Development and validation of an optimized HPLC-UV analytical method for the quantification of 5-methyltetrahydrofolate (Active Folate) in dietary supplements: Application to commercial tablets and capsules
5-MTHF is the active form of folic acid, and it is provided as a dietary supplement in the market. Therefore, in this study we provide a new, simple and economic HPLC-UV analysis method to quantify 5-MTHF in dietary supplement. A new HPLC-UV method was developed and validated for the quantification...
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| Format: | Article |
| Language: | English English |
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Elsevier Inc.
2024
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| Online Access: | http://umpir.ump.edu.my/id/eprint/42946/ http://umpir.ump.edu.my/id/eprint/42946/1/Development%20and%20validation%20of%20an%20optimized%20HPLC-UV%20analytical%20method_ABST.pdf http://umpir.ump.edu.my/id/eprint/42946/2/Development%20and%20validation%20of%20an%20optimized%20HPLC-UV%20analytical%20method.pdf |
| _version_ | 1848826741535014912 |
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| author | Alshishani, Anas Shalash, Marwan Aldalaty, Aya Shaghlil, Lena |
| author_facet | Alshishani, Anas Shalash, Marwan Aldalaty, Aya Shaghlil, Lena |
| author_sort | Alshishani, Anas |
| building | UMP Institutional Repository |
| collection | Online Access |
| description | 5-MTHF is the active form of folic acid, and it is provided as a dietary supplement in the market. Therefore, in this study we provide a new, simple and economic HPLC-UV analysis method to quantify 5-MTHF in dietary supplement. A new HPLC-UV method was developed and validated for the quantification of 5-methyltetrahydrofolate in dietary supplements. The method was optimized in terms of chromatographic conditions, including the mobile phase composition, chromatographic column, and the sample preparation diluent using ascorbic acid. A combination of ammonium acetate buffer with methanol (90:10) was the optimal mobile phase, achieving a retention time of approximately 11.4 min on C18 column with good column performance (N = 6786, Asymmetry = 1.03). The method was validated for selectivity, linearity, precision, accuracy, and robustness. The calibration curve was linear (r2 > 0.999) over the concentration range of 1.20 to 4.80 µg/mL. Precision was demonstrated with RSD% values ranging from 1.38 % to 2.90 % for intra-day precision and 0.96 % to 2.61 % for inter-day precision. Accuracy was confirmed with recovery values between 96.5 % and 110.6 % for spiked samples. To demonstrate the method’s applicability, ten brands of 5-MTHF dietary supplements were collected from local pharmacies. Assay results showed significant variability, with four brands having contents below the acceptable range (90–110 % of labeled contents) and two brands exceeding this range. The stability of 5-MTHF in these products appeared to be formula-dependent, emphasizing the need for regulatory revisions to mandate stability testing for dietary supplements. |
| first_indexed | 2025-11-15T03:49:38Z |
| format | Article |
| id | ump-42946 |
| institution | Universiti Malaysia Pahang |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T03:49:38Z |
| publishDate | 2024 |
| publisher | Elsevier Inc. |
| recordtype | eprints |
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| spelling | ump-429462024-11-19T06:19:09Z http://umpir.ump.edu.my/id/eprint/42946/ Development and validation of an optimized HPLC-UV analytical method for the quantification of 5-methyltetrahydrofolate (Active Folate) in dietary supplements: Application to commercial tablets and capsules Alshishani, Anas Shalash, Marwan Aldalaty, Aya Shaghlil, Lena Q Science (General) QP Physiology RS Pharmacy and materia medica 5-MTHF is the active form of folic acid, and it is provided as a dietary supplement in the market. Therefore, in this study we provide a new, simple and economic HPLC-UV analysis method to quantify 5-MTHF in dietary supplement. A new HPLC-UV method was developed and validated for the quantification of 5-methyltetrahydrofolate in dietary supplements. The method was optimized in terms of