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PAHs in different indoor dusts and their potential cytotoxicity based on two human cell lines

Polycyclic aromatic hydrocarbons (PAHs)in different indoor dusts and their potential cytotoxicity based on two human cell lines

Yuan Kang,Kwai Chung Cheung ?,Ming H.Wong ?

Croucher Institute for Environmental Sciences,Department of Biology,Hong Kong Baptist University,Kowloon Tong,Hong Kong,PR China

a b s t r a c t

a r t i c l e i n f o Article history:

Received 1February 2010Accepted 9April 2010

Available online 7May 2010Keywords:

Air-conditioner ?lter dust MTT assay HepG2cells KERTr cells Hong Kong

In this study,55air-conditioner ?lter dust samples from six different workplaces including commercial

of ?ce,secondary school,shopping mall,hospital,electronic factory and manufacturing plant in Hong Kong were collected for analyses of PAH concentration and cytotoxicity.Chemical analyses showed that the total PAHs ranged from 1.17to 25.5μg/g,with the dust samples from manufacturing plant having the highest concentration.MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrasodium bromide)assay was performed to evaluate the cytotoxicity of organic dust extracts using human hepatocellular liver carcinoma cell line (HepG2)and human skin keratinocyte cell line (KERTr).Each organic dust extract showed marked dose-related response.Dust samples from the manufacturing plant showed the highest cytotoxicity.Curve estimation indicated that power model was ?t for explaining the relationship between the total PAH concentration and lethal concentration 50(LC 50).In addition,a signi ?cant negative correlation was observed between the total PAH concentration and LC 50both on HepG2(r =?0.65,p b 0.01)and KERTr (r =?0.63,p b 0.01)cell lines.Source analyses demonstrated that the PAHs in dust were derived from pyrogenic origins.

?2010Elsevier Ltd.All rights reserved.

1.Introduction

Pollutants in the indoor environment include microbiological organisms (e.g.,bacteria,fungi,and virus),radiation (e.g.,radon gas),chemicals and combustion products (e.g.,pesticides,metals,?ame retardants,polycyclic aromatic hydrocarbons (PAHs))(Cooke,1991).These pollutants adsorb to particulate matter suspended in indoor air which later settles out as indoor dust.

As Hong Kong lies within the subtropical region,most of the indoor environments are air-conditioned in summer or even throughout most of the entire winter.In fact,the dust retained on the air-conditioner ?lter re ?ects the indoor dust and indoor air quality because the air conditioner recirculates the air through the ?lter (Saeed et al.,1998).

PAHs in indoor dust are derived from many origins,including domestic burning,power generation,cooking,tobacco smoking,and parquet ?oor glue (Lohmann et al.,2000;He et al.,2005;Riechelmann et al.,2007).Due to the fact that the indoor environment is protected from environmental degradation,PAHs associated with dust always persist for long periods.Therefore,PAH concentrations in indoor dust are substantially higher than outdoor dust and soil (Chuang et al.,1995).

Due to various toxic persistent organic pollutants contained in dust,organic extracts have been found to exert cytotoxicity (Reyes et al.,2000)and genotoxicity (Sevastyanova et al.,2007)on different cell lines including human skin keratinocytes (KERTr),HepG2cell line and

human diploid lung ?broblast (HEL)cell line.To evaluate cytotoxicity on cell lines,there are many established end point assays such as neutral red assay (Reyes et al.,2000),MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrasodium bromide)assay (Gerlier and Thomasset,1986;He et al.,2008),crystal violet assay (Bellocci et al.,2008)and LDH (lactate dehydrogenase)assay (Burke and Mayer,1974;He et al.,2008).

Compared to other cytotoxicity assays,MTT assay is the most commonly used for the detection of alterations in cellular metabolism,proliferation,or activation (Kim et al.,2008;Amaral et al.,2009).It evaluates the mitochondrial dehydrogenase activity and depends on the degree of cell activation (Mosmann,1983;Kan et al.,1997).

On the other hand,different cell lines show different cytotoxicity responses to the same contaminant (Sevastyanova et al.,2007).The HepG2cell line and KERTr are appropriate candidates to better understand human exposure via dust,because the HepG2cell model could evaluate the potential adverse effects on human health via dust ingestion (Sevastyanova et al.,2007)and the KERTr cell model via dermal contact with dust (Arlian et al.,2008).The human hepatoma cell line HepG2possesses a number of characteristic enzyme pathways of human hepatocytes (Silvers et al.,1994)and is regarded as a “gold standard ”in toxicology.KERTr cells possess xenobiotic detoxi ?cation systems that would also contribute to the toxicity of metabolites transformed by biological systems (Van Pelt et al.,1990).

The objectives of this work were to (1)measure the concentrations of PAHs in different indoor dusts in Hong Kong;(2)evaluate the cytotoxicity of dust organic extracts based on MTT assay;(3)investigate the relationship between the cytotoxicity and concentrations of PAHs contained in dust and (4)to trace the sources of PAHs.

Environment International 36(2010)542–547

?Corresponding authors.Tel.:+852********;fax:+852********.

E-mail addresses:bichung@https://www.doczj.com/doc/7b10791501.html,.hk (K.C.Cheung),mhwong@https://www.doczj.com/doc/7b10791501.html,.hk (M.H.Wong).

