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 Table of Contents  
ORIGINAL ARTICLE
Year : 2021  |  Volume : 2  |  Issue : 1  |  Page : 19-25

Safety study on novel ayurvedic nutritive powder – An experimental study


1 Department of Rasashastra and Bhaishajya Kalpana, SBSAMCH, Mundaragi, Karnataka, India
2 Department of Agadatantra, KAHER, Shri B.M. Kankanawadi Ayurveda Mahavidyalaya, Belagavi, Karnataka, India
3 Department of Rasayana and Vajikarana, KAHER, Shri B.M. Kankanawadi Ayurveda Mahavidyalaya, Belagavi, Karnataka, India
4 Department of Rasashastra and Bhaishajya Kalpana, KAHER, Shri B.M. Kankanawadi Ayurveda Mahavidyalaya, Belagavi, Karnataka, India

Date of Submission18-Feb-2021
Date of Decision28-Feb-2021
Date of Acceptance02-Mar-2021
Date of Web Publication17-Apr-2021

Correspondence Address:
Veena Babu Kupati
Department Rasashastra and Bhaishajya Kalpana, KAHER, Shri B. M Kankanawadi Ayurveda Mahavidyalaya, Belagavi - 590 003, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijaim.ijaim_2_21

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  Abstract 


Introduction: It has been estimated that in India, 65%, i.e., nearly 80 million children under 5 years of age suffer from varying degrees of malnutrition. Most of the children fall into the pit of Malnutrition during the weaning phase. Mothers are expected to make this “weaning Bridge.” This includes utilization of “Supplementary Feeding Programs.” Many weaning foods are available which claims the specific dietary implementation. The development of a nutritive Powder in weaning children is need of hour. This study includes safety of novel Ayurvedic nutritive powder in Wistar rat pups.
Aim: This study aims to evaluate its safety in experimental animals.
Materials and Methods: Ingredients selected were Mudga, Godhuma, Rakta Shali, Ragi, Trikatu, Yastimadhu, and Khanda Sharkara. Safety study of Nutritive Powder was done in 20 days old Wistar Rat pups for 90 days. Parameters selected are HISTOPATHOLOGICAL study and biochemical investigations.
Results: No mortality and abnormal behavioral changes were found in rat pups during experimental study. No remarkable pathological changes were observed. Biochemical Parameters revealed significant decrease in blood sugar, serum urea, and alkaline phosphatase in test group when compared to control.
Conclusion: Nutritive powder when administered in the dose of 40 g/Kg body weight of animal for 90 days. No mortality was seen in any animals of test group. No remarkable pathological changes were seen in histopathological study of liver and kidney. Significant decrease of blood glucose, serum urea serum alkaline phosphatase was seen in test group animals compared to control group animals.

Keywords: Ayurveda, complimentary food, nutritive powder, weaning food


How to cite this article:
Kulkarni V, Hiremath RR, Buduru SP, Kupati VB. Safety study on novel ayurvedic nutritive powder – An experimental study. Indian J Ayurveda lntegr Med 2021;2:19-25

How to cite this URL:
Kulkarni V, Hiremath RR, Buduru SP, Kupati VB. Safety study on novel ayurvedic nutritive powder – An experimental study. Indian J Ayurveda lntegr Med [serial online] 2021 [cited 2021 Oct 25];2:19-25. Available from: http://www.ijaim.com/text.asp?2021/2/1/19/313994




  Introduction Top


Malnutrition is closely linked, either directly or indirectly, to major causes of death and disability worldwide. Worldwide, in 2011, about 101 million children under 5 years of age were underweight and 165 million had stunted growth.[1]

Weaning refers to accustoming the infant to nourishment other than mother's milk. Weaning is a difficult period in an infant's life, because if the food supplements or substitutes are not adequate in quantity and quality the child becomes malnourished. Most of the children fall into the pit of Malnutrition during the weaning and postweaning phase. Some even succumb to it. Jelliffe has suggested “Three Plank Protein Bridge”[2] or the bridge of complementary feeding. Mothers are expected to make this “weaning Bridge” to carry the children across the pit of malnutrition during liquid to solid transition (i.e., weaning period).

  1. Continued breast feeding
  2. Introducing vegetable protein and
  3. Animal protein.


Some mother's do not make a bridge at all and some make a bridge that may collapse into the pit. Hence, a “Safety Net” is needed beneath the bridge. This includes utilization of “Supplementary Feeding Programs.” And also brain development is rapid during first 2 years of life. Brain attains 80% of growth and myelination also completes by 2 years of age; hence, any program aimed at developing the brain should be started as early as possible, before the age of 2.[3],[4]

Considering these facts as background, nutritive powder was developed as per the recommended dietary allowance (RDA) requirement of 1–2 years old children.


