Validation of Horizon Technology Disk Extraction Technology for US EPA Wastewater Method 625.1

Transcription

1 Validation of Horizon Technology Disk Extraction Technology for US EPA Wastewater Method 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 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 Nine different wastewater matrices were evaluated and tested against the criteria listed in Table 6 of method 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.

2 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

3 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

4 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 (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

5 Table 6. Description of the Wastewater Samples Evaluated (9 different matrices) Wastewater Description 1 Synthetic wastewater Prepared following ASTM D 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 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 Table 6 in US EPA Method (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 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 for compliance. Page 5

6 Table 8. Calibration Verification over the Course of Operation Recovery (%) Recovery (%) Recovery (%) Range for Q Pass/Fail Compound 0110_ _ C_02 (%) Acenaphthene Pass Acenaphthylene Pass Anthracene Pass Benzidine Benzo(a)anthracene Pass Benzo(a)pyrene Pass Benzo(b)fluoranthene Pass Benzo(k)fluoranthene Pass Benzo(g,h,i)perylene Pass Benzylbutyl phthalate Pass bis(2-chlorethoxy)methane Pass bis(2-ethylhexyl)phthalate Pass bis(2-chloroisopropyl)ether Pass 4-Bromophenyl-phenylether Pass 2-Chloronaphthalene Pass 4-Chlorophenyl-phenylether Pass Chrysene Pass Dibenz(a,h)anthracene Pass Di-n-butyl phthalate Pass 3,3-Dichlorobenzidine Pass Diethyl phthalate Pass Dimethyl phthalate Pass 2,4-Dinitrotoluene Pass 2,6-Dinitrotoluene Pass Di-n-octyl phthalate Pass Fluoranthene Pass Fluorene Pass Hexachlorobenzene Pass Hexachloro-1,3-butadiene Pass Hexachloroethane Pass Indeno(1,2,3-cd)pyrene Pass Isophorone Pass Naphthalene Pass Nitrobenzene Pass N-Nitrosodi-n-propylamine Pass Phenanthrene Pass Pyrene Pass 1,2,4-Trichlorobenzene Pass 4-Chloro-3-methylphenol Pass 2-Chlorophenol Pass Page 6

7 2,4-Dichlorophenol Pass 2,4-Dimethylphenol Pass 2,4-Dinitrophenol Pass 4,6-Dinitro-2-methylphenol Pass 2-Nitrophenol Pass 4-Nitrophenol Pass Pentachlorophenol Pass Phenol Pass 2,4,6-Trichlorophenol Pass bis(2-chloroethyl)ether 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 Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 50.5 D-195 Pass Pass Benzylbutyl phthalate 94.8 D-140 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass Pass bis(2-chloroisopropyl)ether Pass Pass 4-Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Chrysene Pass Pass Dibenz(a,h)anthracene 60.3 D-200 Pass Pass Di-n-butyl phthalate Pass Pass 3,3-Dichlorobenzidine Pass Pass Diethyl phthalate 108 D-120 Pass Pass Page 7

8 Dimethyl phthalate 102 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 57.5 D-151 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass 4-Chloro-3-methylphenol Pass Pass 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass Pass 2,4-Dimethylphenol Pass Pass 2,4-Dinitrophenol 113 D-173 Pass Pass 4,6-Dinitro-2-methylphenol Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol Pass Pass Pentachlorophenol Pass Pass Phenol Pass Pass 2,4,6-Trichlorophenol Pass Pass bis(2-chloroethyl)ether Pass 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 was used. Although only an indicator of the lower level of the method, they were compared with the MDL values listed in method 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

