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Volume 20, Issue 8
  • ISSN: 1573-4129
  • E-ISSN: 1875-676X

Abstract

Objective

The objective of the present work is to develop and validate a novel, specific, precise, and reliable method for the estimation of in bulk and herbal dosage form using the RP-HPLC method.

Methods

RP-HPLC analysis was performed using a C column of dimension 150×4.6mm, 5 µ. The chromatography system comprised an Agilent 1220 Infinity II LC equipped with a VWD detector and a 1220 Infinity II LC binary pump, wherein the instrument operation was managed through Control Panel software at a flow rate of 0.5 ml/min. Methanol: water in the ratio of 55:45 was used as the mobile phase, and the effluents were analyzed at 275nm. The proposed method was validated for various parameters like linearity, precision, accuracy, robustness, ruggedness, selectivity, limit of detection, limit of quantification, and assay as per the ICH Q2(R1) guidelines.

Results

Linearity was noted over a concentration range of 50-250 µg/ml with a correlation coefficient of 0.999. The limit of detection (LOD) and limit of quantification (LOQ) were determined to be 16.02 and 48.56 µg/mL, respectively. The % RSD for interday and intraday precision studies was less than 2%, which was within the official RSD limit. Recovery analysis performed using marketed formulation was found to be in the range of 97-105%.

Conclusion

The method developed was validated according to the ICH guidelines. Hence, it is evident that the developed method is novel, sensitive, precise, and reliable, and it can be successfully applied to estimate in bulk material and its herbal dosage form.

