Drinking Boiled Tap Water Reduces Human Intake of Nanoplastics and Microplastics. Picie przegotowanej wody z kranu zmniejsza spożycie nanoplastiku i mikroplastiku przez ludzi.

Picie przegotowanej wody z kranu zmniejsza spożycie nanoplastiku i mikroplastiku przez ludzi.

Drinking Boiled Tap Water Reduces Human Intake of Nanoplastics and Microplastics.

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1. Supplemental Texts
2. Materials

Monodisperse fluorescent polystyrene (PS) microspheres (100 mg 10 mL^_1, excitation peak of 540 nm, emission peak of 580 nm) containing three types of functional groups (bare-, carboxyl-, and amino-) and three particle sizes (0.1, 1, and 100 |im), calcium chloride (CaCh; 99.99%), and sodium bicarbonate (NaHCO3; 99.8%) were purchased from Tianjin SaiErQun Technology (Tianjin, China), Shanghai Macklin Biochemical (Shanghai, China), and Shanghai Yuanye Bio-Technology (Shanghai, China), respectively. Fluorescent PS, CaCh, and NaHCO3 solutions were prepared with deionized water, with the final concentrations at 1000, 222, and 336 mg L¹, respectively.

52. Prevention of Sample Contamination and Preparation of Control Samples

To minimize potential cross contamination, laboratory coats and nitrile gloves were worn during sample preparation and analysis. Glass beakers (50 mL) were used for sample preparation and were cleaned and dried at 200 ^oC before use. Before every experimental step, glass beakers were rinsed three times with deionized water and covered by glass lids. Two control samples were also analyzed for each PS nanoplastic heating/boiling group treated on different purposes (Table 1). For one of the control samples, a glass beaker filled with 40 mL deionized water and fluorescent PS microbeads (1 mg L^_1, 1 |im) was heated/boiled for 5 min to induce aggregated PS at 25-100 ^oC. Supernatant samples at 1 mL from a 1 cm depth below the supernatant surface and bottom samples at 1 mL at the bottom of the glass beaker were collected for flow cytometry analysis. The PS precipitation efficiency ranged from 0.7 ± 0.15% to 25 ± 3.2%. For another control samples, a glass beaker filled with 40 mL tap water (180 mg L^_1 of CaCO3) without fluorescent PS added was heated/boiled for 5 min to determine background contamination. Supernatant sample at 1 mL

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was collected from a 1 cm depth below the supernatant surface for flow cytometry analysis and background residual amount was 1.9 ± 1.03 particle pL^_1. All PS concentrations have been blank corrected.

S3.   Sampling, Analysis Methods, and Results for Microplastics

Sampling and Preparation of Microplastics. Microplastics (MPs) including PS sphere (~100 pm), polyethylene fragment (PE, ~150 pm), and polypropylene fiber (PP, ~100 pm) were used to verify the conclusion draw for PS nanoplastics. PS sphere (~100 pm) was purchased from Tianjin SaiErQun Technology (Tianjin, China). A PE plastic bag and a PP mask were placed under direct sunlight in an outdoor setting for 60 d in summer to produce aged PE plastic bag and aged PP mask. The aged PE and PP plastics were fragmented with a triturator, and filtered to obtain PE fragments (~150 pm) and PP fibers (~100 pm). PS sphere, PE fragments, and PP fibers were spiked into 40 mL tap water (180 mg L¹ of CaCO₃) with final MP concentration of 0.00075—0.0028 particle pL^_1 (Table 1). The solutions were heated (25-90 ^oC) and boiled (100 ^oC) for 5 mins. After heating/boiling, the mixed solutions were left to cool for 10 min. Each experiment was repeated three times. Supernatant samples were extracted by a pipette and filtered through 0.22 pm nylon filters. Precipitates were separated for further characterization.

Identification of Microplastics. Recovered PS, PE, and PP MPs in supernatants were identified and counted using a Raman spectroscope (Renishaw; Gloucestershire, UK) linked to a microscope (Leica Microsystems GMS GmbH, Germany). The MP precipitation efficiency was calculated by equation (1) in the main text. A scanning electron microscope (S-4800, Hitachi, Tokyo, Japan) was used to observe the microstructures of the coprecipitates. To preserve the initial binding morphology of NPs and incrustants in water, each 10 pL fresh incrustant sample was titrated on a copper platform, and dried for 1 h in a

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vacuum drying oven at 60 ^oC. Samples on the copper platform were treated with gold spray and observed by the scanning electron microscope.

