Associations between phthalatesexposure and health outcomes of newborns,pregnant women and precocious puberty girls in Taiwan

*Whats Phthalates (PAEs)?They are a family of chemical substances that have been in use for about 50 years, primarily to make polyvinyl chloride (PVC) soft and flexible.Phthalates look like clear vegetable oil.

Physico-chemical properties of Phthalates

CompoundsDimethyl phthalate DMPDiethyl phthalate DEPDi-n-butyl phthalate DBPButyl benzyl phthalate BBzPDi(2-ethylhexyl) phthalate DEHP

Chemical formulaC10H10O4C12H14O4C16H22O4C19H20O4C24H38O4Molecular weight194.19222.23278.35312.35390.62Melting point (C)5.5- 40.5- 35- 35- 47Boiling point (C)282295~302340370386Vapor pressure (mmHg)1 (100C)1.6510-32.710-5610-7110-7Soubility (mg L-1, ) 40001000130.70.3Log Kow 1.62.54.54.97.5Log Koc1.9-2.61.8-2.73.1-4.23.2-4.24.9-6.0MetabolitesMMPMEPMBPMBzPMEHP,MEOHP, MEHHP

Applications of phthalatesPersonal care products

Application of phthalates

PAEs

DEHP

()(PVC)()()()PVC

DBP

(latex adhesive)(cellulose plastic)PVC(PVC)

BBP

(vinyl tile)(food conveyor belt)(carpet tile)(artificial leather)(tarpaulin)(automotive trim)(Weather stripping)(traffic cone)(adhesives)

DEP

DMP

DINP(Di-isononyl phthalate)

PVCPVCPVCPVCPVCPVCPVC

Metabolic pathway of DEHP-Albro, et al., 1982; ATSDR, 1993, Schmid, et al., 1985c.DEHPMEHPHumanPrimarySecondary

ObjectivesPart I: Association between Phthalate Exposure and Thyroid Hormone in Pregnant WomenTo determine the concentrations of urinary phthalate monoesters in pregnant women. To investigate the association between phthalate exposure and thyroid hormone during pregnancy.

ObjectivesPart II: Association between Prenatal Exposure to Phthalates and the Health of NewbornsTo determine the concentrations of amniotic fluid phthalate monoesters in pregnant women. To evaluate the association between prenatal exposure of the fetus to phthalates in amniotic fluid and maternal urine and the health of newborns.

To follow-up our previously established participants in order to determine the correlation between phthalate exposure and thyroid hormones in pregnant women and newborns. ObjectivesPart III: Phthalates may alter thyroid hormones in utero: A follow-up study

Framework

Newborns healthAnogenital distanceGestational age Urine samplePhthalate monoesters Serum &Cord bloodTSH,T3, T4, FT4Phthalate monoestersMaterials and MethodsInformed consent Amniotic fluidPhthalate monoesters Urine samplePhthalate monoestersCreatinineSerum sampleTSH,T3, T4, FT4 Urine samplePhthalate monoestersCreatinine Serum sampleTSH,T3, T4, FT4 QuestionnaireAge, BMI, gestationFood consumptionCosmetic usageFollow up-76 subjects Follow up-65 subjects Interview- 83 subjects alpha fetal protein (AFP) -free -hCG maternal age > 35 yrsundergo amniocentesis

*Measured Parameters Phthalate monoestersmonomethyl phthalate (MMP)monoethyl phthalate (MEP)monobutyl phthalate (MBP)monobenzyl phthalate (MBzP)mono-2-ethylhexyl phthalate (MEHP)SampleUrine/ Amniotic fluidSerumAnalytical methodLC-MS/MS (Agilent 1100/API 3000)

Measured Parameters Physical examinationAnogenital distance (AGD)Birth weightBirth heightGestational ageAnogenital index (AGI)AGI-W= AGD/ birth weight.AGI-L= AGD/ birth length.Thyroid hormonesThyroid stimulating hormone (TSH)Thyroxin (T4)Triiodotyronine (T3) Free thyroxin (FT4)QuestionnairesAge, BMI, gestation age, Smoking, Medical care etc.Food consumptionCosmetic usagePersonal care product usageBuilding characteristics

Table I-1 Demographic characteristics of study participants (n = 76). Characteristics Mean SDAge (years)33.6 3.3Body mass index (BMI)20.9 2.5Duration of gestation (weeks)27.9 2.3Pregnancies and births Number of current pregnancy1.9 1.0 Number of current birth1.5 0.6Smoking status (n/%) Active smoker0/0 Passive smoker14/18.4 Non-smoker62/81.6Alcohol drinker (n/%)0/0New decoration of living/ working place during previous 1 year (n/%) aHome Moving to new decorated house7/9.2 Just decorated4/5.3Workplace Moving to new decorated workplace2/2.6 Just decorated6/7.9Medical care during previous 3 months (n/%) Blood transfusion3/3.9 Intravenous drip3/3.9 Oxygen mask1/1.3 a Moving to new decorated house or workplace means moved into a new building. Just decorated means the place you live or work have decorated recently such as painting or changing the floor.

*Table I-2 Levels of five urinary phthalates monoester in pregnant Taiwanese women and comparison with other studies (g/g creatinine).

Table I-3 Distribution of thyroid hormones in pregnant women. aThe analytic sensitivities of T3, T4, free T4, and TSH were 19.5 ng/dL, 0.42 g/dL, 0.023 ng/dL and 0.014 lU/mL, respectively; the coefficient variations of T3, T4, free T4, and TSH were 2.9%, 4.2%, 3.1%, and 3.0%, respectively.

Hormones aPercentileReference rangeMin5th50th95thMaxT3 (ng/dL)72.686.3132.0209.0246.084.6-202.0T4 (g/dL)4.395.318.8511.213.65.13-14.1Free T4 (ng/dL)0.460.690.931.251.350.93-1.7TSH ( IU/mL)0.220.311.063.45.190.27-4.2

Table I-4 Spearman correlation coefficients a between thyroid hormone, age, BMI, duration of gestation and urinary phthalate monoester levels (n = 75). a Spearman correlation coefficients, *: p < 0.05.

TSHT3T4FT4AgeBMIGestationTSH1T30.1451T40.291*0.709*1FT40.1720.299*0.761*1Age0.2030.307*0.218*0.0081BMI0.0440.3090.1370.0100.1061Gestation0.1060.0180.0380.0570.0290.1311MBP0.0790.2340.248*0.368*0.0820.081*0.098MEP0.0820.0190.0390.0170.0800.095*0.199MEHP-0.0600.0670.1000.0900.1660.0300.030*MBzP0.0800.0840.0340.0070.0640.0270.118MMP0.0780.2590.0890.0070.0380.0060.004

Figure I-1. Correlation between (A) free thyroxine (FT4), (B) thyroxine (T4) and log MBP (ppb) and MBP-c level in pregnant women at the 2nd trimester. AB

