Pb 5 * 5 4.99 3.30 99.8 Se 10 20 1 1.06 13.5 106 Tl 2 30 1 0.810 14.3 81.0 %Recovery *No PQL acceptance limit was found. Preparing for Proposed Revision 3.3 In the proposed U.S. EPA Method 200.9 Revision 3.3, there is a requirement to run an Initial Precision and Recovery (IPR) standard. The proposed IPR currently consists of four aliquots of reagent water spiked at 1 to 5 times the lowest standard and run as samples. The percent recovery and standard deviation are calculated for those four samples (Table 10). There are currently no criteria for this test. Table 10. Data for the proposed initial precision and recovery standard. Average Found (µg/L) Std. Concentration Used (µg/L) Standard Deviation %Recovery As 4.83 5.0 0.15 96.6 Cd 0.293 0.25 0.0096 117 Pb 5.04 5.0 0.10 101 Se 4.94 5.0 0.13 98.7 Tl 5.00 5.0 0.18 99.9 Quality Control Two quality control (QC) checks were analyzed to determine the accuracy of the method. NIST® 1643e Trace Elements in Water is a fresh water reference material. Data and recoveries are listed in Table 11. The ERA PotableWatR™ Inorganic Metals material was used as the independent QC check during the analytical runs. This ERA reference material is also used in round robin U.S. EPA performance testing. The recoveries and data for this experiment and the U.S. EPA round robin experiment are listed in Table 12. The recovered concentrations are within the proficiency testing (PT) performance acceptance limits (PALs) which closely approximate a 95% confidence interval based on historical data. Table 11. Recoveries for NIST® 1643e natural waters reference material using PinAAcle 900T. Element 4 NIST® 1643e (µg/L) Limits (±, µg/L) NIST® Found (µg/L) Std. Dev. (n=2) %Recovery As 60.45 0.70 58.2 0.20 96.2 Cd 6.568 0.073 6.58 0.00080 100 Pb 19.63 0.21 19.5 0.15 99.1 Se 11.97 0.14 10.8 0.12 90.1 Tl 7.445 0.096 6.48 0.010 87.0 Table 12. Recoveries for the determination of metals in ERA PotableWatR™ Inorganic Metals. Certified Value Uncertainty Round Robin %Recovery ERA Found %Recovery (µg/L) (%) Mean (µg/L) Round Robin (µg/L) Found As 25.7 2.5 25.2 98.1 26.8 104 Cd 40.5 0.20 40.1 99.0 37.3 92.0 Pb 24.4 1.2 24.6 101 25.5 105 Se 22.6 1.2 22.5 99.6 22.1 97.6 Tl 7.84 1.4 7.75 98.8 7.35 93.8 Matrix Effects A laboratory fortified blank (LFB) showed good recoveries. One drinking water sample was analyzed with a spike and spike duplicate to check for matrix interferences. The spike recoveries and sample/blank data are shown in Table 13. Table 13. Sample and LFB recoveries and percent difference of duplicate spikes. Spike Lab % Sample Level Blank LFB Recovery Result (µg/L) (µg/L) (µg/L) LFB (µg/L) Sample Sample Spike % Spike Dup % (µg/L) Recovery (µg/L) Recovery As 30 <MDL 29.3 97.5 0.26 32.3 108 31.5 105 Cd 1.5 <MDL 1.42 94.5 0.031 1.49 99.1 1.54 103 Pb 30 <MDL 31.1 104 0.35 28.7 95.8 29.7 98.8 Se 30 <MDL 29.3 97.6 1.5 28.2 89.0 28.2 89.1 Tl 30 0.20 30.5 102 <MDL 30.2 101 33.4 111 Conclusion The PerkinElmer PinAAcle 900T atomic absorption spectrometer is capable of meeting the accuracy and precision requirements for the U.S. EPA and Health Canada regulations. The method detection limit (MDL) for all elements was determined to be well below the practical quantification limit (PQL) required by the method. This detection limit could be lowered even further by using a larger sample volume or through the use of end-capped THGA tubes (Part No. B3000655). Recoveries for two standard reference materials, spiked samples and lab fortified blanks were all within the method’s acceptable limits. References 1.U.S. EPA Method 200.9, Revision 2.2 and 3.0. 2.Environmental Protection Agency, "National Primary Drinking Water Regulations: Arsenic and Clarifications to Compliance and New Source Contaminants Monitoring, Final Rule," Federal Register, vol. 66, no. 14 (January 22, 2001), p. 6981. 3.Environmental Protection Agency, “National Primary Drinking Water Regulations: Inorganic Chemical sampling and analytical requirements,” 40 CFR 141.23. PerkinElmer, Inc. 940 Winter Street Waltham, MA 02451 USA P: (800) 762-4000 or (+1) 203-925-4602 www.perkinelmer.com For a complete listing of our global offices, visit www.perkinelmer.com/ContactUs Copyright ©2011, PerkinElmer, Inc. All rights reserved. PerkinElmer® is a registered trademark of PerkinElmer, Inc. All other trademarks are the property of their respective owners. 009634_01