0160-4120/$–see front matter ?2010Elsevier Ltd.All rights reserved.doi:10.1016/j.envint.2010.04.006

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2.Materials and methods

2.1.Sampling and preparation

Samples of air dust were collected from commercial of?ces located in a commercial building,secondary school,shopping mall,hospital, electronic factory assembling electronic equipment,and from a manufacturing plant which made furniture,toys and textiles,in Hong Kong(Table1).Dust samples were collected from each site using a vacuum cleaner(Black&Decker Dustbuster,USA)on the air-conditioner ?lter.To avoid cross contamination,the vacuum cleaner bag was changed after each sample collection.All samples were?ltered through a stainless-steel sieve(b0.1mm)onto solvent-rinsed aluminum foil to remove debris and large particles.Afterwards,1.5g of the sieved dust was extracted with100ml acetone/dichloromethane/n-hexane(1:1:1, v/v/v)in a Soxhlet apparatus.The extracts were concentrated using a rotary evaporator and were solvent-exchanged to1.5ml n-hexane. Concentrated n-hexane extracts of0.5ml were solvent-exchanged to 0.5ml dimethyl sulfoxide(DMSO),and stored at?20°C before cell assay.Another1ml was stored at?20°C for chemical analyses(Chuang et al.,1995).

2.2.Chemical analyses

For PAH analysis,concentrated extract(1ml)was cleaned up using a sodium sulfate anhydrous/?orisil column and eluted with a mixture of n-hexane and dichloromethane(80ml,7:3,v/v)(U.S.EPA.,1996a).The eluant was evaporated to less than2ml.Deuterated PAHs(ace-naphthene-d10,phenanthrene-d10,chrysene-d12and perylene-d12) were used as internal standards for quanti?cation.GC–MS analysis was performed on Agilent6890N network GC system equipped with5973 inert mass selective detector.The EPA standard method8270C(U.S.EPA., 1996b)was used to determine the following16PAHs:naphthalene (Nap),acenaphthylene(Any),acenaphthene(Ane),?uorine(Fle), phenanthrene(Phe),anthracene(Ant),?uoranthene(Fla),pyrene (Pyr),benz(a)anthracene(B(a)A),chrysene(Chry),benzo(a)pyrene(B (a)P),benzo(b)?uoranthene(B(b)F),benzo(k)?uoranthene(B(k)F), indeno(1,2,3-cd)pyrene(I(cd)P),dibenz(a,h)anthracene(D(ah)A)and benzo(ghi)perylene(B(ghi)P).The peaks of BbF and BkF were extremely close and dif?cult to be distinguished,and therefore these two compounds were combined as one,namely B(b+k)F.

2.3.Quality control

Using the Standard Reference Material(SRM2585,house dust,NIST, USA),it was observed that the recoveries of individual PAH ranged from 79%for pyrene(Pyr)to115%for benzo(a)pyrene(B(a)P).The results were not corrected for the recovery ratios for the SRM.The limit of detection(LOD)of PAHs de?ned as the standard deviation from the mean blank(n=3)ranged from1to6ng/g.2.4.Cell culture and MTT assay

The HepG2cell line(human hepatocellular liver carcinoma cell line) and KERTr cell line(human skin derived keratinocyte),were obtained from the American Type Culture Collection(ATCC,Rockville,USA).HepG2 cells were cultured in Eagle's minimal essential medium supplemented with10%fetal bovine serum according to the procedure provided by ATCC.The KERTr cells were cultured in keratinocyte-serum free medium (Gibco,USA)supplemented with0.05mg/ml bovine pituitary extract (BPE)and35ng/ml epidermal growth factor(EGF).These cells were cultured in a humidi?ed atmosphere with5%CO2at37°C.Mitochondrial capacity to reduce MTT to formazan(Gerlier and Thomasset,1986)was used as a measure of cell viability.At a culture growth between80%and 100%of con?uence,the HepG2cells were trypsinized,counted,and seeded onto96-well tissue culture microtiter plates at a density of2×104 cells/100μl/well.The KERTr cells were seeded onto96-well tissue culture microtiter plates at a density of1×104cells/100μl/well.After24h,the cell culture medium was removed and replaced by100μl culture medium containing?ve different concentrations of each dust organic extract prepared by a2-fold dilution for KERTr cell and a4-fold dilution for HepG2 cell.In each well,the?nal concentration of DMSO was limited to0.5%. After24h incubation,the culture medium was removed and the cells were washed twice with warm PBS.The cells were then incubated with serum free medium containing0.5mg MTT/ml at37°C.The medium was removed and replaced with100μl DMSO after4h.The colored solution was then detected at a wavelength of540nm and at690nm(reference wavelength).0.5%DMSO treated cells were used as the100%viable control.Cell mortality was obtained by the following equation(Mirto et al.,1999):cell mortality=1?(OD sample/OD DMSO),where OD sample is the optical density of sample detection in MTT assay and OD DMSO is the optical density of DMSO control detection in MTT assay.LC50,which is the concentration of contaminants killing50%of the cells,of each dust extract was calculated by using Trimmed Spearman–Karber method with the software provided by US EPA(Trimmed Spearman–Karber program).