Results. Boiling hard water can remove most PS, PE, and PP MPs, and PS, PE, and PP MPs precipitation efficiencies were 95 ± 4%, 81 ± 3%, and 90 ± 3%, respectively, at 100 ^oC. Increasing temperature accelerated the formation of incrustants on spherical, fragmented, and fibrous MP surfaces. MPs continued to be encapsulated by newly formed incrustants (Figure S2) and finally precipitated under gravity, confirming that spherical PS, fragmented PE, and fibrous PP MPs are able to coprecipitate with incrustants in tap water upon boiling. In concluding, the results with NPs in the main text were also applicable to MPs.

S4.   Details of Flow Cytometric Analysis for Supernatant Fluorescent Polystyrene

A Northern Light 3000 flow cytometer (Cytek Biosciences, Fremont, CA) can effectively detect the scattered light signal of fluorescence PS beads larger than 90 nm and allows for determination of PS particle number concentrations. Instrument calibration was performed with Cytek® SpectroFlo QC beads. The fluorescence PS beads were identified based on the 405 nm side scatter (SSC) parameter, and the fluorencence signals in B3 detector through 110 nm and 500 nm green flurescent latex beads (refractive index = 1.59) and nonfluorescent silica (Si) beads with a diameter of 180, 240, and 300 nm (refractive index = 1.43).¹ The minimum forward scatter (FSC) threshold was set to 5000 units to exclude debris from the event count. Set the flow rate to “Low”. Data after 10—15 s of sample acquisition was recorded when the flow rate stabilized. Data was collected at < 1000 events s^_1 and 5000 events were recorded for each sample. Exact measured volume of each sample was recorded by flow cytometry, and then total fluorescence PS concentration of each sample was subsequently extracted from the flow cytometry standard files to calculate the exact fluorescence PS concentration per ^L.

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Flow cytometric signal plots and gating of signal peaks of 1 |im and 0.1 ^m fluorescence PS were shown in Figure S3. Gating was applied to select the fluorescence PS particles. Gating of signal count of selected PS particles was analyzed. Though some background signals were occurred in the PS blank gating plots that overlapped with area of 1 |im and 0.1 |im fluorescent PS (Figures S3a and S3d), the positive signal peaks contributed by the fluorescence PS were distinguishable from the background signals (i.e., Figures S3b and S3e). And quantification of fluorescence PS was done on the B3 detector positive signal area (B3-A+).

S5.   Estimation of Energy Consumption and Carbon Dioxide Emission by Boiling Water through Electric Kettles and Gas Stoves

Electric kettles (electricity) and gas stoves (natural gas) were commonly used to boil water in human daily life. Therefore, the energy consumption and carbon dioxide (CO2) emissions by boiling water through electric kettles and gas stoves were estimated by^{2, 3}

Qi = M x C x (Tf - To) / r|i                                                                                                                                                                                    (1)

FMi = Qi / calorific valuei                                                                                                                                                                                    (2)

CO2 emissionsi =EFi x FMi                                                                                                                                                                                    (3)

Penergy = Qi / Qtotal                                                                                                                                                                                    (4)

Pco2 = CO2 emissionsi / CO2 emissionstotal                                                                                                                                                                                    (5)

where i indicates electric kettle or gas stove; Q is energy required to boil water; M is mass of boiled water (Table S4); C is the specific heat of water which is 4.184 kJ (kg-°C)^_1; Tf and To are the final and the initial water temperatures (100 ^oC and 20 ^oC in the present study), respectively; r| is thermal efficiency of the heating tool (Table S8);² FM is the quantity of the electricity or natural gas used to boil water; calorific value for electricity is 3600 kg kWh^-1 and for natural gas is 36600 kJ m^_3;⁴ EF is the carbon emission factor of the energy, for

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electricity is 673 g kWh^-1,⁵ and for natural gas is 1940 g m^-3;⁶ Penergy is the proportion of energy consumption of electric kettles or gas stoves upon boiling of tap water (Qi, Table S9) in total energy consumption (Qtotal, Table S10); and Pco2 is the proportion of CO2 emissions from consumption of electricity or natural gas upon boiling of tap water (CO2 emissions^ Table S9) in total CO2 emissions from energy consumption (CO2 emissionstotal, Table S10).

