Planta Med 2023; 89(04): 441-453
DOI: 10.1055/a-1876-3009
Natural Product Chemistry & Analytical Studies
Original Papers

Comparative Morpho-micrometric Investigations in Six Indigenous Ocimum Species of India with DOE Based HPTLC Method for Multi-class Component Analysis

1   Pharmanza Herbal Pvt. Ltd., Anand, Gujarat, India
,
Sautik Ghoshal
1   Pharmanza Herbal Pvt. Ltd., Anand, Gujarat, India
,
Chetana Ghule
1   Pharmanza Herbal Pvt. Ltd., Anand, Gujarat, India
,
Krishna Bhatt
2   Department of Quality Assurance, Anand College of Pharmacy, Anand, Gujarat, India
,
Kalpana Patel
2   Department of Quality Assurance, Anand College of Pharmacy, Anand, Gujarat, India
,
1   Pharmanza Herbal Pvt. Ltd., Anand, Gujarat, India
,
Lal Hingorani
1   Pharmanza Herbal Pvt. Ltd., Anand, Gujarat, India
› Author Affiliations

Abstract

The Ocimum genus is one of Indiaʼs prominent botanical classes of traditional medicinal culture comprising medicinally and agronomically important plants. Morphological resemblances, overlapping geographical distribution, and history of traditional nomenclature have necessitated a comprehensive qualitative report for effective quality control and removing the species ambiguity pertaining to this genus. This paper provides detailed morpho-micrometric characteristics used to differentiate between six indigenous Ocimum species of India. Among them, O. gratissimum was distinguished as the only shrub with a fleshy petiole. In green and purple forms, O. tenuiflorum leaves had serrate margins and showed no particular anatomical differences except for the anthocyanins containing epidermal cells of the latter. O. basilicum had glabrous leaves except for the veins, which were puberulous. O. filamentosum had tenuous anther filaments and was the least aromatic while O. africanum had a citrusy odour, which along with the number of xylary rows, size of mesophyll cells, and epidermal cell wall architecture, distinguished it from O. americanum. An HPTLC method was developed using experimental design and validated for quantification of multi-class compounds from terpenoic, phenolic acids, and flavonoids in Ocimum leaves. It was found linear (r 2 > 0.99) with recoveries between 95 – 100% for all compounds. The eluted bands of marker compounds were subjected to HPTLC-MS analysis as a confirmative tool. This is the first anatomical and analytical report of O. filamentosum Forssk. The obtained results could be effectively used for species identification using vegetative characters alone with the anatomical-HPTLC data backing up the former as a rapid and economical tool.

Supporting Information



Publication History

Received: 29 March 2022

Accepted: 27 May 2022

Article published online:
22 August 2022

© 2022. Thieme. All rights reserved.

