# Biodegradability and Acute Aquatic Toxicity of Alcohol Propoxy Sulfates.

Research Report R&D ENV

- 07

- 024 01

- 07

Part 1 of 2

Geoff Russell Ricky Stackhouse Wayne Sorensen

November 2007

Research & Development Lake Charles, Louisiana

## Interoffice Correspondence

To Dave Penney

From Geoff Russell, Ricky Stackhouse, Wayne Sorensen

Date November 30, 2007

**Subject R&D ENV: Biodegradability and Acute Aquatic Toxicity Alcohol Propoxy Sulfates.** 



### Key Words: ALFOTERRA, Alcohol propoxylat es, Alcohol propoxy sulfates, Biodegradation, Aquatic Toxicity, Oil recovery, Emulsification

This report documents Sasol North America Inc.’s biodegradation and aquatic toxicity studies on the new products, ALFOTERRA ® alcohol propoxy sulfate surfactants.

This work shows that the ALFO TERRA ® alcohol propoxy sulfates, based on either C12

- C13, C14

- C15, or C16

- C17 branched / linear primary alcohol, pose little risk to the environment. The ALFOTERRA surfactants are not intended or expected to be released into the environment when used in the recovery of hydrocarbon materials.

Further conclusions drawn from this work include :

1) The rate and extent of biodegradation of the ALFOTERRA® alcohol propoxy sulfates were dependent on the amount of propoxylation and bra nching in the starting alcohol. ALFOTERRA® surfactants with a low proportion ( < 4 moles or < 60%) of propoxylation displayed extensive primary biodegradation and, in some cases, ultimate biodegradation. The surfactants with a higher amount of propoxylati on had limited biodegradation.

2) Acute aquatic toxicities to fish, invertebrates and algae were determined using standard US EPA methods. Based on the toxicity results, the aquatic toxicity of the ALFOTERRA® alcohol propoxy sulfates is similar to that of e quivalent alcohol ethoxy sulfates.

3) Using ALFOTERRA® surfactants for enhanced hydrocarbon recovery (EHR) will not result in

the discharge any material into surface or ground water.

Distribution: K. McWil liams, K. Youngblood, T. Grumbles, V. Stolarski, J. Stokes,

D. Plummer, R. Stackhouse , D. Schär, J. McGrail, Library, LiveLink.ALFOTERRA® Sulfate Report Page 3 of 50



### T able of Contents

I. INTRODUCTION

................................ ................................ ................................ ............................... 7

II. OBJECTIVE S

................................ ................................ ................................ ................................ .... 8

III. CONCLUSIONS

................................ ................................ ................................ ............................... 8

IV. DEFINITION OF MATERIALS

................................ ................................ ................................ ....... 10

4.1 ALFO TERRA® Composition

................................ ................................ ................................ ..... 10

V. BIODEGRADATION STUDY

................................ ................................ ................................ .................. 11 5.1 Biodegradation Test Method

................................ ................................ ................................ ..... 11 5.2 Toxicity and Chemical Oxidation Controls

................................ ................................ ................. 12 5.3 Determination of Organic Carbon by COD values

................................ ................................ ..... 13

5.4 Tes t Materials

................................ ................................ ................................ ............................ 13 5.5 Validity of Biodegradation tests

................................ ................................ ................................ . 13 5.6 Results of the Biodegradation Test (OECD 301 F)

................................ ................................ .... 14

VI. AQUATIC ACUTE TOXICITY STUDIES

................................ ................................ ........................ 16 6.1 Aquatic Acute Toxicity Test Methods and Results

................................ ................................ ..... 16

VII. HISTORICAL BIODEGRADATION AND TOXICITY RESULTS WITH ALFOTERRA®

SURFACTANTS

................................ ................................ ................................ ................................ . 17 7.1 Biodegradation Results from First

- Generation Propoxy

- Sulfate Samples

................................ . 17 7.2 Aquatic Toxicity Results from First

- Generation Propoxy Sulfate Samples

................................ 19

VIII. ENVIRONMENTAL ASSESSMENT OF ALFOTERRA® ALCOHOL PROPOXY SULFATES

..... 20 8.1 Discussion of Biodegradation Results

................................ ................................ ....................... 20 8.2 Discussion of Aquatic Toxicity Results

................................ ................................ ...................... 21 8.3 Discussion of Fate in Use Application

................................ ................................ ....................... 22

IX. SUMMARY

................................ ................................ ................................ ................................ .... 23

X. ACKNOWLEDGEMENTS

................................ ................................ ................................ ............... 23

XI. REFERENCES

................................ ................................ ................................ .............................. 24ALFOTERRA® Sulfate Report Page 4 of 50



### Table of Tables

Table 1. Alcohol feed stocks that could be employed to produce the ALFOTERRA® 123

- XS alcohol propoxy sulfate. The asterisks (*) denotes the alcohol used to produce the samples employed in the following environmental testing.

................................ ................................ ................................ .......... 10

Table 2. Alcohol feed stocks that could be employed to produce the ALFOTERRA® 145

- X alcohol propoxy sulfate. The asterisks (*) denotes the alcohol used to produce the samples employed in the following environmental testing.

................................ ................................ ................................ .......... 11

Tab le 3. Alcohol feed stocks that could be employed to produce the ALFOTERRA® 167

- XS alcohol propoxy sulfate. The asterisks (*) denotes the alcohol used to produce the samples employed in the following environmental testing.

................................ ................................ ................................ .......... 11

Table 4. ALFOTERRA® propoxy sulfate samples included in biodegradation and aquatic testing.

.... 11

Table 5. Amount of ALFOTERRA® propoxylate sulfates in Biod egradation Study and the

............... 13

Table 6. Ultimate Biodegradation of ALFOTERRA® propoxylate sulfates. (Mean of % ThOD and Standard Deviation of the mean) in the OECD 301 F Biodegradation Test

................................ ........ 14

Table 7. Original Aquatic Toxicity Reports.

................................ ................................ ........................ 16

Table 8. Aquatic toxicity results for the ALFOTERRA® propoxylate sulfates.

................................ .... 17

Table 9. Summary of primary and ultimate biodegradation of early

- versions of the ALFOTERRA® propoxy sulfate surfactants in OECD 301A test.

................................ ................................ ................. 18

Table 10. Summary of ultimate biodegradation of ALFOTERRA® surfactants in OECD 301F Test

... 19

Table 11. Acute Aquatic Toxicity of first

- generation ALFOTERRA® surfactants

............................... 19

Table 12. Combined biodegradation results for ALFOTERRA® alcohol propoxy sulfates

................. 20

Table 13. Combined aquatic toxicity resul ts for ALFOTERRA® alcohol propoxy sulfates

................. 21ALFOTERRA® Sulfate Report Page 5 of 50



### Table of Figures

Figure 1. Toxicity control as it relates to biodegradation profile of benzoate % THOD in the presen ce of ALFOTERRA® Sulfates

................................ ................................ ................................ .................. 14

Figure 2. Biodegradation of the ALFOTERRA® sulfates tested

................................ ......................... 16ALFOTERRA® Sulfate Report Page 6 of 50



### Table of Appendices

Appendix A. Biomonitoring Report; Daphnia p ulex & Pimephales promelas . Prepared for Sasol by EE USA. Lab Report # Q

- 506

- 07. Sample ID: ALFOTERRA® 145

- 8S. October 31, 2007

Appendix B. An Evaluation of the Chronic Toxicity of the ALFOTERRA® Samples to Freshwater Alga Selenastrum capricornutum

. Prepared for Sasol by Pacific EcoRisk. October, 2007

Appendix C. Biomonitoring Report; Daphnia pulex & Pimephales promelas

. Prepared for Sasol by

EE USA

. Lab Report # Q

- 6561

- 07

. Sample ID: ALFOTERRA® L167

- 4S

. October 31, 2007

Appendix D. Biomonit oring Report; Daphnia pulex & Pimephales promelas

. Prepared for Sasol by

EE USA

. Lab Report # Q

- 562

- 07

. Sample ID: ALFOTERRA® L167

- 7S

. October 31, 2007

Appendix E. Biodegradation of Early Versions of ALFOTERRA® Propoxy Sulfate Surfactants

Appendix F. Toxicity Reports on Earlier Versions of ALFOTERRA® Propoxy Sulfate Surfactants; Daphnia magna and Pimephales promelas. Prepared for Condea Vista Company by ENSR. Lab Report #8503

- 111

- 006 (015 thru 020). Lab Report #8503

- 106

- 006 (001 thru 008). Augu st 1999

### I. INTRODUCTION

"Tertiary" or "enhanced " recovery techniques are well known in the petroleum industry.