Origin Data

NAMEIDLog MBP (ppb)CreatinineFSHE2ProgTSHT4T3FT4Log MBP-c

E940216US0012.85916254126.90.1219915139.22.747.9972.570.6712.28581328

E940321US0011.6252631320.40.117730165.11.197.93108.30.8462.66480479

E940420US0012.2235165261.80.12060552.091.538.0797.291.012.43252805

E940421US0011.9852001650.50.119200182.70.619.131150.9752.26885973

E940423US0011.5842474543.30.114030101.30.4866.4485.840.7912.27511142

E940428US0011.95480207240.1151174053.410.3684.8180.970.782.68318627

E940513US0012.63396916185.60.10510480117.50.3666.3698.470.7742.36539119

E940518US0011.965558268.30.11043590.82.468.04129.30.8613.04648017

E940523US0012.3774377656.50.136975211.90.9476.7996.910.8172.62538932

E940524US0011.7614294110.40.1884568.53.2310.39132.71.222.74439607

E940524US0021.9524884812.90.10611875770.2167.71050.9372.84189877

E940526US0011.8349551314.70.117220111.21.049.91311.052.66763779

E940530US0011.8920988412.10.12747599.21.046.0892.310.812.80931347

E940601US0011.8058225176.50.11244069.10.5736.63126.30.8041.92216108

E940601US0022.0446067519.70.11244080.50.5068.77128.10.9322.75014053

E940602US0012.14612804132.90.127400114.90.8128.88131.81.012.02260305

E940602US0021.8984325915.10.112060111.91.078.01113.60.9512.89449735

E940602US0031.8810122840.1859070.60.8187.55128.80.8393.27895229

E940604US0011.5543879124.20.12215421570.6099.05127.51.042.63387059

E940606US0012.0999307534.80.1171265596.81.26.3790.890.8622.28403916

E940607US0012.2427301488.10.12913559.851.86.79106.90.7992.29775423

E940609US0011.9646494617.40.116106651311.055.71102.60.7872.72410021

E940613US0011.409296315.40.11590593.40.97.36120.40.832.22177557

E940614US0011.96106345111.80.12785079.750.4598.96135.10.9721.91262164

E940615US0011.6286399122.20.132625224.250.5049.651541.042.28228694

E940618US0011.7271646923.80.113673118.251.19.87131.60.9432.35058774

E940622US0011.9385301275.40.12070777.51.168.41109.20.9972.06115878

E940623US0012.2063528575.40.1012393883.51.366.38115.60.7352.3289815

E940623US0022.29330552310.18750137.40.8556.03130.70.72.80194382

E940627US0011.99537967131.80.125900191.51.577.47107.70.8161.87546426

E940628US0011.6575821536.50.1082602554.70.587.21108.80.792.09528929

E940704US0011.4457499216.60.112130154.31.199.77134.41.052.22564183

E940706US0012.3291443361.90.116570106.61.338.87180.40.8022.53745368

E940708US0011.5723709640.50.10816270145.61.39.5111.71.151.96491594

E940716US0011.238197978.50.1116290107.51.648.75131.40.922.30877905

E940723US0012.1206173746.90.113470103.90.75811.64207.61.032.44944453

E940725US0011.414160049.50.10918550150.41.8311.61147.21.122.43643643

E940725US0021.122000916.60.134090157.61.49.81117.21.162.30245698

E940730US0012.105799139.50.11810630154.63.068.39123.70.8782.509202

E940803US0011.5569544228.40.120920205.90.76411.23149.21.12.10363608

E940811US0012.0979983487.10.11513092.60.80510.48163.11.052.15798019

E940818US0011.6063754329.90.117270176.71.9210.43145.91.062.13070424

E940823US0012.1616583367.90.127700204.80.6439.08132.11.072.32978855

E940824US0011.5767190219.20.16118570115.72.1110.52152.51.012.29341779

E940902US0011.9139084330.50.128800202.70.2317.52122.40.9852.42960859

E940905US0012.1807961260.10.147820441.95.194.39145.10.462.40192165

E940905US0021.40356794250.123230307.10.9039.25139.90.9612.00562793

E940906US0012.5376305841.10.10930160206.61.178.31137.70.8892.92378875

E940923US0011.6032586417.20.10220540160.20.8848.31138.20.7952.36773019

E940928US0011.387990417.10.1116350112.51.6311.17148.50.9832.53673206

E940929US0012.10205118120.50.1694851.730.47810.9192.60.9182.02106414

E941003US0011.5994867540.50.13316980105.11.1210.43158.50.9611.99203172

E941006US0011.5932418134.80.1120160124.42.568.44126.60.82.05166257

E941006US0021.6767974330.60.10422480150.31.179.87159.30.9172.191076

E941007US0012.76327488119.80.1431367045.641.57.02110.80.7382.68481807

E941013US0011.49765853180.12210730761.797.65128.50.8382.24238602

E941020US0011.8939958695.40.128590140.70.9469.17122.20.8971.91444748

E941021US0011.6674779912.30.131258086.22.128.45169.60.7532.57757288

E941101US0012.30777007113.90.137100226.40.4627.62137.80.852.25124635

E941105US0012.1709947146.20.11485649.71.158.29110.41.182.00604733

E941107US0011.9109444164.10.11055085.70.7289.14140.40.9732.10408638

E941117US0012.31699717177.60.1151073054.50.87612.35204.60.9562.06755421

E941203US0012.2741809594.80.127280351.50.5567.06155.50.7662.29737261

E941205US0011.964636729.60.116770165.10.6197.54157.10.7262.49334499

E941207US0011.99659922450.11177053.870.4168.61137.90.9712.34338671

E941213US0012.40973029115.90.114376130.21.6710.23119.71.072.34564686

E941219US0011.6148972212.90.10310278151.20.7679.99133.81.042.50430751

E950103US0011.5174598315.20.19260881.2910.191540.8882.33561624

E950104US0011.9009676243.30.121520159.81.798.93157.10.7882.26447973

E950105US0012.13395384104.80.10219240106.41.7911.83245.80.8972.11359256

E950112US0011.8977919888.70.119180214.51.619.2165.30.9111.94986836

E950112US0021.9444333252.60.112889964.42.2811.7141.71.212.22344757

E950112US0031.377124043.90.11720270170.30.9018.83108.41.072.78605944

E950112US0041.299289337.80.112880154.20.9016.24162.30.7312.40719473

E950121US0011.9382694848.90.117786.7452.179.39140.90.9622.24896062

E950124US0011.6402826375.60.1152221095.10.94813.37224.31.231.76176083

E950126US0011.7209857439.40.19955211.20.87212.47144.21.292.12548952

E950126US0021.5737995836.90.1219501703.8213.59225.31.352.00677322

MBP-FT4

NAMEIDFT4T4Log MBP (ppb)Log MBP-cTSHT3CreatinineFSHE2Prog

E940216US0010.6717.992.859162542.285813282.7472.57126.90.1219915139.2

E940420US0011.018.072.223516522.432528051.5397.2961.80.12060552.09

E940421US0010.9759.131.985200162.268859730.6111550.50.119200182.7

E940423US0010.7916.441.584247452.275111420.48685.8443.30.114030101.3

E940428US0010.784.811.954802072.683186270.36880.97240.1151174053.41

E940513US0010.7746.362.633969162.365391190.36698.47185.60.10510480117.5

E940518US0010.8618.041.965558263.046480172.46129.38.30.11043590.8

E940523US0010.8176.792.377437762.625389320.94796.9156.50.136975211.9

E940524US0011.2210.391.761429412.744396073.23132.710.40.1884568.5

E940524US0020.9377.71.952488482.841898770.21610512.90.1061187577

E940526US0011.059.91.834955132.667637791.0413114.70.117220111.2

E940530US0010.816.081.892098842.809313471.0492.3112.10.12747599.2

E940601US0010.8046.631.805822511.922161080.573126.376.50.11244069.1

E940601US0020.9328.772.044606752.750140530.506128.119.70.11244080.5

E940602US0011.018.882.146128042.022603050.812131.8132.90.127400114.9

E940602US0020.9518.011.898432592.894497351.07113.615.10.112060111.9

E940602US0030.8397.551.881012283.278952290.818128.840.1859070.6

E940604US0011.049.051.554387912.633870590.609127.524.20.1221542157

E940606US0010.8626.372.099930752.284039161.290.8934.80.1171265596.8

E940607US0010.7996.792.242730142.297754231.8106.988.10.12913559.85

E940609US0010.7875.711.964649462.724100211.05102.617.40.11610665131

E940613US0010.837.361.40929632.221775570.9120.415.40.11590593.4

E940614US0010.9728.961.961063451.912621640.459135.1111.80.12785079.75

