Hexane Extractable Material (Oil & Grease) Analysis with Automated Solid Phase Extraction Using the Biotage® Horizon 5000Melissa Lever, Biotage.

IntroductionHexane extractable material (HEM), also known as Oil & Grease extraction is a simple water quality measurement. The measurement can be used to assess pollution for compliance or as a measure of influent contamination to protect a waste treatment plant from levels that might cause malfunction. This measurement is used around the world and regulatory agencies have developed methods such as US EPA 1664B, ISO 11349, and Standard Methods 5520G.1,2,3 We have shown the feasibility of Oil & Grease measurement on a dedicated automated system, the Biotage® Horizon 3100 (previously known as the SPE-DEX® 3100).4 In this work we will demonstrate the performance of a general purpose automated Oil & Grease and semivolatile extraction system, the Biotage® Horizon 5000.

The performance of the system for this application will be demonstrated by showing compliance with the quality control requirement of US EPA Method 1664B for small modifications. A new extraction procedure would be considered a modifica-tion to the method; therefore the laboratory is required to repeat the initial precision and recovery (IPR) study in Section 9.2.2 9 to demonstrate that the modification produces results equivalent to or superior to results produced by this method. If the detection limit is suspected to be affected by the new modification a Method Detection Limit study (MDL) would need to be demonstrated. In this case the same disk and chemistry is used, only the automation has changed. 5

In this study the laboratory confirming the performance of each typical solid phase extraction (SPE) configuration on the Biotage® Horizon 5000 by conducting an IPR study to satisfy the quality control requirements for a method modification.

Instrumentation » Biotage

» Biotage® Horizon 5000 (previously known as the SPE-DEX 5000) Extractor

» Oil and Grease Standard 40 mg

» Pacific® Premium disks (90 mm and 47 mm)

» Atlantic® Fast Flow Pre-Filters (90 mm and 47 mm)

» Dry Disk® Solvent Drying System (SDS)

» DryDisk® Membranes

» Speed-Vap® IV Automated Evaporation System

» Aluminum Pans

» Ohaus® Balance

» Adventurer Pro Model: AV64

» Max: 65 g, d=0.0001, 10 oC/30 oC

Hexane Extractable Material (Oil & Grease) Analysis with Automated Solid Phase Extraction Using the Biotage® Horizon 5000

Melissa Lever, Biotage

Key Words

Hexane extractable material, oil & grease, method 1664B, wastewater

Introduction

Hexane extractable material (HEM), also known as Oil & Grease extraction is a simple water quality measurement. The measurement can be used to assess pollution for compliance or as a measure of influent contamination to protect a waste treatment plant from levels that might cause malfunction. This measurement is used around the world and regulatory agencies have developed methods such as US EPA 1664B, ISO 11349, and Standard Methods 5520G.1,2,3 We have shown the feasibility of Oil & Grease measurement on a dedicated automated system, the Biotage® Horizon 3100 (previously known as the SPE-DEX® 3100).4 In this work we will demonstrate the performance of a general purpose automated Oil &Grease and semivolatile extraction system, the Biotage® Horizon SPE-DEX® 5000.

The performance of the system for this application will be demonstrated by showing compliance with the quality control requirement of US EPA Method 1664B for small modifications. A new extraction procedure would be considered a modification to the method; therefore the laboratory is required to repeat the initial precision and recovery (IPR) study in Section 9.2.2 9 to demonstrate that the modification produces results equivalent to or superior to results produced by this method. If the detection limit is suspected to be affected by the new modification a Method Detection Limit study (MDL) would need to be demonstrated. In this case the same disk and chemistry is used, only the automation has changed. 5

In this study the laboratory confirming the performance of each typical solid phase extraction (SPE) configuration on the Biotage® Horizon SPE-DEX- 5000 byconducting an IPR study to satisfy the quality control requirements for a method modification.