These methods are used to increase production in reservoir s where the formation pressure is no longer h igh enough to push the oil out of the well. One of the most widely used enhanced recovery techniques in the petroleum industry is "waterflooding." In the waterflooding method, an aqueous

- based fluid is pushed through an injection well

into the oil

- rich r egion to force the oil out of the formation. Typically surface

- active agents or surfactants are employed as part of the aqueous fluid to lower the interfacial

tension between t he water and the formation oil. This permits oil droplets to deform,
coalesce, and flow with the flood water toward the producing wells.

Studies conducted at several major universities suggest that alcohol propoxy sulfates are exceptionally good surfactants for the emulsification of crude oils. Thus, these surfactants are being so ught by oil companies pursuing waterflooding methods for inclusion in the aqueous

- based flood fluid

.

Similarly, alcohol propoxy sulfates have also been shown to be useful for the removal of crude oil and organic drilling fluids from drill cuttings. When the drill cuttings are washed in an aqueous fluid containing the alcohol propoxy sulfate surfactant, the residual organic content in the cutting can be reduced to <1%. In some regions of the world, this residual organic content on the cuttings is low eno ugh to allow the cuttings to be discarded at the drilling site instead of having to be land

- filled or otherwise handled as hazardous waste. This is especially advantageous when the drilling site is off

- shore.

For these and other similar applications, Sas ol North America Inc. is interested in commercializing alcohol propoxy sulfates under the trade name ALFOTERRA®.

The alcohol propoxy sulfate is manufactured in three steps. First, primary alcohol (1) is reacted with propylene oxide and catalyst under m oderate temperature and pressure to yield the alcohol propoxylate (2).

O O O O H +

n ( 1 ) ( 2 ) R O H R



### Secondly, the alcohol propoxylate (2) is reacted with SO 3 

### or another appropriate
reagent to form the sulfonic acid of the alcohol propoxylate (3).

S O 3 O O O

( 3 ) S O

O O H O O O H

n ( 2 ) R + R

nALFOTERRA® Sulfate Report Page 8 of 50



### Finally, the sulfonic acid (3) is neutralized with sodium hydroxide to yield the sodium salt of the alcohol propoxy sulfate (4).

+ N a O H O O O

n

( 4 ) S O

O O 

## - N a 

### + O O O

( 3 ) S O

O O H R

n R



### The sodium salt of the alcohol propoxy sulfates h ave not been previously marketed in the US, so a Pre

- Manufacturing Notification (PMN) from the US EPA is required. The PMN will be submitted for a number of alcohol propoxy sulfates with the trade name pattern of ALFOTERRA® 1XX

- YS. The “XX” will be 23, 4 5, or 67 and represents the carbon number of the starting alcohol (C12

- C13, C14

- C15, or C16

- C17, respectively). The “Y” will represent the average number of propylene oxides moles and will range

between 1 and 10. The “S” denotes the product is the sodium

- sulfate derivative. Since the US EPA will estimate the environmental safety of these new products before accepting the PMN, it is prudent to provide in the PMN the biodegradability and aquatic toxicity profiles of these new surfactants. During 1999 an d 2005 initial toxicity and biodegradation studies were conducted on early versions of the ALFOTERRA®

surfactants. 1 

### The current tests were carried out to confirm the earlier results and to provide information on additional versions of the alcohol propoxy sulfates.

II. OBJECTIVES

**•** Determine the biodegradability of alcohol propoxy sulfates in the OECD 301F Manometric Respirometry ”Ready” Biodegradability Test.

**•** Determine the acute aquatic toxicity of alcohol propoxy sulfat es to fish ( Pimephales promelas

– fathead minnows), invertebrates ( Daphnia pulex

– water fleas) and algae ( Selenastrum capricornutum ) using US EPA approved methodologies (EPA

- 821

- R

- 03

- 012,013).

III. CONCLUSIONS

**•** The rat e and extent of biodegradation of the ALFOTERRA® alcohol propoxy sulfates were dependent on the amount of propoxylation and branching in the starting alcohol. ALFOTERRA® surfactants with a low proportion ( < 4 moles or < 60%) of propoxylation displayed ext ensive primary biodegradation and, in some cases, ultimate biodegradation. The surfactants with a higher amount of propoxylation had limited biodegradation.

**•** Acute aquatic toxicities to fish, invertebrates and algae were determined using

standard US EPA m ethods. Based on the toxicity results, the aquatic toxicity of the ALFOTERRA® alcohol propoxy sulfates is similar to that of equivalent alcohol ethoxy sulfates.ALFOTERRA® Sulfate Report Page 9 of 50



### • Using ALFOTERRA® surfactants for enhanced hydrocarbon recovery (EHR) will

not result in the d ischarge any material into surface or ground water.ALFOTERRA® Sulfate Report Page 10 of 50



### IV. DEFINITION OF MATERIALS

4.1 ALFOTERRA® Composition

Sasol will initially offer six A LFOTERRA ® sulfate (structure 4 from Introduction) products to the marke t. Second or third generation products may include other propoxymers from 1

- 10. T he products in the initial offering are as follows:

1. ALFOTERRA ® 123

- 4S

- Four mole propoxy sulfate based on a C12

- 13 branched primary alcohol.

2. ALFOTERRA ® 123

- 8S

- Eight mole propoxy sulfate based on a C12

- 13 branched primary alcohol

3. ALFOTERRA ® 145

- 4S

- Four mole propoxy sulfate based on a C14

- 15 branched primary alcohol

4. ALFOTERRA ® 145

- 8S

- Eight mole propoxy sulfate based on a C14

- 15 branched primary alcohol

5. ALFOTERRA ® 167

- 4S

- Four mole propoxy sulfate based on a C16

- 17 branched primary alcohol

6. ALFOTERRA ® 167

- 7S

- Seven mole propoxy sulfate based on C16

- 17 branched primary alcoho l

There are several alcohols that can be used either alone or blended as starting

materials . It is important for the proposed end

- use application that a certain amount of branching be present in the alcohol portion of the molecule, however the specific ty pe and degree of branching does not appear to be critical. This flexibility allows for the use of several alternative alcohol feed stocks (or blends thereof) in the propoxylate manufacture, based on pricing or availability. Table 1 through 3 lists some possible alcohol feed stocks that may be considered for each propoxylate. An asterisk is used to denote the alcohols employed to make the samples in the testing to follow.

Table 1 . Alcohol feed stocks that could be employed to produce the ALFOTERRA® 123

- X S alcohol

propoxy sulfate . The asterisks (*) denotes the alcohol used to produce the samples employed in the following environmental testing.



### 123 Feedstock C11 C12 C13 C14 C15 C16 C17 Branched

Alcohol 1 0 .5 42 56 1.5 54%

Alcohol 2* 1.5 41 55 1.5 1 94%

Alcohol 3 <1 53 46 <2 50%ALFOTERRA® Sulfate Report Page 11 of 50

Table 2 . Alcohol feed stocks that could be employed to produce the ALFOTERRA® 145

- X alcohol

propoxy sulfate . The asterisks (*) denotes the alco hol used to produce the samples employed in the following environmental testing.