E940615US0011.049.651.628639912.282286940.50415422.20.132625224.25

E940618US0010.9439.871.727164692.350587741.1131.623.80.113673118.25

E940622US0010.9978.411.938530122.061158781.16109.275.40.12070777.5

E940623US0010.7356.382.206352852.32898151.36115.675.40.1012393883.5

E940623US0020.76.032.293305522.801943820.855130.7310.18750137.4

E940627US0010.8167.471.995379671.875464261.57107.7131.80.125900191.5

E940628US0010.797.211.657582152.095289290.58108.836.50.1082602554.7

E940704US0011.059.771.445749922.225641831.19134.416.60.112130154.3

E940706US0010.8028.872.329144332.537453681.33180.461.90.116570106.6

E940708US0011.159.51.572370961.964915941.3111.740.50.10816270145.6

E940723US0011.0311.642.120617372.449444530.758207.646.90.113470103.9

E940725US0011.1211.611.414160042.436436431.83147.29.50.10918550150.4

E940725US0021.169.811.122000912.302456981.4117.26.60.134090157.6

E940730US0010.8788.392.10579912.5092023.06123.739.50.11810630154.6

E940803US0011.111.231.556954422.103636080.764149.228.40.120920205.9

E940811US0011.0510.482.097998342.157980190.805163.187.10.11513092.6

E940818US0011.0610.431.606375432.130704241.92145.929.90.117270176.7

E940823US0011.079.082.161658332.329788550.643132.167.90.127700204.8

E940824US0011.0110.521.576719022.293417792.11152.519.20.16118570115.7

E940902US0010.9857.521.913908432.429608590.231122.430.50.128800202.7

E940905US0010.464.392.180796122.401921655.19145.160.10.147820441.9

E940905US0020.9619.251.403567942.005627930.903139.9250.123230307.1

E940906US0010.8898.312.537630582.923788751.17137.741.10.10930160206.6

E940923US0010.7958.311.603258642.367730190.884138.217.20.10220540160.2

E940928US0010.98311.171.387990412.536732061.63148.57.10.1116350112.5

E940929US0010.91810.92.102051182.021064140.478192.6120.50.1694851.73

E941003US0010.96110.431.599486751.992031721.12158.540.50.13316980105.1

E941006US0010.88.441.593241812.051662572.56126.634.80.1120160124.4

E941006US0020.9179.871.676797432.1910761.17159.330.60.10422480150.3

E941007US0010.7387.022.763274882.684818071.5110.8119.80.1431367045.64

E941013US0010.8387.651.497658532.242386021.79128.5180.1221073076

E941020US0010.8979.171.893995861.914447480.946122.295.40.128590140.7

E941021US0010.7538.451.667477992.577572882.12169.612.30.131258086.2

E941101US0010.857.622.307770072.251246350.462137.8113.90.137100226.4

E941105US0011.188.292.17099472.006047331.15110.4146.20.11485649.7

E941107US0010.9739.141.910944412.104086380.728140.464.10.11055085.7

E941117US0010.95612.352.316997172.067554210.876204.6177.60.1151073054.5

E941203US0010.7667.062.274180952.297372610.556155.594.80.127280351.5

E941205US0010.7267.541.96463672.493344990.619157.129.60.116770165.1

E941207US0010.9718.611.996599222.343386710.416137.9450.11177053.87

E941213US0011.0710.232.409730292.345646861.67119.7115.90.114376130.2

E941219US0011.049.991.614897222.504307510.767133.812.90.10310278151.2

E950103US0010.88810.191.517459832.335616241.2915415.20.1926088

E950104US0010.7888.931.900967622.264479731.79157.143.30.121520159.8

E950105US0010.89711.832.133953842.113592561.79245.8104.80.10219240106.4

E950112US0010.9119.21.897791981.949868361.61165.388.70.119180214.5

E950112US0021.2111.71.944433322.223447572.28141.752.60.112889964.4

E950112US0031.078.831.377124042.786059440.901108.43.90.11720270170.3

E950112US0040.7316.241.299289332.407194730.901162.37.80.112880154.2

E950121US0010.9629.391.938269482.248960622.17140.948.90.117786.745

E950124US0011.2313.371.640282631.761760830.948224.375.60.1152221095.1

E950126US0011.2912.471.720985742.125489520.872144.239.40.19955211.2

E950126US0021.3513.591.573799582.006773223.82225.336.90.121950170

Fig-MBP-FT4

2.859162542.28581328

2.223516522.43252805

1.985200162.26885973

1.584247452.27511142

1.954802072.68318627

2.633969162.36539119

1.965558263.04648017

2.377437762.62538932

1.761429412.74439607

1.952488482.84189877

1.834955132.66763779

1.892098842.80931347

1.805822511.92216108

2.044606752.75014053

2.146128042.02260305

1.898432592.89449735

1.881012283.27895229

1.554387912.63387059

2.099930752.28403916

2.242730142.29775423

1.964649462.72410021

1.40929632.22177557

1.961063451.91262164

1.628639912.28228694

1.727164692.35058774

1.938530122.06115878

2.206352852.3289815

2.293305522.80194382

1.995379671.87546426

1.657582152.09528929

1.445749922.22564183

2.329144332.53745368

1.572370961.96491594

2.120617372.44944453

1.414160042.43643643

1.122000912.30245698

2.10579912.509202

1.556954422.10363608

2.097998342.15798019

1.606375432.13070424

2.161658332.32978855

1.576719022.29341779

1.913908432.42960859

2.180796122.40192165

1.403567942.00562793

2.537630582.92378875

1.603258642.36773019

1.387990412.53673206

2.102051182.02106414

1.599486751.99203172

1.593241812.05166257

1.676797432.191076

2.763274882.68481807

1.497658532.24238602

1.893995861.91444748

1.667477992.57757288

2.307770072.25124635

2.17099472.00604733

1.910944412.10408638

2.316997172.06755421

2.274180952.29737261

1.96463672.49334499

1.996599222.34338671

2.409730292.34564686

1.614897222.50430751

1.517459832.33561624

1.900967622.26447973

2.133953842.11359256

1.897791981.94986836

1.944433322.22344757

1.377124042.78605944

1.299289332.40719473

1.938269482.24896062

1.640282631.76176083

1.720985742.12548952

1.573799582.00677322

Log MBP (ppb)

Log MBP-c

FT4

Urinary MBP

Fig-MBP-T4

2.859162542.28581328

2.223516522.43252805

1.985200162.26885973

1.584247452.27511142

1.954802072.68318627

2.633969162.36539119

1.965558263.04648017

2.377437762.62538932

1.761429412.74439607

1.952488482.84189877

1.834955132.66763779

1.892098842.80931347

1.805822511.92216108

2.044606752.75014053

2.146128042.02260305

1.898432592.89449735

1.881012283.27895229

1.554387912.63387059

2.099930752.28403916

2.242730142.29775423

1.964649462.72410021

1.40929632.22177557

1.961063451.91262164

1.628639912.28228694

1.727164692.35058774

1.938530122.06115878

2.206352852.3289815

2.293305522.80194382

1.995379671.87546426

1.657582152.09528929

1.445749922.22564183

2.329144332.53745368

1.572370961.96491594

2.120617372.44944453

1.414160042.43643643

1.122000912.30245698

2.10579912.509202

1.556954422.10363608

2.097998342.15798019

1.606375432.13070424

2.161658332.32978855

1.576719022.29341779

1.913908432.42960859

2.180796122.40192165

1.403567942.00562793

2.537630582.92378875

1.603258642.36773019

1.387990412.53673206

2.102051182.02106414

1.599486751.99203172

1.593241812.05166257

1.676797432.191076

2.763274882.68481807

1.497658532.24238602

1.893995861.91444748

1.667477992.57757288

2.307770072.25124635

2.17099472.00604733

1.910944412.10408638

2.316997172.06755421

2.274180952.29737261

1.96463672.49334499

1.996599222.34338671

2.409730292.34564686

1.614897222.50430751

1.517459832.33561624

1.900967622.26447973

2.133953842.11359256

1.897791981.94986836

1.944433322.22344757

1.377124042.78605944

1.299289332.40719473

1.938269482.24896062

1.640282631.76176083

1.720985742.12548952

1.573799582.00677322

Log MBP (ppb)

