## Ecological Effects and Endangered Species Risk Assessment **for Choline Chloride**

Choline chloride a quaternary ammonium salt is miscible with water in all proportions and has a measured water solubility of 650 g/L

. Due to the measured and calculated log Kow of

–3.77 and

– 5.15, respectivel y, and the calculated log Koc of 0.37, bio

- and geoaccumulation are not to be expected. Choline chloride was shown to be readily biodegradable according to OECD criteria in a MITI I

- Test within 14 days. In the atmosphere, the substance will be rapidly indirectl y photodegraded b y reaction with OH

- radicals (calculated t 1⁄2 = 6.9 h) based on a 12h

- day

. **Terrestrial Species** The Agency has estimated potential acute toxicity end points for avian and mammalian species (LD 50 

## = 3,129 mg/kg) and calculated potential risk using the Terrestrial Residue Exposure Model (T

- REX) Version 1.4.1 for the orchard spray application of CP Bio, Choline Chloride 20% SP at the maximum rate of 0.282 lbs ai/A (Appendix B ). The T

- REX model using Ken ega values produced low dose

-based values and risk quotients ( RQs ) that were at least one to two orders of magnitude ( 0.00

– 0.06) below the EPA Levels of Concern (LOC) of 0.1 for avian and mammalian endangered species. The possible effect to honey bees is also very

low with an LD 50 

## > 89 ug ai/bee (RQ < 0.09). The Agency also conducted a search (10/0 9/201 5 ) of the Ecological Incident Information System version 2.1.1 (EIIS) and did not find any reports of detrimental exposure of choline chloride to terrestrial species

. This low risk to non

- target terrestrial species suggests that the potential for toxic exposure to endangered/ threatened species is also very

low. The Agency

calculated the chance of individual terrestrial exposure and risk by using the Individu al Effects Chance Model (Version 1.1) which suggest that the chance of effects to a terrestrial species from choline chloride exposure is about 1 in 2,940,000 (p redictor of chance of individual effect using Probit dose

- response curve slope and median lethal estimate Appendix C ) which may

be considered as a highl y unlikely event. The low toxic risk of choline chloride to terrestrial organisms (RQ = 0.0

– 0.6) , the short environmental half

- life (7 days), the low maximum concentration of choline chloride to be applied per season (128.0 g/A), and an understanding of the mode of action of this compound (plant growth regulator) show that there is a very low potential for toxic risk to terrestrial species. Based on the proceeding information, **choline chloride presents a No Effect (NE) to threatened or endangered terrestrial species,**  **if used in accordance with approved labeling**

. **Aquatic Species** Choline chloride is very

water soluble (690 g/L) but exposure to aquatic species is not expected to result in acute toxicity

. The Agency conducted a worst case scenario using the aquatic model FIRST Ver 1.1.1 to assess the potential exposure of runoff and drift from the maximum application of choline chloride formulation to an aquatic area (Appendix A). Using the EPA LOC values for aquatic listed species (0.05), the expected peak estimated environmental concentration of 0. 0 60 ppm , and assuming that 0.282 lbs ai/A, RQ values were calculated at 0.00 showing no acute toxicity risk to aquatic species. The Agency conducted a search (10/ 0 9/201 5 )

## of the Ecological Incident Information System (EIIS, version 2.1), the ‘Aggregate Incident

Reports’ (v. 1.0) database, for ecological incidents involving choline chloride and no aquatic incidents were reported. The Agency also calculated the chance of individual fish and aquatic invertebrate exposure and risk by using the Individual Effects Chance Model (Version 1.1). These calculations suggest that the chance of effects to aquatic invertebrates (food source) or to fish from methyl cho line chloride is about 1 in 418,000,000 (Appendix C ), which may be considered as a highly

unlikely event. Therefore, the Agency concludes **a No E**  **ffect (NE) to aquatic endangered/ threatened species**  because of the lack of potential for direct or indirect tox ic risk to aquatic ecological systems from exposure to choline chloride

