Modernization of Shilajit Processing: Comparative Phytochemical Screening, Antioxidant Potential, and Phenolic Content in Standardized vs. Non- Standardized Extracts


Authors : Komal Kumari; Md Zeyaullah; Preeti S. Saxena; Harsh Jain; V. K. Tripathi

Volume/Issue : Volume 10 - 2025, Issue 8 - August


Google Scholar : https://tinyurl.com/yndcvw8w

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DOI : https://doi.org/10.38124/ijisrt/25aug561

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Abstract : Shilajit, an Ayurvedic medicinal substance, is recognized for its rich humic content and various health benefits, including anti-inflammatory and neuroprotective effects. This study involves a comparative evaluation of phytochemical composition, antioxidant potential, and total phenolic content in standardized and non-standardized Shilajit extracts. The standardized Shilajit was processed through a controlled workflow including extraction, filtration, distillation, and drying, aimed at reducing heavy metal contamination while preserving bioactive compounds. Methodologies employed included qualitative biochemical screening, high-resolution mass spectrometry (HRMS), UV-visible spectrophotometric analysis to determine the E4/E6 ratio, Fourier-transform infrared spectroscopy (FTIR) for functional group identification, and scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy (SEM-EDX) to assess elemental composition and confirm the elimination of heavy metals. The E4/E6 ratio of standardized Shilajit ranged from 2.84 to 2.93, significantly lower than that of non-standardized Shilajit samples, indicating a higher concentration of aromatic components. Additionally, its IR analysis revealed distinct functional groups, whereas non-standardized samples lacked the humic acid band, while SEM-EDX confirmed the absence of heavy metals in all samples. Phytochemical screening indicates the abundance of different compounds, such as flavonoids, phytosterols, terpenoids, phenols, and coumarins, which were further validated by the HRMS study. The phytochemical richness is responsible for significantly higher antioxidant activity (via DPPH and phosphomolybdate assays) and phenolic content in Standardized Shilajit than non-standardized Shilajit samples. These results demonstrate that the standardized production process enhances the safety and therapeutic efficacy of shilajit, supporting its application in health and wellness.

Keywords : VCA-964, Standardized Shilajit, Heavy Metals, Antioxidant, Physiochemical Properties.

References :

