UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON D

. C ., 20460

August 

### 7, 2009
MEMORANDUM OFFICE OF PREVENTION , PESTICIDES AND TOXIC SUBSTANCES

SUBJECT: Supplemental Ethics Review of Chlorpyrifos Human Toxicity St eta/.
FROM: John M. Carley Human Research Ethics Review Officer Office ofPesticide Programs
TO: Anna Lowit, Ph.D. Health Effects Division
RE: Kisicki , J.; Seip, C.; Combs, M. ( 1999) A Rising Dose Toxicological Study to Determine the No-Observable-Eff ect-Level s (NOEL) For Erythrocyte Acetylcholinesterase (AChE) Inhibition and Cholinergic Signs and Symptoms of Chlorpyrifos at Three Dose Levels. Unpublished study prepared by MDS Harris Laboratories under Project No. 21438 and Dow AgroSciences Study No. DR K

0044793-284. 578 p. (MRID 44811 002) Juberg, D

. ; Mattsson, J

. (2008) Updated Dow AgroSciences Response to EPA Query Regarding Two Toxicol ogy Reports. Unpub lish ed document prepared by Dow AgroSciences LLC under Study ID DRJ05282008. 27 p. (MRID 47436401) Carley, J. (2009) Ethics Review ofChlorpyrifos Human Toxicity Study. Memorandum dated May 29, 2009 to Anna Lowit. 11 p.
Kisicki, J. (2009) " Re: Ethics Review ofChlorpyrifos Human Toxicity Study". Unpublished letter dated June, 23 , 2009 to K. Racke of Dow AgroSciences, responding to EPA Ethics Rev iew ofMRID 44811002 dated May 29,2009. 7 p.
Anon. (1998) Attachment to June 23, 2009 J. Kisicki letter to K. Racke with subjects de

- identified. Unpublished Screening Logs dated September 2 1, 1998 and October 6, 1998 , used by MDS-Harris Laboratories in the execution ofthe study reported in MRID 44811002. Submitted to EPA July 1, 2009. 18 p
.
Page 1 of6The last two documents cited above came to EPA after I had completed my May 29, 2009 ethics review of this study. I have reviewed them carefully to assess whether they provide evidence that would lead me to change my earlier conclusion that EPA is forbidden by 40 CFR §26.1704 to rely on the Kisicki et al. study, MRID 44811002, in actions taken under FIFRA or §408 ofFFDCA.
I have not yet seen the final recommendations ofthe HSRB concerning this study. At their meeting on June 24, and at least in part on the basis of assurances in the June 23 , 2009 letter from Dr. Kisicki , they indicated disagreement with my conclusion. Nonetheless, my analysis of the new evidence does not persuade me to change that conclusion.

Supplemental Ethics Review During the HSRB meeting on June 24, 2009, at which the Kisicki et al. study of chlorpyrifos RBC cholinesterase inhibition was discussed, Dow AgroSciences (DAS) submitted to EPA a letter dated June 23, 2009, from Dr. James C. Kisicki ofMSD Pharma Services to Kenneth D. Racke of DAS. Dr. Kisicki was the Study Director for this study, archived under MRIDs 4481 1002, 45144101 , 47429401, and 47436401.
The description of the candidate screening procedure in the Kisicki letter was read to the HSRB during their meeting, and it materially affected their deliberations. The letter as submitted, however, did not include the subject sign-in logs cited as an attachment, which are critical to support the assertions in the letter. At EPA's request, DAS provided the logs to EPA later, after de-identification of subjects.
The central concern of my May 29, 2009 ethics review was whether all participants had provided legally effective informed consent before becoming involved as subjects in**th~** research, as is required by the regulations (21 CFR 50) which governed the conduct ofthis research. I concluded on the basis of information in the study report and the two earlier supplements submitted by DAS that they had not, and therefore that the conduct oft his study was significantly deficient relative to standards prevailing when it was conducted. Dr. Kisicki's letter addressed this concern in two passages. First, in his response # ll, Dr. Kisicki states:
In 1998, when this study was conducted, our internal standard operating procedure (SOP) for screening was as follows : When potential subjects arrived at the research facility, they were given a packet of information about the study. The packet contained a copy ofthe IRB-approved Informed Consent Form. Prior to any screening procedures, the subjects were required to sit and listen to an audio tape of the Informed Consent Form and follow along with the written document. An associate was available to answer any questions. Documentation is available ofthe subject signing a log indicating that they had listened to the audio tape ofthe Informed Consent Form, they understood it, and that they had no further questions

. (See attachment documenting s ubject Page 2 of6listened to the audio recording ofthe Informed Consent Form and the packet given to the study subjects)
Then in discussing compliance with 21 CFR standards, Dr. Kisicki states:
At the time the research was conducted, it was our standard operating procedure to have the subject listen to an audio recording ofthe Informed Consent From and follow along with the written document. However, subjects were not required to sign at that time. Documentation is provided noting subjects listened to the audio recording, they understood it, and they bad no further questions. The subjects were allowed to take the Informed Consent Form with them after they complete the screening process
. The Informed Consent was signed when the subject arrived at the research facility the evening before dosing

. The submitted screening logs, however, were not signed by all the participants in the research. Only 18 of the 47 primary and alternate subjects enrolled in Phase 1 ofthe study, and only 31 ofthe 35 primary and alternate subjects enrolled in Phase 2 of the study signed one or the other ofthe logs provided to EPA. Thus my concern has not been laid to rest.

