British J'ournal of Industrial Medicine 1993;50:623-632

Estimation of individual dermal and respiratoryuptake of polycyclic aromatic hydrocarbons in 12coke oven workers

Joost GM VanRooij, Monika M Bodelier-Bade, Frans J Jongeneelen

AbstractTwelve workers from a coke plant in TheNetherlands participated in an intensive skinmonitoring programme combined with per-sonal air sampling and biological monitoringduring five consecutive eight hour workshifts.The purpose of the study was to make a quan-titative assessment of both the dermal andrespiratory intake of polycyclic aromatichydrocarbons (PAHs). Pyrene was used as amarker compound for both dermal and respi-ratory exposure to PAHs. The biologicalmeasure for the internal exposure to PAHswas urinary 1-OH-pyrene concentration.Measurements on exposure pads at six skinsites showed that mean total skin contamina-tion of the 12 workers ranged between 21 and166 ug pyrene a day. The dermal uptake ofpyrene ranged between 4 and 34 4uglday, whichwas about 200/o of the pyrene contamination onskin. The mean concentration of total pyrenein the breathing zone air of the 12 coke ovenworkers ranged from 0.1 to 5-4 4ug/m3. Themean respiratory uptake of pyrene variedbetween 0*5 and 32*2 ug/day. Based on the esti-mates of the dermal and respiratory pyreneuptake it is concluded that an average 75%(range 2$%/o-95%, n = 12) of the total absorbedamount of pyrene enters the body through theskin. Because of the difference in thepyrene:benzo(a)pyrene ratio between the airsamples and the skin contamination samples,the dermal uptake of benzo(a)pyrene was alsoestimated. This was about 51% of the totalabsorbed amount (range 80%o-92%, n = 12).The total excreted amount of urinary 1-OH-pyrene as a result of exposure to PAHs duringthe five consecutive workshifts varied between

36 and 239 nmol. A multiple regression modelofthe mass balance between pyrene dose (bothdermal and respiratory) and 1-OH-pyreneexcretion confirmed the relevance of the der-mal exposure route. The variation in urinary1-OH-pyrene excretion was determined moreby the dermal pyrene dose than by the respi-ratory dose. The model showed an estimate ofthe percentage of the absorbed amount ofpyrene that is metabolised and excreted as 1-OH-pyrene in urine. For the 12 workers thispercentage varied between 13% and 490/odepending on smoking habits and consump-tion of alcohol. The results of this study indi-cate that among coke oven workers, the skin isthe main route of uptake of PAHs. Preventivemeasures to reduce exposure to PAHs shouldbe focused more on the reduction of dermalcontamination by PAHs than on the reductionofinhaled dose.

(British 'ournal ofIndustrial Medicine 1993;50:623-632)

Coke oven workers are highly exposed to polycyclicaromatic hydrocarbons (PAHs). In coke ovensbituminous coal is destructively distilled to producecoke. The emissions from a coke oven are complexmixtures of gases, liquids, and solid particles. Theairborne particulate matter on top of a coke batterycontains about 15% ofPAHs by weight.'

Epidemiological studies have shown that expo-sures in the coke production industry give rise tolung and skin cancer in humans. The possible agentis coal tar fume, which contains high concentra-tions of PAHs; these are carcinogenic in experi-mental animals.23To control occupational exposure to PAHs,

threshold limit values (TLVs) based on airborneconcentrations are set in many countries. Forexample, the TLV in the United States is based onthe benzene soluble fraction of sampled dust (TLV= 0-2 mg/m3).4 The German technical guiding con-centration (TRK) is based on the benzo(a)pyreneconcentration in air (TRK = 2 ,g/m3).5

Besides the respiratory tract, the skin might also

Department of Toxicology, Faculty of MedicalSciences, University of Nijmegen, Nijmegen, TheNetherlandsJ G M VanRooij, M M Bodelier-Bade, F J Jongeneelen

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Van Rooij, Bodelier-Bade, _Jongeneelen

Table 1 Personal data and characteristics of workers

Respiration volume (l/min)tBody

Personal Functionl Age Height weight Intensive Non-intensivecode work location (y) Alcohol* Smokingt (cm) (kg) activities activities

1 Push side 24 1 0 187 75 34-4 (2 8) 21-6 (5 2)2 Push side 46 1 1 178 86 25-3 (2 8) 18-5 (5 2)3 Supervisor 33 0 1 182 94 28-3 (4 2) 17-8 (3 9)4 Top side 28 1 0 176 78 21-6 (3 5) 19 9 (4-5)5 Supervisor 42 1 0 182 93 37-7 (3 5) 13-8 (4 5)6 Quencher car 59 0 1 180 100 15-5 (3 5) 15-0 (4 5)7 Coke screening 52 1 1 176 84 32-9 (4 9) 10-5 (3-1)8 Pushside 29 1 1 176 70 19-3 (2 8) 14-8 (5 2)9 Miscellaneous 28 0 1 170 80 15-7 (4 0) 13-5 (4 0)10 Miscellaneous 25 0 0 173 68 20-7 (3 5) 12-4 (4 5)11 Coke side 27 0 1 178 87 20-3 (3 5) 8-4 (4 5)12 Coke side 20 0 1 160 52 36-9 (3 5) 26-4 (4 5)