2.5.Statistical analyses

All the statistical tests were performed with SPSS16.0software. Normality of the data was checked by Shapiro–Wilk test.Means of different groups were compared using one-way ANOVA test.For Post Hoc Test in ANOVA,Dunnett's T3method or Least Signi?cant Difference(LSD) method was adopted.Prior to one-way ANOVA test,the homogeneity of variance of independent variable was examined by Levene's test.The high molecular weight(HMW)PAHs and low molecular weight(LMW)PAHs

Table1

The detailed information of the sampling sites.

Sampling sites Sample

numbers

Detailed information about

sampling sites

Commercial of?ce20Located in the commercial building;

commercial area

Secondary school4Residential area

Shopping mall5Commercial area

Hospital16Residential area

Electronic factory6Assembling electronic equipment such

as computer;industrial area Manufacturing plant4All producing furniture,toys and textiles;

the machine in the plant using diesel for

electric power;industrial area Table2

Summary of PAH concentrations in55indoor dust samples in Hong Kong(μg/g).

μg/g Mean±sd Median Minimum Maximum

Nap0.12±0.090.140.020.56

Any0.01±0.0070.020.0030.04

Ane0.02±0.040.010.0020.28

Fle0.03±0.030.030.0050.20

Phe0.77±0.42 1.020.30 2.45

Ant0.03±0.020.040.0080.15

Fla0.86±0.58 1.030.21 3.28

Pyr0.69±0.430.950.20 1.77

B(a)A0.22±0.190.240.050.83

Chry0.54±0.460.700.10 1.94

B(b+k)F0.98±1.15 1.140.08 5.55

B(a)P0.38±0.380.420.11 2.28

I(cd)P0.58±0.980.520.07 5.40

D(ah)A0.06±0.080.060.0020.41

B(ghi)P0.90±1.260.920.08 6.61

a Total PAHs 6.18±5.417.50 1.6427.3

b LMW PAHs0.99±0.54 1.190.41 3.34

c HMW PAHs 5.20±5.04 6.29 1.1725.5

a Total PAHs=the sum of16PAHs.

b LMW PAHs,low molecular weight PAHs having2or3rings(Maertens et al.,2004).

c HMW PAHs,high molecular weight PAHs having4or more rings(Maertens et al.,2004).

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Y.Kang et al./Environment International36(2010)542–547

in each dust sample,and LC 50of dust extracts on the two types of cell lines were compared using paired-sample T -test.Curve estimation was employed to investigate the relationships between PAH concentration and LC 50.The probability value of p b 0.05was set as the level for statistical signi ?cance.

3.Results and discussion

3.1.Distribution and concentration of PAHs in different indoor dusts in Hong Kong The concentrations of individual PAH,total PAHs,low molecular weight (LMW,2–3rings)PAHs,and HMW (high molecular,4–7rings)PAHs in indoor dusts in Hong Kong are listed in Table 2.It has been reported that the total PAH concentration ranged from 0.40to 554μg/g with a mean of 28.3μg/g,low molecular weight (LMW,2–3ring PAHs)PAHs from 0.07to 65.9μg/g with a mean of

4.38μg/g,and high molecular weight PAHs (HMW,4–7ring PAHs)from 0.34to 505μg/g with a mean of 22.8μg/g in settled house dust (SHD)(particle size:less than 150μm)collected from North America,in 18published studies (Maertens et al.,2004).The majority of the SHD samples were collected using the High Volume Small Surface Sampler and some samples were collected from the ?oor using vacuum cleaners.Henceforth,the concentrations of PAHs in the present study fall within the range reported by others.In addition,the total PAH concentrations in the present study are higher than that reported in the house dust collected from ?oors in Kuwait (0.003to 2.92μg/g with a mean of 0.17μg/g;dust particle size:63μm)(Gevao et al.,2007).However,the PAH concentrations are the same level with that found in dust samples collected from carpets of 24houses in Durham,USA (1.25to 1

5.2μg/g with a mean of 4.52μg/g;dust particle size:150μm)(Chuang et al.,1999)and those reported in dust samples collected from air-conditioner ?lters in Jabriya residential area away from oil ?res in Kuwait (mean concentration:4.20μg/g;dust particle size:unknown)(Saeed et al.,1998).The mean PAH concentration in this study is lower than that reported in settled house dust

collected

Fig.1.The PAH concentrations in different indoor dusts in Hong

Kong.

Fig.2.PAH pro ?les in different indoor dusts in Hong Kong.

544Y.Kang et al./Environment International 36(2010)542–547

from ?oors in Ottawa,Canada (1.50to 325μg/g with a mean of 9.53μg/g;particle size:100μm)(Maertens et al.,2008).The dust samples in most of the studies were collected from either the ?oor or the carpet,however,it should be noted that dust from air-conditioner ?lters could re ?ect the indoor dust and indoor air quality because the air conditioner recirculates the air through the ?lter (Saeed et al.,1998).Therefore,the dust from air-conditioner ?lters can provide more information with regard to human exposure to indoor dust than that from the ?oor or carpet.

