Validation of Horizon Technology Disk Extraction Technology for US EPA Wastewater Method 625.1 Zoe Grosser, Alicia Cannon, Michael Ebitson, Melissa Lever, Horizon Technology, Salem, NH USA Nic Rasnake, Jessica Bowker, Allen Fuller, Chris Johnson, ESC Lab Sciences, a subsidiary of Pace Analytical, Mt Juliet, TN USA Key Words 625.1, wastewater, SPE, solid phase extraction, Introduction The US EPA monitors a variety of chemicals in water that may cause harm to humans or wildlife to minimize exposure. Method 625 was developed by the Office of Science and Technology in the Clean Water program to monitor a large suite of semivolatile chemicals in wastewater for compliance with the National Pollution Discharge Elimination System (NPDES). NPDES is a system of permitting that regulates the characteristics of water that is released into a waterway, defined by industrial category. The permitting levels are set depending on the waterway s use. If the waterway is used for recreation or is an important wildlife habitat, the limit may be set lower. The original method was developed in the early 1980 s and has been updated several times since then to allow the use of more modern technology. The latest update has taken place over the last few years and was proposed in a Method Update Rule (MUR) in 2015. 1 The latest version of the method includes a larger suite of analytes (up to 364) and an extensive set of labeled surrogates to better monitor the method performance throughout sample preparation and analysis steps. 2 One of the struggles of both manufacturers and laboratories is how to incorporate new technology into a laboratory or method to make the method easier, more reliable, or more cost effective without concern. Although most methods allow some flexibility, it is difficult to know what is supported by compliance with the existing quality control testing and what might be necessary to incorporate changes in the testing procedure. In the method update rule the EPA included some validation language for methods 624, 608, and 625, defining the scope and validation requirements to support a change in the method approach. An additional document is in draft form to give further guidance on the data required. 3 This process is similar to the Alternate Test Procedure which allows very different new technology, primarily in the analytical step, to be compared to an existing method to show comparability. 4 The validation process does not change the analytical step and makes smaller updates, such as the change from liquid-liquid extraction to solid phase extraction (SPE) in the sample preparation step. It is easier to manage and less costly because one laboratory is required to test nine matrices, rather than nine laboratories. This application note will present the data collected as part of the demonstration of disk solid phase extraction validation for US EPA method 625.1. Nine different wastewater matrices were evaluated and tested against the criteria listed in Table 6 of method 625.1. The analytes chosen for evaluation were from Tables 1 and 2 in method 625.1, with a few extra analytes from Table 3. The analytes from Table 3 are omitted in this note because there was no acceptance criteria for comparison. A full data package was generated and sent to the EPA to ensure we had met the necessary compliance requirements.
Experimental The samples were provided in one liter bottles or poured into 1-liter bottles if a larger composite sample was provided. The samples were processed as a matrix, matrix with a spike and a duplicate matrix with a spike. The samples were extracted using an Atlantic One-pass SPE disk (Horizon Technology) which is a mixed mode disk containing several functionalities. The process was automated with the SPE-DEX 4790 (Horizon Technology) extraction system. The newer SPE-DEX 5000 could be used to automate the procedure. A carbon cartridge (Max Detect, Horizon Technology) was also used to ensure adequate retention of the light end compounds, such as N-Nitrosodi-n-propylamine. See Figure 1 for guidance on the one-pass process. Also see reference 5 which includes a complete description of the extraction protocol. Figure 1. Description of the extraction process, including one pass of the sample through the disk and carbon cartridge and three extraction steps The Fast Flow Disk Holder was used with the samples because they had varying levels of particulate matter and the sample size was 1L. The Fast Flow disk holder uses a 47-mm disk, but allows larger filters to be placed on top to shield the SPE disk from particulate that may cause clogging and keep the flow through the disk fast. The particulate material is retained on the filters and washed with solvent during the elution step, so any material that has been adsorbed on the surface will be included in the extraction. Figure 2 shows a photo of the SPE- DEX 4790 configured with the carbon cartridge and the Fast Flow disk holder. P a ge 2 Figure 2. The SPE-DEX 4790 configured with Fast Flow Disk Holder and Carbon Cartridge. Bottles contain 1-L wastewater samples
The resulting extracts were dried using DryDisk membrane drying system (Horizon Technology) and evaporated to 1 ml with the DryVap In-line Drying and Evaporation System (Horizon Technology). Figure 3 shows the DryVap system with DryDisk glassware holding the drying membranes in front. The dried extract is drawn into the vessel in the back and the evaporation occurs there. Residual water can be seen in a few of the front vessels. The extraction methods used with the SPE-DEX 4790 are shown in Tables 1, 2 and 3. The conditions for operating the DryVap are shown in Table 4. Figure 3. DryVap system with samples in process Table 1. Acid and Neutral Extraction Method Step Solvent Soak Time Dry Time Prewet 1 Acetone 1.00 min 1:00 min Prewet 2 Reagent Water 1:00 min 1:00 min Sample Process Air Dry 3:00 min Rinse 1 Acetone 3:00 min 3:00 min Rinse 2 MeCl 3:00 min 3:00 min Rinse 3 MeCl 2:00 min 2:00 min Rinse 4 MeCl 2:00 min 2:00 min Rinse 5 MeCl 2:00 min 3:00 min Table 2. Ion Exchange Extraction Method Step Solvent Soak Time Dry Time Air Dry 0:00 min Rinse 1 Acetone 0:00 min 3:00 min Rinse 2 1% NH 4 OH 2:00 min 2:00 min Rinse 3 Acetone 3:00 min 2:00 min Rinse 4 MeCl 3:00 min 3:00 min Rinse 5 MeCl 2:00 min 3:00 min Rinse 6 MeCl 2:00 min 3:00 min Rinse 7 MeCl 2:00 min 3:00 min P a ge 3
