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org/estwater Article

A Preliminary Study of Microplastic Abrasion from the Screw Cap

System of Reusable Plastic Bottles by Raman Microspectroscopy
Anja Giese, Jutta Kerpen,* Felix Weber, and Jürgen Prediger

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ABSTRACT: The packaging material of bottled water is

suspected to be a source of microplastics (MP). In this preliminary
study, the screw cap system was examined for its MP release
potential. Therefore, the concentration of MP (≥10 μm) was
Downloaded via 201.103.218.218 on April 10, 2023 at 04:03:10 (UTC).

determined in reusable polyethylene terephthalate (PET) bottles

with polypropylene (PP) caps and polyethylene (PE) seals after
opening and closing once and 11 times. The entire contents of the
bottles were filtered onto a silicon filter after opening. The inside
of the bottles and caps was rinsed with filtered water. The total Si
filter area was analyzed by Raman microspectroscopy. After one
opening, 131 ± 25 microplastic particles (MPP) per liter were
detected. After 11 openings and closings, 242 ± 64 MPP/L were
detected. The increase is caused by a significant increase in the
number of PP particles from 100 ± 27 to 185 ± 52 MPP/L. The concentration of PE and PET particles did not change significantly.
Regardless of polymer type and the number of openings and closings, 80% of the MPP were identified in the smallest size class
investigated (10−50 μm). Using a specially developed method, the concentration of MPP was determined before the screw cap
system was opened and did not differ from the procedural blank.
KEYWORDS: Raman microspectroscopy, microplastic, mineral water, plastic bottles, screw cap system

