Chromatographic Profiling and Cytotoxic studies of Extracts of Buchanania lanzan Spreng and Albizia lebbeck Benth

 

Jyoti Shrivastava1*, T. Ratna Madhuri1, Abhishek Shah2

1Department of Pharmaceutical Chemistry, The Oxford College of Pharmacy, Hongasandra, Bangalore - 560068.

2Era College of Pharmacy, Era University, Sarfarjganj, Lucknow 226003.

*Corresponding Author E-mail: jyotishrivastavapharmacy@gmail.com

 

ABSTRACT:

The various Indian systems of medicine in addition to numerous local health traditions have been successful in managing cancer across the globe using herbs. Keeping all this in mind, the present study has been taken up, to study the cytotoxic potential of B.lanzan and A.lebbeck barks. The objective of the present study is to explore the scientific basis of both plant bark for its anti-carcinogenic utility. Different extracts of plants were screened using the MTT assay technique, and potent extracts were subjected to HPTLC and HPLC to evaluate total phenolic and flavonoid present in crude extracts of plant bark. In our study, we found that the ethanolic fraction of B. lanzan had higher phenolic content 34.92 g/mL than other fractions. While in the case of flavonoids ethyl acetate fraction of A.lebbeck contain the highest concentration 17.00 g/mL. In the MTT assay, B. lanzan ethyl acetate extract (B-EA), ethanolic extract of B. lanzan (BE) and A. lebbeck ethyl acetate extract (L-EA) had shown an IC50 value of 113.9 g/mL, 247.4g/mL, 152.7g/mL, while in A549 cells, B-EA, BE, L-EA had shown 226.9g/mL, 269.1g/mL, 123.3g/mL IC50 values respectively. The effect of samples B-EA, and L-EA was found different in HeLa cell lines from the other two studies. The samples B-EA and L-EA showed the value of 91.9g/mL and 257.1g/mL, respectively. A. lebbeck ethanol extract (LA) did not show significant inhibition in HepG2, A549, and HeLa cells, and sample BE did not show substantial inhibition in HeLa cells. B.lanzan and A. lebbeck have a high potency to treat liver, lung and cervical cancers in vitro, although extensive pharmacological and clinical studies are required to explore inherent mechanisms in future.

 

KEYWORDS: Chromatography, Cytotoxicity, Buchanania lanzan Spreng and Albizia lebbeck Benth.

 

 


INTRODUCTION:

Natural products have been the pillar of chemotherapy as far back as 30 years. The more high-quality parts of them are acquired from plants or micro-organisms, as plant-subsidiary medications.1 Some of the natural origins drugs such as vinblastin, vincristine, topotecan, paclitaxel and other natural antibiotics are the best example of natural origin drugs which are now part of regular clinical treatment.2 According to Cragg et al3 and Mann J2, more than 50 % of anticancer drugs in use today are derived from natural sources.

 

India has an old legacy of common medication. Indian conventional medicine depends on different frameworks comprising Ayurveda, Unani and Siddha. The mentioned common frameworks of Indian medicines are unique in themselves, but their basic principles and applications have a common thread moving among these systems. The current scenario has a keen interest in adopting and studying these traditional methods and explores the potential of these methods on the ground of different healthcare systems. The assessment or standardization of the huge heritage of conventional drug therapy is necessary.4

 

In conventional methods of cancer treatments, herbal drugs are not directly curing the disease, yet they assist with adapting to signs and side effects brought about by the disease. Common signs and symptoms such as anxiety, fatigue, nausea, vomiting, pain, difficulty sleeping, and stress may be lessened by alternative treatments. The traditions vary from place to place. The diversity of herbs is also vast. The various Indian systems of medicine; Ayurveda, Homeopathy, Unani, Siddha and Naturopathy in addition to numerous local health traditions are using herbs successfully in managing cancer across the globe. Keeping all this in mind the present study has been taken up, to study the potential of Buchanania lanzan preng and Albizzia lebbeck Benth so that they can be useful as an anti-cancerous drugs in national and international countries.5

 

Buchanania lanzan preng. (Syn: Buchanania latifolia Roxb, Fam: Anacardiaceae) is a ubiquitous plant in dried parts of India. It is commonly known as Chaar in India and Almondette tree in English. Phytochemical analysis of the plant reveals the presence of all essential components, especially flavonoids and gallic acid in abandoned. Scientific studies on B. lanzan bark have been found to prevent cyclophosphamide-induced genotoxicity and oxidative stress in mice6 while dry fruits have been reported to show immunostimulant and astringent activity7,8.