Table 3. Carbon Extraction Method Step Solvent Soak Time Dry Time Air Dry 0:30 min Rinse 1 Acetone 1:00 min 1:00 min Rinse 2 MeCl 1:00 min 0:04 min Rinse 3 MeCl 1:00 min 0:04 min Rinse 4 MeCl 1:00 min 0:04 min Rinse 5 MeCl 1:00 min 1:00 min Table 4. DryVap System Conditions Parameter Setting Dry Volume 200 ml Heat Power 5 Heat Timer OFF Auto Rinse Mode OFF Nitrogen Sparge 20 psi Vacuum -7 in. Hg The samples were measured using GC/MS. Dilution was performed to check that the samples were being accurately measured and the loading was not distorting the mass spectral quantitation. The samples were analyzed using GC/MS (6890GC/5973MS, Agilent Technologies) at ESC Laboratories in Juliet, TN. The operational conditions are shown in Table 5. Table 5. GC/MS Conditions Injection Amount 1 µl Inlet Temperature 275 o C The standards used for spiking were from Semivolatiles Megamix Standard, EPA 625 and Benzidines Mix (Restek Corp, Bellefonte, PA). The surrogate mix was OLC 03.2 SVOA Mix and N- Nitrosodimethylamine-d6 (Restek Corp.) Reference materials were from Phenova, Denver Colorado and included a standard with varying concentrations of some of the analytes and a certified mix containing a single concentration of each analyte measured. Mode Gas Type Column Conditions Mode Oven Program Pulsed splitless Helium Zebron ZB-Semivolatiles (Phenomenex) Consistent Flow 50 o C hold for 2 min to 325 o C at 25 o C/min hold for 6 min. The samples obtained for analysis are listed in Table 6. They were provided by a large and small publically owned treatment works MS Ions Monitored Scan masses 35-550 (POTW) facility and several environmental laboratories with regular industrial customers. Results and Discussion The validation study was run following the guidance requirements in reference 3. Table 7 shows the types of samples and the total number of samples that must be prepared to demonstrate method validation. Page 4
Table 6. Description of the Wastewater Samples Evaluated (9 different matrices) Wastewater Description 1 Synthetic wastewater Prepared following ASTM D 5905-98 2 Synthetic seawater Prepared from Instant Ocean, a commercially available product closely matching the composition of seawater 3 POTW Influent 1 Geographical coverage of the southern section including residential and treated industrial waste 4 POTW Effluent Effluent from a large treatment plant 5 POTW Effluent plus O&G > 20 mg/l To ensure the criterion is met, the effluent was spiked with 24 mg/l of Oil & Grease Standard 6 Industrial Effluent 1-RC1 PART 446 PAINT FORMULATING POINT SOURCE CATEGORY 7 Industrial Effluent 2-RC2 PART 437 THE CENTRALIZED WASTE TREATMENT POINT SOURCE CATEGORY 8 Industrial Effluent 3-ES PART 432 MEAT AND POULTRY PRODUCTS POINT SOURCE CATEGORY 9 Industrial Effluent 4-Alpha Part 414 - Organic Chemicals. Plastics and Synthetic fibers (OCPSF) Table 7. Analyses required as specified in Method Validation Guidance (3) Number of Number of Analyses Method Application Vendorperformed Study All labs, all matrix types Labs Matrix types Background Analysis IPR- Reagent Water PT Sample MS/MSD MDL Total 1 9 9 4 1 18 7 39 Table 6 in US EPA Method 625.1 (December 2014) lists criterion by analytes for a variety of characteristics to validate that the method applied with changes will meet the requirements of the original method for a variety of challenging matrices representative of those that may be encountered in a commercial laboratory. This table is included in Appendix 1 for easy reference. Nine different matrices were collected for this purpose and measured using solid phase extraction disks and traditional GC/MS measurement. The matrices ranged from simple synthetic wastewater and seawater to POTW influent and effluent and the most challenging wastewater from PART 437 THE CENTRALIZED WASTE TREATMENT POINT SOURCE CATEGORY, which could only be evaporated to five ml after extraction rather than 1 ml. The samples had varying amounts of particulate and all were handled well by the Fast Flow Disk Holder with prefilters to capture the particulate above the adsorbent disk and include it in the extraction. The first column of Table 6 in Method 625.1 is range in % for recovery of the calibration verification standard. The results for this standard during the testing of the extracts is shown in Table 8 and meets the criteria listed in method 625.1 for compliance. Page 5
Table 8. Calibration Verification over the Course of Operation Recovery (%) Recovery (%) Recovery (%) Range for Q Pass/Fail Compound 0110_03 0105_03 0115C_02 (%) Acenaphthene 104 104 100 70-130 Pass Acenaphthylene 99.6 105 98.8 60-130 Pass Anthracene 108 104 99.5 58-130 Pass Benzidine 88.6 104 96.1 Benzo(a)anthracene 98.0 102 95.8 42-133 Pass Benzo(a)pyrene 101 103 87.6 32-148 Pass Benzo(b)fluoranthene 95.7 104 89.4 42-140 Pass Benzo(k)fluoranthene 108 101 83.6 25-146 Pass Benzo(g,h,i)perylene 110 113 104 13-195 Pass Benzylbutyl phthalate 104 105 96.0 43-140 Pass bis(2-chlorethoxy)methane 96.9 102 91.7 52-164 Pass bis(2-ethylhexyl)phthalate 103 107 93.4 43-137 Pass bis(2-chloroisopropyl)ether 97.1 103 97.7 63-139 Pass 4-Bromophenyl-phenylether 110 100 105 70-130 Pass 2-Chloronaphthalene 103 102 99.4 70-130 Pass 4-Chlorophenyl-phenylether 109 102 106 57-145 Pass Chrysene 105 102 97.6 44-140 Pass Dibenz(a,h)anthracene 111 111 99.3 13-200 Pass Di-n-butyl phthalate 107 104 100 52-130 Pass 3,3-Dichlorobenzidine 106 101 101 18-213 Pass Diethyl phthalate 110 105 102 47-130 Pass Dimethyl phthalate 106 104 101 50-130 Pass 2,4-Dinitrotoluene 104 102 98.9 53-130 Pass 2,6-Dinitrotoluene 101 110 102 68-137 Pass Di-n-octyl phthalate 104 105 92.8 21-132 Pass Fluoranthene 107 102 103 47-130 Pass Fluorene 103 103 98.1 70-130 Pass Hexachlorobenzene 112 103 112 38-142 Pass Hexachloro-1,3-butadiene 117 103 118 68-130 Pass Hexachloroethane 104 102 98.2 55-130 Pass Indeno(1,2,3-cd)pyrene 105 111 99.9 13-151 Pass Isophorone 100 104 96.0 52-180 Pass Naphthalene 101 102 97.5 70-130 Pass Nitrobenzene 104 107 100 54-158 Pass N-Nitrosodi-n-propylamine 95.0 105 90.5 59-170 Pass Phenanthrene 101 103 96.6 67-130 Pass Pyrene 100 103 94.9 70-130 Pass 1,2,4-Trichlorobenzene 109 102 105 61-130 Pass 4-Chloro-3-methylphenol 94.4 103 101 68-130 Pass 2-Chlorophenol 98.7 103 94.3 55-130 Pass Page 6