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2025-04-02

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References

  1. AlafnanA. SridharagattaS. SaleemH. KhurshidU. AlamriA. AnsariS.Y. Zainal AbidinS.A. AnsariS.A. AlamriA.S. AhemadN. AnwarS. Evaluation of the phytochemical, antioxidant, enzyme inhibition, and wound healing potential of Calotropis gigantea (L.) dryand: A source of a bioactive medicinal product.Front. Pharmacol.20211270136910.3389/fphar.2021.70136934483902
     [Google Scholar]
  2. KrishnanS. Traditional herbal medicines - A review.2018
     [Google Scholar]
  3. SrinivasanM. SudheerA.R. MenonV.P. Recent advances in Indian herbal drug research guest editor: Thomas Paul Asir Devasagayam ferulic acid: Therapeutic potential through its antioxidant property.J. Clin. Biochem. Nutr.20074092
     [Google Scholar]
  4. PrasathkumarM. AnishaS. DhrisyaC. BeckyR. SadhasivamS. Therapeutic and pharmacological efficacy of selective Indian medicinal plants – A review.Phytomedicine Plus20211210002910.1016/j.phyplu.2021.100029
     [Google Scholar]
  5. DwivediA. PratapS. AwasthiS. GautamP. KadirA. Calotropis gigantea: An in-depth review of its therapeutic potential.J. Pharmacogn. Phytochem.202413271572110.22271/phyto.2024.v13.i2e.14915
     [Google Scholar]
  6. UthirasamyS. ChitraT. MuruganA. ManjulaG. ArulmanickamP. KavithaT. ThinakaranM. Determining the bioactive constituents in Calotropis gigantea leaves by GC-MS, HPLC and FTIR techniques.New Visions Biol. Sci.2021111110.9734/bpi/nvbs/v1/1788C
     [Google Scholar]
  7. MaharR. DixitS. JoshiT. KanojiyaS. MishraD.K. KonwarR. ShuklaS.K. Bioactivity guided isolation of oxypregnane-oligoglycosides (calotroposides) from the root bark of Calotropis gigantea as potent anticancer agents.RSC Advances2016610610421510422610.1039/C6RA23600F
     [Google Scholar]
  8. WangZ.N. WangM.Y. MeiW.L. HanZ. DaiH.F. A new cytotoxic pregnanone from Calotropis gigantea.Molecules200813123033303910.3390/molecules1312303319052526
     [Google Scholar]
  9. M. yuan Wang, Q. Yang, X. X. Yan, Q. L. Wang, J. R. Wang, and Z. nian Wang, Chemical Constituents of Calotropis Gigantea.Chem. Nat. Compd.201753963
     [Google Scholar]
  10. HasballahK. SarongM. RuslyR. FitriaH. MaidaD.R. IqhrammullahM. Antiproliferative activity of triterpenoid and steroid compounds from ethyl acetate extract of Calotropis gigantea root bark against P388 murine leukemia cell lines.Sci. Pharm.20218922110.3390/scipharm89020021
     [Google Scholar]
  11. MishraP. YadavK.S. ShrivastavaP. Comparative qualitative and quantitative phytochemical analysis of Calotropis gigantea and Calotropis procera roots.J. Drug Deliv. Ther.20188417910.22270/jddt.v8i4.1757
     [Google Scholar]
  12. PawarD. GhodkeJ. NasreenS. Antimicrobial activity and HR-LCMS analysis of methanolic extract of Calotropis gigantea.Int. J. Adv. Sci. Res.2022424247
     [Google Scholar]
  13. MutiahR. WidyawaruyantiA. The marker active compound identification of Calotropis gigantea roots extract as an anticancer.Int. J. Pharmacol. Pharm. Sci.3112016
     [Google Scholar]
  14. SenS. SahuN.P. MahatoS.B. Flavonol glycosides from Calotropis gigantea.Phytochemistry19923182919292110.1016/0031‑9422(92)83668‑O1368420
     [Google Scholar]
  15. ThakurS. DasP. ItohT. ImaiK. MatsumotoT. Latex extractables of Calotropis gigantea.Phytochemistry19842392085208710.1016/S0031‑9422(00)84985‑7
     [Google Scholar]
  16. KitagawaI. ZhangR. ParkJ.D. BaekN.I. TakedaY. YoshikawaM. ShibuyaH. Indonesian medicinal plants. I. Chemical structures of calotroposides A and B, two new oxypregnane-oligoglycosides from the root of Calotropis gigantea (Asclepiadaceae).Chem. Pharm. Bull. (Tokyo)19924082007201310.1248/cpb.40.20071423756
     [Google Scholar]
  17. ChaisupasakulP. PekthongD. WangteeraprasertA. KaewkongW. SomranJ. KaewpaengN. ParhiraS. SrisawangP. Combination of ethyl acetate fraction from Calotropis gigantea stem bark and sorafenib induces apoptosis in HepG2 cells.PLoS One2024193e030005110.1371/journal.pone.030005138527038
     [Google Scholar]
  18. CJ. Mohan DasN. PeriakaruppanR. Bioactive compounds of Calotropis gigantea for cancer treatment.Oral Oncology Reports20241010033610.1016/j.oor.2024.100336
     [Google Scholar]
  19. RaviR.G. HarikeshD. ChandrasekharT.R. PramodY.G. PatoleA. Cytotoxic activity of Ethanolic root extract of Calotropis gigantea Linn.Int. J. Drug Dev. Res201134101108
     [Google Scholar]
  20. HabibM.R. karimM.R. Evaluation of antitumour activity of Calotropis gigantea L. root bark against Ehrlich ascites carcinoma in Swiss albino mice.Asian Pac. J. Trop. Med.201141078679010.1016/S1995‑7645(11)60194‑622014733
     [Google Scholar]
  21. PattnaikP.K. KarD. ChhatoiH. ShahbaziS. GhoshG. KuanarA. Chemometric profile & antimicrobial activities of leaf extract of Calotropis procera and Calotropis gigantea.Nat. Prod. Res.201731161954195710.1080/14786419.2016.126634927936921