chromatographic conditions, including the mobile phase composition, chromatographic column, and the sample preparation diluent using ascorbic acid. A combination of ammonium acetate buffer with methanol (90:10) was the optimal mobile phase, achieving a retention time of approximately 11.4 min on C18 column with good column performance (N = 6786, Asymmetry = 1.03). The method was validated for selectivity, linearity, precision, accuracy, and robustness. The calibration curve was linear (r2 > 0.999) over the concentration range of 1.20 to 4.80 µg/mL. Precision was demonstrated with RSD% values ranging from 1.38 % to 2.90 % for intra-day precision and 0.96 % to 2.61 % for inter-day precision. Accuracy was confirmed with recovery values between 96.5 % and 110.6 % for spiked samples. To demonstrate the method’s applicability, ten brands of 5-MTHF dietary supplements were collected from local pharmacies. Assay results showed significant variability, with four brands having contents below the acceptable range (90–110 % of labeled contents) and two brands exceeding this range. The stability of 5-MTHF in these products appeared to be formula-dependent, emphasizing the need for regulatory revisions to mandate stability testing for dietary supplements. Elsevier Inc. 2024-12 Article PeerReviewed pdf en http://umpir.ump.edu.my/id/eprint/42946/1/Development%20and%20validation%20of%20an%20optimized%20HPLC-UV%20analytical%20method_ABST.pdf pdf en http://umpir.ump.edu.my/id/eprint/42946/2/Development%20and%20validation%20of%20an%20optimized%20HPLC-UV%20analytical%20method.pdf Alshishani, Anas and Shalash, Marwan and Aldalaty, Aya and Shaghlil, Lena (2024) Development and validation of an optimized HPLC-UV analytical method for the quantification of 5-methyltetrahydrofolate (Active Folate) in dietary supplements: Application to commercial tablets and capsules. Microchemical Journal, 207 (112040). pp. 1-6. ISSN 0026-265X. (Published) https://doi.org/10.1016/j.microc.2024.112040 https://doi.org/10.1016/j.microc.2024.112040 |
| spellingShingle | Q Science (General) QP Physiology RS Pharmacy and materia medica Alshishani, Anas Shalash, Marwan Aldalaty, Aya Shaghlil, Lena Development and validation of an optimized HPLC-UV analytical method for the quantification of 5-methyltetrahydrofolate (Active Folate) in dietary supplements: Application to commercial tablets and capsules |
| title | Development and validation of an optimized HPLC-UV analytical method for the quantification of 5-methyltetrahydrofolate (Active Folate) in dietary supplements: Application to commercial tablets and capsules |
| title_full | Development and validation of an optimized HPLC-UV analytical method for the quantification of 5-methyltetrahydrofolate (Active Folate) in dietary supplements: Application to commercial tablets and capsules |
| title_fullStr | Development and validation of an optimized HPLC-UV analytical method for the quantification of 5-methyltetrahydrofolate (Active Folate) in dietary supplements: Application to commercial tablets and capsules |
| title_full_unstemmed | Development and validation of an optimized HPLC-UV analytical method for the quantification of 5-methyltetrahydrofolate (Active Folate) in dietary supplements: Application to commercial tablets and capsules |
| title_short | Development and validation of an optimized HPLC-UV analytical method for the quantification of 5-methyltetrahydrofolate (Active Folate) in dietary supplements: Application to commercial tablets and capsules |
| title_sort | development and validation of an optimized hplc-uv analytical method for the quantification of 5-methyltetrahydrofolate (active folate) in dietary supplements: application to commercial tablets and capsules |
| topic | Q Science (General) QP Physiology RS Pharmacy and materia medica |
| url | http://umpir.ump.edu.my/id/eprint/42946/ http://umpir.ump.edu.my/id/eprint/42946/ http://umpir.ump.edu.my/id/eprint/42946/ http://umpir.ump.edu.my/id/eprint/42946/1/Development%20and%20validation%20of%20an%20optimized%20HPLC-UV%20analytical%20method_ABST.pdf http://umpir.ump.edu.my/id/eprint/42946/2/Development%20and%20validation%20of%20an%20optimized%20HPLC-UV%20analytical%20method.pdf |