2,4-Dichlorophenol 101 102 110 64-130 Pass 2,4-Dimethylphenol 103 106 104 58-130 Pass 2,4-Dinitrophenol 104 95.1 116 39-173 Pass 4,6-Dinitro-2-methylphenol 108 102 110 56-130 Pass 2-Nitrophenol 107 102 103 61-163 Pass 4-Nitrophenol 93.7 107 95.5 35-130 Pass Pentachlorophenol 84.4 97.0 105 42-152 Pass Phenol 95.1 101 92.2 48-130 Pass 2,4,6-Trichlorophenol 105 106 113 69-130 Pass bis(2-chloroethyl)ether 94.5 106 90.3 52-130 Pass Before running samples, the laboratory must first demonstrate their capability to use a method with an initial demonstration of compliance, running four spiked reagent water samples through the complete sample preparation and analysis step. The results for recovery accuracy and precision are compared with the range specified for compliance. The range is included in Table 9 (tan background) for easy comparison. Benzidine is not reported due to unpredictable GC results and interferences with heavier matrices of the wastewater samples. Table 9. Initial Demonstration of Compliance Analyte Avg 5x Range X Pass/Fail SD 5x Limit for s (%) Pass/Fail Acenaphthene 92.9 60-132 Pass 8.42 29 Pass Acenaphthylene 89.7 54-126 Pass 8.30 45 Pass Anthracene 97.0 43-120 Pass 8.15 40 Pass Benzo(a)anthracene 94.2 42-133 Pass 7.59 32 Pass Benzo(a)pyrene 95.2 32-148 Pass 8.91 43 Pass Benzo(b)fluoranthene 116 42-140 Pass 9.29 43 Pass Benzo(k)fluoranthene 119 25-146 Pass 10.5 38 Pass Benzo(g,h,i)perylene 50.5 D-195 Pass 3.83 61 Pass Benzylbutyl phthalate 94.8 D-140 Pass 8.03 36 Pass bis(2-chlorethoxy)methane 86.2 49-165 Pass 6.68 32 Pass bis(2-ethylhexyl)phthalate 94.9 29-137 Pass 7.69 50 Pass bis(2-chloroisopropyl)ether 81.6 63-139 Pass 9.26 46 Pass 4-Bromophenyl-phenylether 102 65-120 Pass 9.29 26 Pass 2-Chloronaphthalene 89.0 65-120 Pass 8.29 15 Pass 4-Chlorophenyl-phenylether 99.6 38-145 Pass 9.35 36 Pass Chrysene 101 44-140 Pass 8.59 53 Pass Dibenz(a,h)anthracene 60.3 D-200 Pass 4.69 75 Pass Di-n-butyl phthalate 108 8-120 Pass 8.64 28 Pass 3,3-Dichlorobenzidine 69.8 8-213 Pass 5.69 65 Pass Diethyl phthalate 108 D-120 Pass 8.80 60 Pass Page 7
Dimethyl phthalate 102 D-120 Pass 8.86 110 Pass 2,4-Dinitrotoluene 101 48-127 Pass 9.22 25 Pass 2,6-Dinitrotoluene 103 68-137 Pass 8.49 29 Pass Di-n-octyl phthalate 96.6 19-132 Pass 6.34 42 Pass Fluoranthene 108 43-121 Pass 9.55 40 Pass Fluorene 94.2 70-120 Pass 8.46 23 Pass Hexachlorobenzene 108 8-142 Pass 8.75 33 Pass Hexachloro-1,3-butadiene 89.1 38-120 Pass 11.7 38 Pass Hexachloroethane 69.4 55-120 Pass 9.05 32 Pass Indeno(1,2,3-cd)pyrene 57.5 D-151 Pass 4.65 60 Pass Isophorone 89.5 47-180 Pass 7.33 56 Pass Naphthalene 85.0 36-120 Pass 7.78 39 Pass Nitrobenzene 85.9 54-158 Pass 7.77 37 Pass N-Nitrosodi-n-propylamine 85.9 14-198 Pass 6.42 52 Pass Phenanthrene 95.4 65-120 Pass 7.98 24 Pass Pyrene 87.2 70-120 Pass 7.21 30 Pass 1,2,4-Trichlorobenzene 87.1 57-130 Pass 9.43 30 Pass 4-Chloro-3-methylphenol 91.0 41-128 Pass 6.91 44 Pass 2-Chlorophenol 79.6 36-120 Pass 7.35 37 Pass 2,4-Dichlorophenol 91.1 53-122 Pass 8.86 30 Pass 2,4-Dimethylphenol 94.0 42-120 Pass 7.44 35 Pass 2,4-Dinitrophenol 113 D-173 Pass 11.4 79 Pass 4,6-Dinitro-2-methylphenol 107 53-130 Pass 10.3 122 Pass 2-Nitrophenol 93.2 45-167 Pass 8.15 33 Pass 4-Nitrophenol 76.3 13-129 Pass 8.18 79 Pass Pentachlorophenol 111 42-152 Pass 12.7 52 Pass Phenol 69.9 17-120 Pass 6.27 39 Pass 2,4,6-Trichlorophenol 94.1 52-129 Pass 8.85 35 Pass bis(2-chloroethyl)ether 76.4 43-126 Pass 7.52 65 Pass The criteria were met in all cases, indicating the complete method including the analytical instrument step as operated is under control and further analysis can proceed. The method detection limits (MDLs) were measured using eight low-level spikes of reagent water and carrying them through the entire sample preparation and measurement protocols as samples would be. The MDL was calculated following the equation specified in the method and a student s t value of 2.998 was used. Although only an indicator of the lower level of the method, they were compared with the MDL values listed in method 625.1 to show equivalency. The MDLs determined using SPE are better in most cases than the MDLs in Method 625 determined using liquid-liquid extraction for sample preparation. The one case that stands out in comparison is bis(2-ethylhexyl)phthalate and that may be attributed to contamination. Two of the eight low level spikes were high, influencing the standard deviation for that compound more than for other analytes. Page 8
Table 10. Method Detection Limits (MDLs) Measured using SPE, compared to Method 625.1 Values, using LLE Compound Measured MDL (µg/l) (SPE) MDL from Method 625.1 (µg/l) (LLE) Acenaphthene 0.709 1.90 Acenaphthylene 0.644 3.50 Anthracene 1.12 1.90 Benzo(a)anthracene 0.879 7.80 Benzo(a)pyrene 0.944 2.50 Benzo(b)fluoranthene 1.55 4.80 Benzo(k)fluoranthene 1.49 2.50 Benzo(g,h,i)perylene 2.78 4.10 Benzylbutyl phthalate 0.624 2.50 bis(2-chlorethoxy)methane 0.880 5.30 bis(2-ethylhexyl)phthalate 8.76 2.50 bis(2-chloroisopropyl)ether 1.71 5.70 4-Bromophenyl-phenylether 0.971 1.90 2-Chloronaphthalene 0.764 1.90 4-Chlorophenyl-phenylether 0.985 4.20 Chrysene 0.866 2.50 Dibenz(a,h)anthracene 2.23 2.50 Di-n-butyl phthalate 1.07 2.50 3,3-Dichlorobenzidine 4.94 16.5 Diethyl phthalate 0.976 1.90 Dimethyl phthalate 0.778 1.60 2,4-Dinitrotoluene 1.35 5.70 2,6-Dinitrotoluene 0.886 1.90 Di-n-octyl phthalate 0.895 2.50 Fluoranthene 1.21 2.20 Fluorene 0.894 1.90 Hexachlorobenzene 1.12 1.90 Hexachloro-1,3-butadiene 1.42 0.90 Hexachloroethane 1.77 1.60 Indeno(1,2,3-cd)pyrene 2.33 3.70 Isophorone 1.30 2.20 Naphthalene 1.04 1.60 Nitrobenzene 1.17 1.90 N-Nitrosodi-n-propylamine 1.67 Phenanthrene 1.05 5.40 Pyrene 1.05 1.90 1,2,4-Trichlorobenzene 1.33 1.90 Page 9
4-Chloro-3-methylphenol 1.15 3.00 2-Chlorophenol 0.952 3.30 2,4-Dichlorophenol 0.901 2.70 2,4-Dimethylphenol 0.868 2.70 2,4-Dinitrophenol 2.20 42.0 4,6-Dinitro-2-methylphenol 1.60 24.0 2-Nitrophenol 1.37 3.60 4-Nitrophenol 1.43 2.40 Pentachlorophenol 1.63 3.60 Phenol 0.688 1.50 2,4,6-Trichlorophenol 1.00 2.70 bis(2-chloroethyl)ether 1.55 5.70 The next Tables show the results for the nine different wastewater samples. The tan-colored columns are the criteria presented in Table 6 of Method 625.1. The spike recovery data from the sample is shown in the first column of data. The spike was added at 100 µg/l in each liter of water sample. The relative percent difference between the spike recovery and spike recovery duplicate is calculated using the equation in method 625.1. The tan column criteria are compared with the data from this study and if the data meets the criterion, a pass is indicated in the Pass/Fail column and the color is green. Synthetic wastewater was prepared using the recipe in ASTM-D 5905 98, using a combination of beer, flour, salts and other materials to generate an approximation of a wastewater matrix. Table 11. Synthetic Wastewater Results Recovery (%) Range Pass/Fail RPD (%) RPD (%) Pass/Fail Compound 20x P,Ps(%) 20x Limit Acenaphthene 80.4 47-145 Pass 11.8 48 Pass Acenaphthylene 80.3 33-145 Pass 14.5 74 Pass Anthracene 84.9 27-133 Pass 17.3 66 Pass Benzo(a)anthracene 83.6 33-143 Pass 24.4 53 Pass Benzo(a)pyrene 81.3 17-163 Pass 26.9 72 Pass Benzo(b)fluoranthene 86.6 24-159 Pass 32.5 71 Pass Benzo(k)fluoranthene 88.6 11-162 Pass 22.5 63 Pass Benzo(g,h,i)perylene 88.0 D-219 Pass 27.5 97 Pass Benzylbutyl phthalate 89.8 D-152 Pass 19.3 60 Pass bis(2-chlorethoxy)methane 81.6 33-184 Pass 16.1 54 Pass bis(2-ethylhexyl)phthalate 89.3 36-166 Pass 27.8 76 Pass bis(2-chloroisopropyl)ether 67.1 8-158 Pass 13.0 82 Pass 4-Bromophenyl-phenylether 91.2 53-127 Pass 20.9 43 Pass 2-Chloronaphthalene 77.7 60-120 Pass 14.0 24 Pass 4-Chlorophenyl-phenylether 87.5 25-158 Pass 13.2 61 Pass Page 10