The dust samples from different types of workplaces showed different concentrations of PAHs (Fig.1).The mean concentrations of total PAHs from the commercial of ?ce,secondary school,shopping mall,hospital,electronic factory and manufacturing plant were 5.85±3.19,5.61±2.33,5.53±2.07,3.72±1.82,4.09±2.38and 22.2±6.05μg/g,respectively.The mean concentration of HMW PAHs from the commercial of ?ce,secondary school,shopping mall,hospital,electronic factory and manufacturing plant were 4.87±2.88,4.75±2.11,4.52±1.63,2.96±1.62,3.23±1.93and 20.1±6.81μg/g,respectively.The mean concentrations of LMW PAHs from the commercial of ?ce,secondary school,shopping mall,hospital,electronic factory and manufacturing plant were 0.98±0.42,0.86±0.29,1.01±0.48,0.76±0.31,0.86±0.46and 2.19±0.78μg/g,respectively.Concentrations of total PAHs and HMW PAHs increased in the order of hospital=electronic factory b shopping mall =secondary school=commercial of ?-ce b manufacturing plant,though the difference was not signi ?cant (p N 0.05)due to large standard deviations and different sample sizes among the groups.However,the dust samples of the manufacturing plant contained signi ?cantly higher LMW PAHs (p b 0.05)than the other locations.The HMW PAH concentrations were signi ?cantly higher than the LMW PAH concentrations in all of the indoor dust samples (p b 0.01),indicating the dominance of HMW PAHs in indoor dust.According to our data,the manufacturing plant was heavily contaminated with PAHs.This may be explained by the heavy lubricant used

in production machines and diesel for electrical power,emitting PAHs by thermal degradation of lubricant and combustion of diesel)(Rajput and Lakhani,2009).The hospital dust samples had the lowest level of PAHs among all the samples.There is a paucity of information regarding PAH concentrations in hospital dust.However,Zhao et al.(2008)reported that waste ashes from hospital waste incinerators in China contained very high levels of PAHs (4.16to 199μg/g).Dust samples from the commercial of ?ce,secondary school and shopping mall contained similar PAH concentrations,which also exhibited high variability in each group (Fig.1).This may be due to the special characteristics of these sites (a large number of people and complex anthropogenic activities such as unloading and unpacking of of ?ce furniture,and chemicals).

Fig.2shows the similarities between the PAH pro ?les (distribution patterns of 16PAHs)of the six different types of workplace dust,with Fla,Pyr,B(b +k)F,and B(ghi)P being the most abundant,and Any,Ane and Fle the least abundant.However,there was little difference among all sites.B(b +k)and B(ghi)P accounted for 20%and 21%of the total PAH concentration in the manufacturing plant,and 15%and 18%in the secondary school,respectively.Fla and B(b +k)F accounted for 17%and 15%in the commercial of ?ce,15%and 15%in the shopping mall,and 14%and 15%in the hospital,respectively.Phe and Fla accounted for 15%and 16%in the electronic factory,respectively.In general,the dominant PAHs in the electronic factory belonged to HMW PAHs except for

Phe.

Fig.3.Examples of MTT assay for the six extracts from the commercial of ?ce,secondary school,shopping mall,hospital,electronic factory and manufacturing plant.A:Examples of dose –response curve of the HepG2cells.B:Examples of dose –response curve of KERTr cells.Table 3

LC 50(mg/100μl)of dust extracts on HepG2and KERTr cell lines.Sampling sites

HepG2KERTr LC 50

Min –Max LC 50

Min –Max Commercial of ?ce 0.23±0.160.05–0.700.11±0.070.03–0.24Secondary school 0.25±0.030.20–0.280.13±0.040.08–0.19Shopping mall 0.12±0.050.06–0.190.08±0.050.03–0.16Hospital

0.47±0.340.10–1.020.14±0.080.02–0.26Electronic factory 0.42±0.250.17–0.770.15±0.130.03–0.37Manufacturing plant 0.05±0.050.02–0.120.03±0.010.02–0.04All

0.30±0.26

0.02–1.02

0.12±0.08

0.02–

0.37

Fig.4.Relationship between total PAH concentration and LC 50of dust samples collected from different workplaces in Hong Kong.A:LC 50on HepG2cell line.B:LC 50on KERTr cell line.

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Y.Kang et al./Environment International 36(2010)542–547

3.2.LC50of dust

Fig.3shows the results of HepG2and KERTr MTT assays treated with organic dust extracts.In general,dose–response curves indicated an increase of cell mortality when treated with increased concentration of dust extracts.Table3presents the results of LC50of different workplace dust extracts on HepG2and KERTr cell lines.The LC50of dust extracts on HepG2cells was signi?cantly higher(p b0.01)than that on KERTr cells.It indicated that the dust extracts had higher cytotoxicity on KERTr cells than HepG2cells.This may be explained by the fact that HepG2cells contain a higher activity of detoxi?cation enzymes such as cytochrome P450that are responsible for the metabolism of many foreign compounds than other human cell lines(Turesky et al.,2002).To further evaluate the relative cytotoxicity of dust extracts among sites,statistical differences among the sampling sites were compared.For cytotoxicity on HepG2cell line,dust extracts from the manufacturing plant showed a higher(p b0.05)cytotoxicity than that from the commercial of?ce,secondary school and hospital.However,there was no signi?cant (p N0.05)difference between the manufacturing plant and shopping mall or electronic factory,due to the large standard deviations and the different sample sizes among these groups.In addition,the dust from the shopping mall showed higher cytotoxicity than that from the secondary school and hospital.For the KERTr cell line,dust extracts from the manufacturing plant exerted a lower LC50than that from the commercial of?ce and hospital(p b0.05)indicating that dust samples from the manufacturing plant had a higher cytotoxicity.This indicated that the two types of cell lines showed similar cytotoxicity pro?les when treated with the dust extracts.In addition,there was a signi?cant correlation (r=0.76,p b0.01)between the LC50of dust extracts on HepG2and KERTr cells.