  Selection of Ingredients and Processing Technique Top


Among Pulses, Mudga was selected as it is rich in proteins, possesses laghu guna,[5] grahi karma and is best among all shimbi dhanyas. Among Cereals, Godhuma was chosen for bearing Jeevaniya, Ruchikara[6] and Bramhana Properties. Rakta Shali was selected as it is rich in Iron, Carbohydrates and is best among all shooka dhanya. Ragi was chosen, as it is rich in calcium. Trikatu was included as it is a deepana dravya, to treat mandagni which is prevalent Kapha Pradhana Balyavastha. Since 80% of brain development occurs within 2 years of age, Yastimadhu-Medhya dravya was included.

Among various Processing techniques, roasting technique was chosen, as it reduces antinutrients and improves the flavor, color, texture, digestibility of the starch and overall acceptability of the product.[7]

Every new drug developed needs to be evaluated pre clinically for its safety and efficacy before undergoing clinical study. Hence, safety study was undertaken for 90 days.

Objectives

  • To evaluate safety of novel nutritive powder in experimental animals.



  Materials and Methods Top


The development of nutritive powder was done in the Department of Bhaishajya Kalpana, KAHER Shri B. M. K Ayurevda Mahvaidyalaya, Belagavi.

Ingredients and proportion

Development of ayurvedic nutritive powder

Dry seeds of Mudga, Godhuma, Rakta shali, Ragi [Table 1] were taken separately and cleaned. They were washed under running water separately. Excess water was removed by straining and shade dried by spreading on clean white cotton cloth. Completely dried ingredients were roasted separately on mild flame till they get roasted well and become fragile. Roasted ingredients were powdered separately and sieved (80 no. mesh). Shunti, maricha, Pippali, Yastimadhu, and Khanda sharkara were powdered separately and sieved (80 No. mesh). Powdered drugs were taken together and ground till homogeneous mixture was formed and stored in a clean air tight container [Figure 1].
Table 1: The details of ingredients and quantity of nutritive powder

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Figure 1: (a) Nutritive powder, (b) Nutritive pallets

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Nutritive powder pellets

Nutritive powder was converted into hard Pellets, i.e., palatable form to consume to rat pups [Figure 1]. Nutritive powder with quantity sufficient water was pressed and dried in in hot air oven (70°C) for 4 h 15 min and stored in air tight container.

Experimental study

Wistar rat pups of either sex of 20 days old were procured from animal house, K. L. E. U. Jawaharlal Nehru Medical College, Belagavi, Karnataka. They were exposed to natural day and night cycles with ideal laboratory condition in terms of ambient temperature (22 ± 2°C) and 45%–55% relative humidity with 12/12 h natural light and dark cycle. They were fed with free access of standard pellet diet (Amruta feeds, VRK's Scientist's Choice Laboratory Animal Feed, Baramati, supplied by SaiDurga Feeds and Foods, Bengaluru) and tap water given ad libitum. Experiment was carried out after obtaining permission from Institutional Animal ethics committee” (BMK/IAEC/Res-05/2011).

For study, 20 days old rat pups were selected and study duration is 90 days, as nutritive powder was intended for weaning children.

Grouping

Twenty animals of either sex were randomly divided into two groups [Table 2]; each group contained 10 animals and further divided into subgroups, five female animals and five male animals in test group and six female animals and four male animals in the control group. Animal marking was done by the saturated picric acid as follows: (1) Head, (2) body, (3) tail, (4) limb, (5) both limbs [Figure 2].
Table 2: The groups of experimental study

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Figure 2: (a and b) Marking of the animal, (c) Standard feed, (d) Metabolic cage

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Selection of dose

Test drug was fed in the pellet form orally. Dose in children was calculated on basis of RDA requirements of 1–3 year old children. Pediatric dose calculated was 90 g/day, which was converted into adult human dose based on body weight. Experimental animal's dose was calculated by extrapolating the human dose to animal dose based on the body surface area ration by following Paget and Barnes, 1964 table.[8]

Parameters of experimental study

General observation

Animals of both groups were observed for their food and water intake, general activity, sign of toxicity, gross behavior, and body weight throughout the experimental study [Table 3].
Table 3: Observational signs on experimental animals during study

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Urine investigation

All experimental animals were kept in metabolic cage for 24 h for three times to quantify and analysis of color, pH, leukocytes, nitrite, urobilinogen, protein, blood, specific gravity, ketone, bilirubin, glucose was done by Mission Urine Reagent Strips.

