(MUNNEERIZHIVU)
Thesis Submitted to
The Tamil
Nadu
Dr. M.G.R. Medical University
In partial
fulfilmentfor the award of the degree of
Doctor of Philosophy -July 2018
Dr. M. K
annan
M.D(s).,Ph. D Scholar (part-time) Siddha Central Research Institute (CCRS, Ministry of AYUSH, Govt. of India)
Chennai - 600106
Under the supervision of
Dr. S. Justus Antony M.D(s)., Lecturer - Grade II Dept. of PG Pothu Maruthuvam
“The pleasure, consummation and ecstasy that come along with the successful
accomplishment of any work would be incomplete unless we name the people who
made it possible, whose constant guidance and encouragement served as a beam of
light and crowned our efforts.”
In the first Place, I would like to thank the Almighty for his tremendous grace
for giving me the strength and resilience in traversing the times of distress during the
Project work and enabled me to complete arduous task satisfactorily.
I feel privileged to express my heartfelt gratitude to my Respectable Project
guide Dr. S. Justus Antony,Associate professor, Department of PG Pothu Maruthuvam,
Government Siddha Medical College, Palayamkottai, for his sincere guidance,
constructive criticism, constant encouragement and providing all necessary inputs to
initiate my research work.
I extend my sincere thanks to members of the Doctoral Advisory Committee,
Dr. S. Mohan, Former Director, NIS and Dr. P. Parthiban, Joint-Director, Directorate
of Indian Medicine & Homeopathy.
I thank with immense gratitude from the bottom of my heart to DG, CCRS,
Director in-charge, SCRI, Head of the Department, DCR, SCRI, for providing me
with all the facilities and support during academic period of the study.
I take this opportunity to express my sincere & warm thanks to Dr. G.
Dayanand Reddy, Assistant Director (Pharmacology), Siddha Central Research Institute,
Chennai and Co-guide/ Doctoral advisory committee member of my study.
I express my sincere thanks to Dr. N. Kabilan, Dr. E. M. Manikgantan and
Dr. Rajalakshmi Sivaraman, Department of Siddha, the TN Dr. MGR Medical
University, for their moral support and appropriate help from the university side during
difficult times.
I express my sincere and heartfelt thanks to the Core members of the Institute,
Dr. P. Sathiyarajeswaran, Dr. S. Natarajan, Dr. M. S. Shreedevi, Dr. V. Aarthi,
Dr. G. Kiruthiga, Dr. M. Muthuthamizh, Dr. R. Meena, Mr. N. Kirubakaran and
I thank the Phytochemistry and analysis team Dr. K. Parthasarathy
Dr. S. Suginlal Jabaris and Dr. V. L. Reena for their support for the physiochemical and
analytical studies.
I thank Dr. K. N. Sunil Kumar and S. Brindha from Pharmacognosy
Department for their timely help in botanical authentication.
I express my deep sense of gratitude to Mr. G. Rajesh Kumar in providing
support in data analysis during entire course of the study.
I thank Dr. P. Thenmozhi, Dr. A. Lavanya for their support during thesis
preparation.
I extend my whole hearted thanks to Mr. V. Baskar, Mr. M. Vijayakumar,
Ms. R. Nandhini, Ms. A. Surya, Ms. Gayathri, for their co-operation and support during
the DTP works.
I thank Biochemistry HOD, Dr. R. Ganesan and Pathology HOD,
Dr. V. Ananthalakshmi for their moral support.
I extend my sincere thanks to Mr. D. Radhakrishna Reddy for his help in
review of literatures.
It gives me great pride and immense pleasure to extend my sincere regards and
deepest sense of gratitude to Mrs. V. Indra Johncy, Mr. Harish, Mr. Mohan,
Department of Pharmacy, Siddha Central Research Institute, who have helped in various
levels of the study.
I am thankful to Dr. C. Anbarasi, Dr. L. Juliet, Dr. K. Samraj, Dr. A. Usha,
Dr. R. Susila, Dr. S. Bhavani, Dr. Shyamala Rajkumar, Dr. Eugene Wilson, Dr. S.
Selvarajan for their referral to IGT OPD, in recruiting the IGT patients and constant
follow-up. I am thankful to all laboratory staff of SCRI who had supported me in my
study. I would like to thank all Admin and research staff for their support and constant
final phases of this study and my mother Mrs. M. Gomathi without whom I would not
have come into being, who graced every important moments of my life and stood as Pillars
of Support in all my endeavours with her love and compassion. They are the source of
inspiration always wishing me the best from the core of their heart. I would like to give
special thanks to my Brothers-- M. Paramasivan, M. Subbiah, M. Thangarajan, and
M. Venkateshan, Sisters- In-laws of my Family for their support and Prayers for my
upliftment.