 

Albizzia lebbeck Benth (Shirish, Family: Leguminosae) is a deciduous tree with compound leaves, flat oblong fruits, and round cream-coloured seeds, that grows wild. Albizzia lebbeck is a tree well-known in the Indian subcontinent for its range of uses.9,10 Phytochemical investigations showed that the pod of the Albizzia lebbeck contains 3,5 Dihydroxy 4,7 dimethoxy flavone11,12, and N- Benzoyl L phenyl alaninol13. The beans of the plant contain albigenic acid-a new triterpenoid sapogenin. The plant also contains saponins14,15 macrocyclic alkaloids16 Tannins and flavonols17. An extensive literature review on the plant had put forward strong evidence of anti-asthmatic activity, anti-diabetic, anti-inflammatory, hepatoprotective, antioxidant and anticancer properties in it.

 

Saponins are well-known steroids or glycosides of triterpenoids type, found commonly in a vast variety of plants and their products. These are having potential importance as nutrition for humans and animals. Saponins are known for numerous biological effects. These compounds with structural diversity have been proven to have the potential to stunt the growth of cancerous cells.18. The present study explored the scientific basis for the utility of Buchanania lanzan Spreng bark and Albizzia lebbeck Benth bark for the protection of carcinoma.

 

MATERIALS AND METHODS:

Plant material:

Bark of two plants, 3kg each namely Buchanania lanzan Spreng and Albizzia lebbeck Benth were collected by Dr K. Madhava Chetty, Assistant Professor, Department of Botany, SV University, Tirupati (A.P). The accession/Voucher No. for the specimen is 1123 and 1712 respectively. A certified specimen is kept in the department for future reference.

 

Preparation of crude extract:

The bark Buchanania lanzan Spreng and Albizzia lebbeck Benth was collected and shade dried separately followed by an incubator for two days at 400. The dried bark material was coarsely powdered using an electrical grinder. The powdered material (200gm each) was subjected to extraction with various solvents in the soxhlet apparatus based on the increasing polarity of the solvents separately. The extraction was continued until the defatting of the material had taken place.19

 

To get the homogenous filtrate of extract, it was filtered using Whatman No 1 and cotton wool one after another single time. Then rotary evaporator was used to concentrate the extract to one-tenth at 37-40oC under a vacuum. Complete dry extract was made using a water bath at 40oC. The dry extract was stored at 2-8oC unless used.

 

Determination of Percentage Yield:

The percentage yield of each extract was calculated by using the following formula:

Weight of Extract

Percentage yield = ---------------------------------- x 100

Weight of powder drug Taken

 

Estimation of Phenols and Flavonoids by HPTCL:

High-Performance Thin-Layer Chromatography was performed on silica gel 60F254 (10cm 10cm; 0.25mm layer thickness; Merck). Different extracts were prepared viz aqueous and ethyl acetate of Albizzia lebbeck Benth (LA and LEA) and ethanolic and ethyl acetate extract of Buchanania lanzan Spreng (BE, BEA) in 10ml volumetric flask by taking 250mg of the drug and diluting with water (LA and LEA) and ethyl acetate (BE, BEA) respectively. The final concentration was made is 25mg/ml, which is filtered through a 0.45-micron syringe filter. The filtered solution is further diluted to the strength of 2mg/ml, 4mg/ml, 6mg/ml and 8mg/ml concentrations for HPTLC (CAMAG, Switzerland) analysis.

 

For estimation of phenols, the standard Gallic acid of strength 1mg/ml was spotted on a silica gel 60F254 (Merck, Darmstadt, Germany) TLC plate as a reference compound with other extracts and dried at room temperature. The dried plate was developed by using the solvent system toluene: ethyl acetate: formic acid in the ratio of 7:5:0.1 v/v as mobile phase in a CAMAG- twin-trough glass chamber previously saturated with mobile phase vapour for 20 min.

Similarly, for estimation of flavonoid, standard quercetin of strength 1mg/ml was spotted on a silica gel 60F254 (Merck, Darmstadt, Germany) TLC plate as a reference compound with sample extracts. The dried plate was then developed by using mobile phase toluene: ethyl acetate: formic acid (5:4:0.2 v/v) in a CAMAG- twin-trough glass chamber previously saturated with mobile phase vapour for 20 min.

 

After the development of both the plates, it was dried at 65oC for 2 min. Then phenol estimating plate was scanned at 366nm while the flavonoid estimating plate was scanned at 270nm using camag Scanner 3 (CAMAG, Switzerland) with WinCATS 4 software.

