|
|
Analytical methods for hyaluronan
Černá, Lucie ; Mravec, Filip (referee) ; Pekař, Miloslav (advisor)
This thesis deals with of simple methods for determining the concentration of hyaluronan in aqueous solution and compares the performance of each selected methods such as UV -VIS spectroscopy, IR spectroscopy and the method of determination by alcian blue. These methods were chosen for their simplicity and low cost of the assay. As the most reliable of these three methods, seems to be the method of IR spectroscopy. Determination of the concentration of HA by this method is relatively accurate at concentrations above 0.4 % in deionized water. The method of UV-VIS spectroscopy is based on a linear calibration curve determination, but no characteristic peak was detected, which means that this determination could be influenced by other components of the aqueous solution. Weak ionic strength and molecular weight of HA does not affect the determination. In the method with alcian blue, the results were quite unsatisfactory and does not correspond with the literature. Therefore, this method is not applicable in this form for determining the concentration of HA. Another option for determining the concentration of the HA is ELISA method. These are commercially produced kits suitable for determining low concentrations in low volumes. Its price is higher than the above- mentioned methods.
|
|
|
Methods for determination of hyaluronan
Černá, Lucie ; Venerová, Tereza (referee) ; Pekař, Miloslav (advisor)
This thesis deals with a summary of individual method for determining of hyaluronan concentration, especially in aqueous solutions. Selected methods were compared to their functionality in university workplace. Any functional methods may be used for many purposes in university workplace in research of materials based on hyaluronan. It has been studied usability to assess residual content of hyaluronan in supernatant of gel formed from hyaluronan and surfuctant, in this thesis. We choosed sptectrophotometric methods, especially with UV spectroscopy and IR spectroscopy. Two precipitation methods using alcian blue were also tested. As the most reliable, the least demanding and specific method seems to be the method using IR spectroscopy, thanks to it we are able to determine the concentration of HA from 0.01 %. With this method, it has been found, that if there will be located some amount of HA in supernatant, it will be below the detection limit of assay. The remanining methods were modified to help determine the concentrations of HA, but works only in aqueous solutions. A possible variation for determination of very low concentrations of HA may be the method using ELISA kits. However their cost is higher than the above-mentioned methods.
|
|
|
Methods for determination of hyaluronan
Černá, Lucie ; Venerová, Tereza (referee) ; Pekař, Miloslav (advisor)
This thesis deals with a summary of individual method for determining of hyaluronan concentration, especially in aqueous solutions. Selected methods were compared to their functionality in university workplace. Any functional methods may be used for many purposes in university workplace in research of materials based on hyaluronan. It has been studied usability to assess residual content of hyaluronan in supernatant of gel formed from hyaluronan and surfuctant, in this thesis. We choosed sptectrophotometric methods, especially with UV spectroscopy and IR spectroscopy. Two precipitation methods using alcian blue were also tested. As the most reliable, the least demanding and specific method seems to be the method using IR spectroscopy, thanks to it we are able to determine the concentration of HA from 0.01 %. With this method, it has been found, that if there will be located some amount of HA in supernatant, it will be below the detection limit of assay. The remanining methods were modified to help determine the concentrations of HA, but works only in aqueous solutions. A possible variation for determination of very low concentrations of HA may be the method using ELISA kits. However their cost is higher than the above-mentioned methods.
|
|
|
Analytical methods for hyaluronan
Černá, Lucie ; Mravec, Filip (referee) ; Pekař, Miloslav (advisor)
This thesis deals with of simple methods for determining the concentration of hyaluronan in aqueous solution and compares the performance of each selected methods such as UV -VIS spectroscopy, IR spectroscopy and the method of determination by alcian blue. These methods were chosen for their simplicity and low cost of the assay. As the most reliable of these three methods, seems to be the method of IR spectroscopy. Determination of the concentration of HA by this method is relatively accurate at concentrations above 0.4 % in deionized water. The method of UV-VIS spectroscopy is based on a linear calibration curve determination, but no characteristic peak was detected, which means that this determination could be influenced by other components of the aqueous solution. Weak ionic strength and molecular weight of HA does not affect the determination. In the method with alcian blue, the results were quite unsatisfactory and does not correspond with the literature. Therefore, this method is not applicable in this form for determining the concentration of HA. Another option for determining the concentration of the HA is ELISA method. These are commercially produced kits suitable for determining low concentrations in low volumes. Its price is higher than the above- mentioned methods.
|
guest ::