*Alcohol use during week of measurements; l=yes, O=no.tSmoking of tobacco; l=yes, O=no.tValues are converted to the atmospheric pressure and saturation at body temperature (BTPS). Durations of the work periods (h) withphysically intensive and non-intensive work activities are given in parentheses.

be an important route of body entry of PAHs.Application of tars containing PAHs on to the skinof mice induced local papillomas and skin carcino-mas, and also systemic DNA damage in tissuessuch as the lungs and heart.67 Application of coaltar ointment to the skin of healthy male volunteersresulted in a six to 10-fold increase in the urinary 1-OH-pyrene excretion rate (unpublished data). Thismetabolite of pyrene is increasingly being used andaccepted as a biological indicator of the internaldose of PAHs.8-"2

Occupational dermal contamination with PAHshas been measured with exposure pads (polypropy-lene filter in plaster material) as a pseudoskin moni-toring technique in several work environments.Among road paving workers skin contaminationwith pyrene, after a workshift of eight hours, rangedup to 1400 ng/cm2."3 In the primary aluminiumindustry contamination of skin with pyrene wasfound in the range of 6 to 192 ng/cm2, andbenzo(a)pyrene in the range of 2 to 34 ng/cm2.14 Inthese studies substantial contamination (up to 106ng/cm2) was also detected on exposure pads thatwere pasted underneath "protective" workingclothes. Determination of total PAHs in skin oilshowed a mean contamination of 97 ng/cm2 on theforeheads of roofing workers.'5Among primary aluminium workers we found

that the contamination of skin with PAHs showed abetter correlation with urinary 1-OH-pyrene excre-tion than PAH concentration in breathing zoneair.'4 A quantitative assessment of the dermal intakecould not be made, however.

In the present study among 12 coke oven work-ers, we conducted an intensive skin monitoringprogramme combined with personal air sampling tomake a quantitative assessment of both skin conta-mination and respiratory intake. A model to esti-mate the dermal dose of pyrene based on

measurements of contamination with PAHs isdescribed. A mass balance of input (estimateddermal and respiratory pyrene dose) and output (1-OH-pyrene in urine) is presented.

Study design and methodsThe study was carried out among 12 workers at acoke plant in The Netherlands in September 1990.The 12 workers belong to one crew and representoperating, maintainance, and supervisory person-nel. Table 1 gives the personal data for the workers.The coke plant, which comprises two batteries,

consists of 110 ovens in parallel rows and producesabout 600 000 tonnes of coke a year. Five crews of18 workers and a group of special maintainancepersonnel work in the vicinity of the battery. Thecoal is destructively distilled at a temperature ofaround 1270°C for 18 to 19 hours to produce coke.On average about 47 ovens are worked for eachshift of eight hours. About 95% of the workers wearoveralls during working hours; these are drycleaned after periods of three to five workshifts.Those who work in the vicinity of the ovens areprovided with an air stream helmet. About 60% ofthe workers use it more than four hours a workshift;20% do not use the air stream helmet.

Environmental monitoring to determine contam-ination of skin with PAHs and the concentration ofPAHs in breathing zone air was conducted duringfive consecutive eight hour workshifts (1400-2200).The biological monitoring programme to determinethe internal exposure of the workers to PAHs lastedseven days (the same five workshifts followed by atwo day period off work. Interviews were conduct-ed by a trained interviewer with a questionnaireabout personal characteristics including age,weight, height, smoking habits, and alcohol con-sumption.

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Figure 1 Skin sites where exposure pads are mountedfor themonitoring ofdermal contamination with PAHs.

MONITORING OF SKIN CONTAMINATIONSkin contamination of the workers was measuredwith an exposure pad on six skin sites: the jaw/neck,shoulder, upper arm, wrist, groin, and ankle (fig 1).The exposure pads were round monitoring devices(diameter 60 mm) with a small monitoring area(diameter 18 mm). Flexible polypropylene filtermaterial (Gelmann Sciences) was used as adsorbingmaterial in the exposure pad. The design of theexposure pad has been published elsewhere.'3 Atthe beginning of the shift (1400) pads were pastedon the skin and immediately after work (2200) theywere removed, packed in aluminium foil and storedat -20°C until analysis. The effective monitoringarea of the exposure pad (1 77 cm2) was punchedout and ultrasonically extracted for 10 minutes withdichloromethane (10 ml). After centrifugation(2000 rpm for five minutes), the extract wasanalysed for pyrene and benzo(a)pyrene by highperformance liquid chromatography (columnVydac C18, temperature 40°G) with multiple shiftfluorescence detection (LS-4, Perkin Elmer). Therecoveries of pyrene and benzo(a)pyrene fromexposure pads was 80% and 100% respectively.

MONITORING OF BREATHING ZONE AIRDuring working hours, airborne particulates in thebreathing zone air were collected on a Teflon filter(PTFE filter: diameter 25 mm, pore size 0*5 ,im,Millipore) with samplers that were equipped with acone over the face. In conjunction with a suctionflow of 2 /min (Dupont air samplers, modelsP4000 and S2500), a suction velocity of 1-25 m/swas produced in the face entrance. Three of the 12workers (workers 4, 8, and 12) wore an adsorbent

tube directly behind the sampler to trap the gaseousphase on days 2, 3, and 5. The adsorbent tube con-tains XAD-2 resin as adsorbent (ORBO 43,Supelco, USA).