. Analysis of the Screening Log
Two logs were included in the DAS submission ofJuly 1. First is a ten-page list entitled "Screening List- Full: Screen Date: 21-SEP-1998: Study Number 21438 Group ID: 

### 1." This list includes 107 entries, each consisting ofa Harris Subject ID in the range from 3 through 335 with
or without an associated (redacted) signature. It appears to be complete

- i.e., each page but the last includes 11 entries, but the last page includes only 8, the last ofthem hand-written. The second log is a similar seven-page list entitled "Screening List- Full: Screen Date: 06

-0CT1998: Study N umber 21438 Group 10: 2." This list includes 98 entries, each consisting of a Harris Subject ID in the range from 133 through 579, each one again with or without a (redacted) signature. This list also appears to be complete; the final page contains only 1 0 entries. In no case does the same**Subje~t** ID appear on both the Group 1 and Group 2 lists.
DAS reports that these logs origina lly included the full name, address, and SSN of each candidate. These identifiers have been masked completely, and the signatures have been blacked out. It is, however, still possible to tell which candidates, identified by Subject ID, signed the log

. At the bottom of each page the log bears this language:
Volunteers are to sign beside their name after listening to the Informed Consent Form, if they understand it and have no further questions.
Table 1 below summarizes the numbers of unique Subject IDs appearing either on the screening logs (lines 1-4) or, in line 5, not appearing on the logs provided but included as enrolled primary or alternate subjects in the accounting by SID provided in Appendices 

### II and 

### III to the DAS supp l emental submission ofMay 28,2008 (MRID 47436401.)

Page 3 of61 2 3 4 5 Table 1: Summary Analysis of Screening Logs vs. Enrollment Status

On Log Signed Log Enrolled Phase 1 Phase 2 Total y N N 55 41 96 y y y 18 31 49 y y N i ' 31 22 I J 53 y N y 3 4 7 N n/a y 26 0 26 133 98 231

Neither the study report nor Dr. Kisicki's letter explains how the screening logs were compiled; I assume they incorporated the results of initial recruiting. A high percentage of these candidates (51% for Phase 1 and 42% for Phase 2) were apparently "no-shows"

- they did not sign the log and were not enrolled as either primary or alternate subjects

. 17% of the screened IDs in Phase 1 and 32% in Phase 2 signed the log and were enrolled. These candidates are highlighted in light green in Tables 1-4. A still-significant percentage ofthe IDs on the log (29% for Phase 1 and 22% for Phase 2) signed the log but were not enrolled; these are highlighted in yellow in the tables, and may correspond to the group reported in Appendix 4.24 to the study report (MRID 44811002, pp. 506

-508) as "Subjects not Enrolled on the Study." Candidates who were enrolled as subjects but who did not sign one of the screening logs provided are highlighted in red in the tables. Detailed listings ofboth the Group 1 and Group 2 logs are appended to this review as Tables 2 and 4.
Line 5 in Table 1 reports 26 subjects whose Subject IDs were not included on the September 21 screening log but who were subsequently enrolled in Phase 1. A detailed listing of these 26 enrolled subjects appears in appended Table 3. I note, however, that in Appendices 4.6.1 and 4.6.2 to the study report (MRlD 44811002, pp
. 253-264) all but one of the 20 among these 26 candidates who actually participated in Phase I testing 1 (as subjects 17

- 30 and 32

-36) are reported to have been screened on 30 September, whereas the candidates who participated as subjects 1-16 and 31 were all reported to have been screened on September 23. Thus it appears likely that a second Group 1 screening log was generated and used for screening on September 30-a log which has not yet been submitted to EPA.
The anomaly is Subject 31, reported in Appendices 4.6.1 and 4.6.2 to have been screened on September 23

. SID 343 is linked to Subject 31 in Appendix II to the second DAS supplemental report (MRID 47436401 , p. 23), but SID 343 does not appear on the September 21 screening log used on that date, and is outside the range of SIDs that are on the log. The highest SID**n~mber** on the September 21 log, however, is handwritten; perhaps SID 343 should have

