Qualitative and Quantitative Analysis Method of Recombinant Collagen in Complex Matrix Based on HPLC-MS/MS
The purpose of this study is to achieve the quantitative detection of recombinant type III collagen (rh-COL-III) in dressings with complex matrix. First of all, the marker peptide (GEAGIPGVPGAK) of rhCOL-III was identified with HPLC-MS/MS. Then, a qualitative and quantitative method based on marker...
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2024-11-01
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| author | Jianping Gao Liping Jia Zhenhu Guo Liming Xu Daidi Fan Guifeng Zhang |
| author_facet | Jianping Gao Liping Jia Zhenhu Guo Liming Xu Daidi Fan Guifeng Zhang |
| author_sort | Jianping Gao |
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| description | The purpose of this study is to achieve the quantitative detection of recombinant type III collagen (rh-COL-III) in dressings with complex matrix. First of all, the marker peptide (GEAGIPGVPGAK) of rhCOL-III was identified with HPLC-MS/MS. Then, a qualitative and quantitative method based on marker peptides was established and validated. In order to obtain higher sensitivity, a pretreatment method of liquid, gel, and ointment dressings was optimized. The reference material for quantification was combined using rhCOL-III and blank matrix of each dressing. The results indicated that the relative standard deviation (RSD) of the quantitative method was 2.77%, and the RSD of intraday and interday precision was 2.76% and 2.31%, respectively. The spiking recovery rate was between 80% and 90%. The optimal pretreatment method was Tris-HCl solvent replacement. The optimal trypsin concentration for the dressing solution was 20 μg in 500 μL. The method of preparing standard substances with a blank matrix can effectively eliminate the influence of the matrix effect on the quantitative results. The average spiking recovery rates of 50 μg/mL, 100 μg/mL, and 200 μg/mL in three different dressings ranged from 80% to 120%. The quantitative detection of limit (LOD) of rhCOL-III was 1 ng/mL, 2 ng/g, and 1 ng/g in liquid, ointment, and gel dressings. |
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| institution | Kabale University |
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| language | English |
| publishDate | 2024-11-01 |
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| spelling | doaj-art-ab718fcbd0844cbb8f35741f8266f3c62024-12-27T14:53:24ZengMDPI AGSeparations2297-87392024-11-01111233910.3390/separations11120339Qualitative and Quantitative Analysis Method of Recombinant Collagen in Complex Matrix Based on HPLC-MS/MSJianping Gao0Liping Jia1Zhenhu Guo2Liming Xu3Daidi Fan4Guifeng Zhang5State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, ChinaXi’an Giant Biogene Technology Co., Ltd., Xi’an 710075, ChinaState Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, ChinaNational Institutes for Food and Drug Control, Beijing 102629, ChinaXi’an Giant Biogene Technology Co., Ltd., Xi’an 710075, ChinaState Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, ChinaThe purpose of this study is to achieve the quantitative detection of recombinant type III collagen (rh-COL-III) in dressings with complex matrix. First of all, the marker peptide (GEAGIPGVPGAK) of rhCOL-III was identified with HPLC-MS/MS. Then, a qualitative and quantitative method based on marker peptides was established and validated. In order to obtain higher sensitivity, a pretreatment method of liquid, gel, and ointment dressings was optimized. The reference material for quantification was combined using rhCOL-III and blank matrix of each dressing. The results indicated that the relative standard deviation (RSD) of the quantitative method was 2.77%, and the RSD of intraday and interday precision was 2.76% and 2.31%, respectively. The spiking recovery rate was between 80% and 90%. The optimal pretreatment method was Tris-HCl solvent replacement. The optimal trypsin concentration for the dressing solution was 20 μg in 500 μL. The method of preparing standard substances with a blank matrix can effectively eliminate the influence of the matrix effect on the quantitative results. The average spiking recovery rates of 50 μg/mL, 100 μg/mL, and 200 μg/mL in three different dressings ranged from 80% to 120%. The quantitative detection of limit (LOD) of rhCOL-III was 1 ng/mL, 2 ng/g, and 1 ng/g in liquid, ointment, and gel dressings.https://www.mdpi.com/2297-8739/11/12/339qualitative analysisquantitationrecombinant collagendressingcomplex matrixHPLC-MS/MS |
| spellingShingle | Jianping Gao Liping Jia Zhenhu Guo Liming Xu Daidi Fan Guifeng Zhang Qualitative and Quantitative Analysis Method of Recombinant Collagen in Complex Matrix Based on HPLC-MS/MS Separations qualitative analysis quantitation recombinant collagen dressing complex matrix HPLC-MS/MS |
| title | Qualitative and Quantitative Analysis Method of Recombinant Collagen in Complex Matrix Based on HPLC-MS/MS |
| title_full | Qualitative and Quantitative Analysis Method of Recombinant Collagen in Complex Matrix Based on HPLC-MS/MS |
| title_fullStr | Qualitative and Quantitative Analysis Method of Recombinant Collagen in Complex Matrix Based on HPLC-MS/MS |
| title_full_unstemmed | Qualitative and Quantitative Analysis Method of Recombinant Collagen in Complex Matrix Based on HPLC-MS/MS |
| title_short | Qualitative and Quantitative Analysis Method of Recombinant Collagen in Complex Matrix Based on HPLC-MS/MS |
| title_sort | qualitative and quantitative analysis method of recombinant collagen in complex matrix based on hplc ms ms |
| topic | qualitative analysis quantitation recombinant collagen dressing complex matrix HPLC-MS/MS |
| url | https://www.mdpi.com/2297-8739/11/12/339 |
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