Instrumentation

Biotage Biotage® Horizon 5000 (previously known as the 5000)

Extractor Snip and Pour Oil and Grease Standard Pacific® Premium disks

(90 mm and 47 mm) Atlantic® Fast Flow Pre-Filters (90-mm and 47 mm) Dry Disk® Solvent Drying System (SDS) DryDisk® Membranes Speed-Vap® IV Automated Evaporation System Aluminum Pans

Hexane Extractable Material (Oil & Grease) Analysis with Automated Solid Phase Extraction Using the Biotage® Horizon 5000

Melissa Lever, Biotage

Key Words

Hexane extractable material, oil & grease, method 1664B, wastewater

Introduction

Hexane extractable material (HEM), also known as Oil & Grease extraction is a simple water quality measurement. The measurement can be used to assess pollution for compliance or as a measure of influent contamination to protect a waste treatment plant from levels that might cause malfunction. This measurement is used around the world and regulatory agencies have developed methods such as US EPA 1664B, ISO 11349, and Standard Methods 5520G.1,2,3 We have shown the feasibility of Oil & Grease measurement on a dedicated automated system, the Biotage® Horizon 3100 (previously known as the SPE-DEX® 3100).4 In this work we will demonstrate the performance of a general purpose automated Oil &Grease and semivolatile extraction system, the Biotage® Horizon SPE-DEX® 5000.

The performance of the system for this application will be demonstrated by showing compliance with the quality control requirement of US EPA Method 1664B for small modifications. A new extraction procedure would be considered a modification to the method; therefore the laboratory is required to repeat the initial precision and recovery (IPR) study in Section 9.2.2 9 to demonstrate that the modification produces results equivalent to or superior to results produced by this method. If the detection limit is suspected to be affected by the new modification a Method Detection Limit study (MDL) would need to be demonstrated. In this case the same disk and chemistry is used, only the automation has changed. 5

In this study the laboratory confirming the performance of each typical solid phase extraction (SPE) configuration on the Biotage® Horizon SPE-DEX- 5000 byconducting an IPR study to satisfy the quality control requirements for a method modification.

Instrumentation

Biotage Biotage® Horizon 5000 (previously known as the 5000)

Extractor Snip and Pour Oil and Grease Standard Pacific® Premium disks

(90 mm and 47 mm) Atlantic® Fast Flow Pre-Filters (90-mm and 47 mm) Dry Disk® Solvent Drying System (SDS) DryDisk® Membranes Speed-Vap® IV Automated Evaporation System Aluminum Pans

© Biotage 2019

Hexane Extractable Material (Oil & Grease) Analysis with Automated Solid Phase Extraction | Page 1

Page 2

Ohaus® Balance Adventurer Pro Model: AV64 Max: 65g, d=0.0001, 10oC/30oC

Method Summary

Obtain sixteen 1 liter samples of DI water.

Acidify each sample with hydrochloric acid until pH <2.

Add one Snip and Pour Oil and Grease Standard to each bottle (total concentration of 40 ppm). Extract four samples using the SPE-DEX 5000 with 100-mm Pacific® Premium SPE Disks using the method shown in Table 1.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks using the method shown in Table 2.

Extract four samples using the SPE-DEX 5000 with 100-mm Pacific®Premium SPE Disks with Pre-filters using the methodshown in Table 3.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks with Pre-filters using the methodshownin Table 4.

Dry the extracts using the SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for somemethods, check with your regulatory agency).

Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan. Use the Speed-Vap® IV Evaporation System to evaporate each extract. Weigh each pan and record the weight difference as mg/L of HEM.

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 20 30

9. Elute Sample Container Hexane 22 35 5 Off 1 45 45

10. Elute Sample Container Hexane 22 60 6 Off 1 45 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 45 60

Table 1. SPE-DEX 5000 Extraction Method for 100-mm Pacific Premium Disks (No Pre-Filters)

Page 2

Ohaus® Balance Adventurer Pro Model: AV64 Max: 65g, d=0.0001, 10oC/30oC

Method Summary

Obtain sixteen 1 liter samples of DI water.

Acidify each sample with hydrochloric acid until pH <2.

Add one Snip and Pour Oil and Grease Standard to each bottle (total concentration of 40 ppm). Extract four samples using the SPE-DEX 5000 with 100-mm Pacific® Premium SPE Disks using the method shown in Table 1.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks using the method shown in Table 2.

Extract four samples using the SPE-DEX 5000 with 100-mm Pacific®Premium SPE Disks with Pre-filters using the methodshown in Table 3.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks with Pre-filters using the methodshownin Table 4.

Dry the extracts using the SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for somemethods, check with your regulatory agency).

Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan. Use the Speed-Vap® IV Evaporation System to evaporate each extract. Weigh each pan and record the weight difference as mg/L of HEM.

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 20 30

9. Elute Sample Container Hexane 22 35 5 Off 1 45 45

10. Elute Sample Container Hexane 22 60 6 Off 1 45 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 45 60

Table 1. SPE-DEX 5000 Extraction Method for 100-mm Pacific Premium Disks (No Pre-Filters)

Page 2

Ohaus® Balance Adventurer Pro Model: AV64 Max: 65g, d=0.0001, 10oC/30oC

Method Summary

Obtain sixteen 1 liter samples of DI water.

Acidify each sample with hydrochloric acid until pH <2.

Add one Snip and Pour Oil and Grease Standard to each bottle (total concentration of 40 ppm). Extract four samples using the SPE-DEX 5000 with 100-mm Pacific® Premium SPE Disks using the method shown in Table 1.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks using the method shown in Table 2.

Extract four samples using the SPE-DEX 5000 with 100-mm Pacific®Premium SPE Disks with Pre-filters using the methodshown in Table 3.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks with Pre-filters using the methodshownin Table 4.

Dry the extracts using the SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for somemethods, check with your regulatory agency).

Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan. Use the Speed-Vap® IV Evaporation System to evaporate each extract. Weigh each pan and record the weight difference as mg/L of HEM.

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 20 30

9. Elute Sample Container Hexane 22 35 5 Off 1 45 45

10. Elute Sample Container Hexane 22 60 6 Off 1 45 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 45 60

Table 1. SPE-DEX 5000 Extraction Method for 100-mm Pacific Premium Disks (No Pre-Filters)

Page 2

Ohaus® Balance Adventurer Pro Model: AV64 Max: 65g, d=0.0001, 10oC/30oC

Method Summary

Obtain sixteen 1 liter samples of DI water.

Acidify each sample with hydrochloric acid until pH <2.

Add one Snip and Pour Oil and Grease Standard to each bottle (total concentration of 40 ppm). Extract four samples using the SPE-DEX 5000 with 100-mm Pacific® Premium SPE Disks using the method shown in Table 1.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks using the method shown in Table 2.

Extract four samples using the SPE-DEX 5000 with 100-mm Pacific®Premium SPE Disks with Pre-filters using the methodshown in Table 3.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks with Pre-filters using the methodshownin Table 4.

Dry the extracts using the SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for somemethods, check with your regulatory agency).

Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan. Use the Speed-Vap® IV Evaporation System to evaporate each extract. Weigh each pan and record the weight difference as mg/L of HEM.

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 20 30

9. Elute Sample Container Hexane 22 35 5 Off 1 45 45

10. Elute Sample Container Hexane 22 60 6 Off 1 45 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 45 60

Table 1. SPE-DEX 5000 Extraction Method for 100-mm Pacific Premium Disks (No Pre-Filters)

Page 2

Ohaus® Balance Adventurer Pro Model: AV64 Max: 65g, d=0.0001, 10oC/30oC

Method Summary

Obtain sixteen 1 liter samples of DI water.

Acidify each sample with hydrochloric acid until pH <2.

Add one Snip and Pour Oil and Grease Standard to each bottle (total concentration of 40 ppm). Extract four samples using the SPE-DEX 5000 with 100-mm Pacific® Premium SPE Disks using the method shown in Table 1.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks using the method shown in Table 2.

Extract four samples using the SPE-DEX 5000 with 100-mm Pacific®Premium SPE Disks with Pre-filters using the methodshown in Table 3.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks with Pre-filters using the methodshownin Table 4.

Dry the extracts using the SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for somemethods, check with your regulatory agency).

Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan. Use the Speed-Vap® IV Evaporation System to evaporate each extract. Weigh each pan and record the weight difference as mg/L of HEM.

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 20 30

9. Elute Sample Container Hexane 22 35 5 Off 1 45 45

10. Elute Sample Container Hexane 22 60 6 Off 1 45 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 45 60

Table 1. SPE-DEX 5000 Extraction Method for 100-mm Pacific Premium Disks (No Pre-Filters)

Page 2

Ohaus® Balance Adventurer Pro Model: AV64 Max: 65g, d=0.0001, 10oC/30oC

Method Summary

Obtain sixteen 1 liter samples of DI water.