. **Plants (Aquatic and Terrestrial)**  Concentration levels of choline in plants can range from 28

– 589 ppm . The environmental residue levels of choline chloride after direct application to foliage were calculated from the T

Rex model and show ed a range of 7.93

– 126.90 ppm . According to the assumptions of the EPA FIRST Ver. 1.1.1 aquatic model there is an expected 8% drift from orchard spraying that could result in 0.63

– 10.15 ppm re sidues onto adjacent vegetative sites. These expected environmental concentration to non

- target plants are at or below the background levels found in plants. Choline chloride is non

- toxic to plants and any indirect effects to non
- target plants that could be considered habit for endangered/threatened species is expected to be **No**
**- Effect** 

. **Conclusion** The unique characteristics of this product, choline chloride, the non

- toxic mode of action (plant growth regulator) , biodegradability (7 day half

- life ) , photo degradation (6.9 hours), no bioaccumulate (Log Koc = 0.37) and low potential for exposure (maximum application of 0.282 lbs ai/A 4 times /season) will result in no acute direct or indirect toxic risk to non

- target organisms, including threatened and endangered species. Based on the proceeding information, **choline chloride presents a No Effect (NE) to threatened or endangered terrestrial or**  **aquatic species (plants or animals)**  **, if used in accordance with approved labeling .** 

## Reference **s**

MOE Japan (1999 ). Ministry of Environment. Acute toxicity study of choline chloride on the freshwater alga Pseudokirchneriella subcapitata unpublished study . No. 1998

- 13. BASF AG (2003 ). Department of Product Safety. Laboratory of Ecology . Hemmung der Algenzellvermehrung nach DIN 38412 L9: "78% choline chloride dissolved in water". Unpublished data. Reprint of report No. 09908 (06 Nov 1989). 16 Sept. 2003.

## A**ppendix A.0 Calculate Water Concentrations for Choline Chloride using**  **FIRST Ver 1.1.1**

**MAR 26, 2008**

**Table A.1**  **Water Concentration for Choline Chloride after Application to Stone Fruits at 0.282 lbs/A** 

-------------------------------------- --------------------- -------------------------- ------------------------------ RATE (#/AC) No. APPS & SOIL SOLUBIL APPL TYPE %CROPPED INCORP ONE (MULT) INTERVAL Koc (PPM) (%DRIFT) AREA (IN)

--------------------- ------------------------------------------ ----------------------------------------------- ----- 0.282 (0. 722 ) 4 7 0.4 ******* ABLAST (6.3) 87.0 0.0 FIELD AND RESERVOIR HALFLIFE VALUES (DAYS)

-------------------------------------------------------------------- METABOLIC DAYS UNTIL HYDROLYSIS PHOTOLYSIS METABOLIC COMBINED (FIELD) RAIN/RUNOFF (RESERVOIR) (RES.

- EFF) (RESER.) (RESER.)

--- ---- ----------------------------------------------- ----------------------------------------------------------------- 14 .00 0 14 0.29

- 35.96 14.00 10.08 UNTREATE D WATER CONC ENTRATION ( **PPB** ))

-------------------------------------------------------------------- PEAK DAY (ACUTE) ANNUAL AVERAGE (CHRONIC)



## ---------------------------------------------------------- --------------------------- ------------- ----------

**60.101 1.792** 

## Appendix B.0 T
**- REX Output for Choline Chloride Exposure to Avian and Mammalian** 

**Species after Table B.1**  **Terrestrial Residue Expo sure Model**  **(T**
**- REX) Version 1.4.1**  **Application to Stone F**  **ruit**

**Che mic**  **al N am**  **e: U**  **se F**  **orm ul**  **at ion**  **Ap plic** **at io**  **n R at** **e 0.**  **282 lbs**  **a .i./a**  **cre Ha**  **lf -l**  **if e**  **7 da**  **ys Ap**  **plic at**  **ion Int er** 