I express my hearty thanks to my better half Dr. S. Sonitha, Siddha Physician
for her invaluable help and Sacrifice, and My lovable Children K. S. Akshayaa and K.S.
Sai Adhithyaa, whose love and care were the source of energy during my difficult times.
I express my sincere thanks to Penguin Press and Citizen Printers for their
support in Printing and Binding of the thesis.
Last but not the least, I express my sincere thanks to one and all and to those
whom I might have missed to mention, who gave constant encouragement and help
throughout my academic career.
Thank you one and all.
Chapter Title Page Number
1. Introduction 1
2. Aim & Objective 6
3. Review of Literature 7
4. Scope andPlan of Work 49
5. Materials & Methods 50
6. Result 85
7. Discussion 179
8. Summary and Conclusion 191
9. Recommendations 193
10. Bibliography
11. Annexure
Assessment Forms Table
Certificates -IHEC, CTRI,
Authentication certificates
Plagiarism report
TABLE
NO TITLE
1. Ingredients of Thiripala Chooranam Tablet
2. Preparation of Standard Solutions of concentrations
3. ICP-OES analysis
4. List of Equipments used for evaluation of Tablets
5. Compressibility index& Hausner’s ratio
6. Angle of repose:
7. Weight variation limits
8. Organisms used for Anti-Bacterial Activity
9. Ingredients of Thiripala chooranam tablet
10. Physiochemical Analysis of Individual Herbal Drugs and Thiripala Choornam
Tablets
11. Macro and Micro nutrient
12. Heavy metal Analysis of Thiripala choornam Tablet
13. Microbial analysis of Thiripala chooranam tablet formulation
14. Microbial analysis of Specific Pathogens of Thiripala chooranam tablet
formulation
15. Pesticide residue analysis of Thiripala chooranam tablet formulation
16. Aflatoxin analysis of Thiripala chooranam tablet
17. Organoleptic characters of Thiripala chooranam tablet
18. Bulk characterization of lubricated blends of optimized formulation (F3)
19. FTIR Spectrum of THIRIPALA CHOORANAM
20. FTIR Spectrum THIRIPALA CHOORNAM TABLET
21. Post compression parameters of optimized formulation (F2)
22. Post compression parameters of optimized formulation (F3)
23. Percentage inhibition of test drug Thiripala chooranam tablet on Alpha Amylase
Enzyme Inhibition Assay
25. Percentage inhibition of Thiripala chooranam tablet on α-Glucosidase Enzyme Inhibition Assay
26. IC50 Values for α-Glucosidase Enzyme inhibition by Thiripala chooranam tablet
27. Zone of Inhibition data of Anti-bacterial activity
28. Age Distribution
29. Gender Distribution
30. Educational status
31. Occupation
32. Socio Economic status
33. Family History of Diabetes
34. Distribution of Body Constitution
35. BMI distribution
36. Diet Distribution
37. Duration of treatment in each patient of LTPC – Arm
38. Duration of treatment in each patient of OLSM - Arm
39. Laboratory investigation results of patients in Thiripala Chooranam tablet with
Life style modifications Arm: Arm 1 (LTPC ARM): Before Treatment
40. Laboratory investigation results of patients in Thiripala Chooranam tablet with
Life style modifications Arm: Arm 1 (LTPC ARM): After Treatment:
41. Laboratory investigation results of patients in Only Life style modifications Arm:
Arm 2 (OLSM ARM): Before Treatment
42. Laboratory investigation results of patients in Only Life style modifications Arm:
Arm 2 (OLSM ARM): After Treatment
43. Follow-up Outcome of the Patients Taking Thiripala Chooranam
44. Physical Activity Clause Interval
45. HbA1c analysis
46. The Least Square Means of HbA1c
48. Fasting Blood Sugar (FBS) Before and After between the LTPC arm and OLSM arm
49. OGTT 2 hours
50. Mean reduction of OGTT
51. Mean difference percentage of Reduction of Lipid Profile
52. Baseline Lab Results
53. Post-treatment Lab Results
54. Post-treatment Lab Results
55. Urine Sugar Analysis
56. Responder Analysis of HbA1c
57. Odds Ratio Estimates
58. Table of Treatment
59.
Calculation of Absolute Risk Reduction, Relative Risk Reduction, Relative Risk (ARR,
FIGURE
NO TITLE
1. Thiripala chooranam Tablet
2. Ingredients of Thiripala chooranam tablet: Kadukkai
3. Ingredients of Thiripala chooranam tablet: Nellikkai
4. Ingredients of Thiripala chooranam tablet: Thandrikkai
5. Terminalia chebula
6. Emblica officinalis
7. Terminalia bellerica
8.