Blood investigation

After 90th day, all animals were kept under observation for 24 h on fasting. On 91st day, animals were anesthetized by Diethyl Ether and blood was drawn through retro orbital region by pricking micro capillary tube number 100 mm (Borosilicate glass with both ends open) and collected in two separate labeled vials. Blood was sent to Jeevan laboratory, Belgaum, for biochemical investigations. Biochemical parameters were estimated by serum amount as per the instructions of the manufacturer with Erba test in an auto analyzer.

Hematological investigations

Blood was collected in vial containing EDTA for performing hemoglobin percentage.

Biochemical investigations

Blood was collected in plain vial and serum was separated out for performing biological parameters such as blood glucose (random), serum cholesterol, serum triglycerides, blood urea, serum creatinine, serum alkaline phosphatase, serum glutamate oxaloacetate transferase (SGOT), and serum glutamate-pyruvate transaminase (SGPT).

Histopathological investigations

The organs liver, kidney, spleen, stomach, heart, lungs, thymus, brain, testis, and bone marrow were collected immediately after sacrificing by cervical dislocation [Figure 3] and cleaned of extraneous tissue, weighed and transferred to 10% formalin solution in separate labeled containers. The organs liver, kidney sent for Jeevan laboratory, Belgaum, for histopathological study. The tissues were allowed to remain in it till they were taken up for processing. The slides with sections obtained were scanned in Olympus Corporation, Penta headed microscope (Model BX41TF) under different magnifications [Figure 4]. Changes in the cytoarchitecture were noted. Histopathological studies were conducted as per standards [Table 4].
Figure 3: (a) Retro orbital Blood collection, (b and c) Dissection, (d) Tracing of organs

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Figure 4: Histopathology . kidney and liver

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Table 4: Histopathological report of control and test groups

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Statistical analysis

Student's t-test for unpaired data has been used for analyzing the data generated during the study. A P < 0.05 is considered as statistically significant, P < 0.01 or P < 0.001 is considered statistically highly significant. The level of significance was noted interpreted accordingly using IBM SPSS Statistics for Windows, Version 22.0. Armonk, NY: IBM Corp.


  Results Top


Gross behavior

No mortality was observed in any experimental animals. Sign of respiratory depression, narcosis, and catatonia was not observed in any animal. Male animals of test group found active throughout study than animals of control group and female animals of trail group.

Food consumption and water intake

Food consumption and water intake were found same till the completion of the study.

Observational changes in urine

The parameters such as leukocytes, nitrite, urobilinogen, protein, blood, specific gravity, were found within normal range ketone, bilirubin, glucose was found absent.

The mean value of urine output of 24 h in test group was more than mean value of control group. The difference in mean value of urine output 24 h of test group when compared to control was nonsignificant (P > 0.05) [Table 5].
Table 5: Urine output in ml results of control and test group animals

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The mean value of pH of test group was less than mean value of control group. The difference in mean value of pH of test group when compared to control was nonsignificant (P > 0.05) [Table 6].
Table 6: The pH of urine of control and test group animals

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Weight observation

The mean value of body weight in test group was less than mean value of control group at the end of 30 days, 60 days, and 90 days. The difference in mean value of body weight of treated group when compared to control was nonsignificant (P > 0.05) [Table 7].
Table 7: Weight observation in g of experimental animals

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Organ weight

The mean value of brain weight of treated group was less than mean value of control group. The difference in mean value of brain weight of treated group when compared to control was nonsignificant (P > 0.05) [Table 8].
Table 8: The organ weight in g observation of control and test groups

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The mean value of heart weight of treated group was less than mean value of control group. The difference in mean value of thymus weight of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of left kidney weight of treated group was more than mean value of control group. The difference in mean value of left kidney weight of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of liver weight of treated group was less than mean value of control group. The difference in mean value of liver weight of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of lungs weight of treated group was more than mean value of control group. The difference in mean value of lungs weight of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of the right kidney weight of treated group was less than mean value of control group. The difference in mean value of right kidney weight of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of spleen weight of treated group was less than mean value of control group. The difference in mean value of spleen weight of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of testis weight of treated group was more than mean value of control group. The difference in mean value of testis weight of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of thymus weight of treated group was equal to mean value of control group. The difference in mean value of thymus weight of treated group when compared to control was nonsignificant (P > 0.05).

