Simultaneous Determination of 12 Marker Components in Yeonkyopaedok-san Using HPLC–PDA and LC–MS/MS

Round 1

Reviewer 1 Report

This MS presents an approach based on HPLC-DAD and HPLC-MS/MS for the simultaneous determination of twelve marker components in a traditional Korean herbal formula.

The MS is well written and interesting for researchers working in the filed of herbal medicine.

I only suggests some revision:

• l. 37: I do not understand the ration
• ll. 126-130: please write equations not as inline text
• Figure 1: please remove some chromatogram recorded at the difference wavelenght. The information is redundant. Maybe put it in the SI.
• ll. 169-171: please write large numbers in scientific notation
• Table 1: please write regression eqs in scientific notation (see before)
• conclusions: please rewrite conclusions. What does "most abundant" mean? I think it should be more correct to say that one technique is more sensitive towards the detection of certain analytes with respect to the other

Author Response

Comments and Suggestions for Authors

This MS presents an approach based on HPLC-DAD and HPLC-MS/MS for the simultaneous determination of twelve marker components in a traditional Korean herbal formula.

The MS is well written and interesting for researchers working in the filed of herbal medicine.

 

I only suggests some revision:

 

1. 37: I do not understand the ration

Answer) We appreciate your comment. We removed the phrase “in a 1.75:1.75:1.75:1.75:1.75:1.75:1.75:1.75:1.75:1.75:1.75:1.75 :1.75:1.00 ratio based on dry weight”.

 

1. 126-130: please write equations not as inline text

Answer) We appreciate your comment. We have written using equations.

 

Figure 1: please remove some chromatogram recorded at the difference wavelenght. The information is redundant. Maybe put it in the SI.

Answer) We appreciate your comment. We have modified Figure 1.

 

1. 169-171: please write large numbers in scientific notation

Answer) We appreciate your comment. We have written in scientific notation.

 

Table 1: please write regression eqs in scientific notation (see before)

Answer) We appreciate your comment. We have written regression equations in scientific notation.

 

conclusions: please rewrite conclusions. What does "most abundant" mean? I think it should be more correct to say that one technique is more sensitive towards the detection of certain analytes with respect to the other

Answer) We appreciate your comment. We have deleted a sentence that includes “most abundant”.

Reviewer 2 Report

Yeonkyopaedok-san, also known as Lian Qiao Bai Du San in chinese, is a traditional Chinese medicine for treatment Initial stage of abscesses and carbuncles due to toxic heat. To test the components is a meaningful work. There are several issues the authors need to address to improve the manuscript.

 

 

1 In traditional Chinese medicine, Lian Qiao Bai Du San supports the health of the skin, the gastrointestinal, circulatory, and immune systems. The background and medical application of Yeonkyopaedok-san (YPS) should introduced in detail.

 

2 Whether the 12 marker components have medical effects separately, and which is the mainly active ingredients for medical application? the author should discuss them.

 

3 The title is too complicated; it should be simplified.

 

Author Response

Comments and Suggestions for Authors

Yeonkyopaedok-san, also known as in chinese, is a traditional Chinese medicine for treatment Initial stage of abscesses and carbuncles due to toxic heat. To test the components is a meaningful work. There are several issues the authors need to address to improve the manuscript.

  

1. In traditional Chinese medicine, Lian Qiao Bai Du San supports the health of the skin, the gastrointestinal, circulatory, and immune systems. The background and medical application of Yeonkyopaedok-san (YPS) should introduced in detail.

 Answer) We appreciate your comment. We tried to search for “Lian Qiao Bai Du San” in PubMed, Google Scala, and CNKI to add an introduction to YPS. However, no documents match our search term. Therefore, we regret to inform you that no detailed introduction to “Lian Qiao Bai Du San” is no longer possible.

 

2. Whether the 12 marker components have medical effects separately, and which is the mainly active ingredients for medical application? the author should discuss them.

 Answer) We appreciate your comment. The 12 components commonly have an anti-inflammatory effect [1-10]. In addition, these components are effective against a variety of diseases, so it is not known which components are effective in YPS. Perhaps, a number of components that are characteristic of traditional Chinese medicine are thought to be complementary in their efficacy. In addition, this study focuses on quantitative analysis for quality control of YPS, not on efficacy evaluation of each marker component.