Acidify each sample with hydrochloric acid until pH <2.

Add one Snip and Pour Oil and Grease Standard to each bottle (total concentration of 40 ppm). Extract four samples using the SPE-DEX 5000 with 100-mm Pacific® Premium SPE Disks using the method shown in Table 1.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks using the method shown in Table 2.

Extract four samples using the SPE-DEX 5000 with 100-mm Pacific®Premium SPE Disks with Pre-filters using the methodshown in Table 3.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks with Pre-filters using the methodshownin Table 4.

Dry the extracts using the SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for somemethods, check with your regulatory agency).

Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan. Use the Speed-Vap® IV Evaporation System to evaporate each extract. Weigh each pan and record the weight difference as mg/L of HEM.

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 20 30

9. Elute Sample Container Hexane 22 35 5 Off 1 45 45

10. Elute Sample Container Hexane 22 60 6 Off 1 45 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 45 60

Table 1. SPE-DEX 5000 Extraction Method for 100-mm Pacific Premium Disks (No Pre-Filters)

Page 2

Ohaus® Balance Adventurer Pro Model: AV64 Max: 65g, d=0.0001, 10oC/30oC

Method Summary

Obtain sixteen 1 liter samples of DI water.

Acidify each sample with hydrochloric acid until pH <2.

Add one Snip and Pour Oil and Grease Standard to each bottle (total concentration of 40 ppm). Extract four samples using the SPE-DEX 5000 with 100-mm Pacific® Premium SPE Disks using the method shown in Table 1.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks using the method shown in Table 2.

Extract four samples using the SPE-DEX 5000 with 100-mm Pacific®Premium SPE Disks with Pre-filters using the methodshown in Table 3.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks with Pre-filters using the methodshownin Table 4.

Dry the extracts using the SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for somemethods, check with your regulatory agency).

Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan. Use the Speed-Vap® IV Evaporation System to evaporate each extract. Weigh each pan and record the weight difference as mg/L of HEM.

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 20 30

9. Elute Sample Container Hexane 22 35 5 Off 1 45 45

10. Elute Sample Container Hexane 22 60 6 Off 1 45 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 45 60

Table 1. SPE-DEX 5000 Extraction Method for 100-mm Pacific Premium Disks (No Pre-Filters)

Page 2

Ohaus® Balance Adventurer Pro Model: AV64 Max: 65g, d=0.0001, 10oC/30oC

Method Summary

Obtain sixteen 1 liter samples of DI water.

Acidify each sample with hydrochloric acid until pH <2.

Add one Snip and Pour Oil and Grease Standard to each bottle (total concentration of 40 ppm). Extract four samples using the SPE-DEX 5000 with 100-mm Pacific® Premium SPE Disks using the method shown in Table 1.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks using the method shown in Table 2.

Extract four samples using the SPE-DEX 5000 with 100-mm Pacific®Premium SPE Disks with Pre-filters using the methodshown in Table 3.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks with Pre-filters using the methodshownin Table 4.

Dry the extracts using the SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for somemethods, check with your regulatory agency).

Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan. Use the Speed-Vap® IV Evaporation System to evaporate each extract. Weigh each pan and record the weight difference as mg/L of HEM.

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 20 30

9. Elute Sample Container Hexane 22 35 5 Off 1 45 45

10. Elute Sample Container Hexane 22 60 6 Off 1 45 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 45 60

Table 1. SPE-DEX 5000 Extraction Method for 100-mm Pacific Premium Disks (No Pre-Filters)

Page 2

Ohaus® Balance Adventurer Pro Model: AV64 Max: 65g, d=0.0001, 10oC/30oC

Method Summary

Obtain sixteen 1 liter samples of DI water.

Acidify each sample with hydrochloric acid until pH <2.

Add one Snip and Pour Oil and Grease Standard to each bottle (total concentration of 40 ppm). Extract four samples using the SPE-DEX 5000 with 100-mm Pacific® Premium SPE Disks using the method shown in Table 1.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks using the method shown in Table 2.

Extract four samples using the SPE-DEX 5000 with 100-mm Pacific®Premium SPE Disks with Pre-filters using the methodshown in Table 3.

Extract four samples using the SPE-DEX 5000 with 47-mm Pacific® Premium SPE Disks with Pre-filters using the methodshownin Table 4.