     [Google Scholar]
  22. AkhirJ. TarmanA. Potential and opportunities for utilization of crown flower plants (Calotropis gigantea) in Dusun Bunot Desa Alue Naga Kecamatan Syiah Kuala Banda Aceh.IOP Conf. Ser. Earth Environ. Sci.2020425101200910.1088/1755‑1315/425/1/012009
     [Google Scholar]
  23. KemalaP. IdroesR. KhairanK. RamliM. JalilZ. IdroesG.M. TalleiT.E. HelwaniZ. SafitriE. IqhrammullahM. NasutionR. Green synthesis and antimicrobial activities of silver nanoparticles using Calotropis gigantea from Ie Seu-Um Geothermal Area, Aceh Province, Indonesia.Molecules20222716531010.3390/molecules2716531036014547
     [Google Scholar]
  24. AliE.M. AbdallahB.M. Effective Inhibition of Candidiasis Using an Eco-Friendly Leaf Extract of Calotropis-gigantean-Mediated Silver Nanoparticles.Nanomaterials (Basel)202010342210.3390/nano1003042232121137
     [Google Scholar]
  25. SivapalanS. DharmalingamS. VenkatesanV. AngappanM. AshokkumarV. Phytochemical analysis, anti-inflammatory, antioxidant activity of Calotropis gigantea and its therapeutic applications.J. Ethnopharmacol.202330311596310.1016/j.jep.2022.11596336442758
     [Google Scholar]
  26. ChitmeH.R. ChandraM. KaushikS. Studies on anti-diarrhoeal activity of Calotropis gigantea R.Br. in experimental animals.J. Pharm. Pharm. Sci.200471707515144737
     [Google Scholar]
  27. SombiéE.N. SomdaD.G. KonatéS. YoulO. TraoréT.K. N’doJ.Y.P. Belem-KabréW.L.M.E. OuédraogoG.G. OuédraogoN. HilouA. HPLC analysis and protective effect of fractions from ethanolic extract of Calotropis procera (Ait.) R. Br root bark against diethylnitrosamine induced-hepatic damage in Wistar rats.Trop. J. Nat. Prod. Res.202373462
     [Google Scholar]
  28. BajpaiS. HoodaH. MishraR. Surabhi BajpaiC. SinghR. Comparative analysis of antioxidant properties of extracts of Calotropis procera with different anti-diabetic drugs.Int. J. Herb. Med.20186104
     [Google Scholar]
  29. DuttaM. NezamM. ChowdhuryS. RakibA. PaulA. SamiS.A. UddinM.Z. RanaM.S. HossainS. EffendiY. IdroesR. TalleiT. AlqahtaniA.M. EmranT.B. Appraisals of the Bangladeshi medicinal plant Calotropis gigantea used by folk medicine practitioners in the management of COVID-19: A biochemical and computational approach.Front. Mol. Biosci.2021862539110.3389/fmolb.2021.62539134124140
     [Google Scholar]
  30. ArgalA. SachanR. Anthelmintic activity of Calotropis gigantea roots.J. Pharm. Res.200942826
     [Google Scholar]
  31. DeshmukhP.T. FernandesJ. AtulA. ToppoE. Wound healing activity of Calotropis gigantea root bark in rats.J. Ethnopharmacol.2009125117818110.1016/j.jep.2009.06.00719539020
     [Google Scholar]
  32. ArgalA. PathakA.K. CNS activity of Calotropis gigantea roots.J. Ethnopharmacol.2006106114214510.1016/j.jep.2005.12.02416446065
     [Google Scholar]
  33. KaurK. KumarD. KumarS. Screening of neuropharmacological activities of Calotropis gigantea roots.J. Pharm., Chem. Biol. Sci.231861962014
     [Google Scholar]
  34. SrivastavaS.R. KeshriG. BhargavanB. SinghC. SinghM.M. Pregnancy interceptive activity of the roots of Calotropis gigantea Linn. in rats.Contraception200775431832210.1016/j.contraception.2006.11.01017362713
     [Google Scholar]
  35. JahanN. MushirA. AhmedA. A review on Phytochemical and biological properties of Calotropis gigantea (Linn) R.Br.Discov. Phytomedicine - J. Nat. Prod. Res. Ethnopharmacol.201632152110.15562/phytomedicine.2016.32
     [Google Scholar]
  36. BhavsarS.K. PatilS.N. MurkuteP.S. SuranaS.J. Pharmacological, biological activities and phytochemical constituents of Calotropis gigantea.J. Phytopharmacol.202091616610.31254/phyto.2020.9110
     [Google Scholar]
  37. SahuA. SenS. MishraS.C. Processing and properties of Calotropis gigantea bio-char: A wasteland weed.Mater. Today Proc.2020335334534010.1016/j.matpr.2020.03.024
     [Google Scholar]
  38. AyemeleA.G. WangY. MaL. BuD. XuJ. Turning weeds into feed: Ensiling Calotropis gigantea (Giant milkweed) reduces its toxicity and enhances its palatability for dairy cows.Ecotoxicol. Environ. Saf.202427611629210.1016/j.ecoenv.2024.11629238581911
     [Google Scholar]
  39. Al SulaibiM.A.M. ThiemannC. ThiemannT. Chemical constituents and uses of Calotropis procera and Calotropis gigantea – A review (Part I – The plants as material and energy resources).Open Chem. J.20207111510.2174/1874842202007010001
     [Google Scholar]
  40. HandayaniL. ApriliaS. ArahmanN. BiladM.R. Assessment of fibers from different part of the Calotropis gigantea biomass as a filler of composites foam PVA/PVP.S. Afr. J. Chem. Eng.20244918919810.1016/j.sajce.2024.03.016
     [Google Scholar]
  41. TalithaO. SamanhudiS. SetyawatiA. RahayuM. SakyaA.T. The effect of growth concentration on in vitro shoot multiplication of crown flower (Calotropis gigantea).Plant Breed. Biotechnol.202210424425610.9787/PBB.2022.10.4.244
