Simultaneous Quantification of Eight Marker Components in Traditional Herbal Formula, Haepyoyijin-Tang Using HPLC–PDA
Round 1
Reviewer 1 Report
in the validation of the method, why did not the authors perform
- robustness and
- selectivity>
Author Response
Comments and Suggestions for Authors
in the validation of the method, why did not the authors perform
- robustness and
Answer) We appreciate your comment. In this study, the robustness was performed for the conditions of the mobile phase and the column types. However, it did not show a difference, so it was not represented as data.
- selectivity
Answer) We appreciate your comment. In this study, the selectivity was expressed as relative retention (a) (Table S2). In general, relative retention (a) is often used synonymously with selectivity.
Reviewer 2 Report
The manuscript described two analytical methods for quality control of a herbal preparation containing 12 herbs. Eight chemicals arise from 5 of the 12 herbs have been selected as chemical markers. However, the rationale that justify the choice of the chemical markers are not presented in the manuscript. It seems that the relative weight of the herbs in the preparation is not known in here. Some of the chemicals presence in many different herbs (e.g rosmarinic acid, hesperidin). Glycyrrhizae Radix et Rhizoma present in many different herbal preparation, thus, usually it does not represent a major role in the therapeutic effects of a preparation. I am not sure whether this also apply to HPYJT.
It seems that HPLC-PDA method is already fit for the purpose. It is not clear why the authors would need the LC-MS method in here? In Table 4, the unit of the number are mg/g, I suppose that refers to mg of certain compound in 1 g of sample. It is very strange that the content of the markers in the samples differ by 10 fold using HPLC-PDA and LC-MS. Did the authors forget a dilution factor in the LC-MS results? The manuscript looks more logical when the LC-MS method and the data are removed.
Author Response
Comments and Suggestions for Authors
The manuscript described two analytical methods for quality control of a herbal preparation containing 12 herbs. Eight chemicals arise from 5 of the 12 herbs have been selected as chemical markers. However, the rationale that justify the choice of the chemical markers are not presented in the manuscript. It seems that the relative weight of the herbs in the preparation is not known in here. Some of the chemicals presence in many different herbs (e.g rosmarinic acid, hesperidin). Glycyrrhizae Radix et Rhizoma present in many different herbal preparation, thus, usually it does not represent a major role in the therapeutic effects of a preparation. I am not sure whether this also apply to HPYJT.
It seems that HPLC-PDA method is already fit for the purpose. It is not clear why the authors would need the LC-MS method in here? In Table 4, the unit of the number are mg/g, I suppose that refers to mg of certain compound in 1 g of sample. It is very strange that the content of the markers in the samples differ by 10 fold using HPLC-PDA and LC-MS. Did the authors forget a dilution factor in the LC-MS results? The manuscript looks more logical when the LC-MS method and the data are removed.
Answer) We appreciate your comment. In accordance with opinions of the reviewer, we removed the LC-MS method and the data from the manuscript.
Reviewer 3 Report
The aim of this paper is with the scope of the journal. In this manuscript a new and efficient analytical procedure for the purpose of quality control of simultaneous quantification of eight marker components in traditional herbal formula, Haepyoyijin-Tang (HPYJT) using the two modern and recommended for the purpose of this kind of study chromatographic methods, namely HPLC–PDA and LC- MS/MS were well described.The chromatographic methods have been full checked during validation process in accordance with the International Conference on Harmonisation Guidelines The obtained validation results confirm suitability of the two procedures for the purpose of quality control of investigated traditional herbal prescription frequently used in patients with asthma in Korea.
This manuscript is well prepared. However a small suggestion of reviewer's is:
- to compare the efficacy of the two procedures and general conclusion should be added about them i.e. HPLC-PDA and HPLC-MS/MS on the basis of obtained results of quantitative analysis of the analyzed markers
- to reduce the size of Tables, especially Table 3 to better see results
Author Response
Comments and Suggestions for Authors
The aim of this paper is with the scope of the journal. In this manuscript a new and efficient analytical procedure for the purpose of quality control of simultaneous quantification of eight marker components in traditional herbal formula, Haepyoyijin-Tang (HPYJT) using the two modern and recommended for the purpose of this kind of study chromatographic methods, namely HPLC–PDA and LC- MS/MS were well described.The chromatographic methods have been full checked during validation process in accordance with the International Conference on Harmonisation Guidelines The obtained validation results confirm suitability of the two procedures for the purpose of quality control of investigated traditional herbal prescription frequently used in patients with asthma in Korea.
This manuscript is well prepared. However, a small suggestion of reviewer's is:
1.to compare the efficacy of the two procedures and general conclusion should be added about them i.e. HPLC-PDA and HPLC-MS/MS on the basis of obtained results of quantitative analysis of the analyzed markers
Answer) We appreciate your comment. In accordance with opinions of the reviewer 2, we removed the LC-MS method and the data from the manuscript. Therefore, there is no need to compare the HPLC-PDA and LC-MS/MS.
- to reduce the size of Tables, especially Table 3 to better see results
Answer) We appreciate your comment. Tables 1, 2, 3 are very important data I the method validation of this study and cannot be briefly described any more. Please understand.