Although MTT assay is an in vitro assay that only considers one end point,it provides signi?cant data to establish marked differences in dust samples from different sites in this study.It should be noted that the cytotoxicity of dust extracts in the present study was only caused by the organic fraction.Inorganic partition such as heavy metals could also give rise to cytotoxicity(Mirto et al.,1999).Further investigation is needed to evaluate the cytotoxicity of dust more comprehensively.

3.3.LC50and PAH concentration

The results of curve estimation analysis revealed that power model had a stronger ability to explain the relationship between the LC50and total PAH concentrations than the linear model.For HepG2cell line,the correlation coef?cient of power model(R=?0.65,p b0.01) was higher than that of the linear model(R=?0.44,p b0.01)(Fig.4A).For KERTr cell line, the correlation coef?cient of power model(R=?0.63,p b0.01)was also higher than that of the linear model(R=?0.51,p b0.01)(Fig.4B).According to the data,it seemed that the power model was a better?t for explaining the relationship between the LC50and total PAHs.

The combination of chemical analyses and in vitro bioassays provided an important tool to assess the dust contamination and identify the potential toxic contaminants present.Reyes et al.(2000)found that dust extracts could induce high cytotoxicity (neutral red assay on keratinocytes treated with outdoor air samples from industrial area including PM100and PM10)which was seasonally dependent and particle-size dependent.It revealed that the dust collected during the summer had higher cytotoxicity than that from winter and that greater toxic effects were observed in PM10extracts than in PM100extracts.Lin et al.(2008)reported that the organic extracts of0.18–5.6μm particles had higher cytotoxicity(propidium iodide assay on alveolar macrophages treated with air samples from a heavily traf?cked roadside)than that of 0.018–0.18μm and5.6–18μm particles.However,they had failed to attribute the cytotoxicity potency on alveolar macrophages to the PAHs.

There are limited studies that attempt to isolate and identify cytotoxic compounds from dust samples resulting in cytotoxicity.In the present study,the LC50representing

the

Fig.5.Plot of ratio of I(cd)P/(I(cd)P+B(ghi)P)against Fla/(Fla+Pyr)for indentifying sources of PAHs in different indoor dusts in Hong Kong.A:Pyrogenic and petrogenic source identi?cation.B:Fuel combustion and biomass combustion source identi?cation.

546Y.Kang et al./Environment International36(2010)542–547

cytotoxicity of organic dust extracts on both HepG2cells and KERTr cells was negatively correlated(p b0.01)with the total PAHs contained in dust samples.Although other contaminants such as heavy metals and organochlorine pesticides could also induce cytotoxicity(Kappus and Reinhold,1994;Suzuki et al.,1997),the coef?cient of determination(i.e.,r2values)suggested that PAHs accounted for almost40%of the observed cytotoxicity on both the HepG2(r2=0.42,p b0.01)and KERTr cell lines(r2=0.40,p b0.01).

3.4.PAH source analyses

The scatter plot(Fig.5)shows the ratio of Fla/(Fla+Pyr)and I(cd)P/(I(cd)P+B(ghi)P) in different dust samples.Samples with Fla/(Fla+Pyr)b0.4and I(cd)P/(I(cd)P+B(ghi)P)b 0.2were regarded as originating from petrogenic source,while that with Fla/(Fla+Pyr)N 0.4and I(cd)P/(I(cd)P+B(ghi)P)N0.2as pyrogenic source(Brandli et al.,2006).According to Fig.5A,it was apparent that PAHs in all the samples were derived from pyrogenic sources. In addition,samples with0.4b Fla/(Fla+Pyr)b0.5and0.2b I(cd)P/(I(cd)P+B(ghi)P)b0.5 were regarded as originating from fuel combustion,while that with Fla/(Fla+Pyr)N0.5and I(cd)P/(I(cd)P+B(ghi)P)N0.5from biomass combustion(Brandli et al.,2006).The results showed that two samples from the commercial of?ce,six samples from the hospital and one sample from the secondary school were identi?ed as derived from fuel combustion(Fig.5B). The origins of other samples were dif?cult to identify and may be due to mixed contamination sources(both fuel combustion and biomass combustion)(Soclo et al.,2000).