After sampling, the PTFE filters and the adsor-bent tubes were stored in the dark at - 20°C untilanalysis. The samples were analysed for pyrene andbenzo(a)pyrene within three weeks after sampling.The filters and XAD-2 adsorbents (from both thefront and back section of the adsorbent tubes)were extracted and analysed in a similar way to theexposure pads. The recovery of pyrene andbenzo(a)pyrene on the PTFE filters was 70% and100% respectively. No benzo(a)pyrene was de-tected in extracts of the XAD-2 adsorbents; therecovery of pyrene was 70%.To determine the average daily respiratory vol-

ume of the workers, the workshift was divided intotime with intensive and time with non-intensivephysical activity (see table 1). In both periods therespiratory volume of the workers and the tempera-ture of exhaled air was measured with a portablegas meter (Zentralwerkstatt Gottingen GmbH,Gottingen, Germany) for 10 minutes by themethod of Kofranyi and Michaelis.'6 To be able tocompare the results of the measurements the vol-umes were converted to the atmospheric pressure atthe moment of measurement and to saturation atbody temperature (BTPS).'7

BIOLOGICAL MONITORINGDuring the five workdays, spot urine samples werecollected before (1400) and immediately after theworkshift (2200). In the two day period off workafter the work week urine was collected at the sametimes. The urine samples were stored at - 20°Cuntil analysis. Metabolites in urine were deconju-gated by enzymatic hydrolysis and a solid phasesample clean up was applied. Analysis by HPLCallows the determination of the sum of free andconjugated 1-OH-pyrene in urine at the nmol/llevel. The reproducibility of the analysis of urinary1-OH-pyrene was determined by a repeated analy-sis of 30 randomly selected urine samples and thisgave a coefficient of variation of 12%. A fulldescription of the method is presented elsewhere.'8Urinary 1-OH-pyrene concentrations were cor-rected for creatinine. Correction for very low orhigh creatinine values is not reliable. Therefore,urine samples with a creatinine concentration out-side the range of 4-34 mmol/l were excluded fromthe analyses.

CALCULATIONS OF THE DERMAL DOSEThe absorbed amount of PAHs at a skin regionafter a workshift of eight hours, is assumed to beequal to the total contamination of the skin regionas measured with exposure pads minus the remain-

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der after eight hours:(1) Xabsorbed,j = Xcontainination, - Xrernainderjwhere Xabsorbedj = amount of pyrene absorbed at skinregion; (ng/cm2); Xcontarninationj = pyrene contamina-tion on skin regionj (ng/cm'); Xremainder,j = amount ofpyrene remaining on skin regionj (ng/cm2).The remaining PAHs on the skin after a work-

shift is assumed to be determined by two rateprocesses-namely, the contamination rate and thedermal absorption rate. The differential equationdescribing the remainder of PAHs on a skin region,based on a zero order contamination and a firstorder absorption process, is:

(2) d t,,ain =mi _k- ,* Xremainderj

where mj = pyrene contamination rate at a skinregionj (ng/cm2/h); kj = pyrene absorption rateconstant at a skin regionj (1/h); t = time after start ofexposure (h)The pyrene contamination rate is calculated by

dividing the pyrene contamination of the exposurepad (ng/cm2) by the hours of exposure. The conta-mination of the exposure pad at a skin site, as mea-sured in the skin monitoring programme, is takento represent the dermal contamination of a wholeskin region during the workshift of eight hours.Each skin region represents a percentage of thetotal body area (see table 2).'9 The total body sur-face of a worker is based on weight and height, andcalculated with the formula proposed by Du Boisand Du Bois:20(3) total body area = 71 84 * weight0425 * height0725where total body area is expressed in cm2, weight inkg, and length in cm.

Table 2 presents the mean absorption rate con-stant of four or more fused ring PAHs at variousskin sites as determined after application of coal tar

Table 2 Area ofskin regions and PAH absorption rateconstants used in dermnal dose calculations

Skin site % Of total PAH absorptionof exposure pad Skin region body area* rate constanttJaw/neck Head 6-8 0-065 (0 008)

NeckShoulder Shoulder 22-8 0-135 (0-027)

BackChest

Upper arm Upper arms 9-7 0 070 (0-007)Wrist Forearms 6-7 0 070 (0 007)

Hands 5-6 0 037 (0 005)Groin Hips 27-1 0-053 (0-010)

ThighsAnkle Calves 19-9 0-036 (0 003)

Feet

*Adapted from Popendorf and Lettingwell'9tMean dermal absorption rate constant of four or more fused ringPAHs (1/h) (SEM) as determined in a previous study (unpub-lished data)

ointment to the skin of healthy volunteers (unpub-lished data). A combination of formula (1) and theintegrated form of equation (2) leads to an equa-tion that enables calculation of the absorbedamount of pyrene at a skin region during a work-

(4) Xabsorbed j, T= (mj * T)- m * (1-e -kaj * T))ka,j

shift:where T = duration of workshift (h)The total amount of pyrene absorbed through

n

(5) Dermal dose = E (Xabsorbed j * area)the skin during the workshift is calculated as:where dermal dose = total absorbed amount ofpyrene during the workshift (ng); area, = area ofskin regionj (cm2); n = number of skin regions.

