Supporting Information: Paraben concentrations in maternal urine and breast milk and its association with personal care product use.
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1 Supporting Information: Paraben concentrations in maternal urine and breast milk and its association with personal care product use. Mandy Fisher 1*, Susan MacPherson 1, Joseph M. Braun 2, Russ Hauser 3, Mark Walker 4, Mark Feeley 5, Ranjeeta Mallick 4, René Bérubé 6, Tye E. Arbuckle 1 1Healthy Environments and Consumer Safety Branch, Health Canada, Ottawa, ON 2 Department of Epidemiology, Brown University, Providence, RI Harvard University School of Public Health, Boston, MA, USA. 4The Ottawa Hospital Research Institute, Ottawa, ON, Canada 5Health Products and Food Branch, Health Canada, Ottawa, ON 6Institut National de Santé Publique du Québec, QC, Canada *Corresponding Author: Mandy Fisher Address: 50 Colombine Driveway, Tunney s Pasture. Ottawa Canada. K1A 0K9 mandy.fisher@canada.ca Phone: Number of pages: 12 Number of Figures: 2 Number of Tables: 6 S1
2 Table S1: Validation data of the ID-LC-MS/MS methods for the determination of parabens in urine and milk samples. Analyte Matrix LOD LOQ Calibration Recovery Matrix Intraday range effect Precision Concentration a (µg/l) (µg/l) (µg/l) (%) (%) (%) (µg/l) BP U LOQ to M LOQ to MP U LOQ to M LOQ to PP U LOQ to M LOQ to EP U LOQ to M LOQ to IBP U LOQ to M LOQ to BzP U LOQ to M LOQ to Abbreviations: BP, butylparaben; MP, methylparaben; PP, n-propylparaben; EP, ethylparaben; IBP, isobutylparaben; BzP, benzylparaben; U, urine; M, milk a Concentration of the analyte in the quality control material S2
3 Table S2: Urine Sample Characteristics (n = 542 urine samples collected from 31 women) N % season of sampling spring summer autumn winter Time since last void 0-86 minutes minutes minutes >200 minutes missing 37 Time of Day midnight to 7: :00 to 11: :00 to 17: :00-23: missing 8 Total Volume <40mL to <65 ml to <90 ml >=90mL missing 1 Study Visit T1A (weekday) T1B (weekend day) S3
4 Table S3: Prevalence of Product Use (total # and % of women) in 24-h sampling period (N=31 women) Product Group Abbreviation Examples of products in this category T1a Day1 (n=31) T1a Day2 (n=30) Deodorant and Antiperspirants Baby Lotions, Soaps and other Baby Products Body Soaps Conditioner Eye Makeup n % n % anti Deodorant, antiperspirant babyprod bodysoap condit Face Lotions and Creams face Fragrance and Perfumed Products Hairstyling Products eyemakeup frag hair Baby wipes, diaper ointment, baby body wash, petroleum jelly Body wash, body gel, shower soap Rinse-off conditioner, 2-in-1 shampoo and conditioner, leave-in conditioner Eye shadow, mascara, brow liner Day cream, night lotion, acne cream, eye moisturizer perfume, bubble bath, body mist, air freshener Hairspray, gel, mousse, hair paste Lip Products lip lip balm, lipstick, lip gloss Body Lotions, Creams and Oils General Makeup and Cosmetics lotion Nail Polish and Remover nail Toothpaste and Mouthwash Body butter, belly oil, sunscreen, hand cream makeup Blush, foundation, concealer oral nail polish, nail polisher remover Toothpastes, dental rinse, mouthwash Shampoo shampoo shampoo Facial Soaps, Cleansers and Washes Hand Soaps, Sanitizers and Soap Not Otherwise Specified (NOS) skinclea soap Face cleanser, exfoliator, face masks, eye makeup remover Liquid soap, bar soap, waterless sanitizers, generic soap S4