4.Conclusion

Chemical analyses showed that the dust samples from the manufacturing plant contained the highest concentration of PAHs among all sampling sites.Results of MTT assay indicated that the dust organic extracts from the manufacturing plant also had the greatest cytotoxicity on both HepG2cells and KERTr cells.In addition,the power model revealed that the LC50obtained from MTT assay was negatively correlated(p b0.01)with total PAHs.The results of PAH source analysis demonstrated that PAHs were mostly due to pyrogenic origins. Acknowledgements

The authors would like to thank Dr.Anna O.W.Leung for editing the manuscript and Dr.Homan Leung for?eld sampling.Financial support from the Research Grants Council of the University Grants Committee of Hong Kong(HKBU260209)and Special Equipment Grant(HKBU09) and the Mini-AoE(Area of Excellence,RC/AOE/08-09/01)Fund from Hong Kong Baptist University are gratefully acknowledged. References

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51单片机汇编指令集(附记忆方法)

51单片机汇编指令集 一、数据传送类指令(7种助记符) MOV(英文为Move):对内部数据寄存器RAM和特殊功能寄存器SFR的数据进行传送; MOVC(Move Code)读取程序存储器数据表格的数据传送; MOVX (Move External RAM) 对外部RAM的数据传送; XCH (Exchange) 字节交换; XCHD (Exchange low-order Digit) 低半字节交换; PUSH (Push onto Stack) 入栈; POP (Pop from Stack) 出栈; 二、算术运算类指令(8种助记符) ADD(Addition) 加法; ADDC(Add with Carry) 带进位加法; SUBB(Subtract with Borrow) 带借位减法; DA(Decimal Adjust) 十进制调整; INC(Increment) 加1; DEC(Decrement) 减1; MUL(Multiplication、Multiply) 乘法; DIV(Division、Divide) 除法; 三、逻辑运算类指令(10种助记符) ANL(AND Logic) 逻辑与; ORL(OR Logic) 逻辑或; XRL(Exclusive-OR Logic) 逻辑异或; CLR(Clear) 清零; CPL(Complement) 取反; RL(Rotate left) 循环左移; RLC(Rotate Left throught the Carry flag) 带进位循环左移; RR(Rotate Right) 循环右移; RRC (Rotate Right throught the Carry flag) 带进位循环右移; SWAP (Swap) 低4位与高4位交换; 四、控制转移类指令(17种助记符) ACALL(Absolute subroutine Call)子程序绝对调用; LCALL(Long subroutine Call)子程序长调用; RET(Return from subroutine)子程序返回; RETI(Return from Interruption)中断返回; SJMP(Short Jump)短转移; AJMP(Absolute Jump)绝对转移; LJMP(Long Jump)长转移; CJNE (Compare Jump if Not Equal)比较不相等则转移;