### 145 Feedstock C11 C12 C13 C14 C15 C16 C17 Branched

Alcohol 1 1 62 36 1 57%

Alcohol 2* 1.5 59 39 0.5 95%

Table 3 . Alcohol feed stocks that could be employed to produce the ALFOTERRA® 167

- X S alcohol propoxy sulfate . The asterisks (*) denotes the alcohol used to produce the samples employed in the following environmental testing.



### 167 Feedstock C1 3 C1 4 C1 5 C1 6 C1 7 C1 8 C1 9 Branched

Alcohol 1 * 4 62 33 1 63%

Alcohol 2 5 57 38 ~95%

In order to limit testing cost, aquatic testing was conducted on a subset of the six

ALFOTERRA® sulfates . The samples that were tested are given in Table 4

. It is

believed that the aquatic performance of the three products not tested (ALFOTERRA

123

- 4S, 123

- 8S, and 145

- 4S) or other future generation propoxy sulfates can be extrapolated from the data gathered on the included samples or from literature

references.

Table 4

. ALFOTERRA® propoxy sulfate samples included in biodegradation and aquatic testing.



### Product Biodegradation Aquatic Testing

ALFOTERRA ® 123

- 4S Yes No

ALFOTERRA ® 123

- 8 S Yes No

ALFOTERRA ® 1 45

- 4S Yes No

ALFOTERRA ® 145

- 8 S Yes Yes

ALFOTERRA ® 16 7

- 4 S Yes Yes

ALFOTERRA ® 167

- 7 S Yes Yes

V. BIODEGRADATION STUDY

5.1 Biodegradation Test Method

The biodegradation test was performed on the alcohol propoxylates ALFOTERRA ®

123

- 4S, ALFOTERRA® 123

- 8S, ALFOTERRA® 145

- 4 S, ALFOTERRA® 145

- 8S,

ALF OTERRA® 167

- 4S and ALFOTERRA® 167

- 7S in triplicate. Two 20

- channel Coordinated Environmental Services (CES) Ltd (Cornwell, UK) aerobic electrolytic respirometer units were used following the OECD 301F guidelines 2 

### with slight

modifications that will be discussed later

. Each CES electrolytic respirometer unit

consists of 20 one

- liter vessels fitted with heads /lids containing O 2 

### and CO 2 

### sensors.ALFOTERRA® Sulfate Report Page 12 of 50



### The system measures the amount of CO 2 

### genera ted by means of a cuvette filled with 2% sodium hydroxide and a conductivity probe that determines the change in conductivity of the sodium hydroxide as CO 2 

### is absorbed from the headspace gases of the test vessel. A differential pressure sensor head is de signed to measure the amount

of O 2 

### used by respiring microorganisms. It achieves this by detecting the difference in pressure during the uptake of oxygen and then supplying an equal amount of O 2 

### back to the headspace using an electrolytic cell.

The elect rolytic cell contains a chamber filled with copper sulfate solution. Oxygen is generated in this chamber by passing an electrical charge (initiated by the test vessel head pressure change sensor) between electrodes. It is connected to the headspace of

the sample vessel by a vent. Valves are used to balance the pressure and return generated O 2 

### to the headspace of the test vessel. The data is handled by a PC which continuously records the amounts of CO 2 

### (mg) absorbed and O 2 

### (mg) generated.

The inoculum fo r these tests was derived from the recycled sludge of the City of Lake Charles, Louisiana Domestic Waste Water Plant A. The sludge was aerated for 7

- days prior to use. The inoculum in the test medium was adjusted to a final concentration of 10 mg/L of to tal suspended solids (TSS) which meets the guideline specification of < 30 mg/L TSS. Sodium benzoate (J.T Baker

– 99% purity) was used as the positive

reference test material.

The 1

- liter test vessels containing 900 mL of inoculated medium was mixed prior to receiving the sample solutions . Each flask contained a stir bar which was positioned over a magnetic stirrer to provide vigorous mixing during the test. Stock solution s of 1.0 % (10,000 mg/L) of each test material were made using distilled water. Ta rget

concentration of 100 mg test material was achieved by adding 10 mL from each

ALFOTERRA® stock solution to the inoculated medium . The 1

- liter vessels were then placed in the water bath (22±1°C) of the electrolytic respirometer units. The instrument

u nderwent a 60 minute initialization/equalization before data collection began. CO 2 

### generation and O 2 

### uptake data were recorded by the instrument every six hours during the 28 day test. Blanks were also included in the study for validity purposes. These

bl anks were prepared identical to the samples except that they did not contain the test

materials (i.e. blanks contained inoculum only ).

5.2 Toxicity and Chemical Oxidation Controls

During the definitive test, two types of controls were included to determine the potential of microbiological toxicity and chemical oxidation. The first control type was run on each of the six ALFOTERRA® sulfate samples. This control , a test for toxicity of the sample material on the inoculu m, was done with a concentration of 50 mg of J.T. Baker, sodium benzoate and 100 mg of the ALFOTERRA® sulfate. The second control type was run using only one of the ALFOTERRA® sulfates. In this second control , a sterility

test with 1.0 % sodium azide and 100 mg of ALFOTERRA® 167

- 7S, was included to

evaluate the chemical oxidation influence. The reference chemical was sodium

benzoate. The two control types demonstrated that the test materials were not toxic to the microbial population and that che mical o xidation was irrelevant.ALFOTERRA® Sulfate Report Page 13 of 50



### 5.3 Determination of Organic Carbon by COD values

The organic carbon for the ALFOTERRA® samples w as determined by the HACH DR2000 spectrophotometer chemical oxygen d emand (COD) Low Range Method . The

COD values will be used in the calculation of the theoretical m aximum amount of oxygen uptake from complete oxidation of the test materials (ThOD).

5.4 Test Materials

Table 5 shows the amount of test materials added to the each test vessel and the calculated ThOD which could be produced from the given amount of test material upon complete oxidation.

Table 5

. Amount of ALFOTERRA® propoxylate sulfates in Biodegradation Study and the

calculated theoretical o xygen deman d (ThOD).



### Product mg product COD ThOD (mg)

AF 123

- 4S 100 201.3

AF 123

- 8S 100 203.3

AF

- 145

- 4S 100 228.6

AF 145

- 8S 100 224.6

AF 167

- 4S 100 227.3

AF 167

- 7S 100 232.0

5.5 Validity of Biodegradation tests

The validity of the biodegradation test was confirmed by meeting the following criteria as specified in the OECD 301 F Test Guidelines 2



### .

**•** The delta of the max./min. of replicate %ThOD values at the plateau of the data,

at the end of the test and/or at the end of the 10

- day window, as appropriate, must be less than 20%. The actual range of differences was < 4%.

**•** The extent of biodegradation of the reference compound must reach the pass

level of 60% ThOD on or before day 14. The ref erence compound reach ed 60%

ThOD by day 5

.

**•** The total oxygen uptake in the inoculum blank must not exceed 60 mg/L. The maximum oxygen uptake was < 20 mg/L.

**•** The test material must show no inhibition of the biodegradation of the reference compound. By day 14 the ALFOTERRA’s + R eference benzoate was >25%

ThOD

. 

### The criterion is that the combined ThOD value be >25% in 14 days. Figure 1 shows another way to express inhibition to the biodegradation of the reference standard alone as compared to the effects from the presence of the test material. The lines for the test compounds plus benzoate track above the benzoate indicating the bacterium was capable of degrading benzoate in the presence of the test samples.ALFOTERRA® Sulfate Report Page 14 of 50



### • The sterility control for ALFOTERRA® 167

- 7S d emonstrated that biodegradation and not chemical oxidation was occurring and due to their similar chemical

make

- up, it is assumed the same is true for the other ALFOTERRA® products

.