Maceration microscopy of Kadukkai – Terminalia chebula
fruit rind
9. Maceration microscopy of Nellikkai – Emblica officinalis fruit rind
10.
Maceration microscopy of Thandrikai – Terminalia bellerica fruit
rind
11.
Powder microscopic analysis of Kadukkai – Terminalia chebula fruit
rind
12.
Powder microscopic analysis of f Nellikkai – Emblica
officinalis-dried fruit
13.
Powder microscopic analysis of Thaandrikkai- Terminalia
bellerica pericarp
14. Thiripala chooranam tablet
15. Lignified elements in Thiripala chooranam Tablet
16. Non-lignified elements in Thiripala chooranam Tablet
18.
HPTLC Profiling of Terminalia chebula (KADUKKAI) @ 254
nm
19.
HPTLC Profiling of Terminalia chebula (KADUKKAI) @ 366
nm
20.
HPTLC Profiling of Terminalia chebula (KADUKKAI) @ 520
nm
21. HPTLC Profiling of Emblica officinalis (Nellikkai) @ 254 nm
22. HPTLC Profiling Emblica officinalis (Nellikkai) @ 366 nm
23. HPTLC Profiling Emblica officinalis (Nellikkai) @ 520 nm
24.
HPTLC Profiling Terminalia bellerica (THANRIKKAI) @ 254
nm
25.
HPTLC Profiling Terminalia bellerica (THANRIKKAI) @ 366
nm
26.
HPTLC Profiling Terminalia bellerica (THANRIKKAI) @ 520
nm
27. HPTLC Profiling of Thiripala Choornam Tablet @ 254 nm
28. HPTLC Profiling of Thiripala Choornam Tablet @ 366 nm
29. HPTLC Profiling of Thiripala Choornam Tablet @ 520 nm
30.
UV-VISIBLE ABSORPTION PROFILE- Thiripala
Choornam[TC]
31.
UV-VISIBLE ABSORPTION PROFILE- Thiripala Tablet
[TT]
32. UV-VISIBLE ABSORPTION PROFILE- Thiripala Tablet
33.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Arsenic
34.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Calcium
35.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Cadmium
36.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Copper
37.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Iron
38.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Magnesium
39.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Manganese
40.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Sodium
41.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Lead
42.
Inductively Coupled Plasma – Optical Emission Spectroscopy
studies: Potassium
43. Mass Spectral Studies: Gallic acid
44. Mass Spectral Studies: Quercetin
45. Mass Spectral Studies: Corilagin
47. FTIR Spectrum of Thiripala Chooranam
48. Alpha Amylase Inhibitory Activity of Thiripala Choornam
Tablet:
49.
ALPHA - GLUCOSIDASE ENZYME INHIBITORY
ACTIVITY OF THIRIPALA CHOORANAM TABLET
50. ANTI - BACTERIAL ACTIVITY: Escherichia coli
51. ANTI - BACTERIAL ACTIVITY: Staphylococcus aureus
52. ANTI - BACTERIAL ACTIVITY: Klebsiella pneumonia
53. ANTI - BACTERIAL ACTIVITY: Enterococcus faecalis
54. Age Distribution
55. Gender Distribution
56. Educational Status
57. Occupation Status
58. Socio Economic status
59. Family History of Diabetes
60. Distribution of Body Constitution
61. BMI distribution - ARM 1 (LTPC)
62. BMI distribution - ARM 2 (OLSM)
63. Diet Distribution
64. Physical Activity Chart
65. HbA1c analysis Chart
67. Fasting Blood Sugar (FBS) Before and After between the LTPC arm and OLSM arm
68. Fasting Blood Sugar (FBS) Mean Reduction between the LTPC arm and OLSM arm
69.