9 Table 10. Method Detection Limits (MDLs) Measured using SPE, compared to Method Values, using LLE Compound Measured MDL (µg/l) (SPE) MDL from Method (µg/l) (LLE) Acenaphthene Acenaphthylene Anthracene Benzo(a)anthracene Benzo(a)pyrene Benzo(b)fluoranthene Benzo(k)fluoranthene Benzo(g,h,i)perylene Benzylbutyl phthalate bis(2-chlorethoxy)methane bis(2-ethylhexyl)phthalate bis(2-chloroisopropyl)ether Bromophenyl-phenylether Chloronaphthalene Chlorophenyl-phenylether Chrysene Dibenz(a,h)anthracene Di-n-butyl phthalate ,3-Dichlorobenzidine Diethyl phthalate Dimethyl phthalate ,4-Dinitrotoluene ,6-Dinitrotoluene Di-n-octyl phthalate Fluoranthene Fluorene Hexachlorobenzene Hexachloro-1,3-butadiene Hexachloroethane Indeno(1,2,3-cd)pyrene Isophorone Naphthalene Nitrobenzene N-Nitrosodi-n-propylamine 1.67 Phenanthrene Pyrene ,2,4-Trichlorobenzene Page 9

10 4-Chloro-3-methylphenol Chlorophenol ,4-Dichlorophenol ,4-Dimethylphenol ,4-Dinitrophenol ,6-Dinitro-2-methylphenol Nitrophenol Nitrophenol Pentachlorophenol Phenol ,4,6-Trichlorophenol bis(2-chloroethyl)ether 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 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 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 , 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 Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 88.0 D-219 Pass Pass Benzylbutyl phthalate 89.8 D-152 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass Pass bis(2-chloroisopropyl)ether Pass Pass 4-Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Page 10

11 Chrysene Pass Pass Dibenz(a,h)anthracene 86.6 D-227 Pass Pass Di-n-butyl phthalate Pass Pass 3,3-Dichlorobenzidine 44.3 D-262 Pass Pass Diethyl phthalate 94.6 D-120 Pass Pass Dimethyl phthalate 92.8 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene 93.5 D-152 Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 87.1 D-171 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine 76.2 D-230 Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass 4-Chloro-3-methylphenol Pass Pass 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass Pass 2,4-Dimethylphenol Pass Pass 2,4-Dinitrophenol 132 D-191 Pass Pass 4,6-Dinitro-2-methylphenol 102 D-181 Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol 72.9 D-132 Pass Pass Pentachlorophenol Pass Pass Phenol Pass Pass 2,4,6-Trichlorophenol Pass Pass bis(2-chloroethyl)ether Pass Pass Page 11

12 The synthetic wastewater performed very well and each compound met the criteria listed in method 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 Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 85.4 D-219 Pass Pass Benzylbutyl phthalate 83.1 D-152 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass Pass bis(2-chloroisopropyl)ether Pass Pass 4-Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Chrysene Pass Pass Dibenz(a,h)anthracene 87.4 D-227 Pass Pass Di-n-butyl phthalate Pass Pass 3,3-Dichlorobenzidine 49.4 D-262 Pass Pass Diethyl phthalate 92.6 D-120 Pass Pass Dimethyl phthalate 95.7 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene 90.5 D-152 Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 86.6 D-171 Pass Pass Page 12

13 Indeno(1,2,3-cd)pyrene 86.6 D-171 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine 72.4 D-230 Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass 4-Chloro-3-methylphenol Pass Pass 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass Pass 2,4-Dimethylphenol Pass Pass 2,4-Dinitrophenol 125 D-191 Pass Pass 4,6-Dinitro-2-methylphenol 101 D-181 Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol 88.9 D-132 Pass Pass Pentachlorophenol Pass Pass Phenol Pass Pass 2,4,6-Trichlorophenol Pass Pass bis(2-chloroethyl)ether Pass 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

14 Table 13. POTW Influent from a large treatment facility results Recovery (%) Range Pass/Fail RPD (%) RPD (%) Pass/Fail Compound 20x P,Ps(%) Limit Acenaphthene Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 59.0 D-219 Pass Pass Benzylbutyl phthalate 93.1 D-152 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass Pass bis(2-chloroisopropyl)ether Pass Pass 4-Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Chrysene Pass Pass Dibenz(a,h)anthracene 66.0 D-227 Pass Pass Di-n-butyl phthalate Pass Pass 3,3-Dichlorobenzidine 21.8 D-262 Pass Pass Diethyl phthalate 116 D-120 Pass Pass Dimethyl phthalate 95.7 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene 98.6 D-152 Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 65.2 D-171 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine 84.0 D-230 Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass 4-Chloro-3-methylphenol Pass Pass Page 14