**va l**  **7 da**  **ys Ma**  **xi mu**  **m #**

**A pps**  **./Y ear**  **4 Le**  **ngth of S imu**  **la ti**  **on 1**  **ye ar**  **Va ri**  **ab le ap**  **plic at**  **ion ra te**  **s? no** 

**Bo bw**  **hit e**  **qua il**  **LD 50**  **(m g/k** **g- bw**  **) 3129.**  **00**
**Bo bw**  **hit e**  **qua il**  **LC 50**  **(m g/k** **g-die t)**  **0. 00**  **Ma llar** **d duck**  **NOAE L(**  **mg/k g-**  **bw )**  **0. 00** 

**Bo bw**  **hit e**  **qua il**  **NOAE C**  **(m g/**  **kg-die t)**  **0. 00** 

**3129. 00**  **0. 00**  **0. 00**  **0. 00** 
**Ke na**  **ga Va**  **lu es**  **Short G**  **ra ss**  **126. 90**  **Ta ll G**  **ra ss**  **58. 16**  **Br oa**  **dle af** 

**pla nts**  **71. 38**  **Fruit s/**  **pods**

**/s eed**  **s 7.**  **93 Ar** **thropods 49.**  **70** 

## Endpoi **nt s**  **U pper**  **Bo und**  **Kenaga Re**  **si dues**  **For RQ** **C al**  **cul at**  **io n** 



# Di **etary -ba**  **sed E ECs**  **(p pm**  **) NOAEL**  **(m g/k** **g- bw**  **) NOAE**  **C (m**  **g/ kg-die**  **t)** 

# Ma **mmal s**  **LD 50**  **(m g/k** **g- bw**  **) LC**  **50 (m**  **g/k g-die**  **t) Ch**  **oline c**  **hloride st** **one f** **rui ts**  **CP B**  **io C**  **holine C**  **hloride**



# Av **ia n** 

### Av **ia n Resul**  **ts Avi**  **an Body**  **In ge**  **st ion (Fdry)**  **Inge st**  **io n**  **(F wet** **) %**  **body w gt**  **FI Cl**  **as s**  **Weig ht (g)**  **(g bw**  **/da y)**  **(g/ da**  **y) cons** **um ed**  **(kg-di et**  **/da y)**  **Sma ll**  **20 5**  **23 114**  **2. 28E-02**  **Mid 100**  **13 65**  **65 6.**  **49E-02 La**  **rge 1000**  **58 291**  **29 2.**  **91E-01 20**  **5 5**  **25 5.**  **06E-03 Gra**  **ni vore**  **s 100**  **13 14**  **14 1.**  **44E-02 1000**  **58 65**  **6 6.**  **46E-02**

**Avi an**  **B ody**  **Adj us**  **te d**  **LD50 Weig**  **ht (g) (m**  **g/kg- bw**  **) 20**  **2254. 22**  **100 2869.**  **74 1000**  **4053. 62** 

**sm all**  **mid la**  **rge 20**  **100 1000**  **Short Gra ss**  **144. 53**  **82.42 36.**  **90 Ta**  **ll Gra ss**  **66.24 37.77**  **16. 91**  **Broa dle**  **af**

**p lan**  **ts 81.30**  **46.36 20.**  **76 Fruits**  **/p ods**  **9.03 5.**  **15 2.**  **31 Arth**  **ropods 56.61**  **32.28 14.**  **45 Seeds**  **2.01 1.**  **14 0.**  **51**

**20 100**  **1000 Short Gra**  **ss 0.06**  **0. 03**  **0. 01**  **Ta ll Gra**  **ss 0.03**  **0. 01**  **0. 00**  **Broa dle**  **af**

**p lan**  **ts 0.04**  **0. 02**  **0. 01**  **Fruits /p**  **ods 0.00**  **0. 00**  **0. 00**  **Arth ropods**  **0.03 0.**  **01 0.**  **00 Seeds**  **0.00 0.**  **00 0.**  **00** 