1. INTRODUCTION obtain water from surface waters.9 These data suggest that MP
Microplastics (MP) were found in numerous aquatic matrices, concentrations in bottled waters may be dependent on source,
e.g., in marine water1 and rivers.2 As the ingestion of transport, and treatment.
microplastic particles (MPP) is proven3 and the WHO In addition to the water in the bottle, the packing system
demanded more research on MP in drinking water,4 the was investigated as a source for MP in a few studies.
investigation of MP in beverages is increasingly coming into Schymanski et al. analyzed 118 ± 88 MPP/L (≥5 μm) in
focus. reusable and 14 ± 14 MPP/L in single-use polyethylene
The first studies of MP in drinking water indicate that the terephthalate (PET) bottles by Raman microspectroscopy.
MP concentration in drinking water depends on the source, This concentration was not different from the blank value; 84%
transport, and treatment of the water: Mintenig et al. identified of the MPP was made of polyester (PEST). It was assumed
0.7 MP/m3 (≥20 μm) in drinking water from Lower Saxony that reusable bottles release more microplastics due to abrasion
(Germany).5 Weber et al. detected no MP (≥10 μm) in the because the bottles are exposed to higher levels of stress due to
drinking water of a German city.6 Kirstein et al. also found no their reuse. Schymanski et al. discussed the screw cap system as
MP (≥6.6 μm) at the outlet of a Swedish waterworks.7 Pittroff a possible abrasion source and supposed that MP could enter
et al. determined no significant concentrations of MP (≥5 μm) reusable bottles via the cleaning and refilling process.10
in raw water, drinking water, and tap water from two German In a further study, Oßmann et al. analyzed single-use and
waterworks.8 All four studies investigated drinking water from reusable PET bottles for MP (≥1 μm) by Raman micro-
groundwater or infiltrated groundwater sources.5−8 Kirstein et
al. showed that the transport of drinking water can lead to MP Received: November 9, 2020
emissions (from 19 ± 14 to 809 ± 688 MP/m3).7 Pivokonsky Revised: April 23, 2021
et al. determined concentrations of 14000 ± 1000 and 151000 Accepted: April 26, 2021
± 4000 MP/m3 (≥1 μm) at the outlet of two Czech Published: May 10, 2021
waterworks (30% ≥ 10 μm), despite reduction rates by
treatment of 88% and 40%, respectively. The waterworks
© 2021 The Authors. Published by
American Chemical Society https://doi.org/10.1021/acsestwater.0c00238
1363 ACS EST Water 2021, 1, 1363−1368
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spectroscopy. In single-use bottles, 2689 ± 4371 MP/L were Shore hardnesses of the respective material are as follows: PE-
detected. Newish and older bottles were analyzed separately. LE, D 45; PP, D 70; and semicrystalline PET, D 83. However,
While the MP concentration in newish bottles was comparable PET bottles are made of amorphous PET, which has a
to that of single-use ones, the concentration in older bottles hardness that is lower than that of semicrystalline PET.14 A
was higher (8339 ± 7043 MP/L). In all cases, the predominant brand of mineral water with a low mineral content was chosen
polymer was PET (>80%).11 The results support the to avoid mineral precipitates on the filter.
assumption of Schymanski et al. that the reuse of PET bottles 2.3. Sample Processing. To examine the impact of the
leads to a higher concentration of MP in the beverages.10,11 number of opening and closing, five bottles were opened once
Because MP was also detected in glass bottles, Oßmann et al. and three bottles 11 times, respectively. According to the
assumed the cleaning process as the most probable source for standard DIN 55406,15 a torque of 1.5 Nm was applied for
MP contamination.11 closing. The bottle was turned upside down after every closing,
Both Schymanski et al. and Winkler et al. opened the bottles so the water came in contact with the cap.
once before analysis. The influence of reclosing was not The contents of each bottle were poured into a 1 L glass
investigated.10,11 funnel connected to a tapered custom-made stainless steel
Winkler et al. tested the influence of mechanical stress on funnel to concentrate particles onto a 5 mm diameter area on