15 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass Pass 2,4-Dimethylphenol Pass Pass 2,4-Dinitrophenol 99.8 D-191 Pass Pass 4,6-Dinitro-2-methylphenol 84.4 D-181 Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol 87.7 D-132 Pass Pass Pentachlorophenol Pass Pass Phenol Pass Pass 2,4,6-Trichlorophenol Pass Pass bis(2-chloroethyl)ether Pass 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 Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 44.4 D-219 Pass Pass Benzylbutyl phthalate 73.7 D-152 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass Pass bis(2-chloroisopropyl)ether Pass Pass 4-Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Chrysene Pass Pass Dibenz(a,h)anthracene 51.6 D-227 Pass Pass Di-n-butyl phthalate Pass Pass 3,3-Dichlorobenzidine 13.7 D-262 Pass Pass Page 15

16 Diethyl phthalate 84.3 D-120 Pass Pass Dimethyl phthalate 80.2 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene 80.4 D-152 Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 50.9 D-171 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine 68.3 D-230 Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass 4-Chloro-3-methylphenol Pass Pass 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass Pass 2,4-Dimethylphenol Pass Pass 2,4-Dinitrophenol 95.1 D-191 Pass Pass 4,6-Dinitro-2-methylphenol 84.9 D-181 Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol 60.9 D-132 Pass Pass Pentachlorophenol Pass Pass Phenol Pass Pass 2,4,6-Trichlorophenol Pass Pass bis(2-chloroethyl)ether Pass 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

17 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 Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 40.2 D-219 Pass Pass Benzylbutyl phthalate 88.9 D-152 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass Pass bis(2-chloroisopropyl)ether Pass Pass 4-Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Chrysene Pass Pass Dibenz(a,h)anthracene 48.3 D-227 Pass Pass Di-n-butyl phthalate Pass Pass 3,3-Dichlorobenzidine 17.7 D-262 Pass Pass Diethyl phthalate 95.0 D-120 Pass Pass Dimethyl phthalate 92.5 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene 93.1 D-152 Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 46.5 D-171 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine 75.2 D-230 Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass 4-Chloro-3-methylphenol Pass Pass Page 17

18 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass Pass 2,4-Dimethylphenol Pass Pass 2,4-Dinitrophenol 71.4 D-191 Pass Pass 4,6-Dinitro-2-methylphenol 72.6 D-181 Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol 60.8 D-132 Pass Pass Pentachlorophenol Pass Pass Phenol Pass Pass 2,4,6-Trichlorophenol Pass Pass bis(2-chloroethyl)ether Pass 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 Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 90.0 D-219 Pass Pass Benzylbutyl phthalate 125 D-152 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass bis(2-chloroisopropyl)ether Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Chrysene Pass Pass Dibenz(a,h)anthracene 88.7 D-227 Pass Pass Di-n-butyl phthalate Pass 3,3-Dichlorobenzidine D Diethyl phthalate 108 D-120 Pass Pass Page 18

19 Dimethyl phthalate 93.5 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene 101 D-152 Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 95.2 D-171 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine 88.0 D-230 Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass 4-Chloro-3-methylphenol Pass 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass Pass 2,4-Dimethylphenol ,4-Dinitrophenol D ,6-Dinitro-2-methylphenol 57.4 D-181 Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol D Pentachlorophenol Pass Pass Phenol Pass Pass 2,4,6-Trichlorophenol Pass Pass bis(2-chloroethyl)ether Pass 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

20 Table 17. Centralized Waste Treatment Point Source (437) Results Recovery (%) Range Pass/Fail RPD RPD (%) Pass/Fail Compound 20x P,Ps(%) (%) Limit Acenaphthene Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 67.5 D-219 Pass Pass Benzylbutyl phthalate 104 D-152 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass Pass bis(2-chloroisopropyl)ether Pass Pass 4-Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Chrysene Pass Pass Dibenz(a,h)anthracene 74.0 D-227 Pass Pass Di-n-butyl phthalate Pass Pass 3,3-Dichlorobenzidine 24.2 D-262 Pass Pass Diethyl phthalate 116 D-120 Pass Pass Dimethyl phthalate 101 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene 110 D-152 Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 73.5 D-171 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine 113 D-230 Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass Page 20