 

Calculation: The concentration of the unknown sample was calculated as follows

 

Screening Methods of Anticancer Activity:

To estimate the cytotoxic effect of drugs, the MTT (compound 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide for) Assay was done by the reported method. For the experiment, 32mg/mL stocks were prepared using DMSO. Serial two-fold dilutions were prepared from 320g/mL to 10g/mL using DMEM plain media for treatment. The procedure followed to experiment was as per the previous publications of Shah 2019 et al.20

 

Statistical Analysis - All the data obtained above in the experiments were evaluated statistically by the ANOVA method.

 

RESULTS:

Percentage Yield:

The aqueous extract of Albizzia lebbeck Benth bark showed the highest yield of about 7.3g/200g (3.6%), whereas the petroleum ether extract has shown the lowest yield of 0.5g/200g (0.2%). In comparison, the ethanolic extract of Buchanania lanzan bark showed the highest yield of about 5.55g/200g (2.7%). In contrast, the chloroform extract has shown the lowest yield of 1.8g/200g (0.8%).

 

HPTLC:

While analyzing extracts by HPTLC method, we found that the ethanolic fraction of Buchanania lanzan Spreng had a higher phenolic content fraction i.e. 34.92g/mL than other fractions of Buchanania lanzan Spreng and Albizzia lebbeck Benth. The high concentration of phenol in the ethanolic fraction may be due to the purification and convergence of phenols throughout the fractionation procedure and is probably responsible for its high cytotoxic activity. The aqueous extract of Albizzia lebbeck Benth did not show any trace of phenol in it. (Table 1. Fig 1,2,3). In the process of estimation of flavonoids by HPTLC techniques, we found that the ethyl acetate fraction Albizzia lebbeck Benth contains the highest concentration of flavonoids which is 17.00 g/mL equivalent to g of quercetin per ml of the samples as compared to other crude extracts (Table 2, Fig 4,5,6).

 

Table 1: Total Phenolic content of different extracts of Buchanania lanzan Spreng and Albizzia lebbeck Benth bark

Compound Name

Extracts

Flavonoids as quercetin equivalent (g/mL)

Buchanania lanzan Spreng

B-EA

5.55 g/mL

BE

1.25 g/mL

Albizia lebbeck Benth

LA

0.00 g/mL

L-EA

17.00 g/mL

 

 

Table 2: Total Flavonoid content of different extracts of Buchanania lanzan Spreng and Albizzia lebbeck Benth bark

Compound Name

Extracts

Phenolic as gallic acid equivalent (g/mL)

Buchanania lanzan Spreng

B-EA

4.00 g/mL

BE

34.92 g/mL

Albizia lebbeck Benth

LA

0.00 g/mL

L-EA

2.72 g/mL

 

Fig 1: HPTLC Standard Chromatogram of Gallic Acid

 

Fig 2- HPTLC chromatoplate of BE, BEA, LA, LEA extracts (track3-10) and Gallic acid (track 1, 2, and 11, 12) at 270 nm

 

Fig 3: HPTLC fingerprint of BE, BEA, LA, LEA extracts and Gallic acid at 270nm

 

Fig 4: HPTLC Standard Chromatogram of Quercetin

 

Fig 5: HPTLC Chromatoplate of BE BEA LA LEA extract (track3-10) and Quercetin (track 1,2, and 11,12) at 366nm

 

Fig 6: HPTLC fingerprint of BE, BEA, LA, LEA extracts and Quercetin at 366nm

 

Screening of Anticancer Activity:

The crude extract of Buchanania lanzan Spreng and Albizzia lebbeck Benth bark had shown remarkable potency for anticancer activity on adenocarcinomic human alveolar basal epithelial cell (A549), Human hepatocellular carcinoma cell (HepG2) and human cervical adenocarcinoma carcinoma cell (HeLa). B. lanzan ethyl acetate extract (B-EA), B. lanzan ethanolic extract (BE) and A. lebbeck ethyl acetate extract (L-EA) have shown an IC50 value of 113.9g/mL, 247.4g/mL, 152.7g/mL dose-dependent inhibition in HepG2 cells respectively while samples B-EA, BE, L-EA have shown an IC50 value of 226.9g/mL, 269.1g/mL, 123.3g/mL dose-dependent inhibition in A549 cells respectively. The effect of samples (use extract) B-EA, L-EA found different in HeLa cell lines from the other two studies. The samples B-EA, L-EA had shown an IC50 value of 91.9g/mL, 257.1g/mL dose-dependent inhibition in HeLa cells, respectively (Tabel 3).On the other hand, sample LA did not show significant inhibition in HepG2, A549, HeLa cells, and Sample BE did not show significant inhibition in HeLa cells.