0 20 40 60 80 100 120 140 160

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28

Days% Theoretical Oxygen Benzoate

Benzoate + 123-4S

Benzoate + 123-8S

Benzoate + 145-4S

Benzoate + 145-8S

Benzoate + 167-4S

Benzoate + 167-7S

Figure 1

. Toxicity control as it relates to bi odegradation profile of benzoate % THOD in the presence of ALFOTERRA® Sulfates



### 5.6 Results of the Biodegradation Test (OECD 301 F)

Table 6 displays the results of the biodegradation tests by showing the three

- replicate mean of the percent of the theoretical oxygen demand (%ThOD) produced on each day

as well as the standard deviation of the mean. Figure 2 displays this data graphically using the mean values. Though the ALFOTERRA samples did show some degradation, none of the samples reached the 60% target by the end of the test at day 28.

Table 6

. Ultimate Biodegradation of ALFO TERRA ® propoxylate sulfates . (Mean of % ThOD and Standard Deviation of the mean) in the OECD 301 F Biodegradation Test



### ALFOTERRA®

Product 1 2 3 4 5 6 7

123

- 4S 0.25±0.03 1.07±0.10 2.86±0.19 8.39±0.35 16.50±0.56 18.39±1.38 21.47±1.52


### 123

- 8S 0.32±0.05 0.70 ±0.11 0.93±0.20 1.66±0.13 3.31±0.07 5.10±0.12 8.02±0.10


### 145

- 4S 0.25±0.06 0.71±0.11 1.81±0.42 5.34±0.83 13.96±0.81 17.02±0.92 19.94±1.20


### 145

- 8S 0.62±0.08 1.30±0.13 1.62±0.12 1.83±0.11 2.65±0.13 4.43±0.11 6.43±0.14


### 167

- 4S 0.74±0.04 3.04±0.07 6.20±0.05 10. 13±0.07 14.83±0.03 18.08±0.06 20.1±0.14


### 167

- 7S 0.55±0.01 1.64±0.17 2.28±0.19 3.10±0.16 4.26±0.07 6.26±0.03 9.20±0.20ALFOTERRA® Sulfate Report Page 15 of 50



### ALFOTERRA®

Product 8 9 10 11 12 13 14

123

- 4S 22.79± 1.27 23.51± 1.22 23.97± 1.26 24.42± 1.29 24.99± 1.18 25.76± 0.79 26.73± 0.63


### 123

- 8S 9.75 ± 0.11 11.06± 0.14 11.91± 0.15 12.57± 0.16 13.08± 0.15 13.43± 0.16 13.81± 0.17


### 145

- 4S 21.70± 0.71 22.78± 0.58 23.45± 0.48 24.02± 0.38 24.74± 0.35 25.68± 0.43 26.74± 0.43


### 145

- 8S 8.67± 0.14 10.39± 0.16 12.00± 0.79 13.32± 0.24 14.41± 0.28 15.20± 0.34 16.16± 0.36


### 167

- 4S 22.16± 0 .13 23.85± 0.01 25.62± 0.09 27.23± 0.24 28.98± 0.37 30.64± 0.46 32.31± 0.49


### 167

- 7S 13.31± 0.42 17.16± 0.57 19.97± 0.26 21.70± 0.15 23.00± 0.27 24.20± 0.66 25.42± 0.83



### ALFOTERRA®

Product 15 16 17 18 19 20 21

123

- 4S 27.76± 0.87 28.81± 1.31 29.67± 1.71 30.32± 1.98 30.76± 2.15 31.10± 2.26 31.41± 2.36


### 123

- 8S 14.20± 0.16 14.56± 0.14 14.98± 0.12 15.44± 0.12 15.86± 0.12 16.31± 0.15 16.73± 0.18


### 145

- 4S 27.52± 0.33 28.21± 0.21 28.83± 0.15 29.44± 0.16 30.07± 0.23 30.71± 0.14 31.07± 0.22


### 145

- 8S 17.03± 0.37 17.85± 0.41 18.66± 0.51 19.50± 0.74 20.35± 0 .88 20.98± 0.94 21.63± 0.97


### 167

- 4S 33.75± 0.46 34.83± 0.45 35.68± 0.48 36.39± 0.51 37.07± 0.52 37.62± 0.50 38.32± 0.47


### 167

- 7S 26.56± 0.99 27.58± 1.11 28.49± 1.27 29.36± 1.45 30.22± 1.59 30.90± 1.70 31.63± 1.87



### ALFOTERRA®

Product 22 23 24 25 26 27 28

123

- 4S 31.68± 2.45 31.98± 2.55 32.30± 2.66 32.78± 2.81 33.26± 2.93 33.82± 3.07 34.40± 3.25


### 123

- 8S 17.07± 0.21 17.40± 0.21 17.67± 0.19 17.93± 0.17 18.18± 0.14 18.47± 0.10 18.87± 0.13


### 145

- 4S 31.44± 0.12 31.84± 0.03 32.18± 0.18 32.54± 0.28 32.88± 0.34 33.24± .038 33.59± 0.41


### 145

- 8S 22.13± 0.98 22.66± 0.97 23.13± 0.91 23.64± 0.83 24.19± 0.71 24.59± 0.61 25.23± 0.53


### 167

- 4S 38.93± 0.47 39.56± 0.48 40.05± 0.49 40.51± 0.54 40.97± 0.61 41.28± 0.70 41.87± 0.82


### 167

- 7S 32.19± 2.00 32.83± 2.08 33.40± 2.13 33.97± 2.21 34.54± 2.29 34.96± 2.35 35.64± 2.41ALFOTERRA® Sulfate Report Page 16 of 50

0 10 20 30 40 50 60 70 80 90 100

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28

Days% Theoretical Oxygen 123-4S

123-8S

145-4S

145-8S

167-4S

167-7S

Figure 2 . Biodegradation of the ALFOTERRA® s ulfates tested



### VI. AQUATIC ACUTE TOXICITY STUDIES

6.1 Aquatic Acute Toxicity Test Methods and Results

Aquatic toxicity studies were conducted on three freshwater species; Pimephales promelas (fathead minnow), Daphnia pulex (water flea), and Selenastrum capricornutum (algae). The minnow and water flea tests were conducted by Environmental Enterprise s USA, Inc. (Slidell, LA). The algae tests were performed by Pacific EcoRisk (Martinez, CA). The fathead minnow and Daphnia are, respectively, 96

and 48

- hr acute tests, while the 96

- hr algae test is considered to be a short

- term chronic

test. The acute effect endpoint for fish and Daphnia is the lethal concentration which kills 50% (LC50) of the test organisms. The chronic effect endpoint for S. capricornutum is the concentration that inhibits 50% of algal cell growth (IC50) during

the 96

- hr test period

. Table 7 lists the Appendix Reference number where the original laboratory reports are given, the product, the laboratory that conducted the testing, the laboratory report ID code and the USEPA method followed.

Table 7 . Original Aquatic Toxicity Reports

.