Chrysene 89.1 17-168 Pass 23.8 87 Pass Dibenz(a,h)anthracene 86.6 D-227 Pass 27.0 126 Pass Di-n-butyl phthalate 93.6 1-120 Pass 19.6 47 Pass 3,3-Dichlorobenzidine 44.3 D-262 Pass 8.1 108 Pass Diethyl phthalate 94.6 D-120 Pass 13.5 100 Pass Dimethyl phthalate 92.8 D-120 Pass 15.5 183 Pass 2,4-Dinitrotoluene 93.2 39-139 Pass 16.2 42 Pass 2,6-Dinitrotoluene 90.1 50-158 Pass 17.0 48 Pass Di-n-octyl phthalate 89.2 4-146 Pass 30.9 69 Pass Fluoranthene 87.4 26-137 Pass 18.5 66 Pass Fluorene 81.2 59-121 Pass 8.5 38 Pass Hexachlorobenzene 93.5 D-152 Pass 22.6 55 Pass Hexachloro-1,3-butadiene 75.6 24-120 Pass 16.3 62 Pass Hexachloroethane 49.7 40-120 Pass 2.0 52 Pass Indeno(1,2,3-cd)pyrene 87.1 D-171 Pass 27.3 99 Pass Isophorone 86.3 21-196 Pass 20.3 93 Pass Naphthalene 72.2 21-133 Pass 10.6 65 Pass Nitrobenzene 81.8 35-180 Pass 18.5 62 Pass N-Nitrosodi-n-propylamine 76.2 D-230 Pass 22.3 87 Pass Phenanthrene 84.2 54-120 Pass 15.7 128 Pass Pyrene 81.0 52-120 Pass 16.7 49 Pass 1,2,4-Trichlorobenzene 73.0 44-142 Pass 15.3 50 Pass 4-Chloro-3-methylphenol 87.2 22-147 Pass 20.7 73 Pass 2-Chlorophenol 79.8 23-134 Pass 17.5 61 Pass 2,4-Dichlorophenol 86.3 39-135 Pass 19.7 50 Pass 2,4-Dimethylphenol 92.4 32-120 Pass 21.2 58 Pass 2,4-Dinitrophenol 132 D-191 Pass 38.7 132 Pass 4,6-Dinitro-2-methylphenol 102 D-181 Pass 23.4 203 Pass 2-Nitrophenol 80.0 29-182 Pass 9.9 55 Pass 4-Nitrophenol 72.9 D-132 Pass 16.9 131 Pass Pentachlorophenol 111 14-176 Pass 26.6 86 Pass Phenol 68.7 5-120 Pass 27.5 64 Pass 2,4,6-Trichlorophenol 95.5 37-144 Pass 16.9 58 Pass bis(2-chloroethyl)ether 65.5 12-158 Pass 15.0 108 Pass Page 11
The synthetic wastewater performed very well and each compound met the criteria listed in method 625.1. Synthetic seawater was prepared using Instant Ocean salt mixture, an economical way to prepare an approximate seawater matrix for evaluation. The results are shown in Table 12. Table 12. Synthetic Seawater Results Recovery (%) Range Pass/Fail RPD (%) RPD (%) Pass/Fail Compound 5x P,Ps(%) 5x Limit Acenaphthene 81.6 47-145 Pass 1.76 48 Pass Acenaphthylene 81.7 33-145 Pass 3.05 74 Pass Anthracene 81.7 27-133 Pass 0.99 66 Pass Benzo(a)anthracene 84.2 33-143 Pass 0.25 53 Pass Benzo(a)pyrene 79.8 17-163 Pass 1.06 72 Pass Benzo(b)fluoranthene 83.0 24-159 Pass 10.4 71 Pass Benzo(k)fluoranthene 82.8 11-162 Pass 0.36 63 Pass Benzo(g,h,i)perylene 85.4 D-219 Pass 2.53 97 Pass Benzylbutyl phthalate 83.1 D-152 Pass 1.67 60 Pass bis(2-chlorethoxy)methane 75.1 33-184 Pass 4.24 54 Pass bis(2-ethylhexyl)phthalate 81.7 36-166 Pass 0.21 76 Pass bis(2-chloroisopropyl)ether 67.9 8-158 Pass 0.37 82 Pass 4-Bromophenyl-phenylether 89.2 53-127 Pass 4.36 43 Pass 2-Chloronaphthalene 80.0 60-120 Pass 1.79 24 Pass 4-Chlorophenyl-phenylether 88.8 25-158 Pass 1.13 61 Pass Chrysene 83.9 17-168 Pass 1.24 87 Pass Dibenz(a,h)anthracene 87.4 D-227 Pass 2.84 126 Pass Di-n-butyl phthalate 88.1 1-120 Pass 0.28 47 Pass 3,3-Dichlorobenzidine 49.4 D-262 Pass 12.8 108 Pass Diethyl phthalate 92.6 D-120 Pass 0.14 100 Pass Dimethyl phthalate 95.7 D-120 Pass 1.65 183 Pass 2,4-Dinitrotoluene 92.0 39-139 Pass 1.73 42 Pass 2,6-Dinitrotoluene 95.7 50-158 Pass 3.23 48 Pass Di-n-octyl phthalate 82.7 4-146 Pass 1.81 69 Pass Fluoranthene 88.2 26-137 Pass 0.16 66 Pass Fluorene 81.4 59-121 Pass 0.59 38 Pass Hexachlorobenzene 90.5 D-152 Pass 8.21 55 Pass Hexachloro-1,3-butadiene 76.6 24-120 Pass 5.47 62 Pass Hexachloroethane 54.1 40-120 Pass 4.60 52 Pass Indeno(1,2,3-cd)pyrene 86.6 D-171 Pass 0.22 99 Pass Page 12
Indeno(1,2,3-cd)pyrene 86.6 D-171 Pass 0.22 99 Pass Isophorone 78.4 21-196 Pass 2.06 93 Pass Naphthalene 71.7 21-133 Pass 1.93 65 Pass Nitrobenzene 73.8 35-180 Pass 1.50 62 Pass N-Nitrosodi-n-propylamine 72.4 D-230 Pass 3.50 87 Pass Phenanthrene 82.5 54-120 Pass 0.02 128 Pass Pyrene 79.4 52-120 Pass 0.63 49 Pass 1,2,4-Trichlorobenzene 73.1 44-142 Pass 2.62 50 Pass 4-Chloro-3-methylphenol 84.5 22-147 Pass 3.18 73 Pass 2-Chlorophenol 77.0 23-134 Pass 0.55 61 Pass 2,4-Dichlorophenol 88.7 39-135 Pass 1.97 50 Pass 2,4-Dimethylphenol 85.6 32-120 Pass 0.44 58 Pass 2,4-Dinitrophenol 125 D-191 Pass 1.60 132 Pass 4,6-Dinitro-2-methylphenol 101 D-181 Pass 4.03 203 Pass 2-Nitrophenol 84.4 29-182 Pass 0.55 55 Pass 4-Nitrophenol 88.9 D-132 Pass 6.39 131 Pass Pentachlorophenol 93.7 14-176 Pass 20.2 86 Pass Phenol 44.6 5-120 Pass 40.9 64 Pass 2,4,6-Trichlorophenol 97.7 37-144 Pass 1.21 58 Pass bis(2-chloroethyl)ether 61.2 12-158 Pass 4.08 108 Pass The synthetic seawater passed all the criteria and those results are shown as passing, in green. The next two tables represent typical wastewater influent and effluent from a large POTW facility. The Influent results are shown in Table 13. Page 13
Table 13. POTW Influent from a large treatment facility results Recovery (%) Range Pass/Fail RPD (%) RPD (%) Pass/Fail Compound 20x P,Ps(%) Limit Acenaphthene 89.5 47-145 Pass 10.8 48 Pass Acenaphthylene 83.9 33-145 Pass 10.3 74 Pass Anthracene 88.9 27-133 Pass 10.8 66 Pass Benzo(a)anthracene 87.9 33-143 Pass 12.8 53 Pass Benzo(a)pyrene 90.8 17-163 Pass 12.1 72 Pass Benzo(b)fluoranthene 98.3 24-159 Pass 14.9 71 Pass Benzo(k)fluoranthene 99.7 11-162 Pass 12.8 63 Pass Benzo(g,h,i)perylene 59.0 D-219 Pass 11.6 97 Pass Benzylbutyl phthalate 93.1 D-152 Pass 8.33 60 Pass bis(2-chlorethoxy)methane 81.5 33-184 Pass 15.6 54 Pass bis(2-ethylhexyl)phthalate 105 36-166 Pass 11.7 76 Pass bis(2-chloroisopropyl)ether 73.1 8-158 Pass 19.6 82 Pass 4-Bromophenyl-phenylether 92.2 53-127 Pass 13.1 43 Pass 2-Chloronaphthalene 82.9 60-120 Pass 11.4 24 Pass 4-Chlorophenyl-phenylether 98.5 25-158 Pass 6.6 61 Pass Chrysene 89.8 17-168 Pass 16.4 87 Pass Dibenz(a,h)anthracene 66.0 D-227 Pass 11.2 126 Pass Di-n-butyl phthalate 101 1-120 Pass 1.1 47 Pass 3,3-Dichlorobenzidine 21.8 D-262 Pass 35.5 108 Pass Diethyl phthalate 116 D-120 Pass 0.560 100 Pass Dimethyl phthalate 95.7 D-120 Pass 10.3 183 Pass 2,4-Dinitrotoluene 94.3 39-139 Pass 8.65 42 Pass 2,6-Dinitrotoluene 95.0 50-158 Pass 9.72 48 Pass Di-n-octyl phthalate 91.0 4-146 Pass 13.3 69 Pass Fluoranthene 91.4 26-137 Pass 10.7 66 Pass Fluorene 93.0 59-121 Pass 7.10 38 Pass Hexachlorobenzene 98.6 D-152 Pass 12.4 55 Pass Hexachloro-1,3-butadiene 84.9 24-120 Pass 18.3 62 Pass Hexachloroethane 59.0 40-120 Pass 25.8 52 Pass Indeno(1,2,3-cd)pyrene 65.2 D-171 Pass 12.0 99 Pass Isophorone 84.7 21-196 Pass 16.3 93 Pass Naphthalene 77.8 21-133 Pass 16.7 65 Pass Nitrobenzene 82.1 35-180 Pass 17.7 62 Pass N-Nitrosodi-n-propylamine 84.0 D-230 Pass 16.5 87 Pass Phenanthrene 87.1 54-120 Pass 12.5 128 Pass Pyrene 83.9 52-120 Pass 15.1 49 Pass 1,2,4-Trichlorobenzene 76.6 44-142 Pass 21.6 50 Pass 4-Chloro-3-methylphenol 73.1 22-147 Pass 24.3 73 Pass Page 14
