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. 2019 Nov 28;9:17850. doi: 10.1038/s41598-019-54360-2

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Thin-layer chromatograms of the tobacco samples with and without grey speckled leaves. Note: the left-hand sides of the figures show samples with grey speckles, while the right-hand sides show samples without grey speckles. (a,c) Thin-layer chromatograms without using H₂SO₄-EtOH, (b,d) Thin-layer chromatograms after using 5% H₂SO₄-EtOH. Conditions of (a,b) include a methylene chloride to methanol ratio of 5:1, while those in (c,d) include a methylene chloride: methanol: formic acid ratio of 5:1:0.1.).