Log MBP-c

T4

Urinary MBP

y = -0.0544x + 2.3782R2 = 0.0892

Sheet2

Sheet3

Origin Data

NAMEIDLog MBP (ppb)CreatinineFSHE2ProgTSHT4T3FT4Log MBP-c

E940216US0012.85916254126.90.1219915139.22.747.9972.570.6712.28581328

E940321US0011.6252631320.40.117730165.11.197.93108.30.8462.66480479

E940420US0012.2235165261.80.12060552.091.538.0797.291.012.43252805

E940421US0011.9852001650.50.119200182.70.619.131150.9752.26885973

E940423US0011.5842474543.30.114030101.30.4866.4485.840.7912.27511142

E940428US0011.95480207240.1151174053.410.3684.8180.970.782.68318627

E940513US0012.63396916185.60.10510480117.50.3666.3698.470.7742.36539119

E940518US0011.965558268.30.11043590.82.468.04129.30.8613.04648017

E940523US0012.3774377656.50.136975211.90.9476.7996.910.8172.62538932

E940524US0011.7614294110.40.1884568.53.2310.39132.71.222.74439607

E940524US0021.9524884812.90.10611875770.2167.71050.9372.84189877

E940526US0011.8349551314.70.117220111.21.049.91311.052.66763779

E940530US0011.8920988412.10.12747599.21.046.0892.310.812.80931347

E940601US0011.8058225176.50.11244069.10.5736.63126.30.8041.92216108

E940601US0022.0446067519.70.11244080.50.5068.77128.10.9322.75014053

E940602US0012.14612804132.90.127400114.90.8128.88131.81.012.02260305

E940602US0021.8984325915.10.112060111.91.078.01113.60.9512.89449735

E940602US0031.8810122840.1859070.60.8187.55128.80.8393.27895229

E940604US0011.5543879124.20.12215421570.6099.05127.51.042.63387059

E940606US0012.0999307534.80.1171265596.81.26.3790.890.8622.28403916

E940607US0012.2427301488.10.12913559.851.86.79106.90.7992.29775423

E940609US0011.9646494617.40.116106651311.055.71102.60.7872.72410021

E940613US0011.409296315.40.11590593.40.97.36120.40.832.22177557

E940614US0011.96106345111.80.12785079.750.4598.96135.10.9721.91262164

E940615US0011.6286399122.20.132625224.250.5049.651541.042.28228694

E940618US0011.7271646923.80.113673118.251.19.87131.60.9432.35058774

E940622US0011.9385301275.40.12070777.51.168.41109.20.9972.06115878

E940623US0012.2063528575.40.1012393883.51.366.38115.60.7352.3289815

E940623US0022.29330552310.18750137.40.8556.03130.70.72.80194382

E940627US0011.99537967131.80.125900191.51.577.47107.70.8161.87546426

E940628US0011.6575821536.50.1082602554.70.587.21108.80.792.09528929

E940704US0011.4457499216.60.112130154.31.199.77134.41.052.22564183

E940706US0012.3291443361.90.116570106.61.338.87180.40.8022.53745368

E940708US0011.5723709640.50.10816270145.61.39.5111.71.151.96491594

E940716US0011.238197978.50.1116290107.51.648.75131.40.922.30877905

E940723US0012.1206173746.90.113470103.90.75811.64207.61.032.44944453

E940725US0011.414160049.50.10918550150.41.8311.61147.21.122.43643643

E940725US0021.122000916.60.134090157.61.49.81117.21.162.30245698

E940730US0012.105799139.50.11810630154.63.068.39123.70.8782.509202

E940803US0011.5569544228.40.120920205.90.76411.23149.21.12.10363608

E940811US0012.0979983487.10.11513092.60.80510.48163.11.052.15798019

E940818US0011.6063754329.90.117270176.71.9210.43145.91.062.13070424

E940823US0012.1616583367.90.127700204.80.6439.08132.11.072.32978855

E940824US0011.5767190219.20.16118570115.72.1110.52152.51.012.29341779

E940902US0011.9139084330.50.128800202.70.2317.52122.40.9852.42960859

E940905US0012.1807961260.10.147820441.95.194.39145.10.462.40192165

E940905US0021.40356794250.123230307.10.9039.25139.90.9612.00562793

E940906US0012.5376305841.10.10930160206.61.178.31137.70.8892.92378875

E940923US0011.6032586417.20.10220540160.20.8848.31138.20.7952.36773019

E940928US0011.387990417.10.1116350112.51.6311.17148.50.9832.53673206

E940929US0012.10205118120.50.1694851.730.47810.9192.60.9182.02106414

E941003US0011.5994867540.50.13316980105.11.1210.43158.50.9611.99203172

E941006US0011.5932418134.80.1120160124.42.568.44126.60.82.05166257

E941006US0021.6767974330.60.10422480150.31.179.87159.30.9172.191076

E941007US0012.76327488119.80.1431367045.641.57.02110.80.7382.68481807

E941013US0011.49765853180.12210730761.797.65128.50.8382.24238602

E941020US0011.8939958695.40.128590140.70.9469.17122.20.8971.91444748

E941021US0011.6674779912.30.131258086.22.128.45169.60.7532.57757288

E941101US0012.30777007113.90.137100226.40.4627.62137.80.852.25124635

E941105US0012.1709947146.20.11485649.71.158.29110.41.182.00604733

E941107US0011.9109444164.10.11055085.70.7289.14140.40.9732.10408638

E941117US0012.31699717177.60.1151073054.50.87612.35204.60.9562.06755421

E941203US0012.2741809594.80.127280351.50.5567.06155.50.7662.29737261

E941205US0011.964636729.60.116770165.10.6197.54157.10.7262.49334499

E941207US0011.99659922450.11177053.870.4168.61137.90.9712.34338671

E941213US0012.40973029115.90.114376130.21.6710.23119.71.072.34564686

E941219US0011.6148972212.90.10310278151.20.7679.99133.81.042.50430751

E950103US0011.5174598315.20.19260881.2910.191540.8882.33561624

E950104US0011.9009676243.30.121520159.81.798.93157.10.7882.26447973

E950105US0012.13395384104.80.10219240106.41.7911.83245.80.8972.11359256

E950112US0011.8977919888.70.119180214.51.619.2165.30.9111.94986836

E950112US0021.9444333252.60.112889964.42.2811.7141.71.212.22344757

E950112US0031.377124043.90.11720270170.30.9018.83108.41.072.78605944

E950112US0041.299289337.80.112880154.20.9016.24162.30.7312.40719473

E950121US0011.9382694848.90.117786.7452.179.39140.90.9622.24896062

E950124US0011.6402826375.60.1152221095.10.94813.37224.31.231.76176083

E950126US0011.7209857439.40.19955211.20.87212.47144.21.292.12548952

E950126US0021.5737995836.90.1219501703.8213.59225.31.352.00677322

MBP-FT4

NAMEIDFT4T4Log MBP (ppb)Log MBP-cTSHT3CreatinineFSHE2Prog

E940216US0010.6717.992.859162542.285813282.7472.57126.90.1219915139.2

E940420US0011.018.072.223516522.432528051.5397.2961.80.12060552.09

E940421US0010.9759.131.985200162.268859730.6111550.50.119200182.7

E940423US0010.7916.441.584247452.275111420.48685.8443.30.114030101.3

E940428US0010.784.811.954802072.683186270.36880.97240.1151174053.41

E940513US0010.7746.362.633969162.365391190.36698.47185.60.10510480117.5

E940518US0010.8618.041.965558263.046480172.46129.38.30.11043590.8

E940523US0010.8176.792.377437762.625389320.94796.9156.50.136975211.9

E940524US0011.2210.391.761429412.744396073.23132.710.40.1884568.5

E940524US0020.9377.71.952488482.841898770.21610512.90.1061187577

E940526US0011.059.91.834955132.667637791.0413114.70.117220111.2

E940530US0010.816.081.892098842.809313471.0492.3112.10.12747599.2

E940601US0010.8046.631.805822511.922161080.573126.376.50.11244069.1

E940601US0020.9328.772.044606752.750140530.506128.119.70.11244080.5

E940602US0011.018.882.146128042.022603050.812131.8132.90.127400114.9

E940602US0020.9518.011.898432592.894497351.07113.615.10.112060111.9

E940602US0030.8397.551.881012283.278952290.818128.840.1859070.6

E940604US0011.049.051.554387912.633870590.609127.524.20.1221542157

E940606US0010.8626.372.099930752.284039161.290.8934.80.1171265596.8

E940607US0010.7996.792.242730142.297754231.8106.988.10.12913559.85

E940609US0010.7875.711.964649462.724100211.05102.617.40.11610665131

E940613US0010.837.361.40929632.221775570.9120.415.40.11590593.4

E940614US0010.9728.961.961063451.912621640.459135.1111.80.12785079.75

E940615US0011.049.651.628639912.282286940.50415422.20.132625224.25

E940618US0010.9439.871.727164692.350587741.1131.623.80.113673118.25

E940622US0010.9978.411.938530122.061158781.16109.275.40.12070777.5

E940623US0010.7356.382.206352852.32898151.36115.675.40.1012393883.5

E940623US0020.76.032.293305522.801943820.855130.7310.18750137.4

E940627US0010.8167.471.995379671.875464261.57107.7131.80.125900191.5

E940628US0010.797.211.657582152.095289290.58108.836.50.1082602554.7

E940704US0011.059.771.445749922.225641831.19134.416.60.112130154.3

E940706US0010.8028.872.329144332.537453681.33180.461.90.116570106.6

E940708US0011.159.51.572370961.964915941.3111.740.50.10816270145.6

E940723US0011.0311.642.120617372.449444530.758207.646.90.113470103.9

E940725US0011.1211.611.414160042.436436431.83147.29.50.10918550150.4

E940725US0021.169.811.122000912.302456981.4117.26.60.134090157.6

E940730US0010.8788.392.10579912.5092023.06123.739.50.11810630154.6

E940803US0011.111.231.556954422.103636080.764149.228.40.120920205.9

E940811US0011.0510.482.097998342.157980190.805163.187.10.11513092.6

E940818US0011.0610.431.606375432.130704241.92145.929.90.117270176.7

E940823US0011.079.082.161658332.329788550.643132.167.90.127700204.8

E940824US0011.0110.521.576719022.293417792.11152.519.20.16118570115.7

E940902US0010.9857.521.913908432.429608590.231122.430.50.128800202.7

E940905US0010.464.392.180796122.401921655.19145.160.10.147820441.9

E940905US0020.9619.251.403567942.005627930.903139.9250.123230307.1

E940906US0010.8898.312.537630582.923788751.17137.741.10.10930160206.6

E940923US0010.7958.311.603258642.367730190.884138.217.20.10220540160.2

E940928US0010.98311.171.387990412.536732061.63148.57.10.1116350112.5

E940929US0010.91810.92.102051182.021064140.478192.6120.50.1694851.73

E941003US0010.96110.431.599486751.992031721.12158.540.50.13316980105.1

E941006US0010.88.441.593241812.051662572.56126.634.80.1120160124.4

E941006US0020.9179.871.676797432.1910761.17159.330.60.10422480150.3

E941007US0010.7387.022.763274882.684818071.5110.8119.80.1431367045.64

E941013US0010.8387.651.497658532.242386021.79128.5180.1221073076