 

1. Naveed, M.; Hejazi, V.; Abbas, M.; Kamboh, A.A.; Khan, G.J.; Shumzaid, M.; Ahmad, F.; Babazadeh, D.; FangFang, X.; Modarresi-Ghazani, F.; WenHua, L.; XiaoHui, Z. Chlorgenic acid (CGA): A pharmacological review and call for further research. Biomed Pharmacother 2018, 97, 67–74.
2. Zhou, J.; Sun, Y.Y.; Sun, M.Y.; Mao, W.A.; Wang, L.; Zhang, J.; Zhang, H. Prim-O-gluycosylcimifugin attenuates lipopolysaccharide-induced inflammatory response in RAW 264.7 macrophages. Pharmacogn Mag 2013, 13, 378–384.
3. Mahmoud, A.M.; Hussein, O.E.; Hozayen, W.G.; Bin-Jumah, M.; Abd EI-Twab, S.M. Ferulic acid prevents oxidative stress, inflammation, and liver injury via upregulation of Nrf2/HO-1 signaling in methotrexate-induced rats. Environ Sci Pollut Res Int 2019, doi: 10.1007/s11356-019-07532-6.
4. Li, W.; Wang, Z.; Sun, Y.; Chen, L.; Han, L.; Zheng, Y. Application of response surface methodology to optimise ultrasonic-assisted extraction of four chromones in Radix Saposhnikoviae. Phytochem Anal 2011, 22, 313–321.
5. Mao, Z.; Gan, C.; Zhu, J.; Ma, N.; Wu, L.; Wang, L; Wang, X. Anti-atherosclerotic activities of flavonoids from the flowers of Helichrysum arenarium L. MOENCH through the pathway of anti-inflammation. Bioorg Med Chem Lett 2017, 27, 2812–2817.
6. Yin, L.; Guan, E.; Zhang, Y.; Shu, Z.; Wang, B.; Wu, X.; Chen, J.; Liu, J.; Fu, X.; Sun, W.; Liu, M. Chemical profile and anti-inflammatory activity of total flavonoids from Glycyrrhiza uralensis Fisch. Iran J Pharm Res 2018, 17, 726–734.
7. Hamdan, D.I.; Mahmoud, M.F.; Wink, M.; EI-Shazly, A.M. Effect of hesperidin and neohesperidin form bittersweet orange (Citrus aurantium var. bigaradia) peel on indomethacin-induced peptic ulcers in rats. Environ Toxicol Pharmacol 2014, 37, 907–915.
8. Wang, C.C.; Lai, J.E.; Chen, L.G.; Yen, K.Y.; Yang, L.L. Inducible nitric oxide synthase inhibitors of Chinese herbs. Part 2: Naturally occurring furanocoumarins. Bioorg Med Chem 2000, 8, 2701–2707.
9. Song, J.; Li, N.; Xia, Y.; Gao, Z.; Zou, S.; Kong, L.; Yao, Y.J.; Jiao, Y.N.; Yan, Y.H.; Li, S.H.; Tao, Z.Y.; Lian, G.; Yang, J.X.; Kang, T.G. Arctigenin treatment protects against brain damage through an anti-inflammatory and anti-apoptotic mechanism after needle insertion. Front Pharmacol 2016, 7, 182. Doi: 10.3389/fphar.2016.00182.
10. Hong, M.K.; Hu, L.L.; Zhang, Y.X.; Xu, Y.L.; Liu, X.Y.;He, P.K.; Jia, Y.H. 6-Gingerol ameliorates sepsis-induced liver injury through the Nrf2 pathway. Int Immunopharmacol 2020, 80, 106196. doi: 10.1016/j.intimp.2020.106196.

 

3 The title is too complicated; it should be simplified.

Answer) We appreciate your comment. We have modified the title.

Round 2

Reviewer 2 Report

Lianqiao Baidu pills is the different dosage forms of Lian Qiao Bai Du San in Chinese.

Below are some articles reported Lianqiao Baidu pills about the composition and function. Please discuss the similarities and differences with your YPS.

Quantitative fingerprinting based on the limited-ratio quantified fingerprint method for an overall quality consistency assessment and antioxidant activity determination of Lianqiao Baidu pills using HPLC with a diode array detector combined with chemometric methods. J Sep Sci. 2018 Jan;41(2):548-559.

Micellar electrokinetic chromatography fingerprinting combined with chemometrics as an efficient strategy for evaluating the quality consistency and predicting the antioxidant activity of Lianqiao Baidu pills. J Sep Sci. 2017 Jul;40(13):2838-2848. 

Determination of Baicalin in Lianqiao Baidu pill by HPLC. Liaoning. J. Tradit. Chin. Med. 2013, 40, 756–757.

Improvement of quality standard of Lianqiaobaidu Pill. J. Capit. Med. 2008, 20, 50–52.

Study on Quality Standard for Lianqiaobaidu Pills. Chin. J. Inf. Tradit. Chin Med. 2014, 21, 95–97.