Appendix ALFOTERRA® Lab / ID code Species USEPA Test

A 145

- 8S EE USA Q

- 560l

- 07 Fish EPA / 821 / R

- 02

- 012

A 145

- 8S EE USA Q

- 560

- 07 D aphnia EPA / 821 / R

- 02

- 012

B 145

- 8S Pacific EcoRisk Algae EPA / 821 / R

- 02

- 013

C 167

- 4S EE USA Q

- 561l

- 07 Fish EPA / 821 / R

- 02

- 012

C 167

- 4S EE USA Q

- 561

- 07 Daphnia EPA / 821 / R

- 02

- 012ALFOTERRA® Sulfate Report Page 17 of 50

B 167

- 4S Pacific EcoRisk Algae EPA / 821 / R

- 02

- 013

D 167

- 7S EE USA Q

- 562l

- 07 Fish EPA / 821 / R

- 02

- 012

D 167

- 7S EE USA Q

- 562

- 07 Daphnia EPA / 821 / R

- 02

- 012

B 167

- 7S Pacific EcoRisk Algae EPA / 821 / R

- 02

- 013



### Since the ALFOTERRA ® sulfate samples evaluated in this study are water soluble they

were able t o be used as received. That is, it was not necessary to prepared the Water Accommodation Fraction (WAF) solutions that were used in the ALFOTERRA ® propoxylate study. Table 8 gives the aquatic toxicity LC/IC50 res ults for the ALFOTERRA® propoxy sulfates

.

Table 8

. Aquatic toxicity results for the ALFOTERRA ® propoxylate sulfates

.

ALFOTERRA® Fish 96

- hr

LC 50 (mg/L) **Daphnia** 48

- hr

LC 50 (mg/L) Algae 96

- hr

IC 50 (mg/L)

145

- 8S 1.31 2.89 1.70

167

- 4S 0.49 7.56 0.49

167

- 7S 1.40 15.8 0.89



### VII. HISTORICAL BIODEGRADATION AND TOXICITY RESULTS WITH

ALFOTERRA® SURFACTANTS

Sasol has been working on the development of alcohol propoxy sulfate surfactants for almost 10 years. The end

- use markets (enhance d oil recovery, soil and drill cutting cleaning,) are finally now at the point where commercialization of the surfactants is economically feasible. As a result, Sasol has generated biodegradation and aquatic toxicity data on earlier versions of the ALFOTE RRA sulfate surfactants that are relevant

to the six

- target materials being discussed in this report. This earlier data is presented

below.

7.1 Biodegradation Results from First

- Generation Propoxy

- Sulfate Samples

In 1999, in

- house Ready Biodegradation stu dies were conducted on a two sets of earlier generation ALFOTERRA® type surfactants

. 1 

### OECD 301A methodology was

used. The following surfactants were tested:

12

- 3S

– ALFOTERRA® propoxylate sulfate surfactant produced from a C12

Guerbet alcohol with 3 mo les of propylene oxide

123

- 2S

– ALFOTERRA® propoxylate sulfate surfactant produced from a

branched C12

- 13 alcohol with 2 moles of propylene oxide

123

- 4S

– ALFOTERRA® propoxylate sulfate surfactant produced from a

branched C12

- 13 alcohol with 4 moles of pro pylene oxide

123

- 8S

– ALFOTERRA® propoxylate sulfate surfactant produced from a

branched C12

- 13 alcohol with 8 moles of propylene oxide

145

- 2S

– ALFOTERRA® propoxylate sulfate surfactant produced from a

branched C14

- 15 alcohol with 2 moles of propylene oxi deALFOTERRA® Sulfate Report Page 18 of 50



### 145

- 4S

– ALFOTERRA® propoxylate sulfate surfactant produced from a

branched C14

- 15 alcohol with 4 moles of propylene oxide

The OECD 301A: DOC die

- away test was done as described in the OECD Guideline for Testing of Chemicals adopted July 17,1992. This is a 28

- day aerobic biodegradation

die

- away test on 10

- 30 mg/L water soluble organic compounds (measured as dissolved organic carbon

– DOC) do ne in mineral salts medium inoculated with ≤ 30 mg/L total suspended solids (TSS) from natural sewage/soils or surface waters. Ultimate biodegradation was determined by measuring the DOC concentrations in the test flasks during the test, while Primary bio degradation was measured by using methylene blue active substance (MBAS). The inoculum was derived from activated sludge obtained from the Anderson Mill Municipal Water Reclamation Facility (Austin, TX) and the supernatant from a suspension of 100 g soil in 500 mL of the mineral salts medium. The test was done at a starting concentration of 20 mg DOC/L for the test materials. The summarized results from the study can be found in Table 9

, while the expanded

data c an be found in Appendix E

.

Table 9 . Summary of primary and ultimate b iodegradation of early

- versions of the ALFOTERRA® propoxy sulfate s urfactants in OECD 301A t est

.

ALFOTERRA® Surfactant OECD 301A

% MBAS Loss % DOC Loss

Pass 10

- day

Window 28

- Day

End

Point Pass 10

- day

Window 28

- Day

End

Point

12

- 3S Yes 96,3 No 46.4

123

- 2S Yes 98.2 No 52.1

123

- 4S Yes 91.1 No 28.7

123

- 8S No 46.1 No 10.9

145

- 2S Yes 96.4 No 54.5

145

- 4S Yes 88.9 No 33.9



### In 2005, additional ALFOTERRA® surfac tants were tested used the OECD 301F

method. 2 

### The surfactants studied was as follows:

S23

- 3S

– ALFOTERRA® propoxylate sulfate surfactant produced from a linear

and mono

- methyl branched C12

- 13 alcohol with 3 moles of propylene oxide

S23

- 4S

– ALFOTERRA® pr opoxylate sulfate surfactant produced from a linear

and mono

- methyl branched C12

- 13 alcohol with 4 moles of propylene oxide

S23

- 5S

– ALFOTERRA® propoxylate sulfate surfactant produced from a linear

and mono

- methyl branched C12

- 13 alcohol with 5 moles of pr opylene oxide

S23

- 8S

– ALFOTERRA® propoxylate sulfate surfactant produced from a linear

and mono

- methyl branched C12

- 13 alcohol with 8 moles of propylene oxide

S45

- 2S

– ALFOTERRA® propoxylate sulfate surfactant produced from a linear

and mono

- methyl branch ed C14

- 15 alcohol with 2 moles of propylene oxideALFOTERRA® Sulfate Report Page 1 9 of 50



### The OECD 301F Manometric Respirometry Test was done as described in the OECD Guideline for Testing Chemicals adopted July 17, 1992 with the following minor modifications: (1) reduction in test material c oncentration from 100 mg to 20 mg to reduce possible toxicity to the inoculum, (2) reduction in the pH from 7.4 to 7.0 to improve the efficiency of CO2 removal, and (3) reporting of % ThCO 2 

### (theoretical maximum production of carbon dioxide) instead of % Th O 2 

### (theoretical maximum uptake of oxygen) to improve the accuracy of detecting ultimate biodegradation.

The biodegradation test was done on the ALFOTERRA® surfactants in quadruplicate using two 20 channel Coordinated Environmental Services (CES) Ltd (Corn well, UK) aerobic electrolytic respirometer units following the OECD 301 F guideline with slight modifications (as noted above).

The inoculum for these tests was derived from the recycled sludge from the City of Lake Charles, Louisiana Domestic Waste Wate r Plant A. The sludge was aerated ~ 7 days prior to use. The inoculum in the test medium was adjusted to a final concentration of < 30 mg TSS/L. Sodium benzoate (J.T. Baker

– 99% purity was used as the positive reference material).

The summarized resul ts from the study can be found in Table 10 , while the expanded data can be found in Appendix E.

Table 10 . Summary of u ltimate b iodegradation of ALFOTERRA® s urfactants in OECD 301F Test

ALFOTERRA® Surfacta nt OECD 301A

% ThCO 2 Pass 10

- day

Window 28

- Day

End

Point

S23

- 3S No 60.1

S23

- 4S No 55.5 a

S23

- 5S No 42.4

S23

- 8S No 43.3

S45

- 2S No 64.2 b

a mean of 3 samples

b mean of 2 samples



### 7.2 Aquatic Toxicity Results from F irst

- Ge neration Propoxy Sulfate Samples

Definitive Acute Aquatic Toxicity Tests on Daphnia magna and Pimephales promelas

were done on several first

- generation ALFOTERRA® propoxy sulfate surfactants by ENSR (Ft. Collins, CO) in 1999 (sample description give n in section 7.1) . The methods used for doing these tests were detailed in US EPA Method EPA/600/4

- 90/02/027F

– 1993 and ASTM Method E729

- 96

– 1998. ENSR reports can be found in Appendix F

. The results of acute aquatic toxicity tests are summarized in Table 11

.

