Estimation of Total Carbohydrate content by Phenol Sulphuric acid Method from Eichhornia crassipes (Mart.) Solms

 

Firoj A. Tamboli¹*, Harinath N. More¹, Susmita S. Bhandugare¹, Ankita S. Patil¹,
Namdeo R. Jadhav¹, Suresh G. Killedar²

¹Department of Pharmacognosy, Bharati Vidyapeeth College of Pharmacy,

Near chitranagari, Kolhapur -416013 Maharashtra, India.

²Sant Gajanan Maharaj College of Pharmacy, Mahagaon

Tal- Gadhinglaj Dist- Kolhapur 416503 Maharashtra, India.

 

ABSTRACT:

Eichhornia crassipes is free-floating aquatic macrophytes. For performing normal body function carbohydrate is an essential source for living organisms. Low carbohydrate intake results in muscle breakdown, ketosis, and dehydration. Daily 50-100gms of carbohydrate is recommended. In this study, the total carbohydrate content was detected in the aqueous extract by using phenol- sulphuric acid method. This was a rapid colorimetric method that determines the total carbohydrate present in the sample. The role of sulphuric acid is to convert polysaccharides, oligosaccharides, and disaccharides into monosaccharides. The result showed that the carbohydrate content was 30.50% in the sample solution. So the weed can be utilized as the source of carbohydrate.

 

 

 

INTRODUCTION:

Eichhornia crassipes (water hyacinth), an invasive aquatic weed with a high growth rate almost doubling its population in nearly 5-15 days^1,2,3,4. Its aquatic macrophyte consists of numerous compounds like alkaloids, glycosides, tannins, phenolic acids, and other metabolites with radical scavenging activity. It also have antiviral, antifungal, antitumor and antibacterial activity^5,6. It is observed that it grows at a faster rate in summer at temperature 20-30°C and growth is ceased at a temperature between 8-15°C⁷.

 

It is efficient in taking calcium, magnesium, zinc, copper, boron, etc. It shows a negative impact on the aquatic ecosystem, biodiversity, economy and human well being. The best method to control water hyacinth is to prevent it from entering water bodies. Rather it is difficult to control its growth the best way is to make utilization of weed.^7,8.

 

The reserve carbohydrates are metabolized during respiration and growth which is stood in different parts of plants. It is used by plants for growth and also to get recovered from stress, herbivory, or disturbance^9,10,11. The present study was carried out to determine total carbohydrate content by using the phenol sulphuric acid method. It measures the total carbohydrate present in the sample.

 

MATERIALS AND METHODS:

Collection and Identification of plant material:

The whole plant material (petioles, leaves, roots, flowers) were collected from the Rankala, Kolhapur, India in the month of August. The plant material was washed using distilled water and dried under the shadow. It was grated to form a fine powder. The powder was stored in a airtight container until further use.

 

Extract preparation:

The sample was prepared by using water. The powder of 10gm was weighed and extracted for 24hrs and stirred meanwhile by using 100ml of water. Then it was filtered and used further.

 

Estimation of Total Carbohydrate by proposed Method^12-17:

100mg of glucose was taken in the test tube. To this added 5ml 2.5N HCl and boil on a water bath for 3hrs to hydrolyse. Cooled to room temperature. To this added a sufficient quantity of solid sodium carbonate (Na₂CO₃) till effervescence ceases. It indicates complete neutralization and then filter and make the volume up to 100ml. Pipette out 0.2, 0.4, 0.6, 0.8, 1ml of working standards in the series of test tube. Then pipette out 0.2 ml of the sample solution. Makeup to 1ml with water. Set blank with all reagents without sample.1ml phenol was added to each tube. To this 5ml 96% H₂SO₄ was added and shake well. After 10min shake the content in the tubes and place in a water bath at 25-30°C for 20min In hot acidic medium glucose was dehydrated to hydroxymethyl furfural. This forms a green colored product with phenol. This colour intensity was measured at 490 nm. Finally total amount of carbohydrate was calculated by using following formula.

 

Table 1: Absorbance at 490nm with different concentration of working standard of glucose solution

 

Table 2: Concentration and absorbance of total carbohydrate content observed in sample

 

Calculation^{18, 19, 20}:

The percentage of total carbohydrate content present in Eichhornia crassipes determined as follows:

 

Absorbance corresponds to 0.1 ml of test sample

 = x mg of glucose

 

100 ml of sample solution contain

= x/0.1 *100 mg glucose

= % of total Carbohydrate present.

 

Absorbance of sample was found to be 0.277

Therefore,

                    = (0.03050/0.1) 100

                    = 30.50% total carbohydrate content present      

 

In the experiment conducted here, the total carbohydrate content observed in Eichhornia crassipes was found to be 30.50%.

 

CONCLUSION:

The total carbohydrate content in water hyacinth observed to be 30.50%. The plant Eichhornia crassipes shows good quantity of carbohydrate. The further research on isolation of particular carbohydrate can helps to provide the natural source of carbohydrate. Thus by knowing the percentage of carbohydrate content in water hyacinth, the utilization of it can be made as commercial source of carbohydrate. It is suitable raw material as it is found in large amount and also it has high growth rate.

 

ACKNOWLEDGEMENTS:

The authors are thankful to Dr. H. N. More, Principal, Bharati Vidyapeeth College of Pharmacy, Kolhapur for providing facilities to carry out the work.

 

CONFLICTS OF INTEREST:

The authors declare no conflicts of interest.