OGTT Before and After between the LTPC arm and OLSM arm
70. OGTT Mean Reduction
71. Mean difference percentage of Reduction of Lipid Profile
➢ % - Percentage ➢ & - And
➢ ≥ - Greater than
➢ ≤ - Lesser than
➢ °C - Degree of Celsius
➢ α - Alpha
➢ β – Beta
➢ μL – Microliter
➢ AAS - Atomic Absorption Spectrophotometer
➢ AChE - Acetylcholinesterase
➢ ACP - Acid phosphatase
➢ AGEs - Advanced glycation end products
➢ ALP - Alkaline Phosphatase
➢ AYUSH - Ayurveda Yoga Unani Siddha Homoeopathy
➢ B.Wt - Body Weight
➢ BDL - Below Detection Limit
➢ CCRAS - Central Council for Research in Ayurvedic Sciences
➢ CCRS - Central council of Research in Siddha
➢ CDC - the Centres for Disease Control and Prevention
➢ CFUs - Clinical Forming Units
➢ CRF - Case Report Form
➢ CTRI - Clinical Trial Registry –India
➢ DA - Dopaminergic
➢ DC - Differential Count
➢ DPPH –2, 2- Diphenyl-1-picrylhydrazyl
➢ DMSO – Dimethyl Sulfoxide
➢ DNA - Deoxy Ribonucleic Acid
➢ EDTA - Ethylene Diamine Tetra Acetic Acid
➢ ESR - Erythrocyte Sedimentation Rate
➢ FFA - Free Fatty Acids
➢ GOT - Glutamate Oxaloacetate
➢ GPT - Glutamate Pyruvate Transaminase
➢ g/ml – gram per milliliter
➢ H2 O2 - Hydrogen peroxide
➢ HBA1C – Glycated Hemoglobin
➢ HCl – Hydrochloric acid
➢ HDL – High Density Lipoproteins
➢ HPTLC - High Performance Thin –Layer Chromatography
➢ HPLC - High Performance Layer Chromatography
➢ Hr - Hour
➢ HRBC - Human Red Blood Cell
➢ HNO3 – Nitric Acid
➢ HUVEC - Human Umbilical Vein Endothelial Cells
➢ IHEC - Institutional Human Ethics Committee
➢ IC50 - Inhibitory Concentration At 50%
➢ ICMR - Indian council of Medical Research
➢ ICP -OES – Inductively Coupled Plasma _ Optical emission spectrometry
➢ IEC - Institution Ethics Committee
➢ IFG - Impaired Fasting Glucose
➢ IGT - Impaired Glucose Tolerance
➢ IIT - Indian Institute of Technology
➢ IPD - In Patient Department
➢ i.p - intra peritoneal.
➢ Kg - Kilogram
➢ L - Litre
➢ LDL – Low density Lipoproteins
➢ LPO - Lipid Peroxide Oxidase
➢ LTPC - Thiripala Chooranam tabletwithLife style modifications
➢ MBC - Minimum Bactericidal Concentration
➢ MCC - Microcrystalline Cellulose
➢ MCV - Mean Cell Volume
➢ Mg – Milligram
➢ Mg/ml – Milligram per millilitre
➢ Min - Minutes
➢ Ml / Kg - Millilitre Per Kilogram Body Weight
➢ Mmol/L – Millimoles Per Litre
➢ nm – nanometre
➢ OECD - Organization for Economic Co-Operation and Development
➢ OGTT – Oral glucose tolerance test
➢ OLSM - OnlyLife style modifications
➢ pH – pouvoir hydrogene, “Power of Hydrogen”
➢ PFTE – Poly tetra fluoro ethylene
➢ PLIM – Pharmacopoeial Laboratory for Indian Medicine
➢ Pre-DM - Pre-Diabetes Mellitus
➢ PPBS - Post Prandial Blood Sugar
➢ PPM – Parts per million
➢ PMN - Polymorphs nuclear leukocyte
➢ PNPG - p-nitrophenyl-α-D -glucopyranoside
➢ RDS – Raw Drug Section
➢ ROS - Reactive oxygen Species
➢ UV B - Ultra violet B rays
➢ TAC - Total Antioxidant Capacity
➢ TC – Thiripala Chooranam
➢ Tc -Total Cholesterol
➢ TCHE - Terminalia chebula hydroalcoholic Extract
➢ TLC – Thin layer chromatography
➢ TGL - Triglycerides
➢ TRF - Terminal Restriction Fragments
➢ TT – Thiripala Chooranam tablet
➢ STZ - Streptozotocin
➢ S.D - Standard Deviation
➢ S.E.M - Standard Error of Mean
➢ TNF - Tumour Necrosis Factor
➢ t-BHP - tertiary Butyl Hydro peroxide
➢ UV – Ultra violet
➢ UV B – Ultra Violet B
➢ U/ml – Units per millilitre
➢ V/V - Volume / Volume
➢ W/V - Weight / Volume
➢ W/W - Weight / Weight
➢ WBC - White Blood Cell
Page 1
1. INTRODUCTION
Siddha system is one of the ancient systems of Indian medicine1. This system
focused on leading a healthy life style based on physical, emotional, psychological and
social wellbeing. In AYUSH systems, the promotion of preventive approach to achieve
the goal of being healthy is attained through holistic treatments. In Siddha system,
Kayakalpam plays a major role for well-being2. Kayakalpam herbs of Siddha possess
natural antioxidants which play a vital role in preventing free radical formation and thus
preventing Non-communicable diseases like Pre-diabetes, Diabetes mellitus,
Hypertension, Cancer and so on.