STATISTICAL ANALYSISExposure variables are presented as the geometricmean (GM) because this is considered more repre-sentative of a skewed distribution of data than thearithmetic mean (AM). Data below the detectionlimit were processed as having the numerical valueof half of the detection limit. Correlations wereassessed with Spearman's rank correlation analysis(rs). Multiple regression analysis was carried out tostudy the relation between both dermal and inhala-tory exposure and internal dose. The residuals werechecked for the normal distribution assumption.None of the analyses showed major deviation fromthis assumption. Statistical analyses were per-formed with the SAS computer software package,version 6.0.

ResultsMONITORING OF SKIN CONTAMINATIONThe exposure pads mounted on the jaw/neck andthe wrist showed the highest pyrene contaminationcompared with the other skin sites (shoulder, groin,ankle, and upper arm), on which the pyrene conta-mination was an average threefold lower (see table3). The pyrene contamination on exposure padsmounted underneath protective clothing reached23 ng/cm2. The pyrene contamination on the skinof coke oven workers showed a high dispersal.

Figure 2B shows the interday variation of thepyrene contamination for the exposure pads on thejaw/neck of worker 4 (top side of oven) and worker11 (coke side of oven) during the five workshifts.Based on the measurements on the exposure

pads at six skin sites, total pyrene skin contamina-tion was calculated (see table 4). The mean totalskin contamination of the 12 workers after an expo-sure of eight hours ranged between 21 and 166 ,ug

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Table 3 Pyrene contamination (nglcm2) on exposure pads pasted on to six skin sites of coke oven workers

Skin sitePersonal Functionlcode work location Jawlneck Shoulder Upper arm Wrist Groin Ankle

1 Push side 6-7 (4-9-8-9) 2-2 (0 7-42) 3 0 (20-3 7) 11 1 (58-19-4) 0-5 (0-1-0-7) 1-4 (1-3-1-4)2 Push side 7-2 (2-1-19-0) 2-4 (1-3-34) 1-3 (0-9-2-1) 13-3 (20-29 7) 0-8 (0-7-1-7) 2-3 (0-3-7-3)3 Supervisor 8-6 (5-9-10-6) 2-5 (14-449) 3-3 (1-85-8) 6-1 (20-16-4) 4-7 (2 5-7 3) 1-4 (1-4-1-4)4 Top side 16-5 (120-28-7) 3-0 (2-0-47) 1-9 (0-44-1) 5-2 (20-7-9) 5-7 (2-3-10-7) 3-1 (1-47-6)5 Supervisor 4 2 (2-1-8-9) 3-9 (2-1-18-6) 0 9 (0-31-7) 6-0 (40-11 6) 2-9 (0-7-7 3) 1-4 (1-3-1-4)6 Quencher car 1-5 (0-14-8) 1-1 (0-1-2-8) 0 4 (0 1-1 1) 1-5 (0 7-2 0) 0-7 (0-7-07) 1-7 (1-42-4)7 Coke screening 2 1 (2-1-2-1) 0-8 (0-1-2-8) 0 7 (0-1-2-5) 2-0 (20-2 0) 0-8 (0-7-1-3) 1-4 (1-41-4)8 Pushside 5-5 (3-5-7-2) 1-4 (0-14-8) 3-9 (1-7-6-6) 11-0 (83-16-1) 4-3 (2-3-7-3) 5-6 (44-8-1)9 Miscellaneous 4-9 (0-1-50-9) 1-1 (0-1-3-1) 3 0 (13-9-0) 3-7 (1-7-7-1) 2-7 (1-14-8) 1-6 (1-3-2-5)10 Miscellaneous 10-6 (2-1-29-0) 1 1 (0-1-3-4) 1-8 (0 9-44) 9-4 (20-31-2) 1-4 (0-7-3-8) 1 4 (1-41-4)11 Coke side 15-9 (12 7-18-8) 1-6 (0-1-7-8) 1-4 (0-14-8) 18-8 (15 8-23-5) 15 3 (8-2-23 3) 1-7 (1-3-3-4)12 Coke side 14-0 (8-1-21-2) 3-5 (3-04-1) 4-2 (2-5-8-1) 9-3 (5-7-13-1) 0-9 (0-7-1-6) 2-9 (1-47-5)Overall mean (n = 60) 6-5 (0-1-50-9) 1-9 (0-1-18-6) 1-8 (0-1-9-0) 6-4 (0-7-31-2) 2-1 (0-1-23-3) 2-0 (0-3-8-1)

Results are geometric mean (range); n = 5.

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pyrene (GM, n = 5). The highest pyrene skin con-tamination was found among workers at the cokeside and top side of the ovens; this reached 166 ,ugand 101 ,ug respectively (GM, n = 5).