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  • References

  • 1 Paton AJ, Harley RM, Harley MM. Ocimum: An Overview of Classification and Relationships. In: Hiltunen R, Holm Y, eds. Basil: the genus Ocimum Amsterdam: Harwood Academic Publishers; 1999: 1-36
  • 2 POWO. Plants of the World Online. Facilitated by the Royal Botanic Gardens, Kew. Accessed June 22, 2021 at: http://www.plantsoftheworldonline.org/taxon/urn:lsid:ipni.org:names:21070-1
  • 3 Kumar A, Shukla AK, Shasany AK, Sundaresan V. Systematic Position, Phylogeny, and Taxonomic Revision of Indian Ocimum . In: Shasany A, Kole C, eds. The Ocimum Genome. Compendium of Plant Genomes. Cham: Springer; 2018: 61-72
  • 4 Anonymous The Ayurvedic Pharmacopoeia of India, Part-I, vol.-II. New Delhi: Government of India, Ministry of Health and Family Welfare, Department of Indian System of Medicine & Homeopathy; 1999
  • 5 Chowdhury T, Mandal A, Roy SC, De Sarker D. Diversity of the genus Ocimum (Lamiaceae) through morpho-molecular (RAPD) and chemical (GC–MS) analysis. J Genet Eng Biotechnol 2017; 15: 275-286 DOI: 10.1016/j.jgeb.2016.12.004.
  • 6 Singh D, Chaudhuri PK. A review on phytochemical and pharmacological properties of Holy basil (Ocimum sanctum L.). Ind Crops Prod 2018; 118: 367-382 DOI: 10.1016/j.indcrop.2018.03.048.
  • 7 Brandão LB, Santos LL, Martins RL, Rodrigues ABL, da Costa ALP, Faustino CG, da Silva de Almeida SSM. The potential effects of species Ocimum basilicum on health: A review of the chemical and biological studies. Pharmacogn Rev 2022; 16: 22-26 DOI: 10.5530/phrev.2022.16.4.
  • 8 Zengin G, Ferrante C, Gnapi DE, Sinan KI, Orlando G, Recinella L, Diuzheva A, Jekő J, Cziáky Z, Chiavaroli A, Leone S, Brunetti L, Picot-Allain C, Mahomoodally MF, Angelini P, Covino S, Venanzoni R, Tirillini B, Menghini L. Comprehensive approaches on the chemical constituents and pharmacological properties of flowers and leaves of American basil (Ocimum americanum L). Food Res Int 2019; 125: 108610 DOI: 10.1016/j.foodres.2019.108610.
  • 9 Sharma NK. Ocimum americanum L.-A Plant Remedy Used in Appendicitis by the Tribals of South East Rajasthan. Nelumbo 2003; 45: 203-208 DOI: 10.20324/nelumbo/v45/2003/74213.
  • 10 Ugbogu OC, Emmanuel O, Agi GO, Ibe C, Ekweogu CN, Ude VC, Uche ME, Nnanna RO, Ugbogu EA. A review on the traditional uses, phytochemistry, and pharmacological activities of clove basil (Ocimum gratissimum L.). Heliyon 2021; 7: e08404 DOI: 10.1016/j.heliyon.2021.e08404.
  • 11 Teanpaisan R, Kawsud P, Pahumunto N, Puripattanavong J. Screening for antibacterial and antibiofilm activity in Thai medicinal plant extracts against oral microorganisms. J Tradit Complement Med 2017; 7: 172-177 DOI: 10.1016/j.jtcme.2016.06.007.
  • 12 Soni N, Gill D, Sagar BS, Raheja S, Agrawal S. Ocimum kilimandscharicum: A systematic review. J Drug Deliv Therap 2012; 2: 45-52 DOI: 10.22270/jddt.v2i3.169.
  • 13 Kashyap CP, Ranjeet K, Vikrant A, Vipin K. Therapeutic potency of Ocimum kilimandscharicum Guerke – a review. Global J Pharmac 2011; 5: 191-200
  • 14 Thoppil JE, Minija J, Tajo A, Deena MJ. Microbicidal potential of the essential oil of Ocimum adscendens . J Med Aroma Plant Sci 2000; 22: 256-257
  • 15 Asthana A, Tripathi NN, Dixit SN. Fungitoxic and phytotoxic studies with essential oil of Ocimum adscendens . J Phytopathol 1986; 117: 152-159 DOI: 10.1111/j.1439-0434.1986.tb00639.x.