Dry the extracts using the SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for somemethods, check with your regulatory agency).

Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan. Use the Speed-Vap® IV Evaporation System to evaporate each extract. Weigh each pan and record the weight difference as mg/L of HEM.

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 20 30

9. Elute Sample Container Hexane 22 35 5 Off 1 45 45

10. Elute Sample Container Hexane 22 60 6 Off 1 45 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 45 60

Table 1. SPE-DEX 5000 Extraction Method for 100-mm Pacific Premium Disks (No Pre-Filters) Table 1. Biotage® Horizon 5000 Extraction Method for 90 mm Pacific® Premium Disks (No Pre-Filters).

Page 3

Table 2. SPE-DEX 5000 Extraction Method for 47-mm Pacific Premium Disks (No Pre-Filters)

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 16 35 5 Off 1 45 45

13. Elute Sample Container Hexane 16 35 5 Off 1 45 45

14. Elute Sample Container Hexane 16 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 11 60 2 1 60 60

2. Condition SPE Disk MeOH 11 60 2 1 60 2

Step Sample Flow Rate (#)

6. Load Sample 5

Done Loading Sample Delay (s)

45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 16 35 5 Off 1 20 30

9. Elute Sample Container Hexane 11 35 5 Off 1 45 45

10. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 11 35 5 Off 1 45 45

13. Elute Sample Container Hexane 11 35 5 Off 1 45 45

14. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Page 3

Table 2. SPE-DEX 5000 Extraction Method for 47-mm Pacific Premium Disks (No Pre-Filters)

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 16 35 5 Off 1 45 45

13. Elute Sample Container Hexane 16 35 5 Off 1 45 45

14. Elute Sample Container Hexane 16 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 11 60 2 1 60 60

2. Condition SPE Disk MeOH 11 60 2 1 60 2

Step Sample Flow Rate (#)

6. Load Sample 5

Done Loading Sample Delay (s)

45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 16 35 5 Off 1 20 30

9. Elute Sample Container Hexane 11 35 5 Off 1 45 45

10. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 11 35 5 Off 1 45 45

13. Elute Sample Container Hexane 11 35 5 Off 1 45 45

14. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Method Summary1. Obtain sixteen 1 L samples of DI water.

2. Acidify each sample with hydrochloric acid until pH <2.

3. Add one Oil and Grease 40 mg Standard to each bottle (total concentration of 40 ppm).

4. Extract four samples using the Biotage® Horizon 5000 with 90 mm Pacific® Premium SPE Disks using the method shown in Table 1.

5. Extract four samples using the Biotage® Horizon 5000 with 47 mm Pacific® Premium SPE Disks using the method shown in Table 2.

6. Extract four samples using the Biotage® Horizon 5000 with 90 mm Pacific® Premium SPE Disks with Pre-filters using the method shown in Table 3.

7. Extract four samples using the Biotage® Horizon 5000 with 47 mm Pacific® Premium SPE Disks with Pre-filters using the method shown in Table 4.

8. Dry the extracts using the Dry Disk® SDS with a new DryDisk for each sample (Sodium sulfate and decanting are allowed for some methods, check with your regulatory agency).

9. Label and pre-weigh twenty-four aluminum pans and add the extract of one sample to each pan.

10. Use the Speed-Vap® IV Evaporation System to evaporate each extract.

11. Weigh each pan and record the weight difference as mg/L of HEM.

Page 3

Table 2. SPE-DEX 5000 Extraction Method for 47-mm Pacific Premium Disks (No Pre-Filters)

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 16 35 5 Off 1 45 45

13. Elute Sample Container Hexane 16 35 5 Off 1 45 45

14. Elute Sample Container Hexane 16 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 11 60 2 1 60 60

2. Condition SPE Disk MeOH 11 60 2 1 60 2

Step Sample Flow Rate (#)

6. Load Sample 5

Done Loading Sample Delay (s)

45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 16 35 5 Off 1 20 30

9. Elute Sample Container Hexane 11 35 5 Off 1 45 45

10. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 11 35 5 Off 1 45 45

13. Elute Sample Container Hexane 11 35 5 Off 1 45 45

14. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Page 3

Table 2. SPE-DEX 5000 Extraction Method for 47-mm Pacific Premium Disks (No Pre-Filters)