Application of formula (5) resulted in an esti-mate of the daily dermal dose of pyrene after anexposure of eight hours that ranged between 4 and34 ,ug. The highest mean dermal pyrene doses werefor a worker at the coke side and the worker at thetop side of the oven. These were 34 ug and 22 jugrespectively (GM, n = 5). Thus about 22% of thepyrene contamination on the skin of a workerenters the body (range 20-3%-25-2%, n = 12).Table 4 presents the dermal weekly dose of pyrenefor each worker, which is the sum of five daily skindoses. This varied between 119 and 893 nmolpyrene.

MONITORING OF BREATHING ZONE AIRPersonal air measurements of both gaseous andparticulate pyrene showed an amount of pyrene inthe gas phase equivalent to 29% of the amount ofparticle bound pyrene (GM, range 13%-46%, n =9). The concentration of total pyrene, both particu-late and gaseous, in the breathing zone air of theworkers, of which only particulate pyrene was sam-pled, was calculated assuming that the amount ofpyrene in the gas phase was 29% of the sampledparticulate pyrene.The weekly average concentration of total pyrene

in the breathing zone air of the 12 coke oven work-ers ranged from 0 1 to 5-4 jug/m3 (GM, n = 5).The worker at the top side of the oven had a pyreneconcentration in breathing zone air of 3 0 jug/M3(GM, n = 5). The two coke side workers hadmean pyrene concentrations in breathing zone airof 3-3 and 5-4 jug/m3 (GM, n = 5). Figure 2Cshows the total pyrene concentration in breathingzone air of worker 4 and worker 11 during fiveworkshifts. The concentration for worker 4 variedbetween 1-5 and 7-9 jug/m3, whereas worker 11 had

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Van Rooij, Bodelier-Bade, Jongeneelen

Table 4 Estimates of daily pyrene exposure, pyrene uptake during five days (input), and total urinary 1-OH-pyrene excretionduring seven days (output) for 12 coke oven workers

Daily exposure Input (five days) Output (seven days)

Pyrene; Pyrene; Pyrene; Pyrene; I-OH-Pyrene;skin breathing zone air dermal respiratory excretion in

Personal Function! contamination concentration dose dose urinecode work location (4ug) (pg/rM3) (nmol) (nmol) (nmol)

1 Push side 58-8 0-46 339 66 582 Push side 84-9 1-01 473 102 983 Supervisor 83-4 1-15 474 143 1634 Top side 101-0 3-01 551 362 965 Supervisor 76-1 0-21 492 27 766 Quencher car 27-5 0-29 162 22 587 Coke screening 21-2 0 09 119 12 438 Push side 93-1 1-38 486 125 1029 Miscellaneous 58-3 1-27 342 173 13310 Miscellaneous 60-2 0-82 323 101 3611 Coke side 165-9 3-33 893 238 23512 Coke side 62-6 5-37 354 918 239

a rather constant pyrene concentration in hisbreathing zone air of 33 ,ug/m3 (GM, n = 5).The daily respiratory dose was calculated as:(6) Respiratory pyrene dose = ((pyrpa,, * RF,uwpan)

+ (pyrg., * RFiungsgas) + (PYrpart * RFGI-,.Ct)) * RVwhere pyrp,,, = particulate pyrene concentration

in breathing zone air (Ug/M3); RFl,,, = resorptionfactor of inhaled particulate pyrene that is deposit-ed in the lungs = deposited fraction * particle elu-tion = 0-125 * 1 = 0-125; pyr., = gaseous pyreneconcentration in breathing zone air (ug/m3);RFiunggaS = resorption factor of inhaled gaseouspyrene in the lungs = 0 7; RFGtC,t = resorption fac-tor of inhaled particulate pyrene that is swallowed(gastrointestinal tract) = swallowed fraction *absorption efficiency = 0-625 * 0 40 = 0-25; RV =total respiration volume during an eight hour work-shift (m3).

For the estimation of exposure to particle boundchemicals it is generally assumed that about 25% ofinspired particulates is exhaled, 12-5% is depositedin the lower respiratory tract, and 62.5% is elimi-nated from the lungs and swallowed.2' It is assumedthat the elution of pyrene from the deposited parti-cles in the lower respiratory tract is 100%, and thatof the swallowed fraction of the particle boundpyrene 40% is absorbed in the gastrointestinaltract.2223 A resorption factor of 70% is assumed forinhaled gaseous pyrene.The respiratory volume during the hours of work

is based on the measured respiratory volume of theworkers during physically intensive and non-inten-sive working hours, which varied between 8-4 and37-7 /min (see table 1), calculated with the formu-la:

(7) RV = (WP rv3) + (WPr-i* (rv. )(7)RV= ~1000where WP = workshift with physically intensive(i) or non-intensive (n-i) activities (hours); rv =respiration volume during physically intensive (i) or

non-intensive (n-i) activities (1/hour)The average respiratory volume of the 12 moni-

tored workers during a workshift of eight hoursranged between 6-9 and 15-2 m3 (AM, 10-1 (SD2-7)m3/8h;n =12).The estimated daily respiratory pyrene dose of

the 12 workers varied between 0-5 and 33 3,ug(GM, n = 5). The highest mean respiratory dosesduring a workshift of eight hours were calculatedfor a worker at the coke side of the oven (333 ,ugpyrene (GM, n = 5), and the worker at the top sideof the oven (12 ,ug pyrene (GM, n = 5)).

