This manuscript tries to describe a “rapid designment of compound pH indicator” for high throughput analyses. However, to the best understanding of me, the main point in this manuscript was to build the color ranges of different indicators (and natural pigments) by a new computational method based on Matlab, which does not match the scope of Analytical Methods. Also, I think there are several major problems in this work, which will be detailed below. Therefore, I am afraid I have to reject this manuscript.
1. The authors claimed a new way to design compound pH indicators. However, besides color computation of different indicators, the authors did not mention anything about how to design a “compound pH indicator”. For example, the authors used natural red, methyl orange, bromocresol green, and thymol blue to make the “compound pH indicator” for the bacteria screening. Why did the authors decide to choose these four? Why it has to be four indicators instead of three or two? If I use this method for another application, how should I determine what indicators I should mix to a “compound indicator”? I think this should be a key in the “designment of compound pH indicator”.
2. The authors claimed this method can be applied for high throughput analyses on 96-well plates. However, people usually use 96-well plate for optical analyses, such as UV-vis, fluorescence etc.. If I have limited amount of samples, and I have to measure pH and other molecules by a plate reader. If the compound indicator is mixed with my sample in a 96-well plate, I don’t think it is possible to do afterward optical measurements because of the strong interference from the indicators (chromophores). In this sense, I would say this method is very d...
... middle of paper ...
...eeds to be polished.
2. In the color validation part, the authors claimed accuracy values in percentages. How did the authors obtain these percentage numbers? Just by comparing the computed colors and the true colors by eyes?
3. The highest accuracy of 100% in the text (Page9, Line 51) is not consistent with the number in Figure 2 (97.6%).
4. The author mentioned “workers only need to take few validation experiments for the satisfactive consuming.” How many are “few”? How did the author determine how many validations need to be done?
5. In the sentence of “Organic acids butyrate and acetate”, butyrate and acetate are not acids.
6. The authors did not mention how butanol was measured.
7. Significant figures of numbers are not consistent.
8. In Method part, the authors said “the acids including acetic acid, lactic acid and…”. Did the authors measure all these acids?
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