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 16 35 5 Off 1 45 45

13. Elute Sample Container Hexane 16 35 5 Off 1 45 45

14. Elute Sample Container Hexane 16 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 11 60 2 1 60 60

2. Condition SPE Disk MeOH 11 60 2 1 60 2

Step Sample Flow Rate (#)

6. Load Sample 5

Done Loading Sample Delay (s)

45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 16 35 5 Off 1 20 30

9. Elute Sample Container Hexane 11 35 5 Off 1 45 45

10. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 11 35 5 Off 1 45 45

13. Elute Sample Container Hexane 11 35 5 Off 1 45 45

14. Elute Sample Container Hexane 11 60 6 Off 1 45 60

© Biotage 2019

Hexane Extractable Material (Oil & Grease) Analysis with Automated Solid Phase Extraction | Page 2

Page 3

Table 2. SPE-DEX 5000 Extraction Method for 47-mm Pacific Premium Disks (No Pre-Filters)

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 16 35 5 Off 1 45 45

13. Elute Sample Container Hexane 16 35 5 Off 1 45 45

14. Elute Sample Container Hexane 16 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 11 60 2 1 60 60

2. Condition SPE Disk MeOH 11 60 2 1 60 2

Step Sample Flow Rate (#)

6. Load Sample 5

Done Loading Sample Delay (s)

45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 180 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 16 35 5 Off 1 20 30

9. Elute Sample Container Hexane 11 35 5 Off 1 45 45

10. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 11 60 6 Off 1 45 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 11 35 5 Off 1 45 45

13. Elute Sample Container Hexane 11 35 5 Off 1 45 45

14. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Table 2. Biotage® Horizon 5000 Extraction Method for 47 mm Pacific® Premium Disks (No Pre-Filters).

© Biotage 2019

Hexane Extractable Material (Oil & Grease) Analysis with Automated Solid Phase Extraction | Page 3

Page 4

Table 3. SPE-DEX 5000 Extraction Method for 100 -mm Pacific Premium Disks with Pre-Filters

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 16 60 2 1 60 60

2. Condition SPE Disk MeOH 16 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 480 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 28 35 5 Off 1 25 30

9. Elute Sample Container Hexane 22 35 5 Off 1 50 45

10. Elute Sample Container Hexane 22 60 6 Off 1 50 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 16 60 6 Off 1 50 60

Step Solvent Solvent Vol. Purge Pump N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 16 35 5 Off 1 45 45

13. Elute Sample Container Hexane 16 35 5 Off 1 45 45

14. Elute Sample Container Hexane 16 60 6 Off 1 45 60

Table 3. Biotage® Horizon 5000 Extraction Method for 90 mm Pacific® Premium Disks with Pre-Filters.

© Biotage 2019

Hexane Extractable Material (Oil & Grease) Analysis with Automated Solid Phase Extraction | Page 4

Page 5

Table 4. SPE-DEX 5000 Extraction Method for 47-mm Pacific Premium Disks with Pre-Filters

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 11 60 2 1 60 60

2. Condition SPE Disk MeOH 11 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 300 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 16 35 5 Off 1 23 30

9. Elute Sample Container Hexane 11 35 5 Off 1 48 45

10. Elute Sample Container Hexane 11 60 6 Off 1 48 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 11 60 6 Off 1 48 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 11 35 5 Off 1 45 45

13. Elute Sample Container Hexane 11 35 5 Off 1 45 45

14. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Table 5 shows the initial demonstration of precision and recovery for the 100-mm disks without pre-filters.

Table 5. Four replicates 100-mm Disk, No Pre-Filters

Sample Recovery (mg) Recovery (%) 1 39.2 98.0 2 38.0 95.0 3 38.0 95.0 4 38.4 96.0

Average 96.0 Standard Deviation 1.41

Table 4. Biotage® Horizon 5000 Extraction Method for 47 mm Pacific® Premium Disks with Pre-Filters.

Table 5. Four replicates 90 mm Disk No Pre-Filters. Table 6. Four replicates 90 mm Disk with Pre-Filters.