21 4-Chloro-3-methylphenol Pass Pass 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass 2,4-Dimethylphenol Pass Pass 2,4-Dinitrophenol 21.3 D-191 Pass Pass 4,6-Dinitro-2-methylphenol 70.0 D-181 Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol D Pass Pentachlorophenol Pass Pass Phenol Pass Pass 2,4,6-Trichlorophenol Pass 58 bis(2-chloroethyl)ether Pass 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 Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 69.6 D-219 Pass Pass Benzylbutyl phthalate 82.1 D-152 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass Pass bis(2-chloroisopropyl)ether Pass Pass 4-Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Chrysene Pass Pass Pass/Fai l Page 21

22 Dibenz(a,h)anthracene 76.1 D-227 Pass Pass Di-n-butyl phthalate Pass Pass 3,3-Dichlorobenzidine 4.7 D-262 Pass Pass Diethyl phthalate 89.2 D-120 Pass Pass Dimethyl phthalate 85.6 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene 88.7 D-152 Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 72.5 D-171 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine 69.2 D-230 Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass 4-Chloro-3-methylphenol Pass Pass 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass Pass 2,4-Dimethylphenol Pass Pass 2,4-Dinitrophenol 101 D-191 Pass Pass 4,6-Dinitro-2-methylphenol 88.9 D-181 Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol 71.9 D-132 Pass Pass Pentachlorophenol Pass Pass Phenol Pass Pass 2,4,6-Trichlorophenol Pass Pass bis(2-chloroethyl)ether Pass 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

23 Table 19. Organic Chemicals, Plastics and Synthetic Fibers Category (414) Results Recovery (%) Range Pass/Fail RPD RPD (%) Pass/Fail Compound 20x P,Ps(%) (%) Acenaphthene Pass Pass Acenaphthylene Pass Pass Anthracene Pass Pass Benzo(a)anthracene Pass Pass Benzo(a)pyrene Pass Pass Benzo(b)fluoranthene Pass Pass Benzo(k)fluoranthene Pass Pass Benzo(g,h,i)perylene 45.0 D-219 Pass Pass Benzylbutyl phthalate 101 D-152 Pass Pass bis(2-chlorethoxy)methane Pass Pass bis(2-ethylhexyl)phthalate Pass Pass bis(2-chloroisopropyl)ether Pass Pass 4-Bromophenyl-phenylether Pass Pass 2-Chloronaphthalene Pass Pass 4-Chlorophenyl-phenylether Pass Pass Chrysene Pass Pass Dibenz(a,h)anthracene 53.4 D-227 Pass Pass Di-n-butyl phthalate Pass Pass 3,3-Dichlorobenzidine 40.9 D-262 Pass Pass Diethyl phthalate 122 D Pass Dimethyl phthalate 110 D-120 Pass Pass 2,4-Dinitrotoluene Pass Pass 2,6-Dinitrotoluene Pass Pass Di-n-octyl phthalate Pass Pass Fluoranthene Pass Pass Fluorene Pass Pass Hexachlorobenzene 103 D-152 Pass Pass Hexachloro-1,3-butadiene Pass Pass Hexachloroethane Pass Pass Indeno(1,2,3-cd)pyrene 53.9 D-171 Pass Pass Isophorone Pass Pass Naphthalene Pass Pass Nitrobenzene Pass Pass N-Nitrosodi-n-propylamine 94.1 D-230 Pass Pass Phenanthrene Pass Pass Pyrene Pass Pass 1,2,4-Trichlorobenzene Pass Pass Page 23