自考计算机系统结构试题

一、选择题 1、对汇编语言程序员,下列(A )不是透明的。 A: 中断字寄存器 B: 乘法器 C: 移位器 D: 指令缓冲器 2、假设对A机器指令系统的每条指令的解释执行可直接由B机器的一段微程序解释执行,则A称为(D )。 A: 仿真机 B: 宿主机 C: 虚拟机 D: 目标机 3、 1. 查看下面三条指令:V3←A;V2←V0+V1;V4←V2*V3;假设向量长度小于64,且前后其他的指令均没有相关性,数据进入和流出每个功能部件,包括访问存储器都需要一拍的时间,假设向量的长度为N。三条指令全部采用串行的方法,那么执行的时间是: A: 3N+20 B: 3N+21 C: 3N+22 D: 3N+23 4、某向量处理机有16个向量寄存器,其中V0-V5种分别存放有向量A,B,C,D,E,F,向量的长度是8,向量各元素均为浮点数;处理部件采用两个单功能流水线,加法功能部件时间为2拍,乘法功能部件时间为3拍。采用类似CRAY-1的链接技术,先计算(A+B)*C,在流水线不停的情况下,接着计算(D+E)*F。求此链接流水线的通过时间是多少拍?(设寄存器出入各需1拍) A: 8 B: 9 C: 17 D: 18 5、设有一个4个处理器的MIMD系统,假设在系统中访存取指和取数的时间可以忽略不计;加法与乘法分别需要2拍和4拍;在MIMD 系统中处理器(机)之间每进行一次数据传送的时间为1拍;在MIMD 系统中,每个PE都可以和其它PE有直接的的通路。 求利用此系统计算表达式所需的节拍数。 A: 23 B: 12 C: 11 D: 10 6、以下哪些是周期窃取方式的特点? A : 硬件结构简单 B : 硬件结构复杂 C : 数据输入或输出过程中占用了CPU时间 D : 数据输入或输出过程中不占用CPU时间 7、从下列有关Cache的描述中,选出应填入空格中的正确答案:(1)今有甲、乙两台计算机,甲计算机的Cache存取时间为50ns,主存储器为2us;乙计算机的Cache存储时间为100ns,主存储器为1.2us。设Cache的命中率均为95%,则甲计算机的平均存取时间为__A_ns;乙计算机的平均存取时间为__B_ns。 (2)在Cache中,经常采用直接映象或组相联映象两种方式,在Cache 容量相等的情况下,前者比后者的命中率__C__。 选项 A : 147.5 153.5 155 180 选项 B : 147.5 153.5 155 180 选项 C : 高低相等 8、有研究人员指出,如果在采用通用寄存器指令集结构的计算机里加入寄存器—存储器寻址方式可能提高计算机效率,做法就是用指令ADD R2,0(Rb)代替指令序列LOAD R1,0(Rb) ADD R2,R2,R1假定使用新的指令能使时钟周期增加10%,并且假定只对时钟产生影响,而不影响CPI那么采用新指令,要达到与原来同样的性能需要去掉的LOAD操作所占的百分比?(提示:去掉的是与ADD指令连用的LOAD 指令,假定未采用新指令前LOAD指令占总指令的22.8%)A: 39% B: 36% C: 40% D: 39.8% 9、下述的几个需要解决的问题中,那个是向量处理机所最需要关心 的? A: 计算机指令的优化技术 B: 设计满足运算器带宽要求的存储器 C: 如何提高存储器的利用率,增加存储器系统的容量 D: 纵横处理方式的划分问题 10、一台单处理机可以以标量方式运行,也可以以向量方式运行。 在向量方式情况下,计算可比标量方式快18倍。设某基准程序在此 计算机上运行的时间是T。另外,已知T的25%用于向量方式,其余 机器时间则以标量方式运行。那么在上述条件下与完全不用向量方式 的条件下相比的加速比是: A: 3 B: 3.43 C: 3.33 D: 以上均不正确 11、给定1个采用完全混洗互连网络,并有256个PE的SIMD机器, 加入执行混洗互连函数10次,则原来在PE123中的数据将被送往何 处? A: PE237 B: PE222 C: PE111 D: PE175 12、 设计一种采用加、乘和数据寻径操作的算法,计算表达式 。假设加法和乘法分别需要2个和4个单位 时间,从存储器取指令、取数据、译码的时间忽略不计,所有的指令 和数据已经装入有关的PE。现有一台串行计算机,有一个加法器,一 个乘法器,问最短多少单位时间计算出s? A: 192ns B: 130ns C: 128ns D: 以上结果都不对 13、下列功能,那些一般由硬件实现? A : 第一次关CPU中断 B : 返回中断点 C : 第一次开CPU中断 D : 保存中断点 14、星形网络的网络直径和链路数分别为()和()。 选项 1 : N-1 N/2 2 N(N-1)/2 选项 2 : N-1 N/2 2 N(N-1)/2 16、在计算机系统结构来看,机器语言程序员看到的机器属性是(D)。 A)计算机软件所要完成的功能B)计算机硬件的全部组成 C)编程要用到的硬件知识D)计算机各部件的硬件实 现 17、在提高CPU性能的问题上,从系统结构角度,可以(D)。 A)提高时钟频率B)减少 程序指令条数 C)减少每条指令的时钟周期数D)减少程序指 令条数和减少每条指令的时钟周期数 18、计算机系统结构不包括(C )。 A)主存速度B)机器工作状态C)信息保护 D)数据表示 19、推出系列机的新机器,不能更改的是(A)。 A)原有指令的寻址方式和操作码B)系统 的总线的组成 C)数据通路宽度 D)存储芯片的集成度 20、在系统结构设计中,提高软件功能实现的比例会(C)。 A)提高解题速度 B)减少需要的存储容量 C)提高系统的灵活性 D) 提高系统的性能价格比 21、重叠寄存器技术主要用于解决在RISC系统中因( C )而导致 的问题。 A)JMP指令影响流水线B)CALL 指令的现场保护 C)只有LOAD和STORE指令带来的访问存储器不便D)存储 器访问速度 22、不属于堆栈型替换算法的是(C )。 A)近期最少使用法B)近期最久未用法 C)先进先 出法D)页面失效频率法 23、与全相联映象相比,组相联映象的优点是(B)。 A)目录表小B)块冲突概率低C)命中率高 D)主存利用率高 24、最能确保提高虚拟存储器访主存的命中率的改进途径是(D)。 A)增大辅存容量 B)采用FIFO替换算法并增大页面 C)改用LRU替换算法并增大页面D)改用LRU替换 算法并增大页面数 25、"一次重叠"中消除"指令相关"最好的方法是( A )。 A)不准修改指令B)设相关专用通路C)推后分析下条指令 D)推后执行下条指令 26、在流水机器中,全局性相关是指(D)。 A)先写后读相关B)先读后写相关C)指令相关 D)由转移指令引起的相关 27、下列说法不正确的是(D)。 A)线性流水线是单功能流水线B)动态 流水线是多功能流水线 C)静态流水线是多功能流水线D)动态 流水线只能是单功能流水线 28、16个处理器编号为0、1、…、15,采用单级Cube3互连网络互连, 与13号处理器相连的处理器号是()。 A)2 B)3 C)4 D)5 29、经多级网络串联来实现全排列网络,只能用(C)。 A)多级立方体网络B)多级PM2I网络C)多级混洗交 换网络D)上述任何网络 30、经3级立方体网络对0-7八个端子(0 1 2 3 4 5 6 7)排列, 进行模8移4变换,得到的这八个端子新的排列应当是()。 A)(2 3 4 5 6 7 0 1)B)(4 5 6 7 0 1 2 3) C)(1 2 3 0 5 6 7 4)D)(1 0 3 2 5 4 7 6) 31、虫蚀寻径以流水方式在各寻径器是顺序传送的是(C)。 A)消息B)包C)片 D)字节 32、能实现指令、程序、任务级并行的计算机系统属于(D )。