Page 5

Table 4. SPE-DEX 5000 Extraction Method for 47-mm Pacific Premium Disks with Pre-Filters

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (s)

Sat. Time (s) Soak Time (s) Drain Time (s)

1. Condition SPE Disk Hexane 11 60 2 1 60 60

2. Condition SPE Disk MeOH 11 60 2 1 60 2

Step Sample Flow Rate (#)

Done Loading Sample Delay (s)

6. Load Sample 5 45

Step Dry Time (s) Pump Rate (#)

N2 Blanket

7. Air Dry Disk Timer 300 6 Off

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

8. Elute Sample Container Hexane 16 35 5 Off 1 23 30

9. Elute Sample Container Hexane 11 35 5 Off 1 48 45

10. Elute Sample Container Hexane 11 60 6 Off 1 48 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Drain Time (s)

11. Wash Sample Container MeOH 11 60 6 Off 1 48 60

Step Solvent Solvent Vol. (mL)

Purge Time (s)

Pump Rate (#)

N2 Blanket Sat. Time (s) Soak Time (s) Elute Time (s)

12. Elute Sample Container Hexane 11 35 5 Off 1 45 45

13. Elute Sample Container Hexane 11 35 5 Off 1 45 45

14. Elute Sample Container Hexane 11 60 6 Off 1 45 60

Table 5 shows the initial demonstration of precision and recovery for the 100-mm disks without pre-filters.

Table 5. Four replicates 100-mm Disk, No Pre-Filters

Sample Recovery (mg) Recovery (%) 1 39.2 98.0 2 38.0 95.0 3 38.0 95.0 4 38.4 96.0

Average 96.0 Standard Deviation 1.41

Table 5 shows the initial demonstration of precision and recovery for the 90 mm disks without pre-filters.

Table 6 shows the same information for 90 mm disks with pre-filters.

Page 6

Table 6. Four replicates 100-mm Disk with Pre-Filters

Table 6 shows the same information for 100-mm disks with pre-filters.

Sample Recovery (mg) Recovery (%) 1 38.2 95.5 2 38.1 95.3 3 38.0 95.0 4 38.7 96.7

Average 95.6 Standard Deviation 0.746

Table 7 shows the results for 4 samples using 47-mm disks without pre-filters

Table 7. Four Replicates 47-mm Disks without Pre-Filters

Sample Recovery (mg) Recovery (%) 1 39.3 98.3 2 39.2 98.0 3 39.4 98.5 4 39.7 99.2

Average 98.5 Standard Deviation 0.510

Table 8. Four Replicates 47-mm Disks with Pre-Filters

Sample Recovery (mg) Recovery (%) 1 39.6 99.0 2 39.1 97.8 3 38.4 96.0 4 38.8 97.0

Average 97.5 Standard Deviation 1.27

Table 8 shows the results for 4 samples using 47-mm disks plus pre-filters

© Biotage 2019

Hexane Extractable Material (Oil & Grease) Analysis with Automated Solid Phase Extraction | Page 5

42 4-Nitrophenol 58.5 98.2 43 Dibenzofuran 96.5 96.1 44 2,4-Dinitrotoluene 111.3 101.1 45 Diethyl phthalate 107.0 101.2 46 Fluorene 97.3 96.6 47 4-Chlorophenyl phenyl ether 99.0 95.7 48 4-Nitroaniline 93.8 95.1 49 4,6-Dinitro-2-methylphenol 85.0 98.4 50 NDPA/DPA 90.1 96.3 51 Azobenzene 94.3 97.4 52 4-Bromophenyl phenyl ether 101.8 98.7 53 Hexachlorobenzene 102.5 98.1 54 Pentachlorophenol 100.0 99.1 55 Phenanthrene 102.0 98.5 56 Anthracene 107.8 97.7 57 Carbazole 104.8 101.0 58 Di-n-butyl phthalate 116.3 100.5 59 Fluoranthene 108.5 98.1 60 Benzidine 2.6 63.8 61 Pyrene 101.8 97.9 62 Butyl benzyl phthalate 115.3 98.1 63 Benz(a)anthracene 111.0 97.6 64 3,3'-Dichlorobenzidine 94.6 75.4 65 Chrysene 105.0 99.1 66 Bis(2-ethylhexyl) phthalate 114.5 99.1 67 Di-n-octyl phthalate 11.5 97.0 68 Benzo(b)fluoranthene 128.3 95.5 69 Benzo(k)fluoranthene 106.5 97.6 70 Benzo(a)pyrene 100.5 96.0 71 Indeno(1,2,3-cd)pyrene 139.3 95.0 72 Dibenz(a,h)anthracene 114.8 94.9 73 Benzo(g,h,i)perylene 106.5 90.6