Non-diabetic hyperglycemia that does not satisfy the diagnostic criteria for
diabetes mellitus (DM) is generally known as prediabetes (preDM). The progression
seems to be governed by two processes: 1) a decline in sensitivity to the action of insulin
2) dysfunction and eventual exhaustion of beta-cell function and not producing as much
insulin as required.7 Beta-cell dysfunction starts 10 to 12 years prior to the presentation of
type 2 diabetes3. A preDiabetic state of dysglycemia includes Impaired Fasting Glucose
(IFG) and Impaired Glucose Tolerance (IGT) which has a strong association with insulin
resistance and increased risk of cardiovascular pathology. IGT disease progression forms
onset of Type 2 diabetes mellitus (Glucose ranges from FBS:<126 mg /dL, PPBS:
<200mg/dL and HbA1c: <6.5%) with in few years. So, it is also a risk factor for
mortality4.
The concept of IGT was first introduced by the World Health Organization
(WHO) in 1979. It is generally agreed that all forms of diabetes pass through prediabetes
state before escalating into full-blown diabetes7. Like DM, fasting plasma glucose (FPG)
and oral glucose tolerance tests are both used independently as defining criteria for
preDM5. According to World Health organization, people with IGT show abnormal
two-Page 2 hour post-load plasma glucose values (2-h glucose 7.8–11.0 mmol/l [140–199 mg/dl]).
People with IFG only however, demonstrate an abnormal fasting plasma glucose.
According to ICMR-INDIAB study conducted in 2011 estimated 3.9 million are
in the prediabetes stage in Tamil Nadu. Projections for the whole of India would be 77.2
million preDiabetic patients6. Around 308 million people worldwide are reported to have
‘Impaired Glucose Tolerance’.
Epidemiological data from the Centers for Disease Control and Prevention (CDC)
estimate that 79 million adults aged 20 years and older in the U.S. had prediabetes in
2010, based upon fasting plasma glucose (FPG) and A1C levels14. Its prevalence is on the
rise, with an estimated 470 million people worldwide having prediabetes by 203015. It has
been proposed that 5% to 10% of individuals with prediabetes will progress to diabetes
annually, and about the same number will convert back to normoglycemia15.
Furthermore, patients with prediabetes have a 30% increased risk of developing
Type-2 diabetes mellitus over 4 years and a 70% risk over 30 years16,8. Within a decade
of the initial diagnosis ‘impaired glucose tolerance’ 25% to 75% are estimated to
progress to diabetes17.
In 2007, an estimated 308 million had Impaired Glucose Tolerance (IGT) or about
7.5% of the age group 20 to 79 years. More than 80% live in developing countries18. This
is projected to rise to 418 million, or 8.1% by 2025. Around 10% of people with
combined impaired fasting glucose (IFG) and IGT will progress to diabetes per year. The
progression rate from isolated IGT to diabetes is estimated at 3.7% per year19. The
predicted cumulative 5 to 6 years incidence of development of type 2 diabetes for people
with either IGT or IFG is 20% to 34%, those with both IGT and IFG have a progression
rate of 38% to 65%13. The pooled relative risk for development of new diabetes is 6.02 in
Page 3 to obtain accurate epidemiological data on the prevalence of the condition due to varying
definitions used to classify individuals with elevated glycemic risk20.
It has been observed that glucose measures that define IFG and IGT are able to
identify about 10% of the adults who have prediabetes19. Unfortunately, measures based
on glycated hemoglobin (A1C) levels identify a significantly lower proportion of the
population15. There is a fair amount of evidence to demonstrate that glucose levels lower
than those meeting the current definition of prediabetes may also be associated with
similar risks, particularly in high-risk individuals21. It is well recognized that the risk
of developing type 2 diabetes is higher in men than in women38.
Recently, hemoglobin A1c (HbA1c) has also been used in the diagnosis of
preDM5, 3. It is well-established that the risks of type 2 DM (T2DM) and cardiovascular
disease are significantly increased in preDM subjects.
Pre-diabetic stage is treated in contemporary western medicine with
hypoglycemic drugs. Since lifestyle modifications and poor dietary habits are also
responsible for this disease, earlier evidences have proved that combined effect of diet,
exercise and drugs (Metformin, Acarbose) are effective at preventive progression to DM
from IGT subjects6. According to DPP study intensive lifestyle intervention reduced the
development of diabetes by 58%14. Lifestyle modification is the ideal method of delaying
or preventing diabetes as it also reduces cardiovascular risk profile5. Lifestyle
interventions remain an essential part of management of prediabetes36. But lifestyle
modifications are treated separately and not approached holistically.
Pharmacological studies of the herbal formulations for the treatment of diabetes
indicate that the mechanisms of action of these interventions might be multifactorial.
Medicinal plants enhance the micro-circulation, increase the availability of insulin and
Page 4 Herbs containing polysaccharides restore the function of the pancreatic tissues
causing increased insulin output by the beta-cells. Polysaccharides are the most abundant
and the most diverse materials found on earth which possess potent antioxidant activity
and can protect the liver and kidneys from the injurious effects of diabetes.