Table 11 . Acute Aquatic Toxicity of first

- generation ALFOTERRA® s urfactantsALFOTERRA® Sulfate Report Page 20 of 50

ALFOTERRA® Surfactant **Daphnia magna**

48

- hr. LC 50 mg/L (95% CL) **Pimephales promelas**

96

- hr. LC 50 mg/L (95% CL)

12

- 3S 8 .88 (6.51

– 9.85) 10.99 (10.23

– 11.80)

123

- 2S 2.06 (1.76

– 2.29) 4.65 (4.27

– 5.08)

123

- 8S 2.65 (2.12

– 3.20) 1.68 (1.56

– 1.81)

145

- 2S 2.19 (1.86

– 2.49) 2.31 (2.14

– 2.48)

145

- 4S 2.34 (2.01

– 2.73) 1.62 (1.51

– 1.74)



### VIII. ENVIRONMENTAL ASSESSMENT OF ALFOTERRA® ALCOHOL PROPOXY

SULFATES

The environmental fate and effects of alcohol ethoxy sulfates have been thoroughly studied and have been the subject of a num ber of risk assessments including Environmental and Human Safety of Major Surfactants

– Alcohol Ethoxy Sulfates 3 

### and Human and Environmental Risk Assessment (HERA)

– Alcohol Ethoxysulphates (AES). 4 

### While there is a significant amount of literature data de scribing the environmental fate and effects of alcohol ethoxy sulfates, no literature data was found for alcohol propoxy sulfates. This makes comparisons of the current ALFOTERRA® alcohol propoxy sulfate results difficult.

8.1 Discussion of Biodegradation Results

T able 12 shows the combined biodegradation results of the ALFOTERRA® surfactants from both current and historical testing. The table has the ALFOTERRA® alcohol propoxy sulfates listed by increasing percentage of propoxyl ation. The percent branching of the starting alcohol is also listed for each surfactant.

T able 12

. Combined b iodegradation r esults for ALFOTERRA® a lcohol p ropoxy s ulfates

ALFOTERRA®

Surfactant Approx.

Percent Propoxylation 1 App rox. Percent

Branched Starting Alcohol OECD 301A (Primary Biodeg

– MBAS loss) OECD 301A (Ultimate Biodeg

– DOC loss) OECD 301F (Ultimate Biodeg

– ThCO 2 or ThOD)

145

- 2S 33% 95% 96.4% 54.5%

S45

- 2S 34% 50% 64.2%

123

- 2S 39% 94% 98.2% 52.1%

S23

- 3S 47% 5 0% 60.1%

167

- 4S 48% 63% 41.9%

145

- 4S 49% 95% 88.9% 33.9%

12

- 3S 50% 100% 96.3% 46.4%

S23

- 4S 54% 50% 55.5%

123

- 4S 54% 94% 91.1% 28.7% 34.4%

S23

- 5S 61% 50% 42.4%

167

- 7S 62% 63% 35.6%

145

- 8S 68% 95% 25.2%

S23

- 8S 78% 50% 43.3%

123

- 8S 87% 95% 46.1% 10.9% 18.9%

1 Percentage of PO in propoxylated alcoholALFOTERRA® Sulfate Report Page 21 of 50



### Five of the ALFOTERRA® surfactants ( 12

- 3S, 123

- 2S, 123

- 4S, 145

- 2S and 145

- 4S) with lower amounts of propoxylation ( < 4 moles or < 60%) exceeded 86% primary biodegradation as measured b y MBAS loss within 10 days. This would indicate these substances have lost their anionic surfactant properties and, therefore, have decreased aquatic toxicity.

For ultimate biodegradation, two low

- mole propoxylat e surfactants (S23

- 3S and S45

- 2S) met the ready biodegradability criteria of > 60% in the OECD 301F assay, but did not meet the 10 day window. An additional third surfactant, (S23

- 4S) was close to meeting the criteria with degradation of 55.5%.

The remaining compounds displayed slower and more l imited biodegradation.

The data indicates that the biodegradation profile of the ALFOTERRA® alcohol propoxy sulfates is influenced by both the amount of propoxylation and the amount of branching within the starting alcohol. As the percentage of propoxyla tion increases, the extent of biodegradation decreases. This is consistent with literature published on alkoxylated alcohols. Specifically, Naylor et al. 5 

### determined that biodegradation of alkoxylated alcohols was inversely proportional to the amount of propylene oxide incorporated into the surfactant. In addition, as the amount of branching within the starting alcohol increases, the extent of biodegradation decreases. This effect has been observed before in both the biodegradation of alcohol ethoxylate s and alcohol ethoxy sulfates. 3,6



### From the data, it appears that 4 moles (or < 60%) of propoxylation appears to be the defining point between extensive primary and ultimate biodegradation and slower limited biodegradation.

8.2 Discussion of Aquatic Toxic ity Results

Table 13 shows the combined aquatic toxicity results on the ALFOTERRA® surfactants from both current and historical testing. The table has the ALFOTERRA® alcohol propoxy sulfates listed by increasing p ercentage of propoxylation. The percent branching of the starting alcohol is also listed for each surfactant.

Table 13 . Combined a quatic t oxicity r esults for ALFOTERRA® a lcohol p ropoxy s ulfates

ALFOTERRA®

Surfactant Approx.

Perc ent Propoxylation 1 Approx.

Percent

Branched Starting

Alcohol Algae IC 50 mg/L (95%CL) **Daphnia**

LC 50 mg/L (95%CL) Fathead

Minnow LC 50 mg/L (95%CL)

145

- 2S 33% 95% 2.19 (1.86

– 2.49) 2.31 (2.14

– 2.48)

123

- 2S 39% 94% 2.06 (1.76

– 2.29) 4.65 (4.27

– 5.08)

167

- 4S 48% 63% 0.49 (0.35

– 1.34) 7.56 (6.03

– 9.04) 0.49 (0.44

– 0.56)

145

- 4S 49% 95% 2.34 (2.01

– 2.73) 1.62 (1.51

– 1.74)

12

- 3S 50% 100% 8.88 10.99ALFOTERRA® Sulfate Report Page 22 of 50

(6.51

– 9.85) (10.23

– 11.80)

167

- 7S 62% 63% 0.89 (0.72

– NA) 15.8 (10.7

– 25.6) 1.40 (1.21

– 1.59 )

145

- 8S 68% 95% 1.70 (1.51

– 2.10) 2.89 (2.42

– 3.40) 1.31 (1.23

– 1.39)

123

- 8S 87% 95% 2.65 (2.12

– 3.20) 1.68 (1.56

– 1.81)

1 Percentage of PO in propoxylated alcohol



### Unlike with the biodegradation data, there is no consistent trend with the combin ed aquatic toxicity of the ALFOTERRA® alcohol propoxy sulfates. There does not appear to be any environmentally relevant correlation between aquatic toxicity and amount of propoxylation, amount of branching in the starting alcohol or alkyl chain length. These results obtained for the ALFOTERRA® surfactants are similar to those for equivalent alcohol ethoxy sulfates.