芳烃联合装置

芳烃联合装置

抽提蒸馏塔塔顶产品为非芳烃,作为非芳烃副产品送出装置,塔底产物为富含苯的溶剂,送溶剂回收塔作为进料。抽提蒸馏塔重沸器热源由中压蒸汽提供,通过控制加热蒸汽量来调节热负荷,加热蒸汽分成两股进行控制,主流股(约80%)由定流量控制,次流股流量(约20%)由灵敏板温度与流量串级控制。 溶剂回收塔的作用是实现苯产品与溶剂的分离。溶剂回收塔在减压下操作,塔顶残压由压力控制器控制回收塔蒸汽喷射泵的尾气返回量或氮气吸入量进行调节。溶剂回收塔塔顶产物为苯产品,经白土处理后送往苯检验罐,塔底贫溶剂大部分直接循环使用,少部分去溶剂再生罐进行减压蒸发再生后循环使用。溶剂回收塔重沸器热源由中压蒸汽提供,加热量由重沸器出口凝结水流量进行控制。 溶剂再生罐实际上是一个减压蒸发器,操作压力由压力控制器控制再生罐蒸汽喷射泵的尾气返回量或氮气吸入量进行调节。溶剂再生罐热量由内插式溶剂再生罐加热器提供,加热热源为中压蒸汽,加热量由蒸汽凝结水流量进行调节。再生后溶剂送至贫溶剂泵入口循环使用。溶剂再生罐罐底残渣采用不定期方式排出。 4、对二甲苯装置 对二甲苯装置采用美国UOP的专利工艺技术,主要生产纯度99.8%的对二甲苯(PX)产品,并富产苯、邻二甲苯(OX)、重芳烃等。包括甲苯歧化-烷基转移单元、二甲苯异构化单元、二甲苯精馏单元、吸附分离单元四部分。 甲苯歧化-烷基转移单元采用UOP的TATORAY工艺,选用活性、选择性及稳定性较高的新一代TA-4催化剂,在高温作用下,甲苯和C9A发生歧化和烷基转移反应,生成目的产品苯和二甲苯。可以通过调整甲苯和C9A的比例来实现苯和二甲苯产品的分布。2003年月份催化剂进行了国产化,使用上海石油化工科学研究院自主开发的HAT-97催化剂,该催化剂最大的特点是可以加工3-5%的C10A,并且具有更高的选择性和转化率。 二甲苯异构化单元采用UOP的ISOMAR工艺,选用乙苯异构型I-9K催化剂,在反应过程中建立限定性平衡,通过环烷烃中间体将乙苯最大限度地转化为二甲苯,采用这种催化剂可以从混合二甲苯中获取最高产率的对二甲苯。该催化剂稳定性好,反应压力和氢油比低,不需注氯,减少了系统腐蚀,改善了操作环境。 吸附分离单元采用UOP的PAREX工艺,通过多通道旋转阀实现连续逆流接触,利用分子筛选择吸附PX,再用解吸剂对二乙基苯将PX置换解吸,从而达到分离PX 的目的。选用最新分子筛吸附剂ADS-27,改进吸附系统设备和优化工艺参数,增大了吸附塔的处理能力,对二甲苯单程收率可提高到97%,纯度达到99.80%。 二甲苯精馏单元采用精密分馏工艺,将混合芳烃中的C8A、C9A分离出来,分别作为原料提供给吸附分离和歧化单元,从而将联合装置各单元有机的联合起来。二甲苯塔采用加压操作,操作压力为1.0Mpa(a),利用塔顶和塔底高温物流分别作为其它单元集中供热热源,多余的塔顶汽相通过蒸汽发生器发生1.0Mpa蒸汽,全塔的热量均被利用,节能效果显著。 5、中间原料及溶剂油罐区负责芳烃联合装置的原料、甲苯、溶剂油的收储工作。包括中间原料油罐区、溶剂油罐区、芳烃原料罐区三部分。

计算机基础知识试题(一)及答案

8?在计算机内部,不需要编译计算机就能够直接执行的语言是 ( )。 A.汇编语言 B.自然语言 C.机器语言 D.咼级语言 第一部分 一、单项选择题 1.世界上第一台电子数字计算机取名为()。 A. UNIVAC B.EDSAC C.ENIAC D.EDVAC 2?操作系统的作用是()。 3?个人计算机简称为PC 机,这种计算机属于( 4?目前制造计算机所采用的电子器件是()。 5?—个完整的计算机系统通常包括()。 A.硬件系统和软件系统 B.计算机及其外部设备 C.主机、键盘与显示器 D.系统软件和应用软件 6?计算机软件是指 ()。 A. 计算机程序 B.源程序和目标程序 C.源程序 D.计算机程序及有关 资料 7?计算机的软件系统一般分为()两大部分。 A.系统软件和应用软件B.操作系统和计算机语言 C. 程序和数据D.DOS 和WINDOWS A.把源程序翻译成目标程序 B.进行数据处理 C.控制和管理系统资源的使用 D.实现软硬件的转换 A.微型计算机 B.小型计算机 C.超级计算机 D.巨型计算机 A.晶体管 B.超导体 C.中小规模集成电路 D.超大规模集成电路

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