 

The comparative study of all four extracts revealed that the ethyl acetate extract of B. lanzan Spreng and ethyl acetate extract of A. lebbeck were prominently inhibiting the cell growth of HepG2, A549, HeLa cancerous cell lines with low IC50 values while ethanolic extract of B. lanzan Spreng is only effective against HepG2 and A549 cell lines (Table 3).

 

Table 3: Comparison of % Inhibition (IC50 Value) of the aqueous and ethyl acetate extracts of Albizia lebbeck and ethanol and ethyl acetate extracts for Buchanania lanzan Spreng on A549, HepG2 and HeLa cell lines by MTT assay method.

Compound Name

IC50 g/mL

Cell Line

HEPG-2

A549

HeLa

B-EA

113.9

226.9

91.91

BE

247.4

269.1

IC50 was not calculated due to lesser inhibition

LA

IC50 was not calculated due to lesser inhibition

IC50 was not calculated due to lesser inhibition

IC50 was not calculated due to lesser inhibition

L-EA

152.7

123.3

257.1

 

DISCUSSION:

The well-known, Artemisinin is obtained from Artemisia annua which is a huge success as an antimalarial drug. Further, this molecule has supported to give many new molecules with potent activity and it is worldwide known. The development of novel drugs based on herbal medicines is a leading strategy over the globe. This drug discovery had been there for a long time and going to survive for many more centuries.

 

In continuation of this trend, we selected Buchanania lanzan Spreng21,22 and Albizzia lebbeck Benth23,24,25,26 bark for the evaluation of their cytotoxic potency based on traditional medicinal usages documented in different resources. The crude extracts were prepared by extracting dry bark powders of plants with various solvents, from the all-ethanolic extract of Buchanania lanzan bark aqueous extract of Albizzia lebbeck Benth bark showed the highest yield. In contrast, the chloroform extract of Buchanania lanzan and petroleum ether extract of Albizzia lebbeck Benth bark has shown the lowest yield. Phytochemical screening shows all the essential chemical components in extracts to make it a potent drug.19 For further clarification of its efficacy, we performed the estimation of phenols and flavonoid contents of both the drugs by HPLC and HPTLC27,28 techniques. The result shows that Ethanolic fraction of Buchanania lanzan Spreng was found as a higher phenolic content fraction, i.e. 34.92g/mL.

 

In contrast, the ethyl acetate of fraction Albizzia lebbeck Benth contains the highest concentration of flavonoids 17.00g/mL by HPTLC method. On estimation by HPLC, we found that the aqueous extract of Albizia lebbeck displays the highest content of phenols, 8.47 mcg of gallic acid equivalent and ethyl acetate concentrate of Buchanania lanzan exhibits the highest content of flavonoids, 0.34 mcg equivalent of quercetin. The measurement of aggregates of phenols and flavonoids resolved that the concentrate of Buchanania lanzan and Albizia lebbeck bark has higher contents of phenols in comparison to flavonoid content.19

Pharmacological assays are a concrete way to prove drugs' therapeutic value. In contrast to this, the selected extracts viz ethyl acetate extracts of Buchanania lanzan Spreng and Albizzia lebbeck Benth, the aqueous extract of Albizzia lebbeck Benth and Ethanolic extract of Buchanania lanzan Spreng were subjected to MTT assay on cancerous cell lines i.e. HepG2, A549 and HeLa.29,30 The comparative study of all four extracts is aqueous and ethyl acetate extracts of Albizia lebbeck and ethanol and ethyl acetate extracts of Buchanania lanzan Spreng revealed that ethyl extract of B. lanzan Spreng and ethyl extract of A. lebbeck were prominently inhibiting the cell growth of HepG2, A549, HeLa cancerous cell lines with high IC50 value while ethanolic extract of B. lanzan Spreng is only effective against HepG2 and A549 cell lines.

 

Based on the results got by the different studies, it can be suggested that plant barks contain potent anticancerous bioactive compounds, and it should be evaluated pre clinician and clinical studies for further clarification.

 

CONCLUSION:

The cytotoxic studies and chromatographic profiling of crude extracts of Buchanania lanzan Spreng and Albizia lebbeck Benth gave promising results. However, extensive pharmacological and clinical studies are required to explore its mechanism of action and efficacy in future.