24 4-Chloro-3-methylphenol Pass Pass 2-Chlorophenol Pass Pass 2,4-Dichlorophenol Pass Pass 2,4-Dimethylphenol Pass Pass 2,4-Dinitrophenol 125 D-191 Pass Pass 4,6-Dinitro-2-methylphenol 98.9 D-181 Pass Pass 2-Nitrophenol Pass Pass 4-Nitrophenol 86.0 D-132 Pass Pass Pentachlorophenol Pass Pass Phenol Pass 2,4,6-Trichlorophenol Pass Pass bis(2-chloroethyl)ether Pass 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 Pass Acenaphthylene Pass Anthracene Benzidine Benzo(a)anthracene Pass Benzo(a)pyrene Pass Benzo(b)fluoranthene Benzo(k)fluoranthene Pass Page 24

25 Benzo(g,h,i)perylene Pass Benzylbutyl phthalate bis(2-chlorethoxy)methane bis(2-ethylhexyl)phthalate Pass bis(2-chloroisopropyl)ether 4-Bromophenyl-phenylether Pass 2-Chloronaphthalene 4-Chlorophenyl-phenylether Pass Chrysene Pass Dibenz(a,h)anthracene Pass Di-n-butyl phthalate 3,3-Dichlorobenzidine Diethyl phthalate Pass Dimethyl phthalate Pass 2,4-Dinitrotoluene Pass 2,6-Dinitrotoluene Pass Di-n-octyl phthalate Pass Fluoranthene Fluorene Pass Hexachlorobenzene Pass Hexachloro-1,3-butadiene Hexachloroethane Pass Indeno(1,2,3-cd)pyrene Pass Isophorone Pass Naphthalene Nitrobenzene N-Nitrosodi-n-propylamine Phenanthrene Pass Pyrene Pass 1,2,4-Trichlorobenzene 4-Chloro-3-methylphenol Pass 2-Chlorophenol Pass 2,4-Dichlorophenol Pass 2,4-Dimethylphenol Pass 2,4-Dinitrophenol Pass 4,6-Dinitro-2-methylphenol Pass 2-Nitrophenol Pass 4-Nitrophenol Pass Pentachlorophenol Pass Phenol Pass 2,4,6-Trichlorophenol Pass bis(2-chloroethyl)ether Pass Page 25

26 Table 21. Certified Material Results Page 26 Compound Cert Value µ/l Recovery (%) Acenaphthene Acenaphthylene Anthracene Benzidine Benzo(a)anthracene Benzo(a)pyrene Benzo(b)fluoranthene Benzo(k)fluoranthene Benzo(g,h,i)perylene Benzylbutyl phthalate bis(2-chlorethoxy)methane bis(2-ethylhexyl)phthalate bis(2-chloroisopropyl)ether Bromophenyl-phenylether Chloronaphthalene Chlorophenyl-phenylether Chrysene Dibenz(a,h)anthracene Di-n-butyl phthalate ,3-Dichlorobenzidine Diethyl phthalate Dimethyl phthalate ,4-Dinitrotoluene ,6-Dinitrotoluene Di-n-octyl phthalate Fluoranthene Fluorene Hexachlorobenzene Hexachloro-1,3-butadiene Hexachloroethane Indeno(1,2,3-cd)pyrene Isophorone Naphthalene Nitrobenzene N-Nitrosodi-n-propylamine Phenanthrene Pyrene ,2,4-Trichlorobenzene Chloro-3-methylphenol

27 2-Chlorophenol ,4-Dichlorophenol ,4-Dimethylphenol ,4-Dinitrophenol ,6-Dinitro-2-methylphenol Nitrophenol Nitrophenol Pentachlorophenol Phenol ,4,6-Trichlorophenol bis(2-chloroethyl)ether 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 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

28 References: 1. Method Update Proposed Rule, Federal Register February 19, 2015, page Method 625.1, December 14 revision, can be found in the MUR, February 20, Or downloaded here: 3. Validation of SPE Products and Associated Procedures with Method 625.1, Lemuel Walker, EPA Office of Science and Technology, Protocol for Review and Validation of New Methods for Regulated Organic and Inorganic Analytes in Wastewater Under EPA s Alternate Test Procedure Program, 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 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. AN _01 16 Northwestern Drive, Salem, NH USA Tel: (603) Support-Service@horizontechinc.com

29 Appendix 1 Table 6 from US EPA Method 625.1, December 2014