Reverse phase HPLC method for detection and quantification of lupin seed ?-conglutin
© 2017 Elsevier B.V. A simple, selective and accurate reverse phase HPLC method was developed for detection and quantitation of ?-conglutin from lupin seed extract. A linear gradient of water and acetonitrile containing trifluoroacetic acid (TFA) on a reverse phase column (Agilent Zorbax 300SB C-18)...
| Main Authors: | , , , , |
|---|---|
| Format: | Journal Article |
| Published: |
Elsevier BV
2017
|
| Online Access: | http://hdl.handle.net/20.500.11937/58052 |
| _version_ | 1848760165323505664 |
|---|---|
| author | Mane, S. Bringans, S. Johnson, Stuart Pareek, Vishnu Utikar, Ranjeet |
| author_facet | Mane, S. Bringans, S. Johnson, Stuart Pareek, Vishnu Utikar, Ranjeet |
| author_sort | Mane, S. |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | © 2017 Elsevier B.V. A simple, selective and accurate reverse phase HPLC method was developed for detection and quantitation of ?-conglutin from lupin seed extract. A linear gradient of water and acetonitrile containing trifluoroacetic acid (TFA) on a reverse phase column (Agilent Zorbax 300SB C-18), with a flow rate of 0.8 ml/min was able to produce a sharp and symmetric peak of ?-conglutin with a retention time at 29.16 min. The identity of ?-conglutin in the peak was confirmed by mass spectrometry (MS/MS identification) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The data obtained from MS/MS analysis was matched against the specified database to obtain the exact match for the protein of interest. The proposed method was validated in terms of specificity, linearity, sensitivity, precision, recovery and accuracy. The analytical parameters revealed that the validated method was capable of selectively performing a good chromatographic separation of ?-conglutin from the lupin seed extract with no interference of the matrix. The detection and quantitation limit of ?-conglutin were found to be 2.68 µg/ml and 8.12 µg/ml respectively. The accuracy (precision and recovery) analysis of the method was conducted under repeatable conditions on different days. Intra-day and inter-day precision values less than 0.5% and recovery greater than 97% indicated high precision and accuracy of the method for analysis of ?-conglutin. The method validation findings were reproducible and can be successfully applied for routine analysis of ?-conglutin from lupin seed extract. |
| first_indexed | 2025-11-14T10:11:26Z |
| format | Journal Article |
| id | curtin-20.500.11937-58052 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-14T10:11:26Z |
| publishDate | 2017 |
| publisher | Elsevier BV |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-580522017-11-20T08:58:08Z Reverse phase HPLC method for detection and quantification of lupin seed ?-conglutin Mane, S. Bringans, S. Johnson, Stuart Pareek, Vishnu Utikar, Ranjeet © 2017 Elsevier B.V. A simple, selective and accurate reverse phase HPLC method was developed for detection and quantitation of ?-conglutin from lupin seed extract. A linear gradient of water and acetonitrile containing trifluoroacetic acid (TFA) on a reverse phase column (Agilent Zorbax 300SB C-18), with a flow rate of 0.8 ml/min was able to produce a sharp and symmetric peak of ?-conglutin with a retention time at 29.16 min. The identity of ?-conglutin in the peak was confirmed by mass spectrometry (MS/MS identification) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The data obtained from MS/MS analysis was matched against the specified database to obtain the exact match for the protein of interest. The proposed method was validated in terms of specificity, linearity, sensitivity, precision, recovery and accuracy. The analytical parameters revealed that the validated method was capable of selectively performing a good chromatographic separation of ?-conglutin from the lupin seed extract with no interference of the matrix. The detection and quantitation limit of ?-conglutin were found to be 2.68 µg/ml and 8.12 µg/ml respectively. The accuracy (precision and recovery) analysis of the method was conducted under repeatable conditions on different days. Intra-day and inter-day precision values less than 0.5% and recovery greater than 97% indicated high precision and accuracy of the method for analysis of ?-conglutin. The method validation findings were reproducible and can be successfully applied for routine analysis of ?-conglutin from lupin seed extract. 2017 Journal Article http://hdl.handle.net/20.500.11937/58052 10.1016/j.jchromb.2017.08.025 Elsevier BV restricted |
| spellingShingle | Mane, S. Bringans, S. Johnson, Stuart Pareek, Vishnu Utikar, Ranjeet Reverse phase HPLC method for detection and quantification of lupin seed ?-conglutin |
| title | Reverse phase HPLC method for detection and quantification of lupin seed ?-conglutin |
| title_full | Reverse phase HPLC method for detection and quantification of lupin seed ?-conglutin |
| title_fullStr | Reverse phase HPLC method for detection and quantification of lupin seed ?-conglutin |
| title_full_unstemmed | Reverse phase HPLC method for detection and quantification of lupin seed ?-conglutin |
| title_short | Reverse phase HPLC method for detection and quantification of lupin seed ?-conglutin |
| title_sort | reverse phase hplc method for detection and quantification of lupin seed ?-conglutin |
| url | http://hdl.handle.net/20.500.11937/58052 |