the thread and cap of plastic bottles by opening different the silicon analysis filter (pore width of 10 μm, SmartMem-
single-use PET bottles with a polyethylene (PE) cap 1, 10, and branes GmbH). To seal the filtration apparatus and protect the
100 times. Scanning electron microscopy showed signs of silicon filter, the filter was placed between two silicone mats
abrasion and particles at the thread and cap. Randomly (see Figure 1). The inside of the bottle cap was rinsed with
selected particles were identified as MP by dispersive X-ray
spectroscopy. These results show that the screw cap system is a
potential source of MP in bottled beverages. Winkler et al. did
not investigate how many MPP from the screw cap system
enter the water inside the bottle.12
This preliminary study investigated the screw cap system of
reusable PET bottles as a source of MP. The water of reusable
mineral water bottles was analyzed after one and 11 openings
and closings. To verify the MP concentration before opening
for the first time (after cleaning and filling at the bottling
plant), the MP concentration in the bottle was analyzed
without using the screw cap system.

2. MATERIALS AND METHODS

2.1. Avoidance of Contamination. During sample
preparation, cotton lab coats and cleanroom nitrile gloves
(Simtec, IAB-Reinraum-Produkte GmbH) were worn. All steps
of sample preparation were performed in a laminar flow box
(MSC Advantage 12, Thermo Fisher Scientific Inc.). With the
exception of a squirt bottle made of perfluoroalkoxy alkane
(PFA), fluorinated ethylene propylene (FEP) and ethylene
tetrafluoroethylene (ETFE) (Brand GmbH + Co. KG),
polytetrafluoroethylene (PTFE)-coated tweezers, and silicon
Figure 1. Set-up of the filtration apparatus for transferring samples to
seals, all labware was made of glass or stainless steel. These the analysis filter.
polymers were not considered in the analysis. Filtered
deionized water was used for rinsing steps. It was vacuum
filtrated through two fiberglass filters with pore widths of 0.6 filtered deionized water into the glass funnel. Subsequently, the
μm (MN 85/70 BF, Macherey-Nagel GmbH & Co. KG) and thread of the bottle was rinsed in the same way. The inner
1.2 μm (516-0870, VWR International LLC) and a bottle walls were rinsed with approximately 50 mL of filtered
polycarbonate filter (PC) with a pore size of 8 μm deionized water. The bottle was swayed to remove particles
(TETP04700, Merck KGaA). The PC filter prevents from the walls. The content of the bottle was poured into the
contamination with glass fibers. Preliminary tests showed glass funnel with a rotating motion. The bottle was rinsed a
that the usage of PC filters does not lead to PC contamination second time in the same way. The water in the glass funnel was
of the filtered medium. To detect contamination, blank values filtered. The glass funnel was rinsed with approximately 100
were determined during the main test series (see section 2.4). mL of filtered deionized water.
2.2. Examined Bottle Type. As Winkler et al. showed that After filtration, the filter was transferred to a Petri dish. The
more particles are released from bottles with a discontinuous filter was dried overnight in the laminar flow box. For analysis,
thread than with a continuous thread,12 in this study a bottle the filter was transferred to a glass slide.
with a discontinuous thread was chosen. The PET bottle from 2.4. Blank Values. 2.4.1. Mineral Water Blanks. To
Genossenschaft Deutscher Brunnen (GDB) is the most determine the amount of PET in unopened bottles, a hole was
common reusable bottle in Germany. The 1 L bottle can be melted in the cap of two bottles using a heated metal tip. To
reused up to 25 times.13 The bottle consists of a PET body, a determine the amount of PE and PP, two holes were melted
polypropylene (PP) cap, and a PE seal that is inserted into the into the wall of two different bottles. The second hole was
cap. The cap with the seal is replaced during each filling. The required to allow deaeration while water was poured out of the
1364 https://doi.org/10.1021/acsestwater.0c00238
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Table 1. MPP Detected in the Procedural Blanks