E941020US0010.8979.171.893995861.914447480.946122.295.40.128590140.7

E941021US0010.7538.451.667477992.577572882.12169.612.30.131258086.2

E941101US0010.857.622.307770072.251246350.462137.8113.90.137100226.4

E941105US0011.188.292.17099472.006047331.15110.4146.20.11485649.7

E941107US0010.9739.141.910944412.104086380.728140.464.10.11055085.7

E941117US0010.95612.352.316997172.067554210.876204.6177.60.1151073054.5

E941203US0010.7667.062.274180952.297372610.556155.594.80.127280351.5

E941205US0010.7267.541.96463672.493344990.619157.129.60.116770165.1

E941207US0010.9718.611.996599222.343386710.416137.9450.11177053.87

E941213US0011.0710.232.409730292.345646861.67119.7115.90.114376130.2

E941219US0011.049.991.614897222.504307510.767133.812.90.10310278151.2

E950103US0010.88810.191.517459832.335616241.2915415.20.1926088

E950104US0010.7888.931.900967622.264479731.79157.143.30.121520159.8

E950105US0010.89711.832.133953842.113592561.79245.8104.80.10219240106.4

E950112US0010.9119.21.897791981.949868361.61165.388.70.119180214.5

E950112US0021.2111.71.944433322.223447572.28141.752.60.112889964.4

E950112US0031.078.831.377124042.786059440.901108.43.90.11720270170.3

E950112US0040.7316.241.299289332.407194730.901162.37.80.112880154.2

E950121US0010.9629.391.938269482.248960622.17140.948.90.117786.745

E950124US0011.2313.371.640282631.761760830.948224.375.60.1152221095.1

E950126US0011.2912.471.720985742.125489520.872144.239.40.19955211.2

E950126US0021.3513.591.573799582.006773223.82225.336.90.121950170

Fig-MBP-FT4

2.859162542.28581328

2.223516522.43252805

1.985200162.26885973

1.584247452.27511142

1.954802072.68318627

2.633969162.36539119

1.965558263.04648017

2.377437762.62538932

1.761429412.74439607

1.952488482.84189877

1.834955132.66763779

1.892098842.80931347

1.805822511.92216108

2.044606752.75014053

2.146128042.02260305

1.898432592.89449735

1.881012283.27895229

1.554387912.63387059

2.099930752.28403916

2.242730142.29775423

1.964649462.72410021

1.40929632.22177557

1.961063451.91262164

1.628639912.28228694

1.727164692.35058774

1.938530122.06115878

2.206352852.3289815

2.293305522.80194382

1.995379671.87546426

1.657582152.09528929

1.445749922.22564183

2.329144332.53745368

1.572370961.96491594

2.120617372.44944453

1.414160042.43643643

1.122000912.30245698

2.10579912.509202

1.556954422.10363608

2.097998342.15798019

1.606375432.13070424

2.161658332.32978855

1.576719022.29341779

1.913908432.42960859

2.180796122.40192165

1.403567942.00562793

2.537630582.92378875

1.603258642.36773019

1.387990412.53673206

2.102051182.02106414

1.599486751.99203172

1.593241812.05166257

1.676797432.191076

2.763274882.68481807

1.497658532.24238602

1.893995861.91444748

1.667477992.57757288

2.307770072.25124635

2.17099472.00604733

1.910944412.10408638

2.316997172.06755421

2.274180952.29737261

1.96463672.49334499

1.996599222.34338671

2.409730292.34564686

1.614897222.50430751

1.517459832.33561624

1.900967622.26447973

2.133953842.11359256

1.897791981.94986836

1.944433322.22344757

1.377124042.78605944

1.299289332.40719473

1.938269482.24896062

1.640282631.76176083

1.720985742.12548952

1.573799582.00677322

Log MBP (ppb)

Log MBP-c

FT4

Urinary MBP

y = -0.8073x + 2.6508R2 = 0.1328

Fig-MBP-T4

2.859162542.28581328

2.223516522.43252805

1.985200162.26885973

1.584247452.27511142

1.954802072.68318627

2.633969162.36539119

1.965558263.04648017

2.377437762.62538932

1.761429412.74439607

1.952488482.84189877

1.834955132.66763779

1.892098842.80931347

1.805822511.92216108

2.044606752.75014053

2.146128042.02260305

1.898432592.89449735

1.881012283.27895229

1.554387912.63387059

2.099930752.28403916

2.242730142.29775423

1.964649462.72410021

1.40929632.22177557

1.961063451.91262164

1.628639912.28228694

1.727164692.35058774

1.938530122.06115878

2.206352852.3289815

2.293305522.80194382

1.995379671.87546426

1.657582152.09528929

1.445749922.22564183

2.329144332.53745368

1.572370961.96491594

2.120617372.44944453

1.414160042.43643643

1.122000912.30245698

2.10579912.509202

1.556954422.10363608

2.097998342.15798019

1.606375432.13070424

2.161658332.32978855

1.576719022.29341779

1.913908432.42960859

2.180796122.40192165

1.403567942.00562793

2.537630582.92378875

1.603258642.36773019

1.387990412.53673206

2.102051182.02106414

1.599486751.99203172

1.593241812.05166257

1.676797432.191076

2.763274882.68481807

1.497658532.24238602

1.893995861.91444748

1.667477992.57757288

2.307770072.25124635

2.17099472.00604733

1.910944412.10408638

2.316997172.06755421

2.274180952.29737261

1.96463672.49334499

1.996599222.34338671

2.409730292.34564686

1.614897222.50430751

1.517459832.33561624

1.900967622.26447973

2.133953842.11359256

1.897791981.94986836

1.944433322.22344757

1.377124042.78605944

1.299289332.40719473

1.938269482.24896062

1.640282631.76176083

1.720985742.12548952

1.573799582.00677322

Log MBP (ppb)

Log MBP-c

T4

Urinary MBP

Sheet2

Sheet3

Table I-5. Multivariate linear regression between serum FT4 and T4 levels, and their corresponding urinary phthalate monoesters (FT4: R-square= 0.240; T4: R-square= 0.187)a. a One outlier was excluded because of hypothyroidism (n=75).Multiple regression: FT4 = -0.110, R2=0.24, p|t|Intercept1.2700.0132.3620.001Age0.0240.886-0.3210.177BMI0.0880.5790.3480.120Gestational age-0.1170.598-0.0660.831MBP-0.110

Table II-1 Physical examination of male and female newborns (n = 65).1 The AGD of two girls and boys, respectively, were not available.2 AGI=ano-genital index; AGI-W= AGD/birth weight; AGI-L=AGD/ birth length.3 *: p

Health statusMales (n=33)Females (n=32)P-value3Birth weight (g)3171 (1678-4260)3002(2000-3935)0.156Birth length (cm)49.6 (42.0-56.0)48.3 (42.5-53.5)0.023*Gestational age (weeks)39.0 (35.3-41.7)38.4 (33.6-40.3)0.020*Ano-genital distance (AGD) (mm)123 (10-36)16 (7-23)

Table II-2 Concentrations of five phthalate monoesters in amniotic fluid from female and male infants (ng/mL, n=64)1.1One amniotic fluid sample was failed during analysis. 2ND=not detected, detection limit of MMP, MEP, MBP, MBzP and MEHP were as follow: 1.4, 1.0, 1.4, 1.4, 0.9 ng/mL

Female infantsMin10th50th90thMaxMBP39.345.685.5134.6192.0MEHPND25.024.091.1148.0MEPNDNDND3.96.5MBzPNDNDND84.1233.0MMPNDNDNDND2.92Male infantsMin10th50th90thMaxMBP28.444.381.3127.8145.0MEHPND22.622.1100.6110.0MEPNDNDND4.47.7MBzPNDNDND87.9104.0MMPNDNDNDNDND

Figure II-1 Distribution plot of MBP levels in maternal urine and amniotic fluid (n=59). All the data were log-transformed.It is indicated that current levels of exposure to phthalate in pregnant women may only partly represent in utero fetal exposure

Table II-3 Distribution of health status in females categorized by median levels of phthalate monoester in amniotic fluid (-AF) (n = 31).1Wilcoxon Rank Sum Test, *: p < 0.05;2AGD data were lacking for one female newborn in each group.3AGD data were lacking for two female newborns in the high MEHP-AF group.4Median levels of amniotic fluid MBP in low and high group were 67 and 104 ng/mL, respectively; those of MEHP were 9.5 and 38.8 ng/mL, respectively.

Health status in female infantsMBP-AF2MEHP-AF3Low4(n=15)High4(n=16)P-value1Low4(n=15)High4(n=16)P-value1Birth weight (g)281043931723980.031*291844830614670.527Birth length (cm)47.32.349.21.70.018*48.12.148.62.40.138Gestational age (weeks)38.11.538.71.00.20538.40.938.41.60.591AGD (mm)17.64.013.93.80.024*17.04.114.24.10.109AGI-W (mm/kg)6.21.64.51.50.007*6.01.44.71.80.016*AGI-L (103)3.70.92.80.80.008*3.60.82.90.90.015*

Table II-4. Distribution of health status in boys category by median levels of phthalate monoester in amniotic fluid. (n = 33).1 One male infant of each group in MBP-AF and MEHP-AF, respectively, were lack of AGD.

Health status in male infantsMBPMEHPLow(n=16)High(n=17)P-valueLow(n=16)High(n=17)P-valueBirth weight (g)314648131944690.640321344031315150.815Birth length (cm)49.22.550.02.80.70549.42.449.73.00.971Gestational age (weeks)39.11.038.91.40.39639.10.938.91.50.787AGD (mm)21.26.524.15.70.23423.36.822.25.60.606AGI-W (mm/kg)6.62.17.72.20.2067.42.66.91.70.635AGI-L (103)4.31.24.81.20.3044.71.44.41.00.649

Table II-5 Spearman correlation coefficients1 between birth weight, birth length, gestational age, AGD, AGI-W, AGI-L, and phthalate monoester levels in amniotic fluid in female infants (n = 29)2. 1 *:p< 0.05; #: p

Figure II-2 (A) Linear regressions between MBP in amniotic fluid and AGI-W in females at birth (n=29, R2=0.14, p=0.043); (B) Linear regressions between MBP in amniotic fluid and AGI-L in females at birth (n=29, R2=0.16, p=0.032).Negative associations among amniotic fluid MBP and AGI

Figure III-3 Multiple regression of AGI, MBP in amniotic fluid and gestational age adjustment of gestational age and other phthalate monoesters (amniotic fluid MBP: = 2.73, p = 0.041; gestational age: = 0.899, p = 0.006) .

Table III-1 Concentrations of phthalate monoesters in urine, serum and cord blood at delivery (ng/mL, n=61).aMBP = monobutyl phthalate; MBzP = monobenzyl phthalate; MEP = monoethyl phthalate; MEHP = mono-2-ethylhexyl phthalate; MMP = monomethyl phthalate. bDetection limit (LOD) of phthalate monoesters were: MBP, 1.4; MBzP, 1.4; MEP, 1.0; MEHP, 0.9; MMP, 1.4 ng/mL, respectively. Half of LOD was calculated as the detected value below the LOD.