## Di **etary-b ased**  **R Qs**  **(D ie**  **ta ry**  **-b ased**  **EEC/ LC50**  **or Acute**  **Ch roni**  **c Short Gra**  **ss #DI**  **V/0 !**  **#DI V/0**  **! Ta**  **ll Gra ss**  **#DI V/0**  **! #DI**  **V/0 !**  **Broa dle**  **af**

**p lan**  **ts #DI**  **V/0 !**  **#DI V/0**  **! Fruits**  **/p ods**  **/s eed**  **s #DI**  **V/0 !**  **#DI V/0**  **! Arth**  **ropods #DI**  **V/0 !**  **#DI V/0**  **!** 

### Dose-based EECs **(m g/**  **kg-bw )**  **Avi an**  **C las**  **se s**  **an d**  **Bo dy W**  **ei ght** **s (gra**  **ms )** 



## Do **se- ba**  **sed R**  **Qs (D**  **ose-based EEC/ adj**  **ust ed**  **LD50)**

**RQ s**  **Av ia**  **n Ac**  **ut e R**  **Qs Si**  **ze C**  **la ss (grams)** 

## The application of CP Bio, Choline Chloride 20% SP at the maximum rate of 0.282 lbs ai/A does

not pose an acute risk to nontarget or threatened avian or mammalian species. Risk quotients (RQs) were generated using the acute mammalian LC 50 

## = 3 , 129 mg/kg . The T

- REX model using Kenega values produced low dose

- based values and RQs that were at least one to two orders of magnitude (0.00

– 0.06) below the EPA Levels of Concern (LOC) of 0.1 for avian and mammalian endangered species. **Ma mmal**  **ia n Resul**  **ts**
**Ma mmalian**  **Bo dy**  **Inge st**  **ion ( Fdry)**  **Inge st**  **ion ( Fw**  **et )**  **% body**  **wg t**  **FI Cl**  **as s**  **Weig ht**  **(g bw**  **t/ da**  **y) (g**  **/da y)**  **cons um**  **ed (kg-d**  **iet /da**  **y) 15**  **3 14**  **95 1.**  **43E -02**  **He rbivore**  **s/ 35**  **5 23**  **66 2.**  **31E -02**  **ins ec** **ti vore**  **s 1000**  **31 153**  **15 1.**  **53E -01**  **15 3**  **3 21**  **3. 18E**  **-03 Gra**  **invore s**  **35 5**  **5 15**  **5. 13E**  **-03 1000**  **31 34**  **3 3.**  **40E -02** 
**Ma mmalian**  **Bo dy**  **Adj us**  **te d**  **Ad jus** 

**te d**  **Cl as**  **s Weig**  **ht LD50**  **NOAEL**

**15 6877.**  **01 0.**  **00 He**  **rbivore s/**  **35 5564.**  **24 0.**  **00 ins**  **ec ti**  **vore s**  **1000 2406.**  **70 0.**  **00 15**  **6877. 01**  **0. 00**  **Gra nivore**  **s 35**  **5564. 24**  **0. 00**  **1000 2406.**  **70 0.**  **00**

**15 35**  **1000 Short Gra**  **ss 120.**  **99 83.**  **62 19.**  **39 Ta**  **ll G ra**  **ss 55.**  **45 38.**  **33 8.**  **89 Br** **oa dl**  **eaf**

**plant s**  **68. 06**  **47. 04**  **10. 91**  **Fruit s/**  **pods 7.**  **56 5.**  **23 1.**  **21 Ar** **thropods 47.**  **39 32.**  **75 7.**  **59 Se**  **ed s**  **1. 68**  **1. 16**  **0. 27** 