1 2-Fluorophenol (surr) 67.0 66.1 2 Phenol-d6 (surr) 47.4 70.3 3 Nitrobenzene-d5 (surr) 103.2 89.0 4 2-Fluorobiphenyl (surr) 104.8 92.4 5 2,4,6-Tribromophenol (surr) 131.4 101.4 6 p-Terphenyl-d14 (surr) 113.6 98.3

Page 6

Table 6. Four replicates 100-mm Disk with Pre-Filters

Table 6 shows the same information for 100-mm disks with pre-filters.

Sample Recovery (mg) Recovery (%) 1 38.2 95.5 2 38.1 95.3 3 38.0 95.0 4 38.7 96.7

Average 95.6 Standard Deviation 0.746

Table 7 shows the results for 4 samples using 47-mm disks without pre-filters

Table 7. Four Replicates 47-mm Disks without Pre-Filters

Sample Recovery (mg) Recovery (%) 1 39.3 98.3 2 39.2 98.0 3 39.4 98.5 4 39.7 99.2

Average 98.5 Standard Deviation 0.510

Table 8. Four Replicates 47-mm Disks with Pre-Filters

Sample Recovery (mg) Recovery (%) 1 39.6 99.0 2 39.1 97.8 3 38.4 96.0 4 38.8 97.0

Average 97.5 Standard Deviation 1.27

Table 8 shows the results for 4 samples using 47-mm disks plus pre-filters

Page 6

Table 6. Four replicates 100-mm Disk with Pre-Filters

Table 6 shows the same information for 100-mm disks with pre-filters.

Sample Recovery (mg) Recovery (%) 1 38.2 95.5 2 38.1 95.3 3 38.0 95.0 4 38.7 96.7

Average 95.6 Standard Deviation 0.746

Table 7 shows the results for 4 samples using 47-mm disks without pre-filters

Table 7. Four Replicates 47-mm Disks without Pre-Filters

Sample Recovery (mg) Recovery (%) 1 39.3 98.3 2 39.2 98.0 3 39.4 98.5 4 39.7 99.2

Average 98.5 Standard Deviation 0.510

Table 8. Four Replicates 47-mm Disks with Pre-Filters

Sample Recovery (mg) Recovery (%) 1 39.6 99.0 2 39.1 97.8 3 38.4 96.0 4 38.8 97.0

Average 97.5 Standard Deviation 1.27

Table 8 shows the results for 4 samples using 47-mm disks plus pre-filters

Table 7. Four replicates 47 mm Disk No Pre-Filters. Table 8. Four replicates 47 mm Disk with Pre-Filters.

Table 7 shows the results for 4 samples using 47 mm disks without pre-filters.

Table 8 shows the results for 4 samples using 47 mm disks with pre-filters.

In each case the recovery is better than 95%, well within the acceptable range specified in method 1664B of 83–101%. The standard deviation is well below the 11% limit specified in the method for each of the four configurations tested.

References1. Method 1664, Revision B: n-Hexane Extractable Material

(HEM; Oil and Grease) and Silica Gel Treated n-Hexane Extractable Material (SGT-HEM; Non-polar Material) by Extraction and Gravimetry, available at www.epa.gov, (2010).

2. ISO 11349, Water quality—Determination of low-volatility lipophilic substances—Gravimetric method, First edition 2010-09-15, available at http://www.iso.org/iso/home.htm.

3. Method 5520G, Standard Methods for the Analysis of Water and Wastewater, 21 Edition, available at https://www.standardmethods.org/.

4. Vinson Leung and Michael Ebitson, Modern Hexane-Extractable Material (Oil & Grease) Analysis in Wastewater Samples, AN102-HOR, available from www.biotage.com.

5. David Gallagher, Complying with EPA Method 1664A and 1664B by Automated Solid Phase Extraction Utilizing the SPE-DEX® 4790 with Envision® Platform v1.02, AN025-HOR, available from www.biotage.com.

© Biotage 2019

Hexane Extractable Material (Oil & Grease) Analysis with Automated Solid Phase Extraction | Page 6

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Literature Number: AN118-HOR.V.1

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