Mono-PAEsUrineSerumCord bloodMedian (Range)10-90thMedian (Range)10-90thMedian (Range)10-90thMBPa114(25.4-1830)36.9-550.6158.0(59.6-1080)64.9-413.0256.0(65.2-815)97.4-604.8MEHP40.2(3.6-958)8.4-152.021.0(9.2-99.2)11.7-37.124.7(11.0-665.0)14.2-65.9MEP36.4(NDb-1980)4.6-236.82.8(ND-26.5)ND-6.3ND(ND-9.3)ND-3.4MBzP5.7(ND-218.0)1.9-49.2ND(ND-10.1)ND-2.8ND(ND-26.8)ND-3.6MMP8.3(ND-169)1.7-38.0ND(ND-3.7)ND-2.4ND(ND-13.3)ND-ND

Table III-2 Concentrations of thyroid hormones1 in maternal serum during pregnancy and cord blood at delivery (n=61).1Reference values for thyroid hormones in Taiwan: TSH: 0.27-4.2; T3: 84.6-202.0; T4: 5.13-14.1; FT4: 0.93-1.7. The analytic sensitivities of T3, T4, free T4, and TSH were 19.5 ng/dL, 0.42 g/dL, 0.023 ng/dL and 0.014 lU/mL, respectively. The coefficient variations of T3, T4, free T4, and TSH were 3.0%, 4.1%, 3.2%, and 2.9%, respectively.

Serum Thyroid HormonesFirst-trimesterSecond-trimesterThird-trimesterCord bloodMedianRangeMedianRangeMedianRangeMedianRangeTSH (IU/mL)1.130.01-4.381.060.23-5.192.080.38-6.077.051.63-289T3 (ng/dL)12370.2-23013472.6-24614082.4-27756.335.1-84.6T4 (g/dL)8.263.37-12.19.014.39-13.49.623.60-16.97.663.65-11.7FT4 (ng/dL)0.950.62-1.290.930.46-1.290.990.33-1.311.130.49-1.45

Table III-3 Correlation a coefficients between fetal thyroid hormones and phthalate monoesters in cord blood samples (n=60)a.a Spearman correlation: **p

Figure III-2 Linear correlation between log MBP levels and TSHT4 in cord blood samples (R2=0.081, p=0.028, n=60).

Log MBP-T4-cord

7.66

9.04

5.38

5.44

11.73

5.56

7.23

6.68

6.24

5.89

10.19

8.24

7.67

6.35

7.46

9.09

6.35

7

9.7

8.57

9.01

7.57

10.66

4.16

8.58

8.85

8.56

5.3

10.71

11.36

11.15

10.64

5.83

7.32

5.88

7.88

7.71

7.96

6.83

6

9.53

10.19

8.38

7.3

6.56

7.29

5.27

7.2

8.86

8.29

7.7

8.61

6.26

8.96

8.97

5.65

9.37

5.54

Log MBP (ng/mL)

Thyroxine (T4) (g/dL)

y = -1.4656x + 11.315R2 = 0.055

Log MBP-TSH-cord

0.66181269

0.9380191

0.80955971

0.54157924

1.02938378

0.32221929

1.0211893

0.99078269

0.5390761

0.89597473

0.88705438

1.05115252

1.11327469

1.02653326

0.60959441

1.08062649

1.22427401

1.05918462

0.81090428

0.95808585

1.01452054

0.64542227

1.16106839

0.2121876

0.75511227

0.63042788

0.93043959

0.53402611

0.79588002

0.78175537

0.89762709

1.10311925

0.78532984

0.91115761

1.19617619

0.71096312

0.7176705

0.84818912

1.33984878

0.79448805

0.58771097

1.01828431

1.21404868

0.83058867

0.72181062

1.00475116

0.83122969

0.83122969

1.0733517

0.84509804

1.03941412

0.71600334

0.7363965

0.84010609

1.38632057

0.64836001

1.20384846

0.75358306

Log MBP (ng/mL)

TSH (IU/mL)

y = -0.2124x + 1.3707R2 = 0.060

Log MBP-T4-cord-1

7.66

9.04

5.38

5.44

11.73

5.56

7.23

6.68

6.24

5.89

10.19

8.24

7.67

6.35

7.46

9.09

3.65

6.35

7

9.7

8.57

7.57

9.01

7.57

10.66

4.16

8.58

8.85

8.56

5.3

10.71

11.36

11.15

10.64

5.83

7.32

5.88

7.88

7.71

7.96

6.83

6

9.53

10.19

8.38

7.3

6.56

7.29

5.27

7.2

8.86

8.29

7.7

8.61

6.26

8.96

8.97

5.65

9.37

5.54

Log MBP (ng/mL)

Thyroxine (T4) (g/dL)

Log MBP-TSH-Cord-1

0.66181269

0.9380191

0.80955971

0.54157924

1.02938378

0.32221929

1.0211893

0.99078269

0.5390761

0.89597473

0.88705438

1.05115252

1.11327469

1.02653326

0.60959441

1.08062649

2.4619485

1.22427401

1.05918462

0.81090428

0.95808585

1.21774707

1.01452054

0.64542227

1.16106839

0.2121876

0.75511227

0.63042788

0.93043959

0.53402611

0.79588002

0.78175537

0.89762709

1.10311925

0.78532984

0.91115761

1.19617619

0.71096312

0.7176705

0.84818912

1.33984878

0.79448805

0.58771097

1.01828431

1.21404868

0.83058867

0.72181062

1.00475116

0.83122969

0.83122969

1.0733517

0.84509804

1.03941412

0.71600334

0.7363965

0.84010609

1.38632057

0.64836001

1.20384846

0.75358306

TSH-log

Log MBP (ng/mL)

TSH (IU/mL)

Chart1

35.1594

78.3768

34.701

18.9312

125.511

11.676

75.915

65.3972

21.5904

46.3543

78.5649

92.7

99.5566

67.5005

30.3622

109.4436

106.426

80.22

62.759

77.8156

124.9807

93.1634

33.4594

154.4634

6.7808

48.8202

37.7895

72.9312

18.126

66.9375

68.728

88.085

134.9152

35.563

59.658

92.3748

40.5032

40.2462

56.118

149.3721

37.38

36.8811

106.2817

137.1806

49.421

34.5712

73.7019

35.7306

48.816

104.9024

58.03

84.315

44.772

34.117

62.0032

218.3298

25.1425

149.8263

31.4118

13.9293

TSHxT4

Log MBP in cord blood (ng/mL)

TSH(IU/mL)T4(g/dL)

y = -42.792x + 171.39R2 = 0.081

MBP-TSHxT4

NoLog MBP-cTSHxT4Log TSHxT4

52.7573960335.15941.5460414552

72.624282178.37681.894187528

82.0718820134.7011.5403419903

92.9014583218.93121.2771781436

102.33845649125.5112.0986817898

112.6608654811.6761.0672940863

122.0718820175.9151.8803275964

132.4593924965.39721.8155591543

152.0791812521.59041.3342606885

172.4471580346.35431.6660900271

182.5118833678.56491.8952285621

212.3765769692.71.9670797341

222.4941545999.55661.9980700564

252.3384564967.50051.8293069898

262.434568930.36221.4823332367

282.30319606109.44362.0391903701

302.50242712106.4262.0270477396

312.1139433580.221.9042826576

322.9111576162.7591.7976760149

332.7874604777.81561.8910666704

342.66370093124.98072.0968429528

352.577491893.16341.9692453297

372.873320633.45941.5245181488

382.35410844154.46342.1888255902

402.568201726.78080.831280935

412.021189348.82021.6885995542

442.021189337.78951.5773711457

452.4899584872.93121.8629133594

462.829946718.1261.2583019757

472.117271366.93751.8256694882

492.4517864468.7281.837133706

511.9185545388.0851.9449019587

541.95279244134.91522.1300608815

552.5976951935.5631.5509983898

562.0492180259.6581.7756686898

572.4082399792.37481.9655535111

581.9590413940.50321.6074893365

602.16136840.24621.6047248811

612.5932860756.1181.7491021847

621.8142476149.37212.1742694867

632.8375884437.381.572639297

642.3364597336.88111.5668038657

652.14921911106.28172.0264584924

682.30749604137.18062.137292698

692.6424645249.4211.6939115288

702.3053513734.57121.5387144546

712.0791812573.70191.8674786839

722.5831987735.73061.5530403091

732.3138672248.8161.6885621903

772.65321251104.90242.0207854243

782.5105450158.031.7636525706

792.328379684.3151.9259048443

802.7168377244.7721.6510064951

812.3838153734.1171.5329708355

852.3673559262.00321.7924141041

861.98000337218.32982.3391130169

872.679427925.14251.4004084588

882.52113808149.82632.1755880546

902.557507231.41181.4970928236

921.9642596313.92931.143929292

292.075546961057.4053.0242413597

Data

NoLog MBP-CT4TSH-log

12.757396037.660.66181269

22.62428219.040.9380191

32.071882015.380.80955971

42.901458325.440.54157924

52.3384564911.731.02938378

62.660865485.560.32221929

72.071882017.231.0211893

82.459392496.680.99078269

92.079181256.240.5390761

102.447158035.890.89597473

112.5118833610.190.88705438

142.376576968.241.05115252

152.494154597.671.11327469

182.338456496.351.02653326

192.43456897.460.60959441

212.303196069.091.08062649

222.075546963.652.4619485

232.502427126.351.22427401

242.1139433571.05918462

252.911157619.70.8109042822

262.787460478.570.9580858527

272.663700937.571.2177470741

282.57749189.011.01452054

302.87332067.570.64542227

312.3541084410.661.16106839

332.568201724.160.2121876

342.02118938.580.75511227

372.02118938.850.63042788

382.489958488.560.93043959

392.82994675.30.53402611

402.117271310.710.79588002

422.4517864411.360.78175537

441.9185545311.150.89762709

471.9527924410.641.10311925

482.597695195.830.78532984

492.049218027.320.91115761

502.408239975.881.19617619

511.959041397.880.71096312

532.1613687.710.7176705

542.593286077.960.84818912

551.81424766.831.33984878

562.8375884460.79448805

572.336459739.530.58771097

582.1492191110.191.01828431

612.307496048.381.21404868

622.642464527.30.83058867

632.305351376.560.72181062

642.079181257.291.00475116

652.583198775.270.83122969

662.313867227.20.83122969

702.653212518.861.0733517

712.510545018.290.84509804

722.32837967.71.03941412

732.716837728.610.71600334

742.383815376.260.7363965

782.367355928.960.84010609

791.980003378.971.38632057

802.67942795.650.64836001

812.521138089.371.20384846

832.55750725.540.75358306

Sheet1 (2)

NoLog MBP-CT4TSH-log

12.757396037.660.66181269

22.62428219.040.9380191

32.071882015.380.80955971

42.901458325.440.54157924

52.3384564911.731.02938378

62.660865485.560.32221929

72.071882017.231.0211893

82.459392496.680.99078269

92.079181256.240.5390761

102.447158035.890.89597473

112.5118833610.190.88705438

12...