1 The accounting is necessarily less than complete, since the screening date is reported in Appendices 4.6.1 and 4.6.2 only for the 60 subjects who were ultimately treated, and not for those enrolled as primary subjects but subsequently replaced or for those enrolled as alternate subjects and not used. Page 4 of6been added to the log by hand as well. Most ofthese 26 subjects, however-excepting SID 343/Subject 31, and perhaps others-can probably be accounted for on the screening log used on September 30. · Additional documentation cannot, however, overcome the evidence in hand that three candidates who were screened on September 23 but who did not sign the screening log were subsequently enrolled in Phase I: SID 234 was enrolled and treated as Subject 5, SID 313 was enrolled as Subject 3 but replaced before treatment, and SID 322 was enrolled and treated as Subject 25. Furthermore, four more candidates who were screened on October 13 but who did not sign the screening log were subsequently enrolled in Phase 2: SID 487 was enrolled as Alternate 3 but not treated; SID 495 was enrolled and treated as Subject 46; SID 498 was enrolled as Alternate 1 but not treated; SID 557 was enrolled and treated as Subject 44.
It is clear from this evidence that the SOP described in Dr. Kisicki's June 23 letter was not consistently followed in the conduct of this study. The ethical significance ofthis lapse is less clear.
In my May 29 ethics review I noted that " by the time ofcheck-in

... subjects had given blood and urine samples, provided medical and surgical history , and been subjected to a physical examination at screening, had visited the clinic 48 h before check-in to obtain urine collection containers, and had been collecting their urine for two days."
·J concluded " in my judgment the subjects should all have provided written informed consent at or before the time of screening." They did not, however, sign consent forms until check-in on the evening before treatment.
Dr. Kisicki ' s letter asserts that at the time of screening all candidates received a packet containing the approved consent form, that they were required to follow along as they listened to an audio recording of it, and that they all signed the screening log to acknowledge receipt and understanding ofthe consent form and to indicate that they had no questions about it, all before the first collection of data and samples. On the strength ofthis assurance the HSRB concluded that the delay in signing the consent form until after initiation of data collection was a technical lapse only, without substantive et hical significance

. The HSRB indicated at the meeting that their summary judgment would be that the conduct ofthis research was not "s ignificantly deficient relative to standards prevailing when it was conducted, " and thus that the study is ethically acceptable under the standard of40 CFR §26.1704.
After a careful review the new evidence provided by DAS and MDS Pharma Services, I remain concerned that at least some ofthe participants in this study may have been involved as subjects

- i.e., may have provided information and tissue sam ples for analysis-without first having been properly informed and providing their informed consent.
It is conceivable that all candidates who came to one ofthe screening meetings did, in fact, receive and review the consent form, understand it, and have no unanswered questions, but that they simply failed to sign the screening log. Ifthat's what happened, then it suggests at least carelessness on the part ofthe laboratory staff in carrying out their SOPs and in ensuring compliance with the unambiguous regulatory requirement that "no investigator may involve a Page 5 of6human being as a subject in research covered by t hese regulations unless t he investigator has obtained the legally effective informed consent ofthe subject." (21 CFR §50.20)
I will finalize my ethics review of this study once we have received the recommendations of the HSRB. But based on all the evidence I have seen and·the concerns I have raised in both this supplemental review and in my ethics review ofMay 29, I remain ofthe view that its conduct was significantly deficient relative to standards ofethical research conduct prevailing when it was conducted, and that EPA is forbidden by 40 CFR §26.1704 to rely on it.

Attachment

cc: Paul Lewis, OSA Tom Myers Kelly Sherman William Jordan

Page 6 of6Appendix

Table 2: Dispos it ion of S ubject IDs on Scr eening Log of 2 1 Sep 98
HID/SID is taken from Screening Log. An 'x' in the column headed 

### 'I s /' means a redacted signature appears in the Screening Log opposite the indicated HID/SID

. Original and Final Enrolled Subject Number s linked to SID s are taken from Appendix II to t he second DAS supplemental report (MRID 47436401 , p. 23). Screen Date is taken from Appendices 4

.6 .1 and 4.6.1 to the primary study report (MRID 44811002, pp. 253-264)

Enrolled Subject Screen HID/SID Is/ Number Original Final Date
3 4 8 X 15 17 X 7 7 23-Sep 22 25 32 34 X 21 21 23-Sep 36 45 X 6 6 23-Sep 51 54 65 66 72 X 19 n/a NR 74 78 81 82 83 84 X 86 89 93 101 104 111 112 X 2 2 23-Sep 1 15 X 116 121 X A10 13 2 3