MPP/sample
polymer procedural blank 1 procedural blank 2 procedural blank 3 procedural blank 4 procedural blank 5 Ø ± SD
PP 2 0 2 1 0 1.0 ± 1.0
PET 0 0 1 1 0 0.4 ± 0.5
PE 1 0 1 0 0 0.4 ± 0.5
total MP 3 0 4 2 0 1.8 ± 1.8

other hole. With the melted hole as a pourer, the bottle 3. RESULTS
contents were transferred to an analysis filter as described in 3.1. Procedural Blanks. On average, 1.8 ± 1.8 MPP/
section 2.3. sample were detected. The detected polymers were PP, PET,
2.4.2. Procedural Blanks. Five procedural blanks were and PE (see Table 1).
determined by simulating the steps of the filtration process. A 3.2. Mineral Water Blanks. In the mineral water blanks
closed empty bottle was used to simulate the movements of for PP and PE, 1−8 MPP/filter were detected. The blanks for
rinsing the bottle. The rinsing water for the bottle was poured PET contained 3 and 6 MPP/filter (see Table 2).
directly from the squirt bottle into the glass funnel.
2.5. Raman Microspectroscopy. Analyses were con- Table 2. MPP Detected in the Mineral Water Blanks
ducted with a Raman microspectrometer with a front-
illuminated EMCCD detector (DXR 2xi, Thermo Fisher MPP/sample
Scientific Inc.) and OmnicXi (Thermo Fisher Scientific Inc.). melting hole in the bottle melting hole in the cap
An optical image was generated at 100× magnification. The mineral water mineral water mineral water mineral water
spectroscopic parameters were set as follows: a laser wave- polymer blank 1 blank 2 blank 1 blank 2
length of 785 nm, a standard laser power of 15 mW, an PP 1 7 not considered
exposure time of 0,05 s, and a repetition rate of 50. PET not considered 3 6
Measurements were conducted with a 50 μm slit aperture, a PE 0 1 not considered
10× objective, and a grid of 400 lines/mm. Spectra were
recorded in a range of 50−3250 cm−1 with a resolution of 5 3.3. Microplastic Concentration after Opening and
cm−1. When the spectrum was too indistinct, the particle was Closing a Bottle. After one opening, 131 ± 25 MPP/L were
analyzed at different points and the laser intensity was detected, whereas 11 openings and closing caused a
increased to 20 mW. All particles ≥10 μm on the analysis significantly higher concentration of 242 ± 64 MPP/L. The
filter were analyzed. concentrations of PET- and PE-MPP did not change
Spectra of the bottle materials were recorded in advance. significantly between one and 11 openings. Sixteen ± 12
Measured spectra were compared with the previously recorded PET-MPP/L and 15 ± 7 PE-MPP/L were analyzed after one
spectra and with the library P/N L60001 (S. T. Japan-Europe opening, and 32 ± 22 PET-MPP/L and 25 ± 20 PE-MPP/L
GmbH). Spectra were accepted if the match factor was ≥0.8. were analyzed after 11 openings. The concentration of the
2.6. Statistical Analysis. 2.6.1. Comparison of Proce- predominant PP-MPP increased significantly from 100 ± 27 to
dural and Mineral Water Blanks. Because the mineral water 185 ± 52 MPP/L (see Figure 2).
blanks were determined twice, a comparison of the procedural Regardless of the number of openings and closings and the
and mineral water blanks with statistical parameter tests was type of polymer, approximately 80% of all MPP are in the size
not possible. According to the standard DIN 32645,16 the limit range between 10 and 50 μm (see Figure 3).
of quantification was estimated as
4. DISCUSSION
limit of quantification = yblank + 10 × SDblank 4.1. Procedural Blanks. The number of ≥10 μm MPP in
the procedural blank is small, which shows the effectiveness of
where yblank is the average procedural blank and SDblank is the the measures to prevent contamination. As the blanks capture
standard deviation of the procedural blank. The analysis result contamination from rinsing water as well as from airborne
of a mineral water blank was considered significantly above the particles and contaminated labware, the results of the blanks
procedural blank if it was higher than the limit of are reported as MPP/sample instead of MPP/L. Schymanski et
al. observed a rinsing water blank of 14 ± 13 MPP/L,10 and
quantification.
Oßmann et al. a value of 384 ± 468 MPP/L,11 considering
2.6.2. Comparison of Procedural Blanks and Analysis
sizes down to 5 and 1 μm, respectively. As the number of
Results. The results of the opening/closing test series were
particles increases with a decrease in the particle diameter,8,11
statistically compared with the procedural blank values. The the different blanks are hardly comparable.
MPP numbers were logarithmized on base 10. The variances of 4.2. Mineral Water Blanks. None of the mineral water
both test series were compared with an F-test variance analysis blanks could be distinguished significantly from the procedural
(p = 0.05). In case of the same variances, a one-sided t test (p blank. It can be assumed that no or a negligible number of
= 0.05) was performed to check if the analysis results differ. In MPP (≥10 μm) were present in the bottles before the first
the case of different variances, Welch’s test (p = 0.05) was opening. As only one type of bottle and one brand of water
performed.6 The same procedure was applied to check whether were examined, this conclusion cannot be applied to other
opening 11 times resulted in a significantly higher level of bottles or brands of water with absolute certainty. Because all
particle release than opening once. reusable PET bottles are very similar, it is very likely that MP is
1365 https://doi.org/10.1021/acsestwater.0c00238
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Figure 2. Average concentration and standard deviation of total MPP and single-polymer MPP after one and 11 openings of a bottle.

Figure 3. Average size distribution of the detected MPP after (A) one and (B) 11 openings and its standard deviation.