Table 4 presents the results of the calculations ofthe respiratory weekly dose. For the 12 workers, therespiratory weekly dose of pyrene, which is the sumof the five workshifts, varied between 12 and 918nmol. The top side worker (worker 4) and a cokeside worker (worker 12) had the highest respiratoryweekly doses of 362 and 918 nmol pyrene respec-tively.

BIOLOGICAL MONITORINGFigure 2A shows the rate of 1-OH-pyrene excretionin urine of worker 4 and worker 11 during a periodof seven days. The urinary l-OH-pyrene excretionrate was calculated as:

(8) uOHPrt, = uOHPc0., * ccmat * bwwhere uOHP,at = urinary 1-OH-pyrene excre-

tion rate (nmol/hour); uOHP,cOc = urinary 1-OH-pyrene concentration (nmollmmol creatinine); CG,t= excretion rate constant of creatinine(mmol/hour/kg); bw = body weight (kg).

Creatinine excretion depends on body weightand shows a rather constant excretion rate of25 mg/24 h/kg body weight,24 which is equivalent to0.009 mmol/h/kg body weight. To estimate thetotal amount of 1-OH-pyrene excreted as a result ofa five day period of exposure to PAHs, we deter-mined the area under the curve after correction forbackground 1-OH-pyrene excretion. Table 4 pre-sents the total excreted amount of 1-OH-pyrene for

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Table 5 Multiple regression analysis of the mass balance: pyrene input uinary l-OH-pyrene output*Independent Parameter Standard Partialvariablest estimate error rt F p Value

Dermal pyrene weekly dose (nmol) 0-14 0-019 0-82 53-2 0-0001Respiratory pyrene weekly dose (nmol) 0-13 0-025 0-11 27-3 0-0008Smoking (yes = 1, no = 0) 67-2 12-3 004 30-1 0-0006smoking/alcohol use (both = 1, -45-2 14-6 0-02 9-6 0-015

not both = 0)

Dependent variable: Excreted urinary l-OH-pyrene during seven days, corrected for background excretion (nmol).*n = 12tUse of alcohol and quetelet index were excluded from the model (p > 0-05).

12 coke oven workers over seven days as a result ofexposure to PAHs during five consecutive work-shifts. It ranged from 36 to 239 nmol. The twoworkers at the coke side of the ovens showedthe highest I-OH-pyrene excretion (235 and239 nmol).

A MASS BALANCETo determine the relation between the pyrene dose(both dermal and respiratory) and urinary 1-OH-pyrene excretion, we conducted a multiple regres-

sion analysis on the data (table 5). The regressionanalysis was performed with weekly doses ofpyrene, which is the pyrene exposure during fiveconsecutive workshifts, and the I-OH-pyreneexcretion over seven days to reduce the influence ofinterindividual differences in elimination half life ofpyrene on the relation. Besides the dermal and res-

piratory weekly dose, smoking habits, alcohol con-

sumption during the week of measurements andthe quetelet index (as a measure for fat content)were entered in the regression model. The smokingof cigarettes, drinking of alcohol, and fat contentmay influence urinary 1-OH-pyrene excretion.The multiple regression analysis showed that the

dermal pyrene dose, the respiratory pyrene dose,smoking, and the interaction between smoking andthe drinking of alcohol significantly determine uri-nary 1-OH-pyrene excretion (see table 5). Theeffects of alcohol and fat content were not signifi-cant at p < 0-05 (p = 0-42 and p = 0 09 respec-

tively) and consequently were removed from themodel.

Ninety nine per cent of the variation in urinary1-OH-pyrene excretion was explained by theregression model rather than left to residual error.The estimated parameters of the dermal and respi-ratory pyrene weekly dose were 0-14 (SD 0-019)and 0-13 (SD 0 025) respectively. Based on theparameter estimates it was concluded that for non-smokers about 13% of the total absorbed pyrene

was metabolised and excreted as 1-OH-pyrene inurine. For workers who smoked but did not drinkalcohol, this percentage was on average twofoldhigher (AM = 27%, range 18%-49%, n = 5).For workers who both smoked and drank, it was

estimated that about 21% (AM, range 17%-30%, n

= 3) of the total absorbed pyrene was excreted as1-OH-pyrene in urine.

DiscussionData on PAH contamination of the skin of exposedworkers are limited. It is striking to see the manystudies that have been performed, with PAHsapplied to skin of experimental animals and tohuman skin (in vitro) and the little effort that hasbeen made to assess the skin contamination and todetermine its relevance among exposed workers.Wolff and co-workers studied PAHs in skin oil ofroofing workers."5 We have measured contamina-tion with PAHs among road paving workers andprimary aluminium workers."3'4 The relevance ofthese measurements is shown in the present study,in which monitoring of skin contamination is com-bined with biological monitoring.

