Purification of Flavonoids from Mulberry Leaves via High-Speed Counter-Current Chromatography
Round 1
Reviewer 1 Report
The manuscript was very well written and scientifically sound. I offer no criticisms.
But something to maybe think about. I very much liked your discussion of intermolecular interactions and how this guided your rational selection of the solvent blend. It was very nicely done. Reference 24 which you used in a very nice paper. I would just mention that the parameters bear some resemblance to LSER (linear solvation energy relationship) parameters. Recently, a massive, searchable database has become available:
https://www.ufz.de/index.php?en=31698&contentonly=1&m=0&lserd_data[mvc]=Public/start
When I looked just now, LSER parameters are even available for rutin. This could potentially give you a way to quantify the chemical nature of the solute, and relate this to the solvent.
I emphasize again that the present manuscript if great. This just came to mind and I am passing along to you.
Author Response
The manuscript was very well written and scientifically sound. I offer no criticisms.
But something to maybe think about. I very much liked your discussion of intermolecular interactions and how this guided your rational selection of the solvent blend. It was very nicely done. Reference 24 which you used in a very nice paper. I would just mention that the parameters bear some resemblance to LSER (linear solvation energy relationship) parameters. Recently, a massive, searchable database has become available:
https://www.ufz.de/index.php?en=31698&contentonly=1&m=0&lserd_data[mvc]=Public/start
When I looked just now, LSER parameters are even available for rutin. This could potentially give you a way to quantify the chemical nature of the solute, and relate this to the solvent.
I emphasize again that the present manuscript if great. This just came to mind and I am passing along to you.
Reply:
Thank you for your valuable suggestion. Linear Solution Energy Relationship (LSER) can calculate partition coefficients for various systems. Solubility relationship , solvent systems and rutin polarity can provide basis for the selection of high-speed counter-current chromatography solvent systems. Polar parameters and solubility parameters have the same interpretation of the solvent system. LSER provide E (excess molar refraction), S (polarizability/dipolarity), A (solute hydrogen bond acidity), B and/or B0 (solute hydrogen bond basicity), and L (logarithmic gas-hexadecane partition coefficient). I will continue to calculate LSER parameters of rutin about this article.
Reviewer 2 Report
Insied the mansucript there are some small typing mistakes.
The description is well conducted, but to valorize the conclusions, should be bettere add also, the complete characterization of the extract, to ascertain the validity of the use of rutin as unic standard.
in the manuscript the authors describe a purification procedure which, according to their opinion, is better than those already in use. They extract and purify the leaves, but never indicate any other component besides the rutin. They claim that rutin has a scaffold that is common to many other compounds commonly found in the extracts, but in any case it is necessary to isolate them and characterize them individually. From my point of view it does not seem to me that this method is particularly useful. In fact, although the gel chromatography is longer, it certainly allows to isolate and simultaneously purify the SINGLE components in the extract. I suggest to improve the quality carring out the isolation of the extract components and evaluating their amount.
Author Response
Insied the mansucript there are some small typing mistakes.
The description is well conducted, but to valorize the conclusions, should be bettere add also, the complete characterization of the extract, to ascertain the validity of the use of rutin as unic standard. in the manuscript the authors describe a purification procedure which, according to their opinion, is better than those already in use. They extract and purify the leaves, but never indicate any other component besides the rutin. They claim that rutin has a scaffold that is common to many other compounds commonly found in the extracts, but in any case it is necessary to isolate them and characterize them individually. From my point of view it does not seem to me that this method is particularly useful. In fact, although the gel chromatography is longer, it certainly allows to isolate and simultaneously purify the SINGLE components in the extract. I suggest to improve the quality carring out the isolation of the extract components and evaluating their amount.
Reply:
Thank you for your valuable suggestion. I have made some modifications about typing mistakes. Many compounds can be used as the standard flavonoids, such as quercetin, rutin, etc. We chose rutin as the standard for this article, and rutin has the same maternal nucleus structure as flavonoids. HSCCC is a new high efficiency separation technology, which has a large processing capacity and a short processing time. Flavonoids in mulberry leaves are very few, and the yield obtained by general column method is very small, and it is difficult to detect products. However, HSCCC has the detection function, which can determine the peak time of products and collect products according to the detection spectrum. Finally, the content of flavonoids detected by high performance liquid chromatography(HPLC) is concentrated after HSCCC purification. Other methods can obtain flavonoids that can be as pure as 90%, but the time of purification is too long. By comparing the purification time and final purity, HSCCC has higher research value.