13...

142.376576968.241.05115252

152.494154597.671.11327469

16...

17...

182.338456496.351.02653326

192.43456897.460.60959441

20...

212.303196069.091.08062649

222.075546963.652.4619485

232.502427126.351.22427401

242.1139433571.05918462

252.911157619.70.81090428

262.787460478.570.95808585

272.663700937.571.21774707

282.57749189.011.01452054

29...

302.87332067.570.64542227

312.3541084410.661.16106839

32...

332.568201724.160.2121876

342.02118938.580.75511227

35...

36...

372.02118938.850.63042788

382.489958488.560.93043959

392.82994675.30.53402611

402.117271310.710.79588002

41.7.280.98227123

422.4517864411.360.78175537

43...

441.9185545311.150.89762709

45...

46...

471.9527924410.641.10311925

482.597695195.830.78532984

492.049218027.320.91115761

502.408239975.881.19617619

511.959041397.880.71096312

52...

532.1613687.710.7176705

542.593286077.960.84818912

551.81424766.831.33984878

562.8375884460.79448805

572.336459739.530.58771097

582.1492191110.191.01828431

59...

60...

612.307496048.381.21404868

622.642464527.30.83058867

632.305351376.560.72181062

642.079181257.291.00475116

652.583198775.270.83122969

662.313867227.20.83122969

67...

68...

69...

702.653212518.861.0733517

712.510545018.290.84509804

722.32837967.71.03941412

732.716837728.610.71600334

742.383815376.260.7363965

75...

76...

77...

782.367355928.960.84010609

791.980003378.971.38632057

802.67942795.650.64836001

812.521138089.371.20384846

82...

832.55750725.540.75358306

84...

851.964259637.370.2764618

86...

87...

Sheet2

Sheet3

Table III-4 Multivariate regression between TSH and T4 and their corresponding phthalate monoesters in cord blood (n=60)a

*Possible Hypotheses First, we postulated phthalate exposure may affect fetal thyroid function indirectly via decreased thyroid hormones from their mother. Animal studies have shown that in utero exposure to thyroid hormone affects the regulation of the hypothalamus-pituitary-thyroid axis in newborns (Alonso et al., 2007; Piosik et al., 1997). Our previous study showed decreased FT4 and T4 levels with increasing urinary MBP levels in pregnant women during the second trimester (Huang et al., 2007).Second, phthalate monoesters penetrate the human placenta and may have direct effects on fetal thyroid function.Some toxicological studies have shown that certain phthalates, such as DBP and DEHP, are possible thyroxine antagonists that affect the transportation, secretion, and action of thyroid hormones in adult animals (Hinton et al., 1986; Poon et al., 1997; Sugiyama et al., 2005).Although little information is available about phthalate exposure in utero and its effects on fetal thyroid function, some studies (Huang et al., 2007; Meeker et al., 2007) have shown possible negative effects in humans.

The study on relationship between urinary phthalate monoesters and house dust exposure in precocious puberty girls

Phthalate in house dust(Fromme et al., 2007)

*Exposure route of phthalatesINGESTIONSKIN ABSORPTIONINHALATIONnewlatesttips.info pureautismsolutions.com www.tedmiller.com/?cat=572blogs.babycenter.com Children prone to be higher exposedLower body weightHand to mouth behavior (sucking/ mouthing dust)[Schettler, 2006; Sathyanarayana, 2008; Wittassek, 2010]

Fig 1 The framework of this study

Inclusion criteria:the appearance of secondary sexual characteristics < 8 years oldBreast / Pubic hair development : Tanner breast / pubic hair stage scores Menarche LHRH test: LH, FSH, E2 of plasma (Positive: LH>10mIU/ml)Bone age: advanceAbdominal Ultrasound / MRI Examination

Exclusion criteria:Precocious puberty caused by: Brain tumor/ ovary tumor/Hydrocephalus/ Congenital adrenal hyperplasia (CAH)

Precocious puberty girls (case)

Inclusion criteria:Age from 2 to 8 years non-precocious puberty No secondary sex characteristicsNo breast development No appearance of breast tissue No appearance of pubic hair No menarche

Exclusion criteria:the child who dose not meet any one of the inclusion criteria

Normal girls (control)

Informed consents

Questionnaires

Demographic dataFamily circumstance Medical historyDietary habit

Examination by pediatrician

Collect spot urine sample

LC-MS/MS

MMP, MEP, MBP, MBzP, MEHP, MEOHP, MEHHP

Radioimmunoassay

Kisspeptin 54LH, FSH, E2

Collect blood sample

GC/MS

DMP, DEP, DBP, BBP, DEHP

Collect dust sample (Childs bed, bedroom floor, sofa, living room floor, areas above floor)

Intervention

*Sampling strategy for dust- 5 samplesKitchenbathroomParents roomLiving roomPlayroom

Bed (or sofa)FloorUpper floorMajor activity area(Sleeping room)Sample 1Sample 2Sample 5Minor activity area(Parents room, living room, playroom et al.)Sample 3Sample 4

Dust collection equipmentUsing aluminum-made collector & cellulose filterCovering with aluminum foil and then sealing by parafilm after collecting enough dust sample

*

Distributions of phthalate levels in settled dust (n=965) (g/g dust)*N.D.: non-detection

No. of above detection limit (%)No. of missing data (%)Mean (sd)PercentileMin5th25th50th75th95thMaxDMP47 (4.9)90 (9.3)0.8 (4.4)N.D.N.D.N.D.N.D.N.D.2.1119.6DEP67 (6.9)90 (9.3)2.1 (10.7)N.D.N.D.N.D.N.D.N.D.3.1196.0DBP825 (85.5)137 (14.2)46.0 (93.8)N.D.7.114.123.141.8139.31173.2BBzP575 (59.6)98 (10.2)6.8 (17.2)N.D.N.D.N.D.2.86.223.3312.5DEHP865 (89.6)100 (10.4)1761.0 (2346.5)79.0337.5665.41128.91932.54428.724209.7

*Worldwide comparison of phthalates concentrationND, no data. a 50th and 95th percentiles. b Mean concentration. c 90th percentile.[Adapted from Kolarik, 2008]

StudyCountryNo.DEHP (mg/g dust)BBzP (mg/g dust)DBP (mg/g dust)50th95th50th95th50th95thPresent studyTAIWAN9651.234.430.0030.020.020.14Kolarik et al. 2008Bulgaria1840.997.980.331.569.8530.8Bornehag et al. 2004Sweden3460.774.070.130.600.150.57Pohner et al. 1997Germany2720.452.00NDNDNDNDOie et al. 1997Norway380.64b0.11b0.10bButte et al. 2001Germany2860.742.600.050.320.050.24Becker et al. 2002Germany1990.421.190.010.210.040.16Clausen et al. 2003Denmark230.862.59NDNDNDNDRudel et al. 2003USA1200.340.85c0.040.28c0.020.04cKersten and Reich 2003Germany650.601.600.020.230.050.18Fromme et al. 2004Germany300.701.540.030.220.060.13Becker et al. 2004Germany2520.511.84NDNDNDND

nmean (sd)medianrangeAveragelevels of allsampling siteSampling site at MAJOR roomSampling site at MINOR roombedfloorupper floor levelsbed/sofafloorDMP8750.8 (4.4)0.10.1-119.61900.6 (1.6)0.10.1-18.91830.7 (2.5)0.10.1-32.51291.3 (8.8)0.10.1-119.61860.7 (1.6)0.10.1-12.81870.6 (1.5)0.10.1-14.3DEP8752.1 (10.7)1.00.1-196.01902.3 (8.7)1.00.1-86.61831.7 (7.2)1.00.1-88.21871.2 (3.5)1.00.1-45.41291.0 (1.2)1.00.1-10.61863.8 (19.8)1.00.1-196.0DBP82846.0 (93.8)23.10.1-1173.217825.0 (25.3)17.84.5-212.618367.9 (121.5)33.64.1-984.716845.4 (91.2)20.93.4-834.312838.4 (100.9)17.10.1-956.617150.6 (97.6)30.50.1-1173.2BBzP8676.8 (17.2)2.80.1-312.51865.4 (8.4)2.80.1-82.91839.0 (20.2)4.20.1-185.01833.3 (4.4)2.00.1-40.21295.2 (12.5)2.10.1-120.818610.4 (27.3)3.50.1-312.5DEHP8651761.0 (2346.5)1128.979.0-24209.7187895.4 (509.3)799.579.0-3416.91831757.6 (2339.8)1235.8235.4-23292.31841203.7 (1218.2)807.9119.5-9681.11293196.4 (4360.3)1477.3302.6-24209.71822199.6 (1677.8)1798.2292.1-17218.3

*

DBP level and wooden floor*Significantly higher DBP concentration was found at major activity room and above floor level when using wooden materials for flooring in the room.