Polysaccharides can have number of effects including anti-inflammatory,
immune-stimulating, complement activation, anti-thrombotic, antidiabetic and infection protectant
activity and many more 22,23,24,25,26. Most polysaccharides especially fructans and
mannans are derived from higher plants. They are relatively nontoxic and do not cause
significant side effects, which is a major problem associated with immunomodulatory
bacterial polysaccharides and synthetic compounds 27,28,29,30. This provides a window of
opportunity to prevent or delay the development of type 2 diabetes, and potentially
diminish the risk of cardiovascular and other complications37. Both IGT and IFG are
insulin-resistant states, but they differ in site of insulin resistance39.
However, it is possible to halt or reverse the progression of pre-diabetes, or at
least delay the development of diabetes using Kayakalpa herbs with Siddha lifestyle which regulate blood sugar levels.
Pre - Diabetes can be analogous with Munneerizhivu or piramya rogam in Siddha
system which is caused due to increase in Aiyakuttram. However, bio-combustion
challenge being the concern in the DM is related to Azhal, on slow deterioration of Azhal
leads to DM. Its complications however related to Aiyam. It is indirectly meant that pre-
DM stage, there will be decrease in Azhal and full-blown DM, increase in Aiyam31.
In drugs selection it becomes essential to identify a drug which will reduce the
increased Azhalkutram. Astringent taste regularizes Azhalkutram. Astringent taste
regularizes excessive fluid secreted out of the body. It also checks the disease caused due
Page 5
Thiripala32 Chooranam (TPC) being a Kayakarpam drug in Siddha system of
medicine, having 3 major astringents, regularizes Azhal and reduces the Aiyam.
Thiripala chooranam tablet is a poly herbal Siddha formulation, comprising of three herbs Terminalia chebula, Terminalia bellerica, and Emblica officinalis which is proven as an anti-diabetic drug33. Some studies have investigated the possible
anti-diabetic properties of combination drug Thiriphala chooranam in animal models, one in a
high fructose diet induced and another in alloxan diabetic rats34. The results of these
studies shows that the administration of the extracts reduced the blood sugar level.
Gallic acid is the component present in the composition of Thiripala chooranam
having anti-diabetic activity35. More over the same studies highlighted the anti-oxidant,
and anti-obesity effects of this drug40. These effects are assumed to play a major role to
reduce the chances of transition of IGT patients to established Diabetics. These become
the rationale for the selection of “Thiripala Choornam Tablet” as an interventional drug.
In this study, prediabetics (Munneerizhivu) is treated holistically with the study
drug (Thiripala Choornam tablet) and Siddha Life style modification. So, the above said
poly-herbal formulation, Thiripala Choornam Tablet is more effective, cost effective and
Page 6 2. AIM & OBJECTIVES
To pharmacognostically identify the selected individual plant powders, Kadukkai
(Terminalia chebula), Nellikai (Emblica officinalis), Thandrikai (Terminalia bellerica) for authentication.
To develop the Thiripala Chooranam tablet (Siddha formulation) of the selected plants.
To Standardize individual plant crude powders and Thiripala Chooranam tablet (Siddha
formulation) through analytical tests such as HPTLC, UV-Visible Absorption Profile
Studies, FTIR, ICP- OES, AAS, LC-MS.
To evaluate safety of the trial drug through microbial study, pesticide, aflatoxin and
heavy metal analysis.
To develop and analyze Thiripala Chooranam tablet by Pre - formulation and Post - compression studies as per AYUSH guidelines.
To measure the glycosidic enzyme - inhibiting activity of Thiripala Chooranam tablet
(Siddha formulation) on alpha amylase and alpha glucosidase ( In - Vitro studies).
To evaluate the clinical efficacy of the trial drug in controlling progress to full blown
diabetes with an active control in parallel to prove its efficacy of reducing and
maintaining blood glucose levels in normo - glycemic range and with respect to body
constitution (Udaliyal).
To assess the hypolipidemic efficacy of the trial drug through clinical trial.
To bring about conclusive and demonstrable scientific support for the Thiripala
Chooranam tablet- a traditional Siddha formulation in treating and reversal of Impaired
Page 7
3. REVIEW OF LITERATURE
A detailed and thorough literature survey was carried for the individual
selected plants and Thiripala Chooranam Tablet from Libraries of well-established
institutes (IIT Chennai, CLRI Chennai, The T.N. Dr. M.G.R. Medical University
Chennai, Govt. Siddha Medical College, Chennai and Palayamkottai, National
Institute of Siddha, Chennai, SCRI, Chennai), and various literature sources such as
Medline via Pub Med, Science Direct, and Google Scholar. The collected scientific
reports were presented.