also released from the cap of other types of bottles. Schymanski PET thread during the first opening. In subsequent openings
et al. and Oßmann et al. discussed the cleaning and refilling and closings, the edges may already have been abraded and
processes as a possible source for MP in bottled water. Because therefore fewer particles are produced.
the predominant polymer in reusable PET bottles was PET, Contrary to the results of this study, Schymanski et al. and
they assumed the possibility of release of PET particles from Oßmann et al. identified PET as the most common polymer in
the bottle during the cleaning process.10,11 Alternatively, reusable PET bottles.10,11 There are two possible reasons for
Oßmann et al. discussed contaminated cleaning agents or this finding. First, both studies examined several types of
machines. This assumption was strengthened by the detection bottles. It is unknown if the GDB bottle was one of them. It is
of nonpackaging material in reusable glass bottles.11 For this possible that other bottles show a different abrasion behavior.
study, it can be assumed that all MPP detected in the opening The more likely reason is that the cap and bottle thread in this
and closing test series are released from the screw cap system.
study were rinsed after opening, while Oßmann et al. and
4.3. Particle Release from the Screw Cap System. All
Schymanski et al. did not rinse them.10,11 Apart from the bottle
analysis results are significantly higher than those of the
procedural blank. As the mineral water blank is not and cap threads, there might be another origin of the PP
significantly distinguishable from the procedural blank, it is particles. When the bottles are first opened, the tamper-evident
evident that all analysis results are significantly higher than the ring on the cap breaks at predetermined breaking points. As
mineral water blank. Therefore, the release of MPP from the the caps were rinsed, these particles could have been flushed
screw cap system is proven. Whether this also applies to other onto the filter.
bottle types must be confirmed by examining different bottles. Although PP residues from the cap thread do not necessarily
The concentration of MPP increases with the number of enter the water in the bottle, their release is a possible exposure
openings and closings. However, a high percentage is released pathway for consumers. The external thread of the bottle is
during the first opening. This could be attributed to the fact brought to the mouth for drinking or is used to pour the
that the softer PP cap threads are most abraded by the harder contents of the bottle into glasses, for example.
1366 https://doi.org/10.1021/acsestwater.0c00238
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Figure 4. (A) Spectrum of a MPP (green), compared with the reference spectrum of the bottle cap (white). Despite a strong copper
phthalocyanine spectrum, PP could be identified with the two peaks at 809 and 841 cm−1. (B) Spectra of copper phthalocyanine (red) and PP
(violet).

Schymanski et al. and Oßmann et al. analyzed ≥5 and ≥1 was blue. In addition to the peaks of PP, the particles from the
μm MPP, respectively, and opened each bottle just once.10,11 cap showed a clear spectrum of the pigment copper
To compare their data with those of this study (which analyzed phthalocyanine (see Figure 4). By using reference spectra
>10 μm MPP), only data for MPP ≥10 μm from the other two generated with the original bottle material (see section 2.5),
studies were used. Within this range, Schymanski et al.
MPP from the cap could be clearly identified.
reported 52 MPP/L, representing 44% of their total MPP
amount per liter (118 ± 88 MPP/L),10 and Oßmann et al. 83 The analysis of MPP <10 μm is often not possible without
± 136 MPP/L.11 In this study, 131 ± 25 MPP/L were found. partial measurements of the filter area and extrapolation due to
Even considering the possible influence of rinsing the cap and their high abundance. Brandt et al. showed that analyses of
thread, the results of this study are in a size range comparable filter sub-areas and extrapolation can lead to over- and
to that of the results of Schymanski et al. and Oßmann et al. underestimation.17 In this study, the analysis of MPP ≥10
The observed size distribution of MPP, in which the 10−50 μm was performed by measuring the whole filter area. Errors
μm fraction is the largest, also confirms the results of by extrapolation cannot occur in this study.
Schymanski et al. and Oßmann et al.10,11 Even the fact that
MPP <10 μm were also analyzed in these two studies, all three
results show that the number of MPP increases with a decrease 5. CONCLUSION
in particle diameter. On the basis of the results of Schymanski The release of MP (≥10 μm) from the screw cap system was
et al. and Oßmann et al., it can be assumed that a large number investigated by examining the most commonly used reusable
of MPP <10 μm are released by the screw cap system, although PET bottle brand in Germany. The screw cap system was
this was not investigated in this study.
identified as a source of MP in bottled water. PET from the
4.4. Method Assessment. The method of producing the
mineral water blank value is well suited for determining the bottle, PP from the cap, and PE originating from the seal were
MP concentration in a bottle without opening its screw cap released by opening and closing the cap. PP was the
system. Melting the pouring holes is less likely to produce predominant type of MPP. The release of PP−MPP increases
MPP than cutting or drilling bottles. For the determination of with the number of openings. Independent of the polymer type
the PET blank value, a hole was melted into the cap and the and number of openings and closings, approximately 80% of
seal (made of PP and PE). For the determination of the PE/PP the MPP were in the size range of 10−50 μm. As only one type
blank value, a hole was melted into the bottle wall (made of of bottle was examined, these results need to be verified for
PET). Thus, the MPP released during melting did not
other bottle types in further studies. In addition to the
influence the result.
While PET bottles and seals in caps are often transparent, suspected sources such as release from the bottle walls or
bottle caps are usually colored. If the amount of pigments in a contamination during preparation of the bottles for reuse,
polymer/pigment compound is large, the pigment spectrum further studies should consider the screw cap system as a
could overlay the polymer spectrum. In this study, the PP cap source of MP.
1367 https://doi.org/10.1021/acsestwater.0c00238
ACS EST Water 2021, 1, 1363−1368
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pubs.acs.org/estwater Article