The annual energy required for electric kettles and gas stoves were 0.20-0.47 and 0.25­0.59 GJ individual^-1 year¹ in 2022, respectively (Table S9). According to Statistical Review of World Energy,⁷ energy consumption from different countries ranged from 17-630 GJ year^{- 1} individual^-1 (Table S10). The annual average energy demand for boiling water by electric kettles and/or gas stoves accounted for less than 1% of the annual total energy consumption (Figure S6). Boiling water may not be a burden on energy demands in most countries worldwide.

In addition, annual average CO2 emissions from consumption of electricity and natural gas upon boiling of drinking water were 38-88 and 14—33 kg year¹ individual^-1 in 2022, respectively (Table S9), accounting for less than 4% of the annual total CO2 emissions from energy consumption (Figure S6). Boiling through electric kettles and/or gas stoves could be environmentally friendly household water treatment. In this context, reducing human NMP intake by drinking boiled water is a feasible approach worth promoting worldwide.

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2. Supplementary Tables

Table S1. Conversion Ratios of Number and Mass Concentrations of Fluorescent

Polystyrene (PS) Microsphere.

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^a PS, polystyrene; PE, polyethylene; PP, polypropylene; PET, polyethylene terephthalate; PAN, polyacrylonitrile; AR, alkyd resin; PA, polyamide; PVC, polyvinyl

chloride; PTT, poly-trimethylene terephthalate; N.R., not reported.

^b Little attention has given to tap water NMPs from Africa and Oceania and data are not available. In the present study, surface water from Africa and groundwater from Oceania were considered in the human exposure of NMPs through drinking water consumption.

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(NMP) Precipitation Efficiency upon Boiling.

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^a According to the present study, the tested NMP precipitation efficiencies mainly dominated

by water temperature and hardness instead of NMP properties. Therefore, the precipitation efficiencies in different water hardness upon boiling drawn for polystyrene nanoplastics were considered for assessing the human NMP exposure through boiled water consumption. The precipitation efficiencies were preliminarily divided into 7 categories (Table S5): 0-59 mg L^{_ L} 25 ± 3.2%; 6^79 mg L¹: 33 ± 0.6%; 80-99 mg L¹: 34 ± 0.9%; 10^119 mg L¹: 47 ± 4.2%; 120—179 mg L¹: 77 ± 1.2%; 180-299 mg L¹: 84 ± 2.0%; > 300 mg L¹: 90 ± 1.2%.

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Table S4. Information on the Recommended Daily Intake Rate of Water (L day^-1 individual^-1) Used to Determine Human Nano/Microplastic Consumption of Adults (Male/Female) 19-71 Years Old and Children (Male/Female) 1-18 Years Old.

^a Recommended daily intake rates of boiled and tap water have not been distinguished,^{71, 72} and boiled water is generally obtained by tap water after boiling, thus the intake rate of boiled water is regarded the same as tap water.

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Table S5. Polystyrene Nanoplastic (NP) Precipitation Efficiency in Different Water

Temperature and Water Hardness upon Boiling.

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Table S6. Quantity of Incrustants Formed in Tap Water (180 mg L ¹ of CaCO3, 40 mL) at Different Temperatures.

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Table S7. X-ray Diffraction Standard Cards of Calcium Carbonate (CaCO3) and Magnesium Carbonate (MgCO3).

Calcite (CaCOs) JCPDSNo. 00-047-1743

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Table S10. Global Energy Consumption and Carbon Dioxide (CO2) Emissions from

Energy Consumption. ^a

^a Data is obtained from the Statistical Review of World Energy,⁷ and data of CO2 emissions

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from energy consumption is calculated by dividing total CO2 emissions by the population of the corresponding country.

3. Supplementary Figures

Figure S1. Diagram of supernatant sampling location. (a) Height of 40 mL water column before heating/boiling; (b) Height of water column after heating/boiling, samples in 1cm depth below the surface of the supernatant is near the center of the water column.