Based on the estimates of the individual dermaland respiratory dose of pyrene (table 4) it is con-cluded that 28%-95% (GM 75%, n = 12) of thetotal absorbed amount of pyrene enters the bodythrough the skin. Three major assumptions weremade to obtain these estimates and need somecomment. Firstly, an important aspect of the der-mal dose estimations is whether the amount on theexposure pad indeed reflects the contamination ofthe natural skin. It is possible that the adsorptioncharacteristics of the polypropylene filter in theexposure pad differ from the adsorption character-istics of natural skin. Among workers exposed tocreosote, the exposure pad measurements tend tounderestimate the pyrene contamination on naturalskin (unpublished observations). Secondly, anotherassumption that might underestimate the actualdermal dose is that the absorption of pyrene stopsafter the workers have left their work place. All thecontamination may not, however, be removed by ashower after work. This underestimation is prob-ably limited, however, because in a previous studywe found that the decontamination efficiency of awashing procedure to remove coal tar ointmentfrom skin, with warm tap water and a cleaner fortar and oil-like contamination, was more than 99%(unpublished data). A third source of inaccuracymight be the absorption rate constants used in thecalculations. These are based on the disappearance

629

Van Rooij, Bodelier-Bade, Jongeneelen

Function/work location

Supervisor (n = 5)

Push side (n = 10)

Coke side (n = 8)

Top side (n = 7)

Miscellaneous(n = 13)

Coke screening (n = 7)

Quencher car (n = 5)

Controls (n = 14)

1-OH-Pyrene conc (t,mol/mol creatinine)

Figure 3 Concentrations of l-OH-pyrene in urine of all workers of the coke plant sampled before and afterfive consecutiveworkshifts (geometric mean).

of PAHs from the surface after coal tar ointmentapplication on various regions of the skin ofvolunteers (n = 4); unpublished observations).Although the interindividual differences in theabsorption rate constants were minor (7%) in com-parison with skin site differences (69%), the stan-dard error of the mean PAHs absorption rateconstant at a skin site was rather high (see table 2).The presented dermal pyrene dose estimates basedon variation in absorption rate constants betweenskin regions did not differ significantly from the cal-culated dose based on one overall average absorp-tion rate constant of 0.066/h (results not shown).Guy and Maibach calculated that the dermal dosesof hydrocortisone, malathion, and parathion wereoverestimated two to three times when regional dif-ferences in penetration rate were neglected andonly the penetration rate of the forearm was used inthe calculations.25 Finally, the estimates of the res-piratory dose show that the variation in respirationvolume between the workers may have a greatimpact on the dose. When the estimated respiratorypyrene dose of workers 11 and 12 were compared,the dose of worker 12 was shown to be fourfoldhigher although the mean pyrene breathing zone airconcentration was only a factor of 1 6 higher. Theeight hour respiration volume for worker 12 was152 m3 and for worker 11 only 69 M3n, despitecomparable work activities.

The simultaneous measurement of dermal expo-sure to pyrene, inhalatory pyrene exposure, andtotal body burden (urinary 1-OH-pyrene) enables a

mass balance assessment to control the quality ofboth the dermal and respiratory dose estimates.The presented regression model of the mass bal-ance between both dermal and respiratory pyrenedose and urinary I-OH-pyrene, fits well (the coeffi-cient of determination (r2) = 0 99). The modelalso shows comparable estimates of the fraction ofboth the dermal and respiratory pyrene dose that isexcreted as 1-OH-pyrene in urine (0 14 and 0*13respectively; see table 5). This supports the doseestimates, because it might be expected that thefraction of pyrene, which is metabolised into 1-OH-pyrene and excreted in urine after it has entered thesystemic circulation, is independent of the route ofentry. It also indicates that the dermal pyrene dose,which is on average three times higher than the res-

piratory pyrene dose, determines most of the inter-nal dose. The smoking of cigarettes was found toincrease the urinary 1-OH-pyrene excretion afteroccupational exposure to PAHs. This might becaused by an underestimation of the respiratorydose or by an increased metabolic conversion ofpyrene. The socalled hand-mouth shunt or a

decreased clearance of particle bound pyrene fromthe ciliated surface of the respiratory tract may leadto an underestimation of the respiratory dose of

Urine sample

Before work week

After work week

0 2 3 4 5 6

.IRNMW.

630

1I

Uptake ofpolycyclic aromatic hydrocarbons

workers who smoke cigarettes. The significantinteraction between smoking and the use of alco-hol, however, suggests that the effect of smoking onurinary 1-OH-pyrene excretion is more likely to bea toxicokinetic effect.To determine whether the 12 intensively moni-

tored workers had exposure levels representative ofthe other workers of the plant, we collected spoturine samples of 56 other workers who also workedin the vicinity of the coke ovens. Spot urine samplesof these workers were collected just before and atthe end of five consecutive eight hour workshifts.Control urine samples were available from adminis-trative personnel of the plant (n = 14). Figure 3shows the urinary 1-OH-pyrene concentrations perfunction or work location of the coke oven workersand controls (in p mol/mol creatinine). The 1-OH-pyrene concentrations in urine of the 12 selectedworkers were within the range as measured amongthe other PAH exposed workers of the plant. Theresults show that workers in the vicinity of theovens, especially at the top side, were mostlyexposed to PAHs. About 46% of the workers whowork in the vicinity of the ovens of the coke planthad a urinary 1-OH-pyrene concentration thatexceeded the recently proposed biological exposurelimit of 2-3 pmol/mol creatinine.26 The urinary 1-OH-pyrene concentrations measured among cokeoven workers of this plant ranged between 0-2 and9-3 pmol/mol creatinine. The concentrations mea-sured in another coke plant were of the same range:03-11-33umol/mol creatinine.27Most of the data on exposure of workers to