5 Table S4: Adjusted geometric means (GM) in maternal urine (ug/dl) by Product Use 24h before urine sampling anti frag oral condit shampoo bodysoap soap eyemakeup Product Group No (n=262) Yes (n=267) No (n=476) Yes (n=53 ) No (n=40) Yes (n= 489) No (n=261) Yes (n= 268) No (n=228) Yes (n=301) No (n=171) Yes (n= 358) No (n=304) Yes (n=225 ) No (n=229) Yes (n=230 ) makeup No (n=300) GM (95% CI) 0.16 (0.06,0.42) 0.27 (0.1,0.72) 0.17 (0.07,0.41) 0.96 (0.31,3.04) 0.16 (0.05,0.52) 0.21 (0.08,0.53) 0.15 (0.06,0.39) 0.27 (0.11,0.69) 0.15 (0.06,0.38) 0.27 (0.11,0.69) 0.16 (0.06,0.41) 0.24 (0.09,0.61) 0.27 (0.1,0.73) 0.14 (0.05,0.39) 0.17 (0.07,0.42) 0.29 (0.11,0.76) 0.17 (0.07,0.42) BP EP MP PP % Change p- value GM (95% CI) 2.22 (1.25,3.95) 2.54 (1.43,4.51) 2.25 (1.27,3.99) 3.77 (1.97,7.23) 1.49 (0.79,2.82) 2.48 (1.42,4.33) 2.28 (1.29,4.04) 2.48 (1.4,4.4) 2.24 (1.26,3.98) 2.49 (1.41,4.4) 2.06 (1.15,3.69) 2.55 (1.45,4.5) 2.36 (1.34,4.18) 2.4 (1.35,4.27) 2.26 (1.27,4) 2.55 (1.43,4.54) 2.41 (1.35,4.3) % Chan ge p- value GM (95% CI) (18.49,44.77) (20.66,49.93) (19.19,45.5) 38.9 (22.83,66.29) (15.09,43.59) (20.17,47.17) (19.47,47.1) (19.62,47.44) (18.96,45.85) (20.12,48.1) (18.18,45.08) 31.3 (20.3,48.26) 23.49(15.38, 35.9) (27.86,66.09) 31.6 (20.3,49.18) (18.45,45.44) (22.33,55.73) % Chan ge p- value < GM (95% CI) 4.91 (2.91,8.27) 4.36 (2.59,7.35) 4.53 (2.71,7.56) 5.54 (2.89,10.63 ) 3.53 (1.84,6.75) 4.73 (2.86,7.83) 4.75 (2.8,8.04) 4.5 (2.66,7.61) 4.43 (2.62,7.51) 4.77 (2.84,8.02) 3.89 (2.26,6.71) 5.02 (3,8.41) 3.4 (2.06,5.62) 6.96 (4.17,11.63 ) 4.75 (2.79,8.09) 4.46 (2.59,7.68) 4.54 (2.66,7.77) % Change p-value < S5
6 Yes (n= 229) (0.11,0.76) (1.3,4.22) (15.62,39.99) (2.72,8.24) babyprod No (n=421) (0.08,0.56) (1.25,3.87) (19.59,46.13) (2.67,7.46) Yes (n=108) (0.06,0.5) (1.75,5.81) (19.88,51.4) (2.98,9.45) face No (n=231) (0.08,0.54) (1.53,4.94) (22.95,55.73) (3,8.74) Yes (n= 298) (0.08,0.55) (1.2,3.8) (17.49,41.55) (2.55,7.2) lotion No (n=197) (0.05,0.35) (0.89,2.78) (12.65,30.85) (1.87,5.51) Yes (n=332) < < (0.11,0.72) (1.75,5.35) 1 (25.69,60.73) 1 (3.43,9.68) nail No (n=526) (0.08,0.52) (1.36,4.18) (19.53,46.08) (2.74,7.59) (141.2, < (15.9 Yes (n=3) NA NA NA (0.33,3.31) ) ,259.56) hair No (n=342) (0.09,0.63) (1.3,4.06) (18.42,44.09) (2.66,7.48) Yes (n=187) (0.05,0.38) (1.41,4.53) (21.77,54.1) (2.87,8.48) skinclea No (n=223) (0.08,0.53) (1.77,5.65) (20.16,49.52) (3.09,9.17) Yes (n= 306) (0.08,0.55) (1.09,3.43) (19.07,45.86) (2.46,7.08) lip No (n=371) (0.09,0.62) (1.46,4.57) (19.49,46.49) (2.61,7.35) Yes (n=158) (0.05,0.42) (1.08,3.55) (19.58,49.61) (3.0,9.16) *n=number of samples (i.e.# of samples where oral products were used 24h before) *Separate models were used for each predictor and outcome and specific gravity was included as a covariate in all models. Models were also adjusted study visit, time of urine collection, BMI, job category, season and number of other product categories used were also retained in the model when statistically significant. S6