  • 16 Sumitha KV, Thoppil JE, Shihabudeen S, Bukhari NA, Al-Khattaf FS, Hatamleh AA. Methanol extracts of different species of Ocimum alleviate the peroxide radical-mediated cell injury and redox imbalance in human colon epithelial cells. Physiol Mol Plant Pathol 2022; 117: 101759 DOI: 10.1016/j.pmpp.2021.101759.
  • 17 Anusmitha KM, Aruna M, Job JT, Narayanankutty A, Benil PB, Rajagopal R, Alfarhan A, Barcelo D. Phytochemical analysis, antioxidant, anti-inflammatory, anti-genotoxic, and anticancer activities of different Ocimum plant extracts prepared by ultrasound-assisted method. Physiol Mol Plant Pathol 2022; 117: 101746 DOI: 10.1016/j.pmpp.2021.101746.
  • 18 Ali H, Nguta J, Musila F, Ole-Mapenay I, Matara D, Mailu J. Evaluation of antimicrobial activity, cytotoxicity, and phytochemical composition of Ocimum americanum L. (Lamiaceae). Evid Based Complement Alternat Med 2022; 2022: 6584578 DOI: 10.1155/2022/6484578.
  • 19 Mounira GM, Ahlem Z, Mariem BA, Romdhane M, Okla MK, Al-Hashimi A, Alwase YA, Madnay MM, AbdElgayed G, Asard H, Beemster GTS, AbdElgawad H. Essential oil composition and antioxidant and antifungal activities of two varieties of Ocimum basilicum L. (Lamiaceae) at two phenological stages. Agronomy 2022; 12: 825 DOI: 10.3390/agronomy12040825.
  • 20 Sneha K, Narayanankutty A, Job JT, Olatunji OJ, Alfarhan A, Famurewa AC, Ramesh V. Antimicrobial and larvicidal activities of different Ocimum essential oils extracted by ultrasound-assisted hydrodistillation. Molecules 2022; 27: 1456 DOI: 10.3390/molecules27051456.
  • 21 Thomas V, Nair SV, Ved DK, Shankar D. Controversial identities of medicinal plants in classical literature of Ayurveda. J Ayurveda Integr Med 2020; 11: 565-572 DOI: 10.1016/j.jaim.2019.09.003.
  • 22 Liji KO, Vasudevan CN. Comparative pharmacognostic and phytochemical studies of Ocimum tenuiflorum L. and its substitute Vitex negundo L. Int J Ayurveda Pharma Res 2017; 5: 9-19
  • 23 Jürges G, Sahi V, Rios Rodriguez D, Reich E, Bhamra S, Howard C, Slater A, Nick P. Product authenticity versus globalisation – The Tulsi case. PLoS One 2018; 13: e0207763 DOI: 10.1371/journal.pone.0207763.
  • 24 Kumar A, Rodrigues V, Baskaran K, Shukla AK, Sundaresan V. DNA barcode based species-specific marker for Ocimum tenuiflorum and its applicability in quantification of adulteration in herbal formulations using qPCR. J Herb Med 2020; 23: 100376 DOI: 10.1016/j.hermed.2020.100376.
  • 25 Bhamra S, Heinrich M, Howard C, Johnson M, Slater A. DNA authentication of Tulsi; the cultural and medicinal value of Ocimum species among diasporic South Asian communities in the UK. Planta Medica 2014; 80: P2B13 DOI: 10.1055/s-0034-1394890.
  • 26 Kumar A, Rodrigues V, Mishra P, Baskaran K, Shukla AK, Shasany AK, Sundaresan V. ISSR-Derived Species-Specific SCAR marker for rapid and accurate authentication of Ocimum tenuiflorum L. Planta Medica 2018; 84: 117-122 DOI: 10.1055/s-0043-116853.
  • 27 Khatoon S. Macro-microscopy & planar chromatography – important tools for quality control and identification of adulterants/substitutes of Unani drugs. Int J Adv Pharm Med Bioallied Sci 2015; 3: 58-64
  • 28 Deore SL, Ingole SR, Baviskar BA. Comparative pharmacognostical, phytochemical and biological evaluation of five Ocimum species. Pharmacogn J 2021; 13: 463-474 DOI: 10.5530/pj.2021.13.59.