**15 gra**  **ms 35**  **gra ms**  **1000 gra**  **ms Acu**  **te Chronic**  **Acute Chronic**  **Acute Ch**  **roni c**  **Short Gra ss**  **0. 02**  **#DI V/0**  **! 0.**  **02 #DI**  **V/0 !**  **0. 01**  **#DI V/0**  **! Ta**  **ll G ra**  **ss 0.**  **01 #DI**  **V/0 !**  **0. 01**  **#DI V/0**  **! 0.**  **00 #DI**  **V/0 !**  **Br oa**  **dl eaf** 

**plant s**  **0. 01**  **#DI V/0**  **! 0.**  **01 #DI**  **V/0 !**  **0. 00**  **#DI V/0**  **! Fruit** **s/ pods**  **0. 00**  **#DI V/0**  **! 0.**  **00 #DI**  **V/0 !**  **0. 00**  **#DI V/0**  **! Ar** **thropods 0.**  **01 #DI**  **V/0 !**  **0. 01**  **#DI V/0**  **! 0.**  **00 #DI**  **V/0 !**  **Se ed**  **s 0.**  **00 #DI**  **V/0 !**  **0. 00**  **#DI V/0**  **! 0.**  **00 #DI**  **V/0 !**  

### Do **se-Based EECs (m**  **g/kg-bw )**  **(gra ms**  **) Ma**  **mmalia n C**  **las**

**se s**  **an d Body w**  **ei ght** 

**Me di**  **um mammal La**  **rge mammal**  

## Do **se-b ased**  **R Qs**  **(D ose-**  **based EEC/**  **LD50 or**  **Small mammal**

## Appendix C.0 **Individual Effects Chance Model for Terrestrial and Aquatic Species** 

**Table C.1**  **. Individual Effects Chance Model Version 1.1 for Predicting the Chance of Individual Effects of Choline Chloride to**  **Terrestrial Endangered**  **Species IEC V1.1**  **- Individual Effect Chance Model Version 1.1**  **Predictor of chance of individual effect using Probit**  **dose -**  **response curve slope and median lethal estimate** Enter LC 50 

### or LD 50 

### 3129

Enter desired threshold 0.1 Enter slope of dose

- response 4.5 z score result

- 4.5 Probability associated with z 3.40E

- 06 Chance of individual effect, **~1 in . . .** 2.94E+05



## Table C. **2 . Individual Effects Chance Model Version 1.1 for Predicting**  **the Chance of Individual Effects of Choline Chloride**  **to Aquatic Endangered Species**  **IEC V1.1 -**  **Individual Effect Chance Model Version 1.1 Predictor of chance of individual effect using**  **Probit dose**  **- response curve slope and median lethal estimate**  Enter LC 50 

### or LD 50 

### 100

Enter desired threshold 0.05 Enter slope of dose

- response 4.5 z score result

- 5.85463498 Probability associated with z 2.39E

- 09 Chance of individual effect, **~1 in . . .** 4.18E+08

## Appendix D.0 Calculated RQ Values

**Table D.**  **1 Summary of Specific**  **Ecological Asse**  **ssment Endpoints and Calculated RQ**  **Values for Terrestrial and**  **Aquatic Species Exposed to Choline Chloride after Application to Stone fruits**  **TAXA ME**  **ASURE OF EFFECT Species**  **Toxicity RQ**  **Freshwater Fish Acute**  Rice fish ( Oryzias latipes ) 96 hr LC 50 

### > 100 mg/L 

### 0.00 **Freshwater Invertebrates**  **Acute** Daphnia magna EC 50 

### = 349 mg/L **0.00 Avian**  **Acute** Mallard duck ( Anas platyrhynchos ) a 

### LD 50 

### = 3,129 mg ai/kg **0.00**

**– 0.6**  **Mam mals**  **Acute** Rat (Rattus norvegicus) LD 50 

### = 3,129 mg/kg **0.00**

**– 0.2**  **Insect Acute**  Honey bee ( Apis mellifera ) LD 50 

### > 89 ug ai/bee **< 0. 0**  **9**

a 

### The mammalian acute toxicity data is used as a surrogate for avian oral dose