2-Chlorophenol 80.3 23-134 Pass 20.1 61 Pass 2,4-Dichlorophenol 81.6 39-135 Pass 8.9 50 Pass 2,4-Dimethylphenol 84.1 32-120 Pass 16.5 58 Pass 2,4-Dinitrophenol 99.8 D-191 Pass 9.70 132 Pass 4,6-Dinitro-2-methylphenol 84.4 D-181 Pass 17.5 203 Pass 2-Nitrophenol 86.0 29-182 Pass 15.4 55 Pass 4-Nitrophenol 87.7 D-132 Pass 12.7 131 Pass Pentachlorophenol 105 14-176 Pass 11.0 86 Pass Phenol 78.5 5-120 Pass 27.9 64 Pass 2,4,6-Trichlorophenol 76.5 37-144 Pass 10.5 58 Pass bis(2-chloroethyl)ether 71.2 12-158 Pass 19.9 108 Pass The analytes met all the criteria and Pass is seen in both the recovery and in the relative percent difference between the spike and spike duplicate. The POTW effluent results are shown in Table 14. Table 14. POTW Effluent Results Recovery (%) Range Pass/Fail RPD RPD (%) Pass/Fail Compound 5x P,Ps(%) (%) Limit Acenaphthene 70.9 47-145 Pass 8.21 48 Pass Acenaphthylene 70.5 33-145 Pass 9.34 74 Pass Anthracene 74.5 27-133 Pass 7.71 66 Pass Benzo(a)anthracene 71.0 33-143 Pass 6.40 53 Pass Benzo(a)pyrene 72.0 17-163 Pass 5.43 72 Pass Benzo(b)fluoranthene 77.2 24-159 Pass 4.50 71 Pass Benzo(k)fluoranthene 81.9 11-162 Pass 6.42 63 Pass Benzo(g,h,i)perylene 44.4 D-219 Pass 0.18 97 Pass Benzylbutyl phthalate 73.7 D-152 Pass 9.65 60 Pass bis(2-chlorethoxy)methane 70.8 33-184 Pass 9.32 54 Pass bis(2-ethylhexyl)phthalate 69.4 36-166 Pass 2.17 76 Pass bis(2-chloroisopropyl)ether 62.8 8-158 Pass 12.5 82 Pass 4-Bromophenyl-phenylether 78.4 53-127 Pass 7.28 43 Pass 2-Chloronaphthalene 69.1 60-120 Pass 7.52 24 Pass 4-Chlorophenyl-phenylether 75.1 25-158 Pass 6.78 61 Pass Chrysene 74.7 17-168 Pass 6.06 87 Pass Dibenz(a,h)anthracene 51.6 D-227 Pass 4.43 126 Pass Di-n-butyl phthalate 82.3 1-120 Pass 10.1 47 Pass 3,3-Dichlorobenzidine 13.7 D-262 Pass 40.5 108 Pass Page 15
Diethyl phthalate 84.3 D-120 Pass 11.7 100 Pass Dimethyl phthalate 80.2 D-120 Pass 11.4 183 Pass 2,4-Dinitrotoluene 80.5 39-139 Pass 16.4 42 Pass 2,6-Dinitrotoluene 81.1 50-158 Pass 12.0 48 Pass Di-n-octyl phthalate 71.5 4-146 Pass 3.47 69 Pass Fluoranthene 80.7 26-137 Pass 9.10 66 Pass Fluorene 72.4 59-121 Pass 9.74 38 Pass Hexachlorobenzene 80.4 D-152 Pass 4.15 55 Pass Hexachloro-1,3-butadiene 72.1 24-120 Pass 11.8 62 Pass Hexachloroethane 54.3 40-120 Pass 18.1 52 Pass Indeno(1,2,3-cd)pyrene 50.9 D-171 Pass 2.58 99 Pass Isophorone 73.9 21-196 Pass 9.78 93 Pass Naphthalene 68.3 21-133 Pass 10.8 65 Pass Nitrobenzene 70.8 35-180 Pass 9.42 62 Pass N-Nitrosodi-n-propylamine 68.3 D-230 Pass 15.8 87 Pass Phenanthrene 73.2 54-120 Pass 8.74 128 Pass Pyrene 67.1 52-120 Pass 9.25 49 Pass 1,2,4-Trichlorobenzene 67.9 44-142 Pass 7.59 50 Pass 4-Chloro-3-methylphenol 75.0 22-147 Pass 15.7 73 Pass 2-Chlorophenol 64.0 23-134 Pass 17.7 61 Pass 2,4-Dichlorophenol 70.2 39-135 Pass 8.64 50 Pass 2,4-Dimethylphenol 74.8 32-120 Pass 8.47 58 Pass 2,4-Dinitrophenol 95.1 D-191 Pass 14.8 132 Pass 4,6-Dinitro-2-methylphenol 84.9 D-181 Pass 13.8 203 Pass 2-Nitrophenol 74.4 29-182 Pass 11.0 55 Pass 4-Nitrophenol 60.9 D-132 Pass 17.3 131 Pass Pentachlorophenol 91.2 14-176 Pass 14.4 86 Pass Phenol 57.8 5-120 Pass 13.7 64 Pass 2,4,6-Trichlorophenol 72.5 37-144 Pass 9.69 58 Pass bis(2-chloroethyl)ether 62.2 12-158 Pass 17.6 108 Pass The results pass for all the analytes. At least one of the nine wastewater samples should have a special characteristic, such as more than 20 mg/l of oil & Grease in the sample. To ensure that the sample had this characteristic one POTW effluent sample was spiked with 24 mg/l of oil & grease standard before spiking and extraction of the three 1-L aliquots. Page 16
Table 15. POTW Effluent plus >20 mg/l Oil & Grease Results Recovery (%) Range Pass/Fail RPD RPD (%) Pass/Fail Compound 20x P,Ps(%) % Limit Acenaphthene 83.7 47-145 Pass 4.16 48 Pass Acenaphthylene 80.8 33-145 Pass 3.63 74 Pass Anthracene 89.6 27-133 Pass 6.85 66 Pass Benzo(a)anthracene 84.1 33-143 Pass 6.30 53 Pass Benzo(a)pyrene 87.0 17-163 Pass 6.94 72 Pass Benzo(b)fluoranthene 94.0 24-159 Pass 7.26 71 Pass Benzo(k)fluoranthene 107 11-162 Pass 1.78 63 Pass Benzo(g,h,i)perylene 40.2 D-219 Pass 10.1 97 Pass Benzylbutyl phthalate 88.9 D-152 Pass 2.27 60 Pass bis(2-chlorethoxy)methane 79.2 33-184 Pass 1.86 54 Pass bis(2-ethylhexyl)phthalate 88.8 36-166 Pass 7.12 76 Pass bis(2-chloroisopropyl)ether 65.9 8-158 Pass 16.5 82 Pass 4-Bromophenyl-phenylether 88.1 53-127 Pass 12.7 43 Pass 2-Chloronaphthalene 78.6 60-120 Pass 6.44 24 Pass 4-Chlorophenyl-phenylether 91.3 25-158 Pass 3.74 61 Pass Chrysene 88.6 17-168 Pass 3.89 87 Pass Dibenz(a,h)anthracene 48.3 D-227 Pass 10.5 126 Pass Di-n-butyl phthalate 94.9 1-120 Pass 17.8 47 Pass 3,3-Dichlorobenzidine 17.7 D-262 Pass 27.0 108 Pass Diethyl phthalate 95.0 D-120 Pass 11.1 100 Pass Dimethyl phthalate 92.5 D-120 Pass 0.36 183 Pass 2,4-Dinitrotoluene 87.3 39-139 Pass 3.41 42 Pass 2,6-Dinitrotoluene 88.9 50-158 Pass 5.08 48 Pass Di-n-octyl phthalate 86.0 4-146 Pass 6.73 69 Pass Fluoranthene 98.8 26-137 Pass 6.74 66 Pass Fluorene 84.6 59-121 Pass 5.75 38 Pass Hexachlorobenzene 93.1 D-152 Pass 13.1 55 Pass Hexachloro-1,3-butadiene 84.0 24-120 Pass 0.30 62 Pass Hexachloroethane 55.9 40-120 Pass 8.01 52 Pass Indeno(1,2,3-cd)pyrene 46.5 D-171 Pass 12.6 99 Pass Isophorone 81.4 21-196 Pass 2.78 93 Pass Naphthalene 74.1 21-133 Pass 6.75 65 Pass Nitrobenzene 75.5 35-180 Pass 8.26 62 Pass N-Nitrosodi-n-propylamine 75.2 D-230 Pass 4.86 87 Pass Phenanthrene 82.4 54-120 Pass 9.65 128 Pass Pyrene 81.7 52-120 Pass 5.46 49 Pass 1,2,4-Trichlorobenzene 75.5 44-142 Pass 5.46 50 Pass 4-Chloro-3-methylphenol 76.5 22-147 Pass 6.70 73 Pass Page 17