AUTHOR INFORMATION (10) Schymanski, D.; Goldbeck, C.; Humpf, H.-U.; Fürst, P. Analysis
of microplastics in water by micro-Raman spectroscopy: Release of
Corresponding Author plastic particles from different packaging into mineral water. Water
Jutta Kerpen − Institute for Environmental and Process Res. 2018, 129, 154−162.
Engineering, RheinMain-University of Applied Sciences, (11) Oßmann, B. E.; Sarau, G.; Holtmannspötter, H.; Pischetsrieder,
65022 Wiesbaden, Germany; orcid.org/0000-0002- M.; Christiansen, S. H.; Dicke, W. Small-sized microplastics and
5652-4095; Email: jutta.kerpen@hs-rm.de pigmented particles in bottled mineral water. Water Res. 2018, 141,
307−316.
Authors (12) Winkler, A.; Santo, N.; Ortenzi, M. A.; Bolzoni, E.; Bacchetta,
R.; Tremolada, P. Does mechanical stress cause microplastic release
Anja Giese − Institute for Environmental and Process
from plastic water bottles? Water Res. 2019, 166, 115082.
Engineering, RheinMain-University of Applied Sciences, (13) Genossenschaft Deutscher Brunnen. Mehrweg. https://www.
65022 Wiesbaden, Germany gdb.de/mehrweg/ (accessed 2021-02-05).
Felix Weber − Institute for Environmental and Process (14) Fritsche, H.; Fritsche, C.; Gradl, W.; Kolbinger, J.; Küspert, K.-
Engineering, RheinMain-University of Applied Sciences, H.; Dietmar, M.; Ulrike, R.; Schmiedt, A.; Schwarze, F. Tabellenbuch
65022 Wiesbaden, Germany Kunststofftechnik, 1st ed.; Europa-Fachbuchreihe für Kunststoffberufe,
Jürgen Prediger − Institute for Environmental and Process Verlag Europa-Lehrmittel: Haan-Gruiten, 2016.
Engineering, RheinMain-University of Applied Sciences, (15) Deutsches Institut für Normung e.V. Packmittel - Spezielle
65022 Wiesbaden, Germany Technische Liefer- und Bezugs- sowie Verwendungsbedingungen für ein-
bzw. zweiteilige 28-mm-Schraubverschlüsse aus Polyethylen (PE) bzw.
Complete contact information is available at: aus Polypropylen (PP) und 28-mm-Anrollverschlüsse aus Aluminium
https://pubs.acs.org/10.1021/acsestwater.0c00238 (Al); Beuth Verlag: Berlin, 2015; 55.100 (55406).
(16) Deutsches Institut für Normung e.V. Chemische Analytik -
Notes Nachweis-, Erfassungs- und Bestimmungsgrenze unter Wiederholbedin-
The authors declare no competing financial interest. gungen - Begriffe, Verfahren, Auswertung; Beuth Verlag: Berlin, 2008;

■
71.040.01 (DIN 32645).
(17) Brandt, J.; Fischer, F.; Kanaki, E.; Enders, K.; Labrenz, M.;
ACKNOWLEDGMENTS Fischer, D. Assessment of Subsampling Strategies in Microspectro-
The authors thank Dukagjin Metaj for his groundwork scopy of Environmental Microplastic Samples. Front. Environ. Sci.
regarding the preparation of the samples and the method of 2021, 8, 579676.
filtering the mineral water. The authors also thank René
Langer for his help, when it was needed, and Sebastian Wolff
for his support.

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