 

ACKNOWLEDGEMENT:

The authors are very grateful to Rajiv Gandhi University of Health and Sciences (RGUHS), Bangalore for providing financial assistance. The authors like to thank Dr Paadma M Paarakh, Principal of the oxford college of pharmacy, Hongasandra, Bangalore for providing laboratory facilities to carry out work. Dr Aarti Abhishek Shah for their constant support and guidance on scientific background.

 

CONFLICT OF INTEREST:

None.

 

REFERENCES:

1.     Robert J. Anticancer drug screening with in vitro models. Drugs of the Future. 1997; 22(7):739-46.

2.     Mann J. Natural products in cancer chemotherapy: past, present and future. Nature Reviews Cancer. 2002; 2(2): 143-8. doi.org/10.1038/nrc723

3.     Cragg GM, Newman DJ. Discovery and development of antineoplastic agents from natural sources. Cancer Investigation. 1999;17(2): 153-63. doi.org/10.1076/phbi.39.s1.8.0009

4.     Kapoor, L.D. C.R.C Handbook of Ayurveda Medicinal Plants. 2017, 85. doi.org/10.1201/9781351070997

5.     Mehta SK, Jaiprakash B, Nayeem N. Isolation and phytochemical investigation on leaves of Buchanania lanzan (Chironji). Annals of Biological Research. 2011; 2(3):469-73.

6.     Jain R, Jain SK. Effect of Buchanania lanzan Spreng. bark extract on cyclophosphamide induced genotoxicity and oxidative stress in mice. Asian Pacific Journal of Tropical Medicine. 2012 ;5(3):187-91. doi.org/10.1016/S1995-7645(12)60022-4.

7.     Mehta KS, Mukherjee S, Jaiprakash B. Anti-inflammatory activity of the methanolic extract of Buchanania Lanzan leaves by carrageenan-induced rat paw oedema method. Int J Pharm Sci Rev Res. 2011; 6(2):144-6.

8.     Puri A, Sahai R, Singh KL, Saxena RP, Tandon JS, Saxena KC. Immunostimulant activity of dry fruits and plant materials used in Indian traditional medical system for mothers after child birth and invalids. Journal of Ethnopharmacology. 2000; 71(1-2): 89-92. https://doi.org/10.1016/S0378-8741(99)00181-6

9.     Warokar AS, Ghante MH, Duragkar NJ, Bhusari KP. Anti-inflammatory and antioxidant activities of methanolic extract of Buchanania lanzan Kernel. Indian Journal of Pharmaceutical Education and Research. 2010; 44(4): 363-8.

10.   Kumari A, Kakkar P. Screening of antioxidant potential of selected barks of Indian medicinal plants by multiple in vitro assays. Biomedical and Environmental Sciences. 2008; 21(1): 24-29. https://doi.org/10.1016/S0895-3988(08)60003-3

11.   Pattnaik A, Sarkar R, Sharma A, Yadav KK, Kumar A, Roy P, Mazumder A, Karmakar S, Sen T. Pharmacological studies on Buchanania lanzan Spreng.-A focus on wound healing with particular reference to anti-biofilm properties. Asian Pacific Journal of Tropical Biomedicine. 2013; 3(12): 967-74. https://doi.org/10.1016/S2221-1691(13)60187-2

12.   Kirtikar KR, Basu BD. Indian Medicinal Plants. Indian Medicinal Plants. 1918.

13.   Rashid RB, Chowdhury R, Jabbar A, Hasan CM, Rashid MA. Constituents of Albizzia lebbeck and antibacterial activity of an isolated flavone derivative. Saudi Pharmaceutical Journal. 2003; 11(1/2): 52-6.

14.   Ueda M, Tokunaga T, Okazaki M, Sata NU, Ueda K, Yamamura S. Albiziahexoside: a potential source of bioactive saponin from the leaves of Albizzia lebbeck. Natural Product Research. 2003; 17(5): 329-35. https://doi.org/10.1080/1057563031000072578

15.   Misra LN, Dixit AK, Wagner H. N-demethyl budmunchiamines from Albizzia lebbek seeds. Phytochemistry. 1995; 39(1): 247-9. https://doi.org/10.1016/0031-9422(94)00829-I

16.   Maa YT, Hsiaob SC, Chenb HF, Hsu FL. Tannins from Albizia lebbeck. Phytochemistry. 1997; 46(8): 1451-2. https://doi.org/10.1016/S0031-9422(97)00489-5

17.   Chulet R, Pradhan P, Sarwan SK, Mahesh JK. Phytochemical screening and antimicrobial activity of Albizzia lebbeck. Journal of Chemical and Pharmaceutical Research. 2010; 2(5): 476-84.