- Se p 125 X 11 11 23

-Sep 126 173 X 13 

### nla NRAppendix

Enrolled Subject Screen HID/SID /s/ Number Original Final Date 206 207 225 X 23 23 23-Sep 234 5 5 23-Sep 236 237 X 240 242 X 246 X 248 X 1 1 23-Sep 249 250 251 X 27 n/a NR 252 X 253 255 X 257 258 259 260 X 261 X 262 X 264 267 268 X 270 272 273 X 274 275 X 276 277 X 278 X 279 282 284 X 285 286 X 287 290 X 291 X 292 X 293 X 294 295Appendix

Enrolled S ubj e ct Screen HID/SID lsi Number Original Final Date 296 300 X 30 30 23-Seo 301 X 302 X 9 9 23-Sep 303 X 304 X 305 X 306 307 308 X 26 26 23

-Seo 309 312 X 313 3 n / a 

### NR 314 X 10 10 23-Seo 315 316 317 318 X 319 320 X . A11 n/a NR 322 25 25 23-Sep 323 324 X 24 24 23-Sep 326 X 329 X 16 16 23-Sep 330 X 335 X

### Table 3: Subjects N ot Listed On Screening Log of21 Sept 98 But Enrolled as S ubjects in Phase I Appendix

Original and Final Enrolled Subject Numbers linked to SIDs are taken from Appendix II to the second DAS supplemental report (MRID 47436401 , p. 23). Screen Date is taken from Appendices 4.6.1 and 4

.6.1 to the primary study report (MRID 4481 1002 , pp. 253

-264)

Enrolled Subject Screen HID/SID Number Original Final Date 

### 14 A9 nla 

### NR 23 A4 27 3C>-Seo 332 29 29 30-Seo 343 31 31 23-Seo 349 32 

### n / a 

### NR 384 A1 3 3C>-Seo 385 8 n/a 

### NR 387 33 33 3C>-Seo 390 12 12 30-Seo 391 A2/A10 8 3C>-Seo 395 4 4 30-Seo 399 35 35 3C>-Seo 40 7 14 14 30-Seo 419 15 15 30-Seo 421 A3 20 30-Seo 428 A4 

### 34 30-Seo 432 A 7 nl a 

### NR 433 A6 32 30

-Seo 440 18 18 30-Sep 4

42 28 28 30-Sep 446 

### AS 19 30-Seo 44 7 17 17 30-Seo 462 22 22 

### 3C>-Seo 463 36 

### 36 30-Seo 470 A8 

### n / a 

### NR 472 20 

### n / a 

### NRAppendix

Table 4 : Disposition of S ubject IDs on Screening Log of 6 Oct 98
HID/SID taken from Screening Log. A n "x" in the column headed 

### " I

s /" indicates that a redacted signature appears in the Screening Log opposite the indicated HID/SID

. Original and Final Subject Numbers associated with SIDs are taken from Appendi x II to the second DAS supplemental re port (MRID 47436401 , p. 23). Screen Date taken f rom Appendices 4

.6 .1 and 4.6.1 to primary study report (MRID 4481 1002, pp. 253-264)
Enrolled Sub ject Screen HID/SID /s/ Number Date Original Final 133 219 X 222 X 54 54 13

-0ct 235 281 X 52 52 1 3

-0ct 339 350 X A4 58 1 3

-0ct 359 363 X 370 406 X 448 X 56 56 13-0ct 449 X 468 X A11 n/a NR 469 X A9 n/a NR 476 477 X A5 n/a NR 478 479 480 X 481 484 485 X 45 4 5 13

-0ct 487 A3 n / a 

### NR 488 489 X 51 51 13

-0ct 490 X A2 n/a NR 491 492 493 X 40 40 13-0ct 494 495 46 46 13-0ct 496 X 59 59 13

-0ct 497 498 A1 n/a 

### NR 499 X 48 48 13

-0ct 501Appendix

Enrolled Subject Screen H

I D/SID /s/ Number Original Final Date 502 503 504 507 X 509 X 43/A8' 47 13-0ct 510 X 511 X 513 X 514 515 516 X 517 518 X 55 55 13-0ct 519 521 X 522 X 41 41 13-0ct 523 X 526 X A? 43 13-0ct 528 529 X 530 X 531 X 47 n/a NR 532 534 X 535 X 58 n/a NR 536 537 X 37 n/a NR 538 X 539 X 38 38 13-0ct 5

40 541 542 543 544 X 545 X 60 60 13-0ct 546 547 548 X 549 X 49 49 13-0ct 550 X 55 1 552 553 X 42 n/a NR 554 555 X A6 42 13-0ctAppendix

Enrolled Subject Screen HID/SID /s/ Number Date Original F inal 556 557 44 44 13-0ct 558 X 39 39 13-0ct 559 560 X 53 53 13-0ct 561 X 562 563 X 565 566 568 X 57 57 13-0ct 569 X 50 50 13-0ct 571 X A8 n/a NR 572 X 578 579 X A10 37 13-0ct