3.1 ETHNOBOTANICAL REVIEW
3.1.1 PLANT PROFILE (Terminalia chebula Retz)
Botanical Source : Terminalia chebula Retz.
Family : Combretaceae
Synonyms : Buceras chebula (Retz.) Lyons, Myrobalanus chebula (Retz.)
Gaertn.
Parts Used : Dried pericarp
Taxonomic classification
Kingdom : Plantae
Division : Tracheophyta
Class : Magnoliopsida
Order : Myrtales
Family : Combretaceae
Genus : Terminalia
Species : Terminalia chebula
Vernacular Names
Sanskrit : Kayastha
Hindi : Harra
Kannda : Halle
Malayalam : Kattuka
English : Chebulic myrobalan
Tamil : Kadukkai
Page 8
Distribution
The plant is found throughout India chiefly in deciduous forests. It occurs
abundantly in North India. Its range extends southwards at 300 to 900 m altitude41.
Description of drug
Fruit is a hard, stony drupe, greenish yellow in colour, odourless, ovate,
longitudinally wrinkled, 3.5 to 4 cm in length, 1.5 to 2 cm wide and has 5 to 6 ridges
(longitudinal ribs). In some, the basal portion is narrower and somewhat elongated on
tapering42.
Habit
Moderate sized or large tree48.
Chemical Constituents
Fruit
Tannins (20-40%)43, which on hydrolysis give chebulic acid and D-galloyl
glucose, chebulagic acid, chebulinic acid, ellagic acid and gallic acid; a tannin
terchebin44, an ellagitannin terchebulin45, syringic acid46, gallic acid (1.21%)47.
Traditional Medicinal Uses
Jaundice, eye diseases, bleeding disorders, laxative, ascites48.
PHYTOCHEMICAL INVESTIGATIONS
➢ Terminalia chebula contains the triterpenes arjun glucoside 1,
arjungenin and the chebulosides 1&2. Other constituents contain
tannins up to 30%, chebulic acid 3-5%, chebulinic acid 30%, tannic
acid 20-40%, ellagic acid, 2,4-chebulyi–β-D-glucopyranose, gallic
acid, ethyl gallate, punicalagin, terflavin A, terchebin, anthraquinone,
flavonoids like luteolin, rutins, and quercetin etc.
PHARMACOLOGICAL ACTION
➢ The water-soluble fraction of T. chebula (WFTC) was effective against
systemic and local anaphylaxis. Shin et al. showed that the injection of
WFTC with doses of 0.01–1.0 g/kg inhibited anaphylactic shock
100%. When WFTC was pretreated at concentrations ranging from
dose-Page 9
dependently. In addition, this study showed that WFTC increased
anti-dinitrophenol and IgE-induced tumor necrosis factor- (TNF-) α
production49.
➢ T. chebula has pharmacological activities relevant to dementia therapy.
Different extracts from T.chebula have exhibited
concentration-dependent inhibitory activities on AChE and BChE. T. chebula with a
high content of phenolic constituents exhibits strong antioxidant and
neuroprotective properties in vitro and in vivo. Hence T. chebula
proves to be beneficial in the treatment of Alzheimer’s disease 50.
➢ Anti-ulcerogenic activity: Animals pretreated with doses of 200 and
500 mg/kg, Terminalia chebula hydroalcoholic extract showed
significant reduction in lesion index, total affected area and percentage
of lesion in comparison with control group (p<0.05 and p<0.01) in the
aspirin, ethanol and cold restraint stress-induced ulcer models.
Similarly extracts increased mucus production in aspirin and
ethanol-induced ulcer models. At doses of 200 and 500 mg /kg Terminalia
chebula extract showed antisecretory activity in pylorus ligated model which lead to a reduction in the gastric juice volume, free acidity, total
acidity and significantly increased gastric pH. This activity thus lends
pharmacological credence to the suggested use of the plant as a natural
remedy in the treatment or management of ulcer 51,52.
➢ Neuroprotective activity: The methanol and water extracts of
Terminalia chebula exhibit neuroprotective activities against H202-induced toxicity toward PCl 2 cells and are potential candidates for the
treatment of H202-induced neurodegenerative disease. The effective
neuroprotective activity of the water extract is consequence of its "OH
and HP2 scavenging activities, its greatest extraction yield and its total
phenolic and tannin content 53,54.