Identification of Lian qiao bai du Pill by Thin Layer Chromatography. J. Chin. Mod. Med. 2010, 17, 111.

Simultaneous Determination of Forsythin, Chrysophanol and Emodin in Lianqiao Baidu Pills by Micellar Electrokinetic Chromatography‐electrospray Ionization Mass Spectrometry. Chem. 2010, 73, 814–818.

Ministry of Public Health of China. National Standard for Chinese Patent Drugs. People's Health Press, Beijing 1989.

Author Response

Lianqiao Baidu pills is the different dosage forms of Lian Qiao Bai Du San in Chinese.

Below are some articles reported Lianqiao Baidu pills about the composition and function. Please discuss the similarities and differences with your YPS.

 Answer) We appreciate your comment. In general, most traditional Chinese medicine, traditional Koran medicine, and Kampo medicine are called Tang (湯), Pill (丸), and San (散) depending on the formulation method. Therefore, we reviewed the references provided by the reviewer. As a result, we found a huge difference in the references. In other words, “San” and “Pill” are different in Yeonkyopaedok-san (YPS). The YPS used in our study consists of 14 different medicines (Forsythiae Fructus, Lonicerae Flos, Schizonepetae Spica, Saposhnikoviae Radix, Osterici seu Notopterygii Radix et Rhizoma, Araliae Continentalis Radix, Bupleuri Radix, Cnidii Rhizoma, Aurantii Fructus Immaturus, Platycodonis Radix, Poria Sclerotium, Glycyrrhizae Radix et Rhizoma, Menthae Herba, and Zingiberis Rhizoma Recens). However, YPP used in China consists of 19 medicinal herbs (Forsythiae Fructus, Lonicerae Flos, Violae Herba, Trichosanthis Radix, Scutellariae Radix, Coptidis Rhizoma, Rhellodendri Cortex, Rhei Radix et Rhizoma, Sohporae Radix, Schizonepetae Spica, Saposhnikoviae Radix, Angelicae Dahuricae Radix, Osterici seu Notopterygii Radix et Rhizoma, Ephedrae Herba, Menthae Herba, Bupleuri Radix, Angelicae Gigantis Radix, Paeoniae Radix, and Glycyrrhizae Radix et Rhizoma, Menthae Herba) [1-5]. These two herbal formulas are called “San” and “Pill” similarly, but they are completely different herbal formula. Therefore, it is not suitable to discuss the YPP in this study.

1. Study on Quality Standard for Lianqiaobaidu Pills. Chin. J. Inf. Tradit. Chin Med. 2014, 21, 95–97.
2. Quantitative fingerprinting based on the limited-ratio quantified fingerprint method for an overall quality consistency assessment and antioxidant activity determination of Lianqiao Baidu pills using HPLC with a diode array detector combined with chemometric methods. J Sep Sci. 2018 Jan;41(2):548-559.
3. Micellar electrokinetic chromatography fingerprinting combined with chemometrics as an efficient strategy for evaluating the quality consistency and predicting the antioxidant activity of Lianqiao Baidu pills. J Sep Sci. 2017 Jul;40(13):2838-2848.
4. Determination of Baicalin in Lianqiao Baidu pill by HPLC. Liaoning. J. Tradit. Chin. Med. 2013, 40, 756–757.
5. Simultaneous Determination of Forsythin, Chrysophanol and Emodin in Lianqiao Baidu Pills by Micellar Electrokinetic Chromatography‐electrospray Ionization Mass Spectrometry. Chem. 2010, 73, 814–818.

Round 3

Reviewer 2 Report

I think the explain is  far-fetched.

Although Lianqiao Baidu pills  consists of 19 medicinal herbs, and YPS consists of 14 medicinal herbs. Are the Yeonkyopaedok-san (YPS),  Lianqiao Baidu pills, and Lianqiao Baidu San treatment for the same diseases or symptoms?

Others has been indentified the quantitative fingerprinting of  Lianqiao Baidu pills, and reported its function. The studies are similar, please  discuss the similarities and differences in your manustript.

Author Response

Comments and Suggestions for Authors

I think the explain is far-fetched.

Although Lianqiao Baidu pills consists of 19 medicinal herbs, and YPS consists of 14 medicinal herbs. Are the Yeonkyopaedok-san (YPS), Lianqiao Baidu pills, and Lianqiao Baidu San treatment for the same diseases or symptoms?

Others has been indentified the quantitative fingerprinting of Lianqiao Baidu pills, and reported its function. The studies are similar, please discuss the similarities and differences in your manustript.

 Answer) We appreciate your comment. We added to the previous study of “Lianqiao Baidu pill” in Introduction section.