  1. Wilson E, Rajamanickam GV, Dubey GP, Klose P, Musial F, Saha FJ, Rampp T, Michalsen A, Dobos GJ. Review on shilajit used in traditional Indian medicine. J. of Ethnopharmacol. 2011 Jun 14;136(1):1-9.https://doi.org/10.1016/j.jep.2011.04.033
  2. Agarwal SP, Khanna R, Karmarkar R, Anwer MK, Khar RK. Shilajit: a review. Phytother. Res.: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 2007 May;21(5):401-5.https://doi.org/10.1002/ptr.2100
  3. Pingali U, Nutalapati C. Shilajit extract reduces oxidative stress, inflammation, and bone loss to dose-dependently preserve bone mineral density in postmenopausal women with osteopenia: A randomized, double-blind, placebo-controlled trial. Phytomed. 2022 Oct 1; 105:154334.https://doi.org/10.1016/j.phymed.2022.154334
  4. Musthafa MS, Ali AR, Ali AR, Mohamed MJ, War M, Naveed MS, Al-Sadoon MK, Paray BA, Rani KU, Arockiaraj J, Balasundaram C. Effect of Shilajit enriched diet on immunity, antioxidants, and disease resistance in Macrobrachium rosenbergii (de Man) against Aeromonas hydrophila. Fish & shellfish immunol. 2016 Oct 1;57:293-300.https://doi.org/10.1016/j.fsi.2016.08.033
  5. Lawley S, Gupta RC, Goad JT, Canerdy TD, Kalidindi SR. Anti-inflammatory and anti-arthritic efficacy and safety of purified shilajit in moderately arthritic dogs. J Vet Sci Anim Husb. 2013;1(3):302. doi: 10.15744/2348-9790.1.302
  6. Ghosal S, Singh SK, Kumar Y, Srivastava R, Goel RK, Dey R, Bhattacharya SK. Anti‐ulcerogenic activity of fulvic acids and 4′‐methoxy‐6‐carbomethoxybiphenyl isolated from shilajit. Phytother. Res. 1988 Dec;2(4):187-91.https://doi.org/10.1002/ptr.2650020408
  7. Kloskowski T, Szeliski K, Krzeszowiak K, Fekner Z, Kazimierski Ł, Jundziłł A, Drewa T, Pokrywczyńska M. Mumio (Shilajit) as a potential chemotherapeutic for the urinary bladder cancer treatment. Sci. Rep. 2021 Nov 19;11(1):22614. doi.org/10.1038/s41598-021-01996-8
  8. Durg S, Veerapur VP, Thippeswamy BS, Ahamed SM. Antiepileptic and antipsychotic activities of standardized Śilājatu (Shilajit) in experimental animals. Anc. Sci. of Life. 2015 Oct 1;35(2):110-7. DOI: 10.4103/0257-7941.171675
  9. Mishra RK, Verma HP, Singh N, Singh SK. Male infertility: lifestyle and oriental remedies. J. of Sci. Res. 2012; 56:93-101.
  10. Mosavi S, Tabarrai M, Tansaz M, Salehinia H, Grylka S, Rahmani A, Shahali S. Effects of oral Shilajit tablets on sexual function and sexual quality of life among reproductive-aged women: a triple-blind randomized clinical trial. Tradit. Med. Res. 2023;8(11):66. http://DOI:10.53388/TMR20230305002
  11. Das A, Datta S, Rhea B, Sinha M, Veeraragavan M, Gordillo G, Roy S. The human skeletal muscle transcriptome in response to oral Shilajit supplementation. J. of Med. Food. 2016 Jul 1;19(7):701-9. https://doi.org/10.1089/jmf.2016.0010
  12. Keller JL, Housh TJ, Hill EC, Smith CM, Schmidt RJ, Johnson GO. The effects of Shilajit supplementation on fatigue-induced decreases in muscular strength and serum hydroxyproline levels. J. of the Int. Soc. of Sports Nutr. 2019 Dec;16:1-9. https://doi.org/10.1186/s12970-019-0270-2
  13. Agarwal SP, Anwer K, Khanna R, Ali A, Sultana Y. Humic acid from Shilajit: A physico-chemical and spectroscopic characterization. J. of the Serb. Chem. Soc. 2010;75(3):413-22. https://doi.org/10.2298/JSC090316006A
  14. Ghosal, S., Lal, J., Singh, S. K., Goel, R. K., Jaiswal, A. K., & Bhattacharya, S. K. (1991). The need for formulation of Shilajit by its isolated active constituents. Phytotherapy research, 5(5), 211-216. https://doi.org/10.1002/ptr.2650050505
  15. Tripathi, Y. B., Saha, S., Ojha, A., Singh, V. K., Kumar, S., Gautam, A., Tripathi, S., Dubey, V. S. K., & Tripathi, P. (2024). Novel parameters to define bio‑efficacy of Shilajit resin and its validation. International Journal of Pharmaceutical Sciences and Research, 15(8), 2434–2444. https://doi.org/10.13040/IJPSR. 0975‑8232
  16. Kangari P, Roshangar L, Iraji A, Talaei-Khozani T, Razmkhah M. Accelerating effect of Shilajit on osteogenic property of adipose-derived mesenchymal stem cells (ASCs). J. of Orthop. Surg. and Res. 2022 Sep 24;17(1):424. https://doi.org/10.1186/s13018-022-03305-z
  17. Shaikh JR, Patil M. Qualitative tests for preliminary phytochemical screening: An overview. Int. J. of chem. studies. 2020 Mar 1;8(2):603-8. I: https://doi.org/10.22271/chemi.2020.v8.i2i.8834
  18. Muniyandi JM, Lakshman K. Preliminary studies of phytochemical investigation on coastal medicinal plants of Boloor, Mangalore. INDO AMERICAN J. OF PHARM. SCI. 2018 Feb 1;5(2):1309-15. DOI: 10.5281/zenodo.1196309
  19. Singh V, Kumar R. Study of phytochemical analysis and antioxidant activity of Allium sativum of Bundelkhand region. Int. J. of Life-Sciences Sci. Res. 2017 Nov;3(6):1451-8. http://DOI:10.21276/ijlssr.2017.3.6.4
  20. Baliyan S, Mukherjee R, Priyadarshini A, Vibhuti A, Gupta A, Pandey RP, Chang CM. Determination of antioxidants by DPPH radical scavenging activity and quantitative phytochemical analysis of Ficus religiosa. Mol. 2022 Feb16;27(4):1326.https://doi.org/10.3390/molecules27041326
  21. Shah P, Modi HA. Comparative study of DPPH, ABTS and FRAP assays for determination of antioxidant activity. Int. J. Res. Appl. Sci. Eng. Technol. 2015 Aug;3(6):636-41.
  22. Sachett A, Gallas-Lopes M, Conterato GM, Herrmann AP, Piato A. Antioxidant activity by FRAP assay: in vitro protocol. Protocols [Internet]. 2021. DOI: 10.17504/protocols.io.btbpnimn
  23. Jan S, Khan MR, Rashid U, Bokhari J. Assessment of antioxidant potential, total phenolics and flavonoids of different solvent fractions of Monotheca buxifolia fruit. Osong publ. health and res. perspect. 2013 Oct 1;4(5):246-54.https://doi.org/10.1016/j.phrp.2013.09.003
  24. Rege AN, Juvekar P, Juvekar AR. In vitro antioxidant and anti-arthritic activities of Shilajit. Int J Pharm Pharm Sci. 2012;4(2):650-3.
  25. Crizel RL, Siebeneichler TJ, Zandoná GP, Hoffmann JF, Ferreira CD. Phenolic Compound Extraction, Identification, and Health Aspects: Part II. Colored Cereals: Properties, Processing, Health Benefits, and Industrial Uses. 2025 Mar 28:255.
  26. Wang LA. GraphRobot. Online). https://www.graphrobot.com. 2019.
  27. Zykova MV, Schepetkin IA, Belousov MV, Krivoshchekov SV, Logvinova LA, Bratishko KA, Yusubov MS, Romanenko SV, Quinn MT. Physicochemical characterization and antioxidant activity of humic acids isolated from peat of various origins. Mol. 2018 Mar 24;23(4):753.https://doi.org/10.3390/molecules23040753
  28. Anwer MK, Agarwal SP, Ali A, Sultana Y. Molecular complexes of aspirin with humic acid extracted from shilajit and their characterization. . J. of Incl. Phenom. and Macrocycl. Chem. 2010 Jun;67:209-15. http://doi:10.1007/s10847-009-9699-2
  29. Khanna R, Witt M, Anwer MK, Agarwal SP, Koch BP. Spectroscopic characterization of fulvic acids extracted from the rock exudate Shilajit. Org. Geochem. 2008 Dec 1;39(12):1719-24. https://doi.org/10.1016/j.orggeochem.2008.08.009
  30. Cheng G, Niu Z, Zhang C, Zhang X, Li X. Extraction of humic acid from lignite by KOH-hydrothermal method. Appl. Sci. 2019 Mar 31;9(7):1356. https://doi.org/10.3390/app9071356
  31. Wongsa P, Phatikulrungsun P, Prathumthong S. FT-IR characteristics, phenolic profiles and inhibitory potential against digestive enzymes of 25 herbal infusions. Sci. Rep. 2022 Apr 22;12(1):6631. https://doi.org/10.1038/s41598-022-10669-z
  32. Coates J. Interpretation of infrared spectra, a practical approach. Encycl. of analytical chem.  2000 Sep 15;12:10815-37.
  33. Yaqoob Z, Batool SA, Khan A, Hussain R, Raza MA, Alqahtani MS, Abbas M, Almubarak HA, Alqahtani FF, Rehman MA. Characterization and medicinal applications of Karakoram shilajit; angiogenesis activity, antibacterial properties and cytotoxicity. Mater. Res. Express. 2023 Oct 31;10(10):105403. http://DOI10.1088/2053-1591/ad04bd
  34. Zhu K, Prince RL. Calcium and bone. Clin. biochem. 2012 Aug 1;45(12):936-42. https://doi.org/10.1016/j.clinbiochem.2012.05.006
  35. Salhan M, Kumar B, Tiwari P, Sharma P, Sandhar HK, Gautam M. Comparative anthelmintic activity of aqueous and ethanolic leaf extracts of Clitoria ternatea. Int J Drug Dev Res. 2011 Jan;3(1):62-9.
  36. JothiMuniyandi M, Jayachitra A. PHYTOCHEMICAL INVESTIGATION OF BARLERIA LONGIFLORA LINN. F. IN WESTERN GHATS MADURAI. Res. J of Life Sci., Bioinform., Pharma. and Chem. Sci. http://doi:10.26479/2019.0503.57
  37. Bioquest AA. Quest Graph™ IC50 Calculator. AAT Bioquest Available at: https://www. aatbio. com/tools/ec50-calculator (Accessed, May 2025). 2025.