8.3 Discussion of Fate in Use Application

As mentioned previously, the intended use of ALFOTERRA® surfactants is in enhanced hydrocarbon reco very (EHR) operations. ALFOTERRA® alcohol propoxy sulfates may be used in various aspects of improving recovery of hydrocarbon materials (oil, natural gas, etc.) from underground reservoirs. Many factors may influence the flow of hydrocarbons from a well . Conditions such as permeability, hydrocarbon concentration, porosity, temperature, presence of water, pressure of the hydrocarbon containing formation, and formation damage due to well treatment and drilling operations can all have an impact on the prod uction and economic potential of a hydrocarbon reservoir.

Before use, ALFOTERRA® surfactants will be blended with other chemicals, such

as polymers, salts, other surfactants, alcohols and water (predominate ingredient) to produce a water

- flood or carrier fluid that will be injected into the hydrocarbon reservoir. The role of the alcohol propoxy sulfate surfactant is to reduce the interfacial surface

tension between the oil and the carrier fluid in order to increase the amount of hydrocarbon that can be f lushed/pulled out of the reservoir. Typically, only low concentrations (0.1

- 1.0% as active) of ALFOTERRA® surfactants are required in these operations.

Ultimately, both the hydrocarbon material and the carrier fluid are pumped from the

reservoir as a mixture. It is then necessary to separate the water from the other organic materials (hydrocarbon material and other organics from original carrier fluid formulation). The water is normally separated using either chemical or physical

methods. It is esti mated the separated water will only contain trace levels (< 10 mg/L) of ALFOTERRA® alcohol propoxy sulfate. This low level of surfactant is a result of two processes. First, after the initial injection of the ALFOTERRA® containing water flood,

additiona l water may be injected to improve hydrocarbon recovery. This will result in a dilution of the ALFOTERRA concentration in the recovered hydrocarbon/water mixture. Second, a significant portion of the surfactant is expected to bind to the subsurface

forma tion. The amount of adsorption will be dependant on the composition of the formation. Adsorption losses are predicted to range from 50% for shale formations toALFOTERRA® Sulfate Report Page 23 of 50



### 100% for dolomite formations. The separated water is then normally reused to make

additional carrier fluid, or injected into a disposal well if reuse is not possible.

As a result, ALFOTERRA® surfactants will not be released into surface or ground water and the predicted exposure concentration (PEC) will be zero.

IX. SUMMARY

The rate and extent of biodegradation of the ALFOTERRA® alcohol propoxy sulfates were dependent on the amount of propoxylation and branching in the starting alcohol. ALFOTERRA® surfactants with a low proportion ( < 4 moles or < 60%) of propox ylation displayed extensive primary biodegradation and, in some cases, ultimate biodegradation. The surfactants with a higher amount of propoxylation had limited biodegradation. The aquatic toxicity of the ALFOTERRA® alcohol propoxy sulfates is

similar t o that of equivalent alcohol ethoxy sulfates.

During Enhanced Hydrocarbon Recovery, no level of ALFOTERRA® surfactants will be discharge into surface or ground water and the application will, therefore, not produce any adverse effects to the environment

.

X. ACKNOWLEDGEMENTS

The authors acknowledge Rebe kah Cohen for skillful assistance with the biodegradation test, and Rocco Di Modugno and the staff at Sasol Italy for producing the samples.ALFOTERRA® Sulfate Report Page 24 of 50



### XI. REFERENCES

1. Nielsen, A.M. and G. Russell. 2006. Biodegradability and Aquatic Toxicity of ALFOTERRA® Surfactants. Internal Sasol North America Research Report (attached to PMN as supporting document).

2 . OECD. 1993. OECD Guidelines f or the Testing of Chemicals

– OECD 301F Manometric Respirometry Test

. Organization of Economic Cooperation and Development. Paris

3

. Arthur D. Little, Inc. 199 1

. Environmental and Human Safety of Major Surfactants

. Volume 1. Anionic Surfactants. Part 2. Alcohol Ethoxy Sulfates. Final report to the Soap and Detergent Association

4. HERA . 2004. Human & Environmental Risk Assessment on Ingredients of European Household Cleaning Products

- Alcohol Ethoxy sulphates (AES)

5. Naylor, C.G., F.J. Castaldi and B .J. Hayes. 1988. Biodegradation of nonionic surfactants containing propylene oxide. J. Am. Oil Chem. Soc. 65:1669

- 1676 as reported in Talmage, S.S. 1994. Environmental and Human Safety of Major Surfactants : Alcohol Ethoxylates and Alkylphenol Ethoxylates

. Lewis Publishers, Boca Raton, F L

6

. Talmage, S.S. 1994. Environmental and Human Safety of Major Surfactants : Alcohol Ethoxylates and Alkylphenol Ethoxylates

. Lewis Publishers, Boca Raton, F LALFOTERRA® Sulfate Report Page 25 of 50



# Biodegradability and Acute Aquatic Toxicity of Alcohol Prop oxy Sulfates.

Research Report R&D ENV

- 07

- 024 01

- 07

Part 2 of 2 Appendices

Geoff Russell Ricky Stackhouse Wayne Sorensen

November 2007

Research & Development Lake Charles, LouisianaALFOTERRA® Sulfate Report Page 26 of 50



### Appendix A

. Biomonitoring Report **Daphnia pulex & Pimephales promelas**

.

Prepared for Sasol by EE USA Lab Report # Q

- 506

- 07 Sample ID: ALFOTERRA® 145

- 8 S
October 31, 200 7ALFOTERRA® Sulfate Report Page 27 of 50



### Appendix B

. An Evaluation of the Chronic Toxicity of the AL FOTERRA® Samples to Freshwater Alga **Selenastrum capricornutum**

.

Prepared for Sasol by Pacific EcoRisk. October, 2007ALFOTERRA® Sulfate Report Page 28 of 50



### Appendix C

. Biomonitoring Report. **Daphnia pulex & Pimephales promelas**

.

Prepared for Sasol by EE USA Lab Report # Q

- 6 561

- 07 Sampl e ID: ALFOTERRA® L167

- 4 S
October 31, 200 7ALFOTERRA® Sulfate Report Page 29 of 50



### Appendix D. Biomonitoring Report. **Daphnia pulex & Pimephales promelas**

.