2-Chlorophenol 70.9 23-134 Pass 5.95 61 Pass 2,4-Dichlorophenol 76.5 39-135 Pass 7.19 50 Pass 2,4-Dimethylphenol 81.8 32-120 Pass 6.10 58 Pass 2,4-Dinitrophenol 71.4 D-191 Pass 5.05 132 Pass 4,6-Dinitro-2-methylphenol 72.6 D-181 Pass 6.21 203 Pass 2-Nitrophenol 84.8 29-182 Pass 1.38 55 Pass 4-Nitrophenol 60.8 D-132 Pass 12.2 131 Pass Pentachlorophenol 84.9 14-176 Pass 15.7 86 Pass Phenol 60.0 5-120 Pass 0.38 64 Pass 2,4,6-Trichlorophenol 81.6 37-144 Pass 5.19 58 Pass bis(2-chloroethyl)ether 66.7 12-158 Pass 0.52 108 Pass The results show excellent compliance with the established criteria, even with the added oil & grease. Table 16 shows a more complicated industrial effluent from the Paint Formulating Point Source Category (446). Heavy non-target matrix caused internal standard failures. Dilution was needed to meet method criteria for the internal standard. Table 16. Paint Formulating Point Source (446) Category Results Recovery (%) Range Pass/Fail RPD RPD (%) Pass/Fail Compound 250x P,Ps(%) (%) Limit Acenaphthene 95.0 47-145 Pass 7.74 48 Pass Acenaphthylene 89.3 33-145 Pass 10.2 74 Pass Anthracene 94.7 27-133 Pass 3.80 66 Pass Benzo(a)anthracene 93.8 33-143 Pass 0.10 53 Pass Benzo(a)pyrene 107 17-163 Pass 3.43 72 Pass Benzo(b)fluoranthene 78.3 24-159 Pass 3.27 71 Pass Benzo(k)fluoranthene 115 11-162 Pass 5.30 63 Pass Benzo(g,h,i)perylene 90.0 D-219 Pass 14.6 97 Pass Benzylbutyl phthalate 125 D-152 Pass 16.7 60 Pass bis(2-chlorethoxy)methane 108 33-184 Pass 20.0 54 Pass bis(2-ethylhexyl)phthalate 196 36-166 6.50 76 Pass bis(2-chloroisopropyl)ether 8-158 82 4-Bromophenyl-phenylether 109 53-127 Pass 5.15 43 Pass 2-Chloronaphthalene 92.8 60-120 Pass 4.76 24 Pass 4-Chlorophenyl-phenylether 113 25-158 Pass 5.25 61 Pass Chrysene 110 17-168 Pass 0.16 87 Pass Dibenz(a,h)anthracene 88.7 D-227 Pass 9.30 126 Pass Di-n-butyl phthalate 132 1-120 4.71 47 Pass 3,3-Dichlorobenzidine D-262 108 Diethyl phthalate 108 D-120 Pass 11.3 100 Pass Page 18
Dimethyl phthalate 93.5 D-120 Pass 11.2 183 Pass 2,4-Dinitrotoluene 97.5 39-139 Pass 12.0 42 Pass 2,6-Dinitrotoluene 99.1 50-158 Pass 5.14 48 Pass Di-n-octyl phthalate 104 4-146 Pass 0.29 69 Pass Fluoranthene 105 26-137 Pass 3.23 66 Pass Fluorene 98.3 59-121 Pass 2.32 38 Pass Hexachlorobenzene 101 D-152 Pass 12.0 55 Pass Hexachloro-1,3-butadiene 100 24-120 Pass 14.6 62 Pass Hexachloroethane 72.5 40-120 Pass 13.7 52 Pass Indeno(1,2,3-cd)pyrene 95.2 D-171 Pass 15.8 99 Pass Isophorone 76.9 21-196 Pass 10.5 93 Pass Naphthalene 93.9 21-133 Pass 10.1 65 Pass Nitrobenzene 94.0 35-180 Pass 57.4 62 Pass N-Nitrosodi-n-propylamine 88.0 D-230 Pass 8.65 87 Pass Phenanthrene 96.0 54-120 Pass 7.68 128 Pass Pyrene 97.3 52-120 Pass 4.55 49 Pass 1,2,4-Trichlorobenzene 97.1 44-142 Pass 13.4 50 Pass 4-Chloro-3-methylphenol 804 22-147 14.3 73 Pass 2-Chlorophenol 78.9 23-134 Pass 15.1 61 Pass 2,4-Dichlorophenol 88.7 39-135 Pass 4.09 50 Pass 2,4-Dimethylphenol 32-120 58 2,4-Dinitrophenol D-191 132 4,6-Dinitro-2-methylphenol 57.4 D-181 Pass 19.3 203 Pass 2-Nitrophenol 64.7 29-182 Pass 23.4 55 Pass 4-Nitrophenol D-132 131 Pentachlorophenol 52.8 14-176 Pass 24.4 86 Pass Phenol 34.3 5-120 Pass 37.9 64 Pass 2,4,6-Trichlorophenol 116.8 37-144 Pass 8.62 58 Pass bis(2-chloroethyl)ether 84.2 12-158 Pass 23.1 108 Pass The samples had to be diluted by 250x to yield reasonable results for recovery. The precision was not as good as cleaner samples, but still met the criteria for most of the analytes. The next industrial wastewater sample was from the Centralized Waste Treatment Point Source Category (437). This sample could not be evaporated below 5 ml and was marked and shipped for analysis at the higher volume Page 19
Table 17. Centralized Waste Treatment Point Source (437) Results Recovery (%) Range Pass/Fail RPD RPD (%) Pass/Fail Compound 20x P,Ps(%) (%) Limit Acenaphthene 102 47-145 Pass 2.34 48 Pass Acenaphthylene 90.4 33-145 Pass 9.29 74 Pass Anthracene 106 27-133 Pass 5.02 66 Pass Benzo(a)anthracene 98.7 33-143 Pass 3.80 53 Pass Benzo(a)pyrene 99.6 17-163 Pass 5.68 72 Pass Benzo(b)fluoranthene 105 24-159 Pass 8.36 71 Pass Benzo(k)fluoranthene 115 11-162 Pass 0.35 63 Pass Benzo(g,h,i)perylene 67.5 D-219 Pass 6.55 97 Pass Benzylbutyl phthalate 104 D-152 Pass 2.47 60 Pass bis(2-chlorethoxy)methane 97.1 33-184 Pass 3.91 54 Pass bis(2-ethylhexyl)phthalate 106 36-166 Pass 3.67 76 Pass bis(2-chloroisopropyl)ether 86.2 8-158 Pass 11.2 82 Pass 4-Bromophenyl-phenylether 104 53-127 Pass 9.55 43 Pass 2-Chloronaphthalene 87.1 60-120 Pass 8.81 24 Pass 4-Chlorophenyl-phenylether 106 25-158 Pass 7.88 61 Pass Chrysene 105 17-168 Pass 3.90 87 Pass Dibenz(a,h)anthracene 74.0 D-227 Pass 4.81 126 Pass Di-n-butyl phthalate 110 1-120 Pass 6.24 47 Pass 3,3-Dichlorobenzidine 24.2 D-262 Pass 27.3 108 Pass Diethyl phthalate 116 D-120 Pass 5.33 100 Pass Dimethyl phthalate 101 D-120 Pass 3.01 183 Pass 2,4-Dinitrotoluene 105 39-139 Pass 3.35 42 Pass 2,6-Dinitrotoluene 111 50-158 Pass 8.16 48 Pass Di-n-octyl phthalate 103 4-146 Pass 4.09 69 Pass Fluoranthene 110 26-137 Pass 8.72 66 Pass Fluorene 99.6 59-121 Pass 6.89 38 Pass Hexachlorobenzene 110 D-152 Pass 9.10 55 Pass Hexachloro-1,3-butadiene 99.6 24-120 Pass 4.09 62 Pass Hexachloroethane 61.6 40-120 Pass 16.1 52 Pass Indeno(1,2,3-cd)pyrene 73.5 D-171 Pass 4.54 99 Pass Isophorone 100 21-196 Pass 2.71 93 Pass Naphthalene 96.0 21-133 Pass 2.49 65 Pass Nitrobenzene 94.2 35-180 Pass 0.79 62 Pass N-Nitrosodi-n-propylamine 113 D-230 Pass 2.55 87 Pass Phenanthrene 97.2 54-120 Pass 7.33 128 Pass Pyrene 90.9 52-120 Pass 8.40 49 Pass 1,2,4-Trichlorobenzene 90.2 44-142 Pass 5.68 50 Pass Page 20
4-Chloro-3-methylphenol 76.1 22-147 Pass 8.65 73 Pass 2-Chlorophenol 87.1 23-134 Pass 9.48 61 Pass 2,4-Dichlorophenol 17.0 39-135 35.1 50 Pass 2,4-Dimethylphenol 107 32-120 Pass 4.01 58 Pass 2,4-Dinitrophenol 21.3 D-191 Pass 30.7 132 Pass 4,6-Dinitro-2-methylphenol 70.0 D-181 Pass 4.88 203 Pass 2-Nitrophenol 95.9 29-182 Pass 2.82 55 Pass 4-Nitrophenol D-132 12.0 131 Pass Pentachlorophenol 108 14-176 Pass 6.97 86 Pass Phenol 121 5-120 Pass 3.20 64 Pass 2,4,6-Trichlorophenol 67.4 37-144 Pass 58 bis(2-chloroethyl)ether 80.1 12-158 Pass 12.2 108 Pass Although this wastewater seemed to be the most difficult to prepare, the results were very good and only a few analytes were troublesome. The next table shows the results for a wastewater sample from the meat and poultry products category. Table 18. Meat and Poultry Products Point Source Category (432) Results Recovery (%) Range Pass/Fail RPD RPD (%) Compound 5x P,Ps(%) (%) Limit Acenaphthene 77.0 47-145 Pass 2.40 48 Pass Acenaphthylene 71.1 33-145 Pass 2.64 74 Pass Anthracene 77.5 27-133 Pass 2.41 66 Pass Benzo(a)anthracene 78.1 33-143 Pass 0.14 53 Pass Benzo(a)pyrene 67.7 17-163 Pass 2.31 72 Pass Benzo(b)fluoranthene 84.0 24-159 Pass 1.39 71 Pass Benzo(k)fluoranthene 82.1 11-162 Pass 2.43 63 Pass Benzo(g,h,i)perylene 69.6 D-219 Pass 4.34 97 Pass Benzylbutyl phthalate 82.1 D-152 Pass 0.64 60 Pass bis(2-chlorethoxy)methane 73.2 33-184 Pass 0.73 54 Pass bis(2-ethylhexyl)phthalate 89.9 36-166 Pass 17.9 76 Pass bis(2-chloroisopropyl)ether 66.0 8-158 Pass 3.69 82 Pass 4-Bromophenyl-phenylether 86.3 53-127 Pass 3.97 43 Pass 2-Chloronaphthalene 74.0 60-120 Pass 3.17 24 Pass 4-Chlorophenyl-phenylether 82.6 25-158 Pass 1.43 61 Pass Chrysene 86.8 17-168 Pass 0.25 87 Pass Pass/Fai l Page 21