 

REFERENCE:

1.     Omondi, E.A., Ndiba, P.K. & Njuru, P.G. Characterization of water hyacinth (E. crassipes) from Lake Victoria and ruminal slaughterhouse waste as co-substrates in biogas production. SN Appl. Sci. 2019; 1, 848.

2.     Tao R, Zeng R, Xiao-Yan Y, Zhang S, Yang ZH. Water Hyacinth (Eichhornia crassipes) Biomass as a Biofuel Feedstock by Enzymatic Hydrolysis. Bio Resources.2016; 11(1):2372–2380.

3.     Degaga AH. Water Hyacinth (Eichhornia crassipes) Biology and its Impacts on Ecosystem, Biodiversity, Economy and Human Wellbeing. J. Life Sci. Biomed. 2018;8(6): 94-100.

4.     BVVS Surya Kiran, Bishnupada Biswal, G Nagarjuna Reddy, LK Kanthal, N Sridhar. Evaluation of anthelmintic activity of Eichhornia crassipes roots. Research J. Pharmacology and Pharmacodynamics. 2013; 5(3): 183-184.

5.     Ali H. et al. The world's worst aquatic plant as a safe cancer medicine Antitumor activity on melanoma induced mouse by Eichhornia crassipes: in vivo studies. J Pharm Res. 2009; 2(1):365-6.

6.     Shanab S.M. et al. Allelopathic Effects of water hyacinth (Eichhornia crassipes). PlusOne. 2010; 5:1-8.

7.     Owens, C.S and Madsen, J.D. Low temperature limits of waterhyacinth. Journal of Aquatic Plant Management. 1995; 33: 63– 68.

8.     B.S. Krishnamoorthy, N. Nattuthurai, Arunkumar K., Revathi T., S. Kamatchi. Preliminary Studies on Phytochemicals of Solvent Extracts of Eichhornia crassipes. Research J. Pharm. and Tech. .2014; 7(6):  635-637.

9.     Navarro L and Phiri G. Water hyacinth in Africa and the Middle East. A survey of problems and solutions. International Development Research Centre, Ottawa (CA). 2000.

10.  Hammuel C, Anyim B P, Raplong H H, Inekwe U V, Batari M L. Phytochemical Screening and In Vitro Antibacterial Acitivity of Methanol and Aqueous Extracts of Acalypha Racemosa Leaves. Res. J. Pharmacognosy & Phytochem. 2014; 6(3): 118-121.

11.  Praveen Kumar Uppala, K. Atchuta Kumar, Sujit Kumar Patro, Murali Krishna. B. Experimental Evaluation of Antidepressant activity of Aqueous and Chloroform Leaf and Shoot Extracts of Eicchornia crassipes Linn in Mice. Asian J. Res. Pharm. Sci. 2015; 5(3): 139-144.

12.  Madsen JD. Seasonal biomass and carbohydrate allocation in a southern population of Eurasian watermilfoil. J Aquat Plant Manage. 1997; 35:15–21.

13.  Nielson S.S.: Phenol-Sulfuric Acid method for total carbohydrates. Food Science Text Series. 2010; pp.47-53.

14.  Quantitative Evaluation of Carbohydrate Levels in Different Natural Foodstuffs by UV-Visible Spectrophometer. Asian J. Pharm. Ana. 2012; 2(1): 10-11.

15.  V.S. Asha, Ch. Madhu, Kavya. Mannem, V. Prateesh Kumar. Quantitative Evaluation of Carbohydrate Levels in Seeds for Home Use by UV-Visible Spectrophotometer. Asian J. Pharm. Ana. 2012; 2(3):71-72.

16.  Kavya Mannem, Ch. Madhu, V.S. Asha, V. Prateesh Kumar. Quantitative Evaluation of Carbohydrate Levels in Green Leafy Vegetables for Home use by UV-Visible Spectrophotometer. Asian J. Pharm. Ana.2012; 2(3): 79-80.

17.  Ch. Madhu, J. Swapna, R. Sneha Spandana, I. Niharika, K. Rohith Raj, B. Kalyan, M. Raja Rajeshwari, T. Prathyusha. Quantitative Evaluation of Carbohydrate Levels in Seeds for Home use By UV-Visible Spectrophotometer. Asian J. Pharm. Ana. 2012; 2(4): 104-105.

18.  Vavilala Prateesh Kumar, Ch. Madhu, Kavya Mannem, V.S. Asha, A. Sambasiva Rao, M. Sravan Prasad. Quantitative Evaluation of Carbohydrate Levels in Fruits by UV-Visible Spectrophotometer. Asian J. Pharm. Tech. 2012; 2(3):  99-100.

19.  Ch. Madhu, J. Swapna, R. Sneha Spandana, I. Niharika, K. Rohith Raj, B. Kalyan, M. Rajarajeshwari, T. Prathyusha. Quantitative Evaluation of Carbohydrate Levels in Seeds for Home use by UV-Visible Spectrophotometer. Asian J. Pharm. Tech. 2012; 2(3):  110-111.

20.  Bokov D.O., Kulaeva I.R., Potanina O.G., Sergunova E.V., Bondar A.A., Evgrafov A.A., Antsyshkina A.M., Krasnyuk I.I.. Carbohydrates determination in the Snowdrops (Galanthus L.) herbal pharmaceutical substances by TLC and UV-Spectrophotometry. Research J. Pharm. and Tech. 2020; 13(1):243-249.

 

 

Received on 09.06.2020                    Modified on 04.07.2020

Accepted on 19.07.2020                   ©AJRC All right reserved