Prepared for Sasol by EE USA Lab Report # Q

- 562

- 07 Sample ID: ALFOTERRA® L167

- 7 S
October 31, 200 7ALFOTERRA® Sulfate Report Page 30 of 50



### Appendix E

. Biodegradation of Early Ver sions of ALFOTERRA® Propoxy Sulfate Surfactants

Primary Biodegradation of ALFOTERRA® Surfactants in the OECD 301A Test



### ALFOTERRA®

Surfactant % MBAS Loss

1 6 9 10 13 15 17 22 28

12

- 3S

± Std. Dev. 4.7

± 1.4 86.3

± 0.7 87.9

± 0.2 88. 9

± 1.9 88.8

± 0.1 89.1

± 2.8 95.2

± 0.3 --

-- 96.3

± 0.4

123

- 2S

± Std. Dev. 5.1

± 0.5 95.8

± 0.2 97.4

± 0.2 97.6

± 0.3 97.2

± 1.5 98.4

± 0.1 97.5

± 0.2 98.3

± 0.1 98.2

± 0.2

1 23

- 4S

± Std. Dev. 10.1

± 5.5 87.6

± 2.2 86 .9

± 0.4 86.8

± 0.3 88.6

± 0.2 91.5

± 0.6 89.0

± 0.6 --

-- 91.1

± 0.6

123

- 8S

± Std. Dev. 5.3

± 1.7 35.8

± 4.2 46.0

± 3.2 39.9

± 5.6 45.5

± 0.9 51.6

± 2.5 45.3

± 1.3 --

-- 46.1

± 0.3

1 45

- 2S

± Std. Dev. 8.3

± 1.8 97.2

± 0.2 96.7

± 0.1 97.6

± 0.1 97.8

± 0.3 98.6

± 0.2 --

-- --

-- 96.4

± 0.1

145

- 4S

± Std. Dev. 4.7

± 1.41 86.4

± 0.7 87.9

± 0.2 88.9

± 1.9 88.8

± 0.1 88.8

± 2.5 88.1

± 2.5 --

-- 88.9

± 2.3



### Ultimate Biodegradation of ALFOTERRA® Surfactants i n the OECD 301A Test



### ALFOTERRA®

Surfactant % DOC Loss

1 3 6 9 10 13 15 17 22 28

12

- 3S

± Std. Dev. 13.8

± 7.7 1.7

± 2.4 5.8

± 0.5 13.8

± 2.2 24.4

± 1.2 25.5

± 2.5 35.4

± 2.7 40.0

± 0.1 45.7

± 2.5 46.4

± 4.9

123

- 2S

± Std. Dev. 8.4

± 11.9 20.8

± 1.6 29.2

± 0.1 38.3

± 1.0 39.6

± 3.5 43.0

± 0.6 47.3

± 2.8 53.7

± 2.1 56.7

± 1.34 52.1

± 1.41

123

- 4S

± Std. Dev. 3.9

± 1.3 1.4

± 1.6 8.8

± 2.8 16.1

± 0.4 17.4

± 1.1 17.1

± 0.2 21.5

± 2.1 28.0

± 0 .6 29.9

± 3.3 28.7

± 2.5

123

- 8S

± Std. Dev. 0

± 0 0

± 0 1.3

± 1.8 0

± 0 10.9

± 4.2 5.7

± 1.3 4.7

± 0.8 14.0

± 0.8 15.1

± 2.3 10.9

± 1.6

14 5

- 2S

± Std. Dev. 2.4

± 0.2 21.6

± 0.5 28.8

± 0.5 37.8

± 2.2 42.8

± 0.2 4 3.3

± 0.8 45.2

± 2.9 51.5

± 0.8 54.6

± 3.1 54.5

± 3.5

145

- 4S

± Std. Dev. 2.7

± 0.9 2.9

± 1.3 13.3

± 3.5 18.2

± 0.8 24.8

± 0.1 26.3

± 2.1 28.1

± 4.2 34.7

± 0.6 36.2

± 1.2 33.9

± 1.3

Acetate

± Std. Dev. 88.9

± 9. 5 97.4

± 0.8 96.5

± 0.5 98.5

± 1.5 96.5

± 0.3 99.2

± 1.3 97.6

± 0.9 98.3

± 2.5 96.5

± 1.1 100

± 0ALFOTERRA® Sulfate Report Page 31 of 50



### Ultimate Biodegradation of ALFOTERRA® Surfactants in the OECD 301F Test



### ALFOTERRA®

Surfactants %ThCO 2



### 1 2 3 4 6 8 10 12 14 16 18 20 22 24 26 28

S45

- 2 S (sample 1)

± Std. Dev. 0.1

± 0.4 4.8

± 0.3 16.3

± 0.2 23.8

± 0.2 36.8

± 0.4 47.4

± 0.3 52.0

± 0.2 54.5

± 0.3 56.1

± 0.5 56.4

± 0.1 57.7

± 0.2 58.3

± 0.9 59.2

± 1.0 60.0

± 0.8 61.6

± 0.6 62.3

± 0.5

S45

- 2S (sample 2)

± St d. Dev. 0

± 0 5.3

± 0.7 16.5

± 1.2 24.5

± 1.2 37.7

± 0.6 47.8

± 0.4 52.5

± 0.4 55.5

± 0.6 57.3

± 0.7 58.8

± 0.3 60.5

± 0.3 61.2

± 0.6 62.5

± 0.8 63.8

± 1.3 65.3

± 1.6 66.1

± 1.8

S23

- 3S

± Std. Dev. 0

± 0 3.4

± 0.1 11.3

± 0.3 19.6

± 0.5 33. 4

± 0.2 41.1

± 2.3 45.1

± 2.6 49.2

± 2.0 51.2

± 1.7 53.1

± 1.5 54.7

± 0.8 56.0

± 1.1 56.8

± 1.0 58.0

± 1.0 59.6

± 1.0 60.1

± 0.8

S23

- 4S (sample 1) 0 2.2 8.7 14.5 24.4 29.7 33.3 35.6 36.4 37.7 41.6 43.9 43.7 45.0 46.2 46.5

S23

- 4S (sample 2)

± Std. Dev. 0.1

± 0.4 2.3

± 0.7 8.3

± 0.4 14.8

± 1.3 26.9

± 2.6 33.8

± 0.1 37.9

± 1.3 41.4

± 2.0 43.2

± 2.7 43.8

± 3.5 45.6

± 3.1 46.6

± 2.9 46.9

± 3.2 47.7

± 3.4 49.6

± 2.9 50.0

± 2.9

S23

- 4S (sample 3)

± Std. Dev. 0.2

± 0.8 2.3

± 0. 5 8.6

± 0.3 15.5

± 1.1 31.6

± 4.2 43.3

± 4.5 49.7

± 5.9 52.6

± 6.4 55.0

± 5.6 56.8

± 5.1 57.6

± 3.9 60.8

± 3.7 61.4

± 3.5 64.3

± 8.2 66.6

± 8.7 69.9

± 10.3

S23

- 5S 0.7 1.0 2.8 4.4 12.1 19.4 23.9 28.3 30.9 31.7 34.3 34.8 36.3 38.6 41.7 42.4

S23

- 8S

± Std. Dev. 0

± 0 1.3

± 0.7 4.1

± 0.3 8.3

± 0.1 20.8

± 2.0 26.8

± 2.2 30.3

± 2.4 33.7

± 2.6 35.6

± 2.4 36.6

± 2.7 38.2

± 2.4 39.2

± 2.5 40.3

± 3.0 41.3

± 3.1 42.9

± 3.2 43.3

± 3.3

Sodium Benzoate

± Std. Dev. 2.6

± 2.7 39.2

± 2. 3 50.9

± 3.6 62.6

± 4.9 78.1

± 5.9 83.9

± 5.4 89.6

± 4.8 91.0

± 4.3 93.2

± 4.0 95.4

± 3.8 97.0

± 3.4 97.9

± 3.2 98.4

± 3.3 98.8

± 3.3 99.0

± 3.4 99.1

± 3.4ALFOTERRA® Sulfate Report Page 32 of 50



### A ppendix F

. Toxicity Reports on Earlier Versions of ALFOTERRA® Propoxy Sulfate Surfactants. **Daph nia magna and Pimephales promelas.** 

Prepared for Condea Vista Company by ENSR Lab Report # 8503

- 111

- 006 (015 thru 020) Lab Report #8503

- 106

- 006 (001 thru 008) August 1999ALFOTERRA® Sulfate Report Page 33 of 50ALFOTERRA® Sulfate Report Page 34 of 50ALFOTERRA® Sulfate Report Page 35 of 50ALFOTERRA® Sulfate Report Page 36 of 50ALFOTERRA® Sulfate Report Page 37 of 50ALFOTERRA® Sulfate Report Page 38 of 50ALFOTERRA® Sulfate Report Page 39 of 50ALFOTERRA® Sulfate Report Page 40 of 50ALFOTERRA® Sulfate Report Page 41 of 50ALFOTERRA® Sulfate Report Page 42 of 50ALFOTERRA® Sulfate Report Page 43 of 50ALFOTERRA® Sulfate Report Page 44 of 50ALFOTERRA® Sulfate Report Page 45 of 50ALFOTERRA® Sulfate Report Page 46 of 50ALFOTERRA® Sulfate Report Page 47 of 50ALFOTERRA® Sulfate Report Page 48 of 50ALFOTERRA® Sulfate Report Page 49 of 50ALFOTERRA® Sulfate Report Page 50 of 50