Dibenz(a,h)anthracene 76.1 D-227 Pass 4.02 126 Pass Di-n-butyl phthalate 89.1 1-120 Pass 4.82 47 Pass 3,3-Dichlorobenzidine 4.7 D-262 Pass 22.3 108 Pass Diethyl phthalate 89.2 D-120 Pass 1.49 100 Pass Dimethyl phthalate 85.6 D-120 Pass 0.36 183 Pass 2,4-Dinitrotoluene 84.7 39-139 Pass 3.53 42 Pass 2,6-Dinitrotoluene 86.5 50-158 Pass 1.48 48 Pass Di-n-octyl phthalate 83.6 4-146 Pass 1.02 69 Pass Fluoranthene 86.1 26-137 Pass 2.32 66 Pass Fluorene 77.1 59-121 Pass 4.31 38 Pass Hexachlorobenzene 88.7 D-152 Pass 0.95 55 Pass Hexachloro-1,3-butadiene 74.4 24-120 Pass 2.60 62 Pass Hexachloroethane 55.5 40-120 Pass 1.25 52 Pass Indeno(1,2,3-cd)pyrene 72.5 D-171 Pass 3.71 99 Pass Isophorone 76.3 21-196 Pass 3.34 93 Pass Naphthalene 71.1 21-133 Pass 0.34 65 Pass Nitrobenzene 73.5 35-180 Pass 2.07 62 Pass N-Nitrosodi-n-propylamine 69.2 D-230 Pass 4.89 87 Pass Phenanthrene 80.0 54-120 Pass 2.38 128 Pass Pyrene 75.1 52-120 Pass 0.06 49 Pass 1,2,4-Trichlorobenzene 71.8 44-142 Pass 0.63 50 Pass 4-Chloro-3-methylphenol 76.5 22-147 Pass 1.11 73 Pass 2-Chlorophenol 68.9 23-134 Pass 3.63 61 Pass 2,4-Dichlorophenol 76.6 39-135 Pass 0.74 50 Pass 2,4-Dimethylphenol 77.5 32-120 Pass 1.31 58 Pass 2,4-Dinitrophenol 101 D-191 Pass 4.41 132 Pass 4,6-Dinitro-2-methylphenol 88.9 D-181 Pass 2.82 203 Pass 2-Nitrophenol 77.8 29-182 Pass 1.43 55 Pass 4-Nitrophenol 71.9 D-132 Pass 10.3 131 Pass Pentachlorophenol 91.7 14-176 Pass 1.01 86 Pass Phenol 55.0 5-120 Pass 9.28 64 Pass 2,4,6-Trichlorophenol 78.2 37-144 Pass 0.61 58 Pass bis(2-chloroethyl)ether 63.9 12-158 Pass 2.22 108 Pass This matrix was successfully spiked and measured for all the analytes of interest. The final effluent measured was from the organic chemicals, plastics and synthetic fibers category (414). The results are shown in Table 19. Page 22
Table 19. Organic Chemicals, Plastics and Synthetic Fibers Category (414) Results Recovery (%) Range Pass/Fail RPD RPD (%) Pass/Fail Compound 20x P,Ps(%) (%) Acenaphthene 94.1 47-145 Pass 7.31 48 Pass Acenaphthylene 89.8 33-145 Pass 6.24 74 Pass Anthracene 97.6 27-133 Pass 4.74 66 Pass Benzo(a)anthracene 98.0 33-143 Pass 4.17 53 Pass Benzo(a)pyrene 102 17-163 Pass 0.00 72 Pass Benzo(b)fluoranthene 119 24-159 Pass 2.67 71 Pass Benzo(k)fluoranthene 117 11-162 Pass 7.70 63 Pass Benzo(g,h,i)perylene 45.0 D-219 Pass 4.71 97 Pass Benzylbutyl phthalate 101 D-152 Pass 2.37 60 Pass bis(2-chlorethoxy)methane 91.9 33-184 Pass 2.31 54 Pass bis(2-ethylhexyl)phthalate 120 36-166 Pass 10.1 76 Pass bis(2-chloroisopropyl)ether 82.1 8-158 Pass 4.79 82 Pass 4-Bromophenyl-phenylether 102 53-127 Pass 4.04 43 Pass 2-Chloronaphthalene 89.3 60-120 Pass 5.75 24 Pass 4-Chlorophenyl-phenylether 104 25-158 Pass 2.55 61 Pass Chrysene 101 17-168 Pass 2.37 87 Pass Dibenz(a,h)anthracene 53.4 D-227 Pass 2.72 126 Pass Di-n-butyl phthalate 119 1-120 Pass 5.99 47 Pass 3,3-Dichlorobenzidine 40.9 D-262 Pass 23.5 108 Pass Diethyl phthalate 122 D-120 7.86 100 Pass Dimethyl phthalate 110 D-120 Pass 2.85 183 Pass 2,4-Dinitrotoluene 109 39-139 Pass 0.70 42 Pass 2,6-Dinitrotoluene 101 50-158 Pass 7.64 48 Pass Di-n-octyl phthalate 89.8 4-146 Pass 8.55 69 Pass Fluoranthene 102 26-137 Pass 13.0 66 Pass Fluorene 97.1 59-121 Pass 4.98 38 Pass Hexachlorobenzene 103 D-152 Pass 6.60 55 Pass Hexachloro-1,3-butadiene 95.4 24-120 Pass 0.84 62 Pass Hexachloroethane 61.3 40-120 Pass 8.10 52 Pass Indeno(1,2,3-cd)pyrene 53.9 D-171 Pass 0.97 99 Pass Isophorone 93.9 21-196 Pass 0.45 93 Pass Naphthalene 89.1 21-133 Pass 2.57 65 Pass Nitrobenzene 89.6 35-180 Pass 4.85 62 Pass N-Nitrosodi-n-propylamine 94.1 D-230 Pass 1.03 87 Pass Phenanthrene 96.1 54-120 Pass 3.69 128 Pass Pyrene 94.9 52-120 Pass 1.99 49 Pass 1,2,4-Trichlorobenzene 82.6 44-142 Pass 12.1 50 Pass Page 23
4-Chloro-3-methylphenol 89.3 22-147 Pass 6.96 73 Pass 2-Chlorophenol 94.1 23-134 Pass 1.11 61 Pass 2,4-Dichlorophenol 99.3 39-135 Pass 1.78 50 Pass 2,4-Dimethylphenol 106 32-120 Pass 5.38 58 Pass 2,4-Dinitrophenol 125 D-191 Pass 5.50 132 Pass 4,6-Dinitro-2-methylphenol 98.9 D-181 Pass 4.72 203 Pass 2-Nitrophenol 98.4 29-182 Pass 0.34 55 Pass 4-Nitrophenol 86.0 D-132 Pass 0.48 131 Pass Pentachlorophenol 119 14-176 Pass 2.38 86 Pass Phenol 1559 5-120 3.40 64 Pass 2,4,6-Trichlorophenol 86.6 37-144 Pass 13.0 58 Pass bis(2-chloroethyl)ether 80.4 12-158 Pass 4.65 108 Pass The analytes met the criteria except for two. Diethyl phthalate was very close to the acceptable range and may be higher due to low level contamination observed in the native matrix. In the case of phenol the native matrix appeared to contain large amounts of phenol, which were not subtracted out in the recovery calculation and likely swamped the concentration of the spike if subtraction had been attempted. Overall, the wastewater matrices were successfully spiked and measured recoveries were within the range specified. The relative percent difference (RPD) between the matrix spike and duplicate sample showed excellent agreement when compared to the limit allowed. The agreement was generally well below the limit, sometimes more than an order of magnitude better. The last validation requirement is to measure a certified reference material for all of the analytes being validated for this method. We investigated this with help from Phenova and found that any one performance test material will only contain 60% of the analytes to be in compliance with NELAC.6 This is to provide a more challenging material to labs being tested that will have a different selection of analytes over the course of time. Because of the random selection of analytes in each solution approximately 7 test materials would be required to cover the full suite of analytes, increasing our expense and laboratory testing workload. We developed a different approach that would provide the same data at a more reasonable expense which included one performance test matrix and one certified material matrix. The performance test matrix had 60% of the analytes of interest present at varying concentrations. The certified material contained all of the analytes at one concentration. Tables 20 and 21 show the results for the certified materials. Table 20. WP Base Neutrals and Acids in Water (Phenova, Inc.) Results Recovery Cert Acceptance Pass/Fail Compound 1x Limits (%) Acenaphthene 78.9 35.6-122 Pass Acenaphthylene 75.4 35.1-125 Pass Anthracene Benzidine Benzo(a)anthracene 84.2 46.6-123 Pass Benzo(a)pyrene 77.2 35.6-129 Pass Benzo(b)fluoranthene Benzo(k)fluoranthene 91.6 22.1-155 Pass Page 24