  • 29 Joshi A, Pathak AK, Tailang M. Comparative pharmacognostical characterization of selected species of Ocimum . J Drug Deliv Thera 2014; 4: 72-80 DOI: 10.22270/jddt.v4i2.785.
  • 30 Parida S, Mohapatra BK, Mahalik G. Anatomical study of six Ocimum species: The valuable method used in Indian Systems of Medicines (ISM). Int J Ayur Medi 2020; 11: 278-283 DOI: 10.47552/ijam.v11i2.1507.
  • 31 Ahirwar PK, Pathak J, Soni P, Tripatrhi MK. Pharmacognostical standardization and HPTLC fingerprint of various species of Ocimum leaf. J Emer Techno Inno Res 2019; 6: 295-302 DOI: 10.13140/RG.2.2.15830.91207.
  • 32 Gurav TP, Dholakia BB, Giri AP. A glance at the chemo-diversity of Ocimum species: Trends, implications, and strategies for the quality and yield improvement of essential oil. Phytochem Rev 2021; 2: 1-35 DOI: 10.1007/s11101-021-09767-z.
  • 33 Azzazy MF. Micromorphology of pollen grains, trichomes of sweet Basil, Egypt. Adv Compl Alter Medi 2019; 5: 427-433
  • 34 Fernandes VF, de Almeida LB, Feijó EVRdaS, Silva DdaC, de Oliveira RA, Mielke MS, Costa LCdoB. Light intensity on growth, leaf micromorphology and essential oil production of Ocimum gratissimum . Revista Brasileira de Farmacognosia 2013; 23: 419-424 DOI: 10.1590/S0102-695X2013005000041.
  • 35 Homa K, Barney WP, Ward DL, Wyenandt CA, Simon JE. Morphological characteristics and susceptibility of basil species and cultivars to Peronospora belbahrii. Hort Sci 2016; 51: 1389-1396 DOI: 10.21273/HORTSCI09778-16.
  • 36 Nassar MA, El-Sagai MU, Azoz SN. Anatomical and phytochemical studies on Ocimum basilicum L. plant (Lamiaceae). Inter J Adv Rese 2014; 2: 204-226
  • 37 Ingole SN. Study of some anatomical characters in Ocimum americanum L. (Lamiaceae). Vidyabharati Inter Interdisc Res J 2012; 1: 61-68
  • 38 Deogade MS, Prasad KS. Standardization of wild Krushna tulasi (Ocimum tenuiflorum Linn) leaf. Inter J Ayur Medi 2019; 10: 52-61 DOI: 10.47552/ijam.v10i1.1172.
  • 39 Siya M, Mary AP. Studies on pharmacognosy, phytochemical and antimicrobial activity of Ocimum sanctum L. World J Sci Res 2017; 2: 01-12
  • 40 Sarma DS, Babu AV. Pharmacognostic and phytochemical studies of Ocimum americanum . J Chem Pharma Res 2011; 3: 337-347
  • 41 Patel KG, Patel VG, Patel KV, Gandhi TR. Validated HPTLC method for quantification of myricetin in the stem bark of Myrica esculenta Buch. Ham. Ex D. Don, Myricaceae. JPC – J Planar Chromat 2010; 23: 326-331 DOI: 10.1556/JPC.23.2010.5.4.
  • 42 Javanmardi J, Khalighi A, Kashi A, Bais HP, Vivanco JM. Chemical characterization of basil (Ocimum basilicum L.) found in local accessions and used in traditional medicines in Iran. J Agric Food Chem 2002; 50: 5878-5883 DOI: 10.1021/jf020487q.
  • 43 Vieira RF, Simon JE. Chemical characterization of basil (Ocimum spp.) found in the markets and used in traditional medicine in Brazil. Econ Bot 2000; 54: 207-216 DOI: 10.1007/BF02907824.
  • 44 Girme A, Bhoj P, Saste G, Pawar S, Mirgal A, Raut D, Chavan M, Hingorani L. Development and validation of RP-HPLC method for Vicenin-2, Orientin, Cynaroside, Betulinic Acid, Genistein, and major eight bioactive constituents with LC-ESI-MS/MS profiling in Ocimum genus. J AOAC Int 2021; 104: 1634-1651 DOI: 10.1093/jaoacint/qsab067.