18.   George, F.; Zohar, K.; Harinder, P.S.; Makkar.; Kaluas, The biological action of saponins in animal systems: a review. B. British J. of Nutrition. 2002: 587-605. https://doi.org/10.1079/BJN2002725

19.   J.Shrivastava, T. Madhuri. Phytochemical analysis and HPLC estimation of phytoconstituents of Buchanania lanzan Spreng. Advance Pharmaceutical Journal. 2019; 4(5): 113-120. https://doi.org/10.31024/apj.2019.4.5.1

20.   Shah AA, Gourishetti K, Nayak Y. Osteogenic activity of resveratrol in human fetal osteoblast cells. Pharmacognosy Magazine. 2019; 15(64): 250. https://doi.org/10.4103/pm.pm_619_18

21.   Joshi H, Pagare M, Patil L, Kadam V. Invitro antioxidant activity of ethanolic extract of leaves of Buchanania lanzan Spreng. Research J. Pharm. and Tech. 2011; 4(6): 920-4. https://doi.org/10.5958/0974-360X

22.   Joshi H, Pagare M, Patil L, Kadam V. Cardioprotective Effect of Ethanolic Extract of Buchanania lanzan Spreng. Against Isoproterenol Induced Myocardial Infarction in Rats: A Biochemical, Electrocardiographic Evaluation. Group. 2012; 1(8.58): 0-375.

23.   Argade PA, Bhutkar MA, Magdum CS. Albizzia lebbeck extract mediated synthesis and characterization of Zinc oxide Nanoparticle. Asian Journal of Pharmaceutical Research. 2019; 9(1): 1-6. https://doi.org/10.5958/2231-5691.2019.00001.7

24.   Argade PA, Bhutkar MA, Magdum CS. Albizzia lebbeck extract mediated synthesis of Zinc Oxide Nanoparticles and study of its In-vitro Anti-diabetic and Anti-oxidant activity. Asian Journal of Pharmacy and Technology. 2019; 9(2): 93-8. https://doi.org/10.5958/2231-5713.2019.00016.3

25.   Velraj M, Vijayalakshmi A, Jayakumari S, Ramamoorthy S, Ravichandiran V, Srikanth J. Antidepressant-Like Effects of the ethanolic extract of Albizzia lebbeck (Linn) leaves in animal models of depression. Research Journal of Pharmacognosy and Phytochemistry. 2010; 2(1): 30-3. https://doi.org/10.5958/0975-4385

26.   Shah UD, Shah MB, Saluja AK. A Pharmacognostical study on Albizzia lebbeck bark. Research Journal of Pharmacognosy and Phytochemistry. 2010; 2(3): 241-5.

27.   Jain V, Jain T, Saraf S, Saraf S. HPTLC method for routine quality control of Ayurvedic formulation Drakshadi gutika. Asian Journal of Pharmaceutical Analysis. 2013; 3(4): 111-4. https://doi.org/10.5958/22315675

28.   Satyanarayana L, Naidu SV, Rao MN, Ayyanna C, Kumar A. The Estimation of Raltigravir in Tablet dosage form by RP-HPLC. Asian Journal of Pharmaceutical Analysis. 2011; 1(3): 56-8.

29.   Pavithra S, Banu N. A First Report on the Antiproliferative activity of Sodium Copper Chlorophyllin from Endangered Medicinal Plant Rhinacanthus nasutus on HepG2 and HeLa Cell Lines. Research Journal of Pharmacy and Technology. 2017; 10(1): 325. https://doi.org/10.5958/0974-360X.2017.00066.X

30.   Selvam R, Anandhi D, Saravanan D, Revathi K. Antioxidant Properties of Punica grantum Fruit rind extract against liver Hepatocellular Carcinoma studied in HEPG2 Cell Line. Research J. Pharm. and Tech. 2019; 12(10): 4719-23. https://doi.org/10.5958/0974-360X.2019.00813.8

 

 

 

 

 

Received on 26.09.2022 Modified on 18.03.2023

Accepted on 20.07.2023 RJPT All right reserved

Research J. Pharm. and Tech 2024; 17(1):103-108.

DOI: 10.52711/0974-360X.2024.00016