PAHs at other coke plants are confined to air con-centrations of PAHs, generally benzo(a)pyrene.The mean concentration of benzo(a)pyrene inbreathing zone air of the 12 coke oven workers var-ied between 0.01 and 13-9 ,ug/m3 (data not shown),which is within the range measured in otherstudies.' 27-31The particulate pyrene concentrations in breath-

ing zone air measured in this study are comparablewith those measured in a previous study amongcoke oven workers of another coke plant in TheNetherlands.27 Data on gaseous pyrene in breathingzone air samples of coke oven workers are scarce.In the present study, the percentage of gaseouspyrene in total pyrene varied between 11% and30% (GM 21%, n = 9). Bjorseth and coworkersreported a mean 5% in stationary air samples at thebattery top.' Regression analysis of thebenzo(a)pyrene concentration in air (GM of mea-surements during five workshifts in pg/m3n) and theend of week urinary 1-OH-pyrene concentration(umol/mol creatinine) of the 12 workers and con-trols gave the relation: (end-of-week 1-OH-pyrene)= 0.54 + 1-34 * (benzo(a)pyrene in air). With thisformula the urinary 1-OH-pyrene concentration

was estimated for workers, when exposed to theGerman technical guiding concentration of 2,pgbenzo(a)pyrene/m'. This gave a biological exposurelimit of 3-2 pumol 1-OH-pyrene/mol creatinine(95% confidence interval 2-4-4 1), which is similarto the value of 2-3 pmol/mol as proposed byJongeneelen.'6

Data on dermal contamination with PAHs forcoke oven workers of other coke plants are notavailable. Among the coke oven workers the meandaily pyrene contamination on skin ranged between21 and 166,pg/day. This is lower than the meanpyrene skin contamination as estimated for primaryaluminium workers (about 400 pg/day)'4. Thepyrene contamination on exposure pads worn bythe coke workers is comparable with those worn byroad paving workers, except for the pyrene contam-ination on the wrist pad, which is on average threetimes higher among road paving workers"3. Table 3shows that the pyrene contamination on exposurepads worn by coke oven workers who have a com-parable work location and work activities, varies inextent and dispersal (compare for example, workers1 1 and 12). Apparently, both the extent and disper-sal of PAH contamination on skin of workersexposed to PAHs are not only determined by workenvironment, but also by function and personal fac-tors. Personal factors that might influence skin con-tamination by PAHs are personal hygiene,individual working methods, the use of protectiveclothing, and the frequency of laundering.The suitability of pyrene as a marker compound

for PAHs in air samples is shown in a previousstudy. Particulate pyrene correlated well with thesum of 11 PAHs (r = 0-88, n = 47)27. In our pre-sent study, a significant correlation between pyreneand benzo(a)pyrene in both breathing zone air sam-ples and skin exposure samples was found. TheSpeannan correlation coefficients were 0-94 (n =58) and 0-73 (n = 378) respectively; however, theratio pyrene: benzo(a)pyrene (wt/wt) was 2 1 (GM,range 0-6-16-2) in breathing zone air samples and6-3 (GM, range 0-15-165) in skin contaminationsamples. Because of the difference in the ratiopyrene: benzo(a)pyrene between breathing zone airsamples and skin contamination samples, we alsoestimated the dermal and respiratory uptake ofbenzo(a)pyrene by applying the presented formu-lae. This showed that the percentage of the totalabsorbed amount of benzo(a)pyrene that enters thebody through skin ranged from 8%-92% (GM =51%, n = 12).The correlation between the PAH concentration

in breathing zone air and the estimated PAHuptake by skin was rather low. For pyrene theSpearman correlation coefficient was 0 59 (p =0-045, n = 12) and for benzo(a)pyrene 0 55 (p =0-067, n = 12). Because of the high uptake of

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Van Rooij, Bodelier-Bade, Jongeneelen

PAHs through the skin we suggest that exposure isassessed by a biological monitoring method, whichgives a more accurate assessment of the internalPAH exposure of a worker than breathing zone airmeasurements. This study confirms that the mea-surement of 1-OH-pyrene in urine as an indicatorof internal PAH exposure, is a powerful biologicalmonitoring method.

In summary, we have found that among cokeoven workers, the uptake of PAHs through skin isvery relevant in terms of the internal dose. Forpyrene an average 75% (range 28%-95%) of thetotal dose was absorbed through skin. Forbenzo(a)pyrene it was 51% (range 8%-92%). Ourresults indicate that preventive measures to reduceexposure to PAHs should be focused more on thereduction of dermal contamination with PAHs thanon the reduction of the inhaled dose.

We gratefully thank P Jetten for his contribution tothis study. We also thank the coke oven workersand the medical officer of the coke plant, P J MHopmans, MD (Occupational Health Service"Zeeland", Vlissingen, The Netherlands), for theirsupport and contribution. This study is financiallysupported by the European Community on Coaland Steel contract 7280/01/002.

Requests for reprints to: Dr J G M VanRooij,Department of Toxicology, Faculty of MedicalSciences, University of Nijmegen, PO Box 9101,6500 HB Nijmegen, The Netherlands.

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Accepted 5 October 1992.

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