Shilajit, an Ayurvedic medicinal substance, is recognized for its rich humic content and various health benefits, including anti-inflammatory and neuroprotective effects. This study involves a comparative evaluation of phytochemical composition, antioxidant potential, and total phenolic content in standardized and non-standardized Shilajit extracts. The standardized Shilajit was processed through a controlled workflow including extraction, filtration, distillation, and drying, aimed at reducing heavy metal contamination while preserving bioactive compounds. Methodologies employed included qualitative biochemical screening, high-resolution mass spectrometry (HRMS), UV-visible spectrophotometric analysis to determine the E4/E6 ratio, Fourier-transform infrared spectroscopy (FTIR) for functional group identification, and scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy (SEM-EDX) to assess elemental composition and confirm the elimination of heavy metals. The E4/E6 ratio of standardized Shilajit ranged from 2.84 to 2.93, significantly lower than that of non-standardized Shilajit samples, indicating a higher concentration of aromatic components. Additionally, its IR analysis revealed distinct functional groups, whereas non-standardized samples lacked the humic acid band, while SEM-EDX confirmed the absence of heavy metals in all samples. Phytochemical screening indicates the abundance of different compounds, such as flavonoids, phytosterols, terpenoids, phenols, and coumarins, which were further validated by the HRMS study. The phytochemical richness is responsible for significantly higher antioxidant activity (via DPPH and phosphomolybdate assays) and phenolic content in Standardized Shilajit than non-standardized Shilajit samples. These results demonstrate that the standardized production process enhances the safety and therapeutic efficacy of shilajit, supporting its application in health and wellness.

Keywords : VCA-964, Standardized Shilajit, Heavy Metals, Antioxidant, Physiochemical Properties.

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