AJAC  Vol.5 No.2 , January 2014
Quantitative Determination of Nitidine from Roots and Plant Tissue Culture Extracts of Toddalia asiatica (Linn.) Using HPTLC
Abstract: Toddalia asiatica Linn. is an important medicinal plant belonging to the family Rutaceae. The plant is well known for its antimalarial activity, which has been attributed to the presence of benzophenanthridine alkaloid nitidine in the roots of plants. A simple, rapid, sensitive, accurate, repeatable and robust HPTLC method has been developed and validated for the quantitative determination of nitidine in the dried roots and plant tissue culture extracts of T. asiatica. Nitidine was estimated at 332 nm by densitometry using Silica gel 60 F254 as stationary phase and chloroform:methanol (7:1, v/v), and as mobile phase. Linearity was observed in the concentration range of 25 -200 ng/spot for nitidine. The limit of detection and limit of quantitation were found to be 0.026 and 0.086 ng/spot respectively for nitidine. Developed method was validated according to the ICH guidelines with respect to precision, accuracy, specificity and robustness. The technique has been applied for the first time for the estimation of nitidine in roots and plant tissue culture extracts of T. asiatica. Statistical analysis data indicate the accuracy and reliability of the method.
Cite this paper: C. Praveena and C. Veeresham, "Quantitative Determination of Nitidine from Roots and Plant Tissue Culture Extracts of Toddalia asiatica (Linn.) Using HPTLC," American Journal of Analytical Chemistry, Vol. 5 No. 2, 2014, pp. 65-69. doi: 10.4236/ajac.2014.52010.

[1]   R. K. Molmoori, H. C. Rodda, K. Asres and C. Veeresham, “Toddalia asiatica (Linn.) Lam.—A Comprehensive Review,” Pharmacognosy Reviews, Vol. 2, No. 4, 2008, pp. 386-397.

[2]   D. M. N. Gakunju, E. K. Mberu, S. F. Dossaji, A. I. Gray, R. D. Waigh, P. G. Waterman and W. M. Watkins, “Potent Antimalarial Activity of the Alkaloid Nitidine, Isolated from a Kenyan Herbal Remedy,” Antimicrobial Agents and Chemotherapy, Vol. 39, No. 12, 1995, pp. 2606-2609.

[3]   F. W. Muregi, A. Ishih, T. Miyase, T. Suzuki, H. Kino, T. Amanod, G. M. Mkoji and M. Terada, “Antimalarial Activity of Methanolic Extracts from Plants Used in Kenyan Ethnomedicine and Their Interactions with Chloroquine (CQ) against a CQ-Tolerant Rodent Parasite, in Mice,” Journal of Ethnopharmacology, Vol. 111, No. 1, 2007, pp. 190-195.

[4]   J. M. Nyangulu, S. L. Hargreaves, S. L. Sharples, S. P. Mackay, R. D. Waigh, O. Duval, E. K. Mberu and W. M. Watkins, “Antimalarial Benzo[c]phenanthridines,” Bioorganic and Medicinal Chemistry Letters, Vol. 15, No. 8, 2005, pp. 2007-2010.

[5]   M. A. Rashid, K. R. Gustafson, Y. Kashman, J. H. Cardellina II, J. B. Mc Mohan and M. R. Boyd, “Anti-HIV Alkaloids from Toddalia asiatica,” Natural Product Research, Vol. 6, No. 2, 1995, pp. 153-156.

[6]   V. K. Tandon and R. B. Chhor, “Current Status of AntiHIV Agents,” Current Medicinal Chemistry—Anti-Infective Agents, Vol. 4, No. 1, 2005, pp. 3-28.

[7]   H. Iwasaki, H. Oku, R. Takara, H. Miyahira, K. Hanashiro, Y. Yoshida, et al., “The Tumor Specific Cytotoxicity of Dihydronitidine from Toddalia asiatica Lam.,” Cancer Chemotherapy Pharmacology, Vol. 58, No. 4, 2006, pp. 451-459.

[8]   V. K. Singh and J. N. Govil, “Standardization of Herbal/ Ayurvedic Formulations: Recent Progress in Medicinal Plants,” 3rd Edition, Vol. 24, Stadium Press LLC, Houston, 2006, pp. 25-44.

[9]   H. C. Andola and V. K. Purohit, “High Performance Thin Layer Chromatography (HPTLC): A Modern Analytical Tool for Biological Analysis,” Nature and Science, Vol. 8, No. 10, 2010, pp. 58-61.

[10]   V. Sharma, A. P. Gupta, P. Bhandari, R. C. Gupta and B. Singh, “A Validated and Densitometric HPTLC Method for the Quantification of Withaferin-A and Withanolide-A in Different Plant Parts of Two Morphotypes of Withania somnifera,” Chromatographia, Vol. 66, No. 9-10, 2007, pp. 801-804.

[11]   K. Ashraf, M. Mujeeb, A. Ahmad, M. Amir, Md. N. Mallick and D. Sharma, “Validated HPTLC Analysis Method for Quantification of Variability in Ccontent of Curcumin in Curcuma longa L (turmeric) Collected from Different Geographical Region of India,” Asian Pacific Journal of Tropical Biomedicine, Vol. 2, No. 2, 2012, pp. S588-S584.

[12]   P. Srivastava, N. Tiwari, A. K. Yadav, V. Kumar, K. Shanker, R. K. Verma, M. M. Gupta, A. K. Gupta and S. P. Khanuja, “Simultaneous Quantification of Withanolides in Withania somnifera by a Validated High-Performance Thin-Layer Chromatographic Method,” Journal of AOAC International, Vol. 91, No. 5, 2008, pp. 1154-1161.

[13]   P. B. Shinde, P. D. Aragade, M. R. Agrawal, U. A. Deokate and S. S. Khadabadi, “Simultaneous Determination of Withanolide A and Bacoside A in Spansules by HighPerformance Thin-Layer Chromatography,” Indian Journal of Pharmaceutical Sciences, Vol. 73, No. 2, 2011, pp. 240-243.

[14]   V. Y. Thakker, V. N. Shah, U. D. Shah and M. P. Suthar, “Simultaneous Estimation of Gallic Acid, Curcumin and Quercetin by HPTLC Method,” Journal of Advanced Pharmacy Education and Research, Vol. 1, 2011, pp. 70-80.

[15]   R. K. Molmoori, H. C. Rodda and C. Veeresham, “Production of Nitidine from Callus Cultures of Toddalia asiatica,” International Journal of Pharmaceutical Sciences and Nanotechnology, Vol. 3, No. 2, 2010, pp. 1028-1033.

[16]   International Conference on Harmonization (ICH), “Validation of Analytical Procedures; Methodology,” Q2A, Geneva, 1994, pp. 1-5.

[17]   International Conference on Harmonization (ICH), “Validation of Analytical Procedures; Methodology,” Q2B, Geneva, 1996, pp. 3-9.