Benzo(g,h,i)perylene 78.3 37.6-131 Pass Benzylbutyl phthalate bis(2-chlorethoxy)methane bis(2-ethylhexyl)phthalate 77.8 34.3-131 Pass bis(2-chloroisopropyl)ether 4-Bromophenyl-phenylether 84.7 41.5-127 Pass 2-Chloronaphthalene 4-Chlorophenyl-phenylether 83.3 39.2-126 Pass Chrysene 89.5 44.5-128 Pass Dibenz(a,h)anthracene 82.7 35.2-141 Pass Di-n-butyl phthalate 3,3-Dichlorobenzidine Diethyl phthalate 87.2 17.6-140 Pass Dimethyl phthalate 84.8 10.0-144 Pass 2,4-Dinitrotoluene 89.4 40.6-125 Pass 2,6-Dinitrotoluene 94.5 40.6-121 Pass Di-n-octyl phthalate 92.0 28.8-144 Pass Fluoranthene Fluorene 85.1 43.8-136 Pass Hexachlorobenzene 97.3 44.4-121 Pass Hexachloro-1,3-butadiene Hexachloroethane 61.2 10.0-111 Pass Indeno(1,2,3-cd)pyrene 71.2 29.2-126 Pass Isophorone 74.1 36.5-124 Pass Naphthalene Nitrobenzene N-Nitrosodi-n-propylamine Phenanthrene 84.2 46.7-123 Pass Pyrene 79.9 41.0-131 Pass 1,2,4-Trichlorobenzene 4-Chloro-3-methylphenol 82.1 38.0-124 Pass 2-Chlorophenol 71.5 28.8-120 Pass 2,4-Dichlorophenol 77.2 32.4-119 Pass 2,4-Dimethylphenol 71.3 27.6-124 Pass 2,4-Dinitrophenol 125 10.0-136 Pass 4,6-Dinitro-2-methylphenol 109 32.0-136 Pass 2-Nitrophenol 76.3 32.8-122 Pass 4-Nitrophenol 89.1 10.0-136 Pass Pentachlorophenol 97.1 33.5-134 Pass Phenol 43.8 10.0-136 Pass 2,4,6-Trichlorophenol 86.1 37.1-122 Pass bis(2-chloroethyl)ether 66.1 26.7-124 Pass Page 25
Table 21. Certified Material Results Page 26 Compound Cert Value µ/l Recovery (%) Acenaphthene 50 83.3 Acenaphthylene 50 80.6 Anthracene 50 79.9 Benzidine 50 57.4 Benzo(a)anthracene 50 87.7 Benzo(a)pyrene 50 84.2 Benzo(b)fluoranthene 50 94.1 Benzo(k)fluoranthene 50 88.9 Benzo(g,h,i)perylene 50 78.4 Benzylbutyl phthalate 50 89.3 bis(2-chlorethoxy)methane 50 77.8 bis(2-ethylhexyl)phthalate 50 91.2 bis(2-chloroisopropyl)ether 50 77.4 4-Bromophenyl-phenylether 50 89.8 2-Chloronaphthalene 50 77.5 4-Chlorophenyl-phenylether 50 85.6 Chrysene 50 93.3 Dibenz(a,h)anthracene 50 81.7 Di-n-butyl phthalate 50 96.0 3,3-Dichlorobenzidine 50 69.9 Diethyl phthalate 50 93.9 Dimethyl phthalate 50 91.5 2,4-Dinitrotoluene 50 93.6 2,6-Dinitrotoluene 50 93.5 Di-n-octyl phthalate 50 97.0 Fluoranthene 50 92.8 Fluorene 50 85.6 Hexachlorobenzene 50 91.8 Hexachloro-1,3-butadiene 50 66.0 Hexachloroethane 50 65.7 Indeno(1,2,3-cd)pyrene 50 79.9 Isophorone 50 74.1 Naphthalene 50 74.0 Nitrobenzene 50 78.1 N-Nitrosodi-n-propylamine 50 77.8 Phenanthrene 50 86.9 Pyrene 50 84.3 1,2,4-Trichlorobenzene 50 72.2 4-Chloro-3-methylphenol 50 83.3
2-Chlorophenol 50 73.5 2,4-Dichlorophenol 50 78.4 2,4-Dimethylphenol 50 70.2 2,4-Dinitrophenol 50 116 4,6-Dinitro-2-methylphenol 50 110 2-Nitrophenol 50 78.0 4-Nitrophenol 50 92.0 Pentachlorophenol 50 107 Phenol 50 45.4 2,4,6-Trichlorophenol 50 86.9 bis(2-chloroethyl)ether 50 74.0 The recoveries of compounds from both materials is very good and covers the full suite of analytes in the validation process. It meets the requirement of demonstrating recovery on a known material from a provider skilled in providing certified testing materials. Conclusion US EPA method 625 is an important method for evaluation of water pollution or clean-up. It allows a full suite of analytes to be evaluated at one time using GC/MS. Sample preparation is an important part of the process and disk solid phase extraction can provide advantages in using less solvent and providing good extraction across the suite of analytes considered while minimizing exposure. Method 625.1, currently awaiting publication in the Federal Register as part of the Method Update Rule of February 2015 (MUR) signed by the EPA Administrator on Dec 15, 2016, includes a new validation process for changes to the method such as the substitution of SPE disks for liquid-liquid extraction. The process is similar to the Alternative Test Procedure (ATP) process, but with fewer requirements and the vendor can provide the work. The validation procedure outlined in the MUR and clarified by the Guidance document was followed to show compliance with the requirements in Table 6 of method 625.1. The work was done in conjunction with an accredited environmental laboratory. A variety of wastewater samples was used to challenge the method with complex matrices with differing analytes and matrix components that may interfere with good adsorption and release of the analytes. The results show excellent performance of the One-Pass disk coupled with a carbon cartridge for capture of the analytes for all but the most difficult wastewater samples. Solid phase extraction disks are another tool for the environmental laboratory to consider when evaluating their workflow for increased efficiency and safety. Other things can be considered once analytical performance is demonstrated, such as the ability to use less solvent, and reducing the need for evaporation and subsequent solvent recollection. Automation can also enhance reproducibility. Excellent duplicate agreement was shown here, even for the more difficult samples. Overall, analytical performance meets the criteria required and other favorable factors can be included in the decision making process to bring newer technology into today s modern laboratory. Page 27
References: 1. Method Update Proposed Rule, Federal Register February 19, 2015, page 8946. 2. Method 625.1, December 14 revision, can be found in the MUR, February 20, 2014. Or downloaded here: https://nepis.epa.gov/exe/zypdf.cgi/p100lvhc.pdf?dockey=p100lvhc.pdf 3. Validation of SPE Products and Associated Procedures with Method 625.1, Lemuel Walker, EPA Office of Science and Technology, 2016. 4. Protocol for Review and Validation of New Methods for Regulated Organic and Inorganic Analytes in Wastewater Under EPA s Alternate Test Procedure Program, https://www.epa.gov/sites/production/files/2016-03/documents/chemical-newmethod-protocol_feb-2016.pdf. 5. February 2016, W.R.Jones, A. Cannon, M. Ebitson, D. Gallagher, and Z. Grosser, New Method US EPA 625 with Solid Phase Extraction for Challenging Wastewaters, AN091 (2014) available from www.horizontechinc.com. Acknowledgement: There are several groups who generously contributed wastewater samples for this project and we would like to thank: Rogers and Callcott Environmental Enersolv Analytical Laboratory Alpha Analytical MWRA, Deer Island, MA Treatment Plant City of Lawrence, MA Wastewater Treatment Plant A big thank you also goes to Ty Garber of Phenova, Inc. who helped us figure out a scheme to measure every analyte against a certified material. His ability to think outside the box was amazing and much appreciated. www.horizontechinc.com AN1171706_01 16 Northwestern Drive, Salem, NH 03079 USA Tel: (603) 893-3663 Email: Support-Service@horizontechinc.com
Appendix 1 Table 6 from US EPA Method 625.1, December 2014