     [Google Scholar]
  42. GanesanV. ShanmugamV. AlagumalaiV. KaliyamoorthyB. DasO. MisraM. Optimisation of mechanical behaviour of Calotropis gigantea and Prosopis juliflora natural fibre-based hybrid composites by using Taguchi-Grey relational analysis.Composites Part C: Open Access20241310043310.1016/j.jcomc.2024.100433
     [Google Scholar]
  43. KharatK.R. KharatA.S. The Calotropis gigantea methanolic extract induces apoptosis in human breast carcinoma cells.Iran. J. Med. Sci.201944648349231875083
     [Google Scholar]
  44. KumarD. KumarA. KumarV. RajA. RaiR.R.M. BaliyanV. KumarN. A comprehensive review on analytical method development using RP-HPLC and recent advances in pharmaceutical applications.J. Res. Appl. Sci. Biotechnol.202322536010.55544/jrasb.2.2.9
     [Google Scholar]
  45. PrathapB. SrinivasaG.H. ArthanariswaranP. AkalankaD. JohnsonP. A review - Importance of RP-HPLC in analytical method development.Int. J. Novel Trends Pharm. Sci.2013315
     [Google Scholar]
  46. GanjiwaleS.V. DewaniA.P. ChandewarA.V. A comprehensive overview Of HPLC method development and validation.Int. J. Pharma Sci.202421
     [Google Scholar]
  47. ShindeS. RajurkarV. Advances in high-performance liquid chromatography (HPLC) method development and validation: A comprehensive review.J. Drug Deliv. Biother.202412768610.61920/jddb.v1i02.138
     [Google Scholar]
  48. KhanH. Analytical method development in pharmaceutical research: Steps involved in HPLC method development.Asian J. Pharm. Res.20177320310.5958/2231‑5691.2017.00031.4
     [Google Scholar]
  49. Shivaji BhamareK. Balasaheb GangurdeS. BhamareN.S. TaroY.V. BargudeA.J. LadeH.R. DhavaleP. A review on phytochemical and biological properties of Calotropis gigantea.Int. J. Res. Publ. Rev. J.202341593
     [Google Scholar]
  50. GaneshV. Poorna BasuriP. SahiniK. NaliniC.N. Retention behaviour of analytes in reversed‐phase high‐performance liquid chromatography — A review.Biomed. Chromatogr.2023377e548210.1002/bmc.548235962484
     [Google Scholar]
  51. RaposoF. Ibelli-BiancoC. Performance parameters for analytical method validation: Controversies and discrepancies among numerous guidelines.Trends Analyt. Chem.202012911591310.1016/j.trac.2020.115913
     [Google Scholar]
  52. SanapG.S. ZarekarN.S. PawarS.S. Review on method development and validation.Int. J. Pharm. Drug Anal.20175177
     [Google Scholar]
  53. SpínolaV. Llorent-MartínezE.J. CastilhoP.C. Determination of vitamin C in foods: Current state of method validation.J. Chromatogr. A2014136921710.1016/j.chroma.2014.09.08725441066
     [Google Scholar]
  54. PumJ. A practical guide to validation and verification of analytical methods in the clinical laboratory.Adv. Clin. Chem.20199021528110.1016/bs.acc.2019.01.00631122610
     [Google Scholar]
  55. HPLC troubleshooting: Why signal-to-noise ratio determines limit of detection.2022Available from: https://www.thermofisher.com/blog/analyteguru/hplc-troubleshooting-tips-why-signal-to-noise-ratio-determines
  56. SadaphalP. DhamakK. Review article on High-Performance Liquid Chromatography (HPLC) method development and validation.Int. J. Pharm. Sci. Rev. Res.202274232910.47583/ijpsrr.2022.v74i02.003
     [Google Scholar]
  57. ChauhanV. GroverP. BhardwajM. KumarS. NagarajanK. Development and validation of fast and sensitive RP-HPLC stability-indicating method for quantification of piroxicam in bulk drug.J. Chromatogr. Sci.2024bmae02110.1093/chromsci/bmae02138704244
     [Google Scholar]
  58. SoniaK. DrN. Development and validation of HPLC and UV-Visible spectrophotometric method for the pharmaceutical dosage form and biological fluid – Review.Eur. J. Biomed. Pharm. Sci.20163382
     [Google Scholar]
  59. TransfiguracionJ. MenaJ.A. AucoinM.G. KamenA.A. Development and validation of a HPLC method for the quantification of baculovirus particles.J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.20118791616810.1016/j.jchromb.2010.11.01121123120
     [Google Scholar]
  60. Assay.Available from: https://en.wikipedia.org/wiki/Assay
/content/journals/cpa/10.2174/0115734129343858241007073450
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Establishment and Validation of a Robust Reversed-Phase HPLC Method for the Determination of Calotropis gigantea in Bulk Material and Marketed Product
Current Pharmaceutical Analysis 20, 920 (2024); https://doi.org/10.2174/0115734129343858241007073450
/content/journals/cpa/10.2174/0115734129343858241007073450
/content/journals/cpa/10.2174/0115734129343858241007073450
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  • Article Type:     Research Article
Keyword(s): Calotropis gigantea; estimation; ICH guidelines; method development; RP-HPLC; validation

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