Results of biochemical parameters

The mean value of serum alkaline phosphatase of treated group was less than mean value of control group. The difference in mean value of serum alkaline phosphatase of treated group when compared to control was significant (P < 0.05) [Table 9].
Table 9: The results of biochemical parameters of experimental animals

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The mean value of serum cholesterol of treated group was less than mean value of control group. The difference in mean value of serum cholesterol of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of blood glucose (random) of treated group was less than mean value of control group. The difference in mean value of blood glucose (random) of treated group when compared to control was significant (P < 0.05).

The mean value of serum creatinine of treated group was more than mean value of the control group. The difference in mean value of serum creatinine of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of blood urea of treated group was less than mean value of control group. The difference in mean value of blood urea of treated group when compared to control was significant (P < 0.05).

The mean value of SGOT of treated group was more than mean value of control group. The difference in mean value of SGOT of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of SGPT of treated group was less than mean value of control group. The difference in mean value of SGPT of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of serum triglycerides of treated group was less than mean value of control group. The difference in mean value of serum triglycerides of treated group when compared to control was nonsignificant (P > 0.05).

The mean value of hemoglobin in test group was more than mean value of control group. The difference in mean value of hemoglobin pH of test group when compared to control was nonsignificant (P > 0.05) [Table 10].
Table 10: The Hemoglobin in g percentage of control and test groups

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  Discussion Top


Experimental study was conducted with protocol approved by institutional animal ethics committee and planned to assess safety of nutritive powder on experimental animals for the duration of 90 days. Test group animals were compared with control group. In this study, all quantitative data were analyzed by paired and un-paired t-test.

All the ingredients selected for the development of nutritive powder are either food ingredients, deepaneeya drugs, balya drugs. These are already using in routine day-to-day life. Nutritive powder was given orally for 90 days.

Mortality was not observed in experimental animals of both groups for 90 days. Sign of toxicity such as fur deterioration, mucous discharge, diarrheal sign, and bloody mucus was not seen. All animals found active throughout the experimental study.

Weight of organs brain, thymus, heart, lungs, liver, spleen, kidney, and testis was recorded. No significant difference in organs weight was seen in test group animals compared to control group. Histopathology study revealed comparable results, mild congestion was observed may be due to anesthesia (Ether). Blood glucose (48.00 ± 18.97) was found significantly decrease in test group as compared to control group (76.50 ± 10.07). It may be due to hypoglycemic activity of Trikatu when administered orally for long duration in pups. Alkaline phosphatase (261.00 ± 59.47) was found significantly decrease in test group as compared to control group (365.17 ± 13.23). Serum urea (41.67 ± 3.08) was found significantly decrease in test group as compared to control group (46.50 ± 1.87). However, both the values are within normal rage.

Hb % of experimental animals of control group was found within the normal range and nonsignificant increase compared to the control group.

Impaired food and water intake is also one of important sign of toxicity. In this study, diet and water intake of all experimental animals was found similar.

Insignificant decrease in body weight of test group animals was observed compared to the control group. This may be due to increased metabolism, quantity of Trikatu might be high leading to higher Deepana-Pachana effect this might have burnt body tissues leading to decrease in body weight of Animals.


  Conclusion Top


Nutritive powder when administered in the dose of 40 g/Kg body weight of animal for the duration of 90 days. No mortality was seen in any animals of test group. No remarkable changes observed in histopathological study reports. However, combination of nutritive powder with administered for 90 days reduced body weight, blood glucose, serum urea serum alkaline phosphatase was seen in rat pups of test group compared to the control group.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
WHO. Essential Nutrition Actions: Improving Material, Newborn, Infant and Young Child Health and Nutrition. Geneva: World Health Organization; 2013.  Back to cited text no. 1
    
2.
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5.
The Ayurvedic Pharmacopoeia of India. Vol. 3, New Delhi: Govt of India, Ministry of Health and Family Welfare Dept of Indian System of Medicine; 2000. p. 123-4.  Back to cited text no. 5
    
6.
Suri R. Bhojanakutuhalam. Mysore: M/s Tech Prints; 2012. p. 12-3.  Back to cited text no. 6
    
7.
Temesgen M. Nutritional status of Ethiopian weaning and complementary foods: A review. Open Access Sci Rep 2013;2:621. [doi: 10.4172/scientificreports].  Back to cited text no. 7
    
8.
Ghosh MN. Fundamentals of Experimental Pharmacology. 3rd ed. Kolkatta: Hilton and Co.; 2005. p. 192.  Back to cited text no. 8
    


    Figures

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    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6], [Table 7], [Table 8], [Table 9], [Table 10]



 

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