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Figure S2. Polystyrene, polyethylene, and polypropylene microplastics (PS, PE, and PP MPs) and incrustants coprecipitates during boiling of tap water (180 mg L^_1 of CaCO3). (a) PS, PE, and PP MP precipitation efficiencies in the supernatant at 25-100 ^oC; (b) Scanning electron microscopic of PS and incrustant coprecipitates at 25-100 ^oC; (c) Elemental mapping of PE and incrustant coprecipitates at 25-100 °C, green indicates “C” element, yellow indicates “O” element, and red indicates “Ca” element; and (d) Scanning electron microscopic of PP and incrustant coprecipitates at 25-100 ^oC.

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Figure S3.

a)

PS-blank

b)

Standard 1 |jm

0      1.0M 2.0M 3.0M 4.0M

FSC-A

0      1.0M    2.0M    3.0M 4.0M

FSC-A

2.0 1.50 ! 1.00

10 10 10 10 ComD-B3-A

d)

PS-blank

10 10 10 ComD-B3-A

tO 10      10

CamD-B3-A

e)

Standard 0.1 pm

10 10 10 FSC-A

10 10 ComD-B3-A

c)

PS-water 1 pm

0       1.0M     2.0M     3.0M 4 0M

FSC-A

10 10 10 Como-BS-A

f)

PS-water 0.1 pm

10 10 10

FSC-A

Comp-B3-A-

4.53

Comp-B3-A*

95.5

10 10 10 Como-B3-A

Flow cytometric signal plots and gating of signal peaks of 1 pm and 0.1 pm fluorescence polystyrene (PS). (a) Supernatant of boiled water (180 mg L^_1 of CaCO3) without 1 pm PS (PS-blank); (b) PS standard solution (1 pm, 1mg L^_1; Standard 1 pm); (c) Supernatant of PS polluted boiled water (1 pm, 1mg L^_1, 180 mg L¹ of CaCO3; PS-water 1 pm); (d) Supernatant of boiled water (180 mg L^_1 of CaCO3) without 0.1 pm PS (PS-blank); (e) PS standard solution (0.1 pm, 1mg L^_1; Standard 0.1 pm); and (f) Supernatant of PS polluted boiled water (0.1 pm, 1mg L^_1, 180 mg L^_1 of CaCO3; PS-water 0.1 pm). B3-A+ indicated fluorescence positive signal, and the quantification of fluorescence PS was analyzed on the positive signal area.

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10          20           30           40           50           60           70

2Theta (degree)

Figure S4. Morphology and composition of incrustants in different conditions. (a)

Scanning electron microscopic (SEM) images of bare-polystyrene (PS, 1 |im, 1 mg L^_1) and incrustant coprecipitates formed in tap water at different temperatures (180 mg L¹ of CaCO3, 40 mL, 25-100 ^oC); (b) SEM images of bare-PS (1 |im, 1 mg L^_1) and incrustant coprecipitates in different water hardness upon boiling (6^300 mg L^_1 of CaCO3, 100 ^oC); (c) SEM images of bare-PS and incrustant coprecipitates in different PS concentrations (1 |im, 0-5 mg L^_1) upon boiling of tap water (180 mg L^_1 of CaCO3, 100 ^oC); and (d) SEM images and (e) X-ray diffraction patterns of bare-, carboxyl-, and amino- PS and incrustant coprecipitates upon boiling of tap water (1 and 0.1 ^m, 1 mg

L¹, 180 mg L¹ of CaCO3, 100 ^oC).

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NMP-incrustant

in boiled water

Encapsulated NMPs

co-precipitation Aggregate & precipitate

Figure S5. Mechanism governing the coprecipitation of nano/microplastics (NMPs) and calcium carbonate (CaCO3) incrustants upon boiling.

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CO

NMP

Boiling

Interracial nucleation

HoCO

Boiled is Safer

Tap water-------- ► HCO

CaCOi nuclei

Ca²

Crystal growth

NMP separation

Figure S6. Proportion of energy consumption and carbon dioxide (CO2) emissions from boiling water using electric kettles (electricity) and gas stoves (natural gas).

(a) Annual average energy demand to boil water by electric kettle and/or gas stove accounted for lower than 1% of the annual total energy consumption; (b) annual average CO2 emissions from consumption of electricity and/or natural gas upon boiling water accounted for lower than 4% of the annual total CO2 emissions from energy consumption.

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a)

b)

Proportion of energy consumption (%)

1.2

1.0

0.8

0.6

0.4

0.2

0.0

o Electricity

Natural gas

• to
• ⁴

0)

CN

o 3

O

**-

o

8 ²

tr

o

1. -1

• ¹

fill

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