7 Table S5: Adjusted geometric mean urinary concentrations according to timing of most recent personal care product use among pregnant women in the P4 Study. BP EP MP PP Product Group N GM (95% CI) p-value GM (95% CI) p-value GM (95% CI) p-value GM (95% CI) p-value anti frag oral condit shampoo bodysoap 0-6 Hours (0.13,0.94) (1.39,4.72) (24.59,62.76) (3.65,11.21) Hours (0.09,0.59) (1.13,3.73) (17.07,41.71) (2.05,5.95) > 24 Hours (0.08,0.55) (0.96,3.29) (14.57,37.16) (2.46,7.55) No Use (0.05,0.33) 2.95(1.61,5.41) 33.45(21.12,52.98) 5.38(3.11,9.31) 0-6 Hours (0.46,8.63) (1.89,9.15) (29.91,120.65) (3.44,19.51) Hours (0.25,2.56) (2,7.5) (20.88,62.8) (2.59,9.88) > 24 Hours (0.05,1.22) (1.01,6.04) (15.57,80.91) (1.63,12.85) No Use (0.07,0.36) 2.23(1.27,3.94) 28.93(18.72,44.71) 4.48(2.69,7.46) 0-6 Hours (0.1,0.56) (1.43,4.46) (21.5,51.04) (3.43,9.72) Hours (0.08,0.44) (1.45,4.48) (19.39,45.34) (2.51,6.98) > 24 Hours (0.03,0.38) (0.87,3.57) (15.27,51.6) (1.67,7.55) No Use (0.02,0.31) 0.86(0.43,1.72) 18.92(10.37,34.53) 3.13(1.49,6.6) 0-6 Hours (0.11,0.74) (1.68,5.57) (25.21,63.77) (4.23,12.83) Hours (0.08,0.48) (1.12,3.65) (14.25,35.22) (1.87,5.5) > 24 Hours (0.04,0.28) (0.99,3.35) (12.72,32.76) (2.31,7.21) No Use (0.08,0.55) 2.73(1.5,4.96) 40.64(25.62,64.48) 5.28(3.04,9.17) 0-6 Hours (0.12,0.79) (1.77,5.7) (28.04,68.75) (4.7,13.9) Hours (0.09,0.53) (1.26,4.01) (15.85,38.06) (2.03,5.83) > 24 Hours (0.04,0.3) (1.11,3.7) (14.1,35.98) (2.27,7.07) No Use (0.07,0.45) 2.27(1.26,4.1) 35.58(22.63,55.94) 4.83(2.79,8.35) 0-6 Hours (0.1,0.62) (1.79,5.83) (26.37,65.26) (4.77,14.21) Hours (0.08,0.47) (1.27,4.07) (16.91,40.97) (2.24,6.49) > 24 Hours (0.03,0.25) (0.66,2.47) (10.89,31.92) (2.11,7.87) No Use (0.08,0.51) 2.25(1.24,4.09) 31.25(19.66,49.65) 3.75(2.14,6.56) soap 0-6 Hours (0.05,0.44) (1.2,3.94) (29.32,74.47) (4.64,14.11) S7
8 eyemakeup makeup babyprod face lotion nail hair 6-24 Hours (0.03,0.24) (1.25,4) (24.82,61.08) (3.32,9.68) > 24 Hours (0.03,0.26) (0.84,3.02) (13.1,36.93) (1.26,4.43) No Use (0.13,0.97) 2.72(1.53,4.84) 23.25(14.98,36.1) 3.65(2.17,6.14) 0-6 Hours (0.17,1.18) (1.5,5.1) (22.71,60.65) (3.53,11.52) Hours (0.13,0.77) (1.3,4.24) (16.18,40.99) (2.45,7.42) > 24 Hours (0.13,1.04) (1.04,3.71) (16.85,47.84) (3.1,10.98) No Use (0.05,0.26) 2.38(1.32,4.29) 31.72(19.96,50.42) 4.17(2.4,7.24) 0-6 Hours (0.13,0.92) (1.41,4.77) (16.47,45.58) (3.37,11.17) Hours (0.13,0.9) (1.7,5.57) (13.7,36.81) (2.56,8.13) > 24 Hours (0.17,1.83) (1.84,7.41) (15.86,54.03) (2.79,12.31) No Use (0.05,0.28) 1.91(1.06,3.42) 35.95(22.21,58.18) 4.08(2.33,7.14) 0-6 Hours (0.04,0.32) (1.59,5.73) (16.41,47.28) (2.43,8.71) Hours (0.06,0.51) (1.57,5.39) (21.04,57.48) (2.74,9.08) > 24 Hours (0.04,0.53) (0.84,3.29) (15.84,50.92) (1.36,5.59) No Use (0.09,0.53) 2.33(1.32,4.1) 29.73(19.16,46.14) 4.83(2.9,8.06) 0-6 Hours (0.07,0.48) (1.13,3.7) (20.6,51.33) (3.32,9.86) Hours (0.07,0.42) (1.2,3.84) (15.74,38.06) (2.26,6.43) > 24 Hours (0.05,0.36) (1.44,5.08) (20.87,57.29) (3.29,11.15) No Use (0.1,0.63) 2.78(1.53,5.04) 34.31(21.69,54.29) 4.52(2.62,7.81) 0-6 Hours (0.11,0.71) (2.05,6.33) (32.23,79.27) (4.85,14.22) Hours (0.11,0.66) (1.86,5.64) (23.02,55.45) (2.86,8.13) > 24 Hours (0.08,0.64) (1.29,4.34) (15.13,41.29) (2.06,6.98) No Use (0.03,0.24) 1.07(0.6,1.93) 16.07(9.97,25.89) 2.73(1.54,4.85) 0-6 Hours 3 NA NA 1.01(0.31,3.24) (135.5,1196.4) (15.6,251.9) Hours 0 NA NA NA NA NA NA NA NA > 24 Hours 0 NA NA NA NA NA NA NA NA No Use (0.08,0.45) 2.38(1.36,4.16) 29.59(19.21,45.57) 4.52(2.72,7.5) 0-6 Hours (0.06,0.41) (1.45,4.87) (31.09,81.68) (4.69,15.07) Hours (0.04,0.23) (1.18,3.83) (17.32,43.49) (2.01,6.09) S8
9 skinclea lip > 24 Hours (0.08,0.65) (1.28,4.69) (17.81,51.39) (1.76,6.4) No Use (0.1,0.59) 2.41(1.37,4.27) 28.33(18.3,43.85) 4.75(2.82,8) 0-6 Hours (0.06,0.41) (1.07,3.61) (23.84,60.6) (3.39,10.38) Hours (0.08,0.51) (0.98,3.23) (17.43,42.85) (2.12,6.19) > 24 Hours (0.05,0.48) (0.95,3.65) (19.59,59.1) (2.77,10.66) No Use (0.08,0.52) 3.63(1.98,6.67) 28.29(17.7,45.22) 4.88(2.78,8.56) 0-6 Hours (0.05,0.37) (1.09,3.77) (21.21,57.15) (3.3,10.92) Hours (0.06,0.45) (1,3.37) (18.21,47.4) (2.82,8.91) > 24 Hours (0.06,0.7) (0.82,3.11) (14.4,43.53) (2.58,10.01) No Use (0.09,0.51) 2.72(1.53,4.83) 29.88(19.29,46.28) 4.2(2.5,7.06) * Separate models for each predictor and outcome and all models were adjusted for specific gravity. Additional covariates including study visit, time of urine collection, BMI, job category, season and number of other product categories used were also retained in the model when statistically significant. S9
10 Table S6: Adjusted % change of methylparaben (MP) and propylparaben (PP) breast milk concentrations according to the number of personal care product categories used in the last 24 hours. Total Personal Care Products Used (Frequency in last 24 hours) % Change MP % Change PP (95% CI) (95% CI) Low Users Reference Reference (0 4 product categories) Medium Users 100% 43% (5-7 product categories) (-84%,2442%) (-85%,1287%) High Users 260% 382% (8 14 product categories) (-75%,5014%) (-55%,5058%) *Separate models for each predictor and outcome. *The total number of product categories used in the last 24 hours by participants were summed and women were categorized into three product user groups corresponding to the bottom 43% (low users, 0 4 product categories, n=21), the middle 31% (medium users, 5-7 product categories, n=15), and top 27% (high users, 8 14 product categories, n=13) S10
11 Figure S1: Product Category Use by Low users (0 5 product categories), medium users (6-8 product categories), and high users (10 14 product categories) within a 24 hr period. Percentage of Users Hour Product Category Use High Medium Low oral bodysoap shampoo skinclea lotion face condit anti eyemakeup soap hair makeup S11
12 Figure S2: Least squares adjusted GM by time since use (0-6h, 6-24h, >24h, No Use) S12
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