Simple, efficient and economical methods for isolation and estimation of novel isoflavone using RP-HPLC
The study was undertaken to develop a simplified procedure for the isolation of bioactive isoflavone from Iris kashmiriana, using a direct method of isolation, avoiding the use of chromatographic techniques. The compound was isolated by commercially viable procedure. The extraction of powdered drug...
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Elsevier
2017-01-01
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| Series: | MethodsX |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016117300079 |
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| author | Afroze Alam Kamlesh Kumar Naik Navneet Kumar Upadhaya Shailendra Kumar K.L. Dhar |
| author_facet | Afroze Alam Kamlesh Kumar Naik Navneet Kumar Upadhaya Shailendra Kumar K.L. Dhar |
| author_sort | Afroze Alam |
| collection | DOAJ |
| description | The study was undertaken to develop a simplified procedure for the isolation of bioactive isoflavone from Iris kashmiriana, using a direct method of isolation, avoiding the use of chromatographic techniques. The compound was isolated by commercially viable procedure. The extraction of powdered drug (500 g) was done with petroleum ether (60–80) using a Soxhlet apparatus (24 h run). The petroleum ether extract (gums and resins 2.13 g) was obtained and the marc (400 g) was subjected to extraction with 95% methanol using a Soxhlet apparatus (24 h run). The methanolic extract (5 g) was subjected to successive fractionation with toluene, chloroform and ethyl acetate and n- butanol. On the basis of phytochemical analysis, the glycoside was present in n- butanol fraction. The n-butanol fraction (1.5 g) was taken in dried methanol, passed through activated animal charcoal and subjected to acid hydrolysis. The isoflavone (250 mg), was obtained after the usual process of separation. The purity of the compound was checked by analyzing TLC (Thin Layer chromatography) and melting point. Further, the chemical method was used to characterize the compound by shift reagents using UV spectroscopy. The quantitative estimation of isoflavone was done using RP-HPLC and was found to be 98.9% pure.
• The “previously undescribed” isoflavone was isolated by modifying approach of solvent/solvent extraction, fractionation and acid hydrolysis.
• The spectroscopic characterization was equaly done by IR, 1HNMR, 13CNMR, Mass spectrometry.
• 98.9% purity was achieved using RP-HPLC with simple solvent (Methanol and Water 55: 45). |
| first_indexed | 2024-12-10T05:04:46Z |
| format | Article |
| id | doaj.art-25239191d05a4e04b1179090958db2b6 |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-12-10T05:04:46Z |
| publishDate | 2017-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-25239191d05a4e04b1179090958db2b62022-12-22T02:01:16ZengElsevierMethodsX2215-01612017-01-014C12813310.1016/j.mex.2017.02.001Simple, efficient and economical methods for isolation and estimation of novel isoflavone using RP-HPLCAfroze Alam0Kamlesh Kumar Naik1Navneet Kumar Upadhaya2Shailendra Kumar3K.L. Dhar4School of Pharmaceutical Sciences, Shoolini University, Bajhol, Solan, 173229, Himachal Pradesh, IndiaNandha College of Pharmacy, Perundurai Main Road, Erode, Tamil Nadu, 638052, IndiaSchool of Pharmaceutical Sciences, Shoolini University, Bajhol, Solan, 173229, Himachal Pradesh, IndiaGovernment Pharmacy Institute, Agam Kuan, Patna, 800007, Bihar, IndiaSchool of Pharmaceutical Sciences, Shoolini University, Bajhol, Solan, 173229, Himachal Pradesh, IndiaThe study was undertaken to develop a simplified procedure for the isolation of bioactive isoflavone from Iris kashmiriana, using a direct method of isolation, avoiding the use of chromatographic techniques. The compound was isolated by commercially viable procedure. The extraction of powdered drug (500 g) was done with petroleum ether (60–80) using a Soxhlet apparatus (24 h run). The petroleum ether extract (gums and resins 2.13 g) was obtained and the marc (400 g) was subjected to extraction with 95% methanol using a Soxhlet apparatus (24 h run). The methanolic extract (5 g) was subjected to successive fractionation with toluene, chloroform and ethyl acetate and n- butanol. On the basis of phytochemical analysis, the glycoside was present in n- butanol fraction. The n-butanol fraction (1.5 g) was taken in dried methanol, passed through activated animal charcoal and subjected to acid hydrolysis. The isoflavone (250 mg), was obtained after the usual process of separation. The purity of the compound was checked by analyzing TLC (Thin Layer chromatography) and melting point. Further, the chemical method was used to characterize the compound by shift reagents using UV spectroscopy. The quantitative estimation of isoflavone was done using RP-HPLC and was found to be 98.9% pure. • The “previously undescribed” isoflavone was isolated by modifying approach of solvent/solvent extraction, fractionation and acid hydrolysis. • The spectroscopic characterization was equaly done by IR, 1HNMR, 13CNMR, Mass spectrometry. • 98.9% purity was achieved using RP-HPLC with simple solvent (Methanol and Water 55: 45).http://www.sciencedirect.com/science/article/pii/S2215016117300079Iris kashmerianaIsoflavoneRP-HPLCSpectroscopic chracterizationQuantitative estimation |
| spellingShingle | Afroze Alam Kamlesh Kumar Naik Navneet Kumar Upadhaya Shailendra Kumar K.L. Dhar Simple, efficient and economical methods for isolation and estimation of novel isoflavone using RP-HPLC MethodsX Iris kashmeriana Isoflavone RP-HPLC Spectroscopic chracterization Quantitative estimation |
| title | Simple, efficient and economical methods for isolation and estimation of novel isoflavone using RP-HPLC |
| title_full | Simple, efficient and economical methods for isolation and estimation of novel isoflavone using RP-HPLC |
| title_fullStr | Simple, efficient and economical methods for isolation and estimation of novel isoflavone using RP-HPLC |
| title_full_unstemmed | Simple, efficient and economical methods for isolation and estimation of novel isoflavone using RP-HPLC |
| title_short | Simple, efficient and economical methods for isolation and estimation of novel isoflavone using RP-HPLC |
| title_sort | simple efficient and economical methods for isolation and estimation of novel isoflavone using rp hplc |
| topic | Iris kashmeriana Isoflavone RP-HPLC Spectroscopic chracterization Quantitative estimation |
| url | http://www.sciencedirect.com/science/article/pii/S2215016117300079 |
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