Sampling site*50th (25th-75th)p-valueUsing wooden floorNo wooden floorMajor bed19.1 (12.0-26.6)17.3 (11.6-29.4)0.74Major floor38.1 (25.3-92.5)30.6 (17.9-52.5)0.02Upper floor33.3 (18.6-62.9)18.8 (12.1-31.5)

Wax using and phthalates concentration*Regular waxing was associated with higher BBzP and DEHP levels in dusts collected from the floor.

Using waxNo using waxpn50th (25th-75th)n50th (25th-75th)BBzP89.0 (6.7-11.3)1233.9 (1.0-8.1)0.04DEHP82209.2 (1575.1-2809.4)1231105.0 (640.5-1743.4)0.01

Parental occupations and DEHP levels*DEHP

Sampling site*With parental occupantNo parental occupantpn50th (25th-75th)n50th (25th-75th)Major bed14707.4 (611.9-1233.9)173805.1 (543.8-1131.0)0.64Major floor151379.8 (670.2-1811.2)1681232.3 (736.1-1857.2)0.90Minor bed153030.9 (1409.0-7054.5)1141450.8 (813.5-3019.9)0.02Minor floor151801.3 (1156.0-2582.5)1671795.1 (1183.0-2894.8)0.82Upper floor151058.9 (654.1-2632.1)169798.3 (486.2-1351.2)0.09Average level of five sites152107.6 (1643.4-2456.9)1781446.9 (1054.7-1985.3)0.03

*

Worldwide comparison of phthalate metabolites concentration*

MMPMEPMBPMBzPMEHPMEHHPMEOHP1932-125.615.558.65.2 8.848.945.23286-11-91.341.439.45.12--CDC, 20053936-111.4585.131.133.44.4123.333.6CDC, 20051632-28163-64.519.214.92.911.4Sathyanarayana et al., 20086Premature newborns----10029623419Calafat et al., 2004906.4-9.2

Correlation of concentrations between dust-borne phthalates in different place and their corresponding metabolites of childs urine *p

Correlation of concentrations between dust-borne phthalates and their corresponding metabolites of childs urine *p

Dietary pattern and phthalates metabolites*p

*

SubjectsThe present case-control study was carried out from 2006 to 2009.

71 central precocious puberty (CPP) girls were recruited from the policlinic of Pediatric Endocrinology in National Cheng Kung University Hospital and 29 normal similar age-range girls were recruited as control group from elementary school or kindergarten.

Table The levels of seven PAEs metabolite in case and control (g/g-creatinine)Wilcoxon rank sum test; Geometric mean (Range) The analytical results of urine samples

Case

(N=71)

Control

(N=29)

P-value

MMP

7.74 (0.83-64.3)

5.66 (0.94-31.3)

0.168

MEP

26.9 (1.21-379)

12.0 (1.06-337)

0.001

MBP

93.6 (22.3-451)

77.9 (20.5-275)

0.230

MBzP

7.72 (1.14-84.3)

4.44 (0.95-50.4)

0.007

MEHP

12.8 (4.03-69.3)

9.02 (3.07-29.6)

0.048

Case

(N=39)

Control

(N=9)

MEHHP

65.5 (24.5-291)

31.1 (12.3-202)

0.002

MEOHP

60.8 (21.9-244)

29.1 (13.8-177)

0.001

(g/L)

MMP

MEP

MBP

MBzP

MEHP

MEHHP

MEOHP

70

2.6-11.5

5.57

18.54

67.00

5.60

8.77

64.05

59.98

29

2.1-8.4

3.26

6.90

44.78

2.55

5.33

27.53

25.73

99

2.1-11.5

4.76

13.88

59.54

4.45

7.58

55.05

51.01

328

6-11

-

91.3

41.4

39.4

5.12

-

-

CDC, 2005

393

6-11

1.45

85.1

31.1

33.4

4.41

23.3

33.6

CDC, 2005

163

2-28

163

-

64.5

19.2

14.9

2.9

11.4

Sathyanarayana et al., 2008

6

Premature newborns

-

-

-

-

100

2962

3419

Calafat et al., 2004

90

6.4-9.2

(mg/kg)

DMP

DEP

DBP

BBP

DEHP

100

0.12

(0.12-12.1)

1.00 (0.08-124)

30.2 (6.13-1173)

3.81

(1.00-313)

1521 (374-7436)

101

0.12 (0.12-32.5)

1.00 (0.08-88.2)

34.2 (4.67-985)

3.50

(1.00-185)

1229 (294-23292)

100

0.12 (0.12-2.12)

1.00

(0.08-86.6)

16.2 (4.49-153)

2.50

(1.00-82.9)

658 (253-3417)

101

0.12 (0.12-12.6)

1.00 (0.08-45.4)

21.9 (3.39-834)

2.13

(1.00-14.7)

905 (162-9681)

46

0.12 (0.12-5.34)

1.00 (0.08-10.6)

18.3 (4.99-320)

2.59

(1.00-24.0)

1411 (303-23829)

38

-

10

100

110

640

ie et al., 1997

23

-

-

-

-

858

Clausen et al.,2003

30

1.5

29.7

47.0

29.7

(5-816)

703

(231-1763)

Fromme et al., 2004

120

-

4.98 (ND-111)

20.1 (ND-352)

45.5 (3.87-1310)

340 (16.7-7700)

Rudel et al., 2003

65

-

-

47

19

600

Kersten and Reich, 2003

346

-

0

(0-2425)

150

(0-5446)

135 (0-45549)

770

(0-40459)

Bornehag et. al., 2005

()

ResultsTable The kisspeptin-54 levels, basal hormones and physical characteristics from part of precocious puberty and normal girls in this study.

Normal girls

(N=11)

Precocious puberty girls (N=26)

P-value

LHRH test positive (N=11)

Treatment with Leuplin; Menarche (N=15)

Kisspeptin-54* (pmol/l)

1.95 (1.69-2.18)

2.14 (1.71-2.61)

2.29 (1.98-2.86)

0.02

Fig The association between Kisspeptin-54 and ln MBP-Cr (g/g-Cr).R2= 0.288;p= 0.001Results

Results (Intervention strategies):

PVC

()

creatinine()(ng/mL)

P&

Pre-post MEP

19.2

(2.04-622)

71.5

(10.3-282)

-

-

-

-

0.08#

Pre-post MBP

-

-

130

(16.6-846)

16.8

(1.70-150)

114

(16.5-846)

19.2

(1.7-163)

0.01*,1 0.018*,2

Pre-post MEHHP

-

-

-

-

73.6

(5.98-895)

20.3

(7.05-108)

0.042*

Pre-post MEOHP

-

-

-

-

27.1

(0.71-483)

8.21

(1.51-53.4)

0.079#

Pre-post MECCP

-

-

-

-

93.7

(21.6-722)

21.1

(5.93-27.8)

0.013*

&: Wilcoxon Rank Sum Test#: p

creatinine() (ug/g-creatinine)

P&

Pre-post MMP

-

-

3.44

(0.02-12.4)

1.81

(0.22-5.38)

-

-

0.079#

Pre-post MEP

-

-

-

-

38.5

(0.51-450)

5.71

(1.14-61.0)

0.072#

Pre-post MBP

53.5

(7.15-247)

16.6

(3.21-158)

-

-

-

-

0.066#

&: Wilcoxon Rank Sum Test

#: p

DEHPDBPBBzPDEHPMBPMEHHPMEOHPDBPDEHP/MEP MEHPMEHHPMEOHPDEPDEHPDEPDEHP()MBPkisspeptinDBPkisspeptin

?

AcknowledgementsWe are grateful for the financial support of Taiwan National Science Council and deeply appreciative of the subjects participated in this study.Livestock Research Institute, Council of AgricultureMain-Lian Lee, Ming-Chih TengDepartment of PediatricsYen-Yin Chou, Shio-Jean Lin Department of Obstetrics and GynecologyPao-Lin Kao, and other gynecologistsResearch Center of Environmental Trace Toxic SubstancesAll stuffs

***Phthalates are widely used as plasticizer to increase the polymers flexibility , elongation or ease of processing.The mostly commercial used phthalates including these five types.Due to their lipophilic property, they are easily to bioaccumulate in human being.They will become metabolites in our body including these types, and DEHP will have second metabolites.**This slide show the application of phthalates.They are widely used in childrens toys, personal care products, medical devices, building materials, and food packaging.Almost everything in our daily life. **This slide show the application of phthalates.They are widely used in childrens toys, personal care products, medical devices, building materials, and food packaging.Almost everything in our daily life. *General population may exposed to phthalates via food intake, inhalation, dermal contact with cosmetics and personal care products.

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