➢ Anti-convulsant activity: The ethanolic extract of Terminalia chebula
significantly reduced the duration of seizures induced by maximal
electroshock. The ethanol extract in doses of 200 and 500 mg/ kg
Page 10
same doses also protected animals from pentylenetetrazole-induced
tonic seizures and significantly delayed the onset of tonic seizures
produced by picrotoxin55. The ethanolic extract of Terminalia chebula
(EETC) possess anticonvulsant activity since it reduced the duration of
seizures produced by maximal electroshock and delayed the latency
of seizures produced by pentylenetetrazole and picrotoxin55. This
provides a pharmacological justification for the traditional use of the
plants fruits in the management of epilepsy in some rural parts of
India.
➢ Anti-oxidant activity: Terminalia chebula are new potential sources
of natural antioxidants for food and nutraceutical products. The
methanol extract of Terminalia chebula had the greatest total
triterpenoid content and exhibited good antioxidant activity in the
HRP-luminol-H202 assay56. The water extract appeared to have the
greatest total phenolic and tannin content and showed good antioxidant
activities in both CuS04-Phen-Vc-HP2 and luminol - H202 assays. The
95% ethanol extract exhibited good antioxidant activity in the
pyrogallol-luminol assay. Thus, the three extracts present various
levels of ROS scavenging efficiency due to differences between the
mechanisms of the four ROS chemiluminescence systems. Aqueous
extract of natural herb, Terminalia chebula inhibitsxanthine / xanthine
oxidase activity and is also an excellent scavenger of DPPH radicals.
It is concluded that the aqueous extract of T. chebula acts as a potent antioxidant and since it can protect cellular organelles from the
radiation-induced damage, it may be considered as a probable
radioprotector57. Protective effects of an aqueous extract of Terminalia
chebula fruit on the tert-butyl hydroperoxide (t-BHP)-induced oxidative injury was observed in cultured rat primary hepatocytes and
rat liver58,59. It has stronger antioxidant activity than alpha-tocopherol.
HPLC analysis with diode array detection indicated the presence of
hydroxybenzoic acid derivatives, hydroxycinnamic acid derivatives,
flavonol aglycones and their glycosides, as main phenolic
Page 11
➢ On comparison with the typical aqueous extraction method the
extraction efficiency was highest for microwave treatment followed by
ultrasonication61. The study revealed a 17 .6% increase in the
yield of phenolics and a 14% increase in the tannin content of the
microwave extracts. A 20.6% increase in the antioxidant activity of
tthe microwave extract was also obtained. The sonication extracts
showed an increase of 0.6, 5 and 9.69% in the yield of phenolics,
tannins and antioxidant activity, respectively.
➢ Hepatoprotective activity: Terminalia chebula extract was found to
prevent the hepatotoxicity caused by the administration of rifampicin
(RIF), isoniazid (INH) and pyrazinamide (PZA) combination in
sub-chronic model in mice (12 weeks)62.
➢ Cardioprotective activity: Terminalia chebula extract pretreatment
was found to ameliorate the effect of isoproterenol on lipid peroxide
formation and retained the activities of the diagnostic marker enzymes
m - isoproterenol induced myocardial damage in rats63. Its pericap has
also been reported to have cardioprotective activity in isolated frog
heart model64.
➢ Cytoprotective activity: Ethanolic extract of Terminalia chebula
fruit exhibited significant cytoprotective effect against UV B-induced
oxidative damage. These observations were attributed to the
inhibitory effect of the Terminalia chebula extract on the age dependent shortening of the telomere length as shown by the
Southern Blots of the Terminal Restriction Fragments (TRFs) of
DNA extracted from sub-culture passages. Cytoprotective effect on
oxidative stress and inhibitory effect on cellular aging of its fruits
Page 12
➢ Antidiabetic and retino-protective activity: The anti-diabetic
property of medicinal plants and its relationship with their
antioxidant potential have long been established66. The methanolic
extract of Terminalia chebula, Terminalia bellerica, Emblica
officinalis and their combination named 'Triphala' was found to inhibit lipid peroxide formation and scavenge hydroxyl and superoxide
radicals in the diabetic rats confirming their antidiabetic potential67.
Anti-diabetic potential of Terminalia chebula fruits was effective on streptozotocin (STZ) -induced experimental diabetes in rats. Oral
administration of ethanolic extract of the fruits (200 mg/kg body
weight/rat/day) for 30 days significantly reduced the levels of blood
glucose, glycosylated hemoglobin and stimulated insulin in diabetic
rats. Also, a decrease in the number of secretory granules of β-cells
was observed in the STZ induced diabetic rats and these pathological
abnormalities were normalized after treatment with T. chebula
extract68.
➢ Terminalia chebula methanolic extract (TCE) containing 2.7%
chebulic acid showed preventive effects against the formation
of advanced glycation end products (AG Es) and endothelial cell
dysfunction69. When the effects of TCE on AGE formation and
on protein crossing linking by glycation with D-threose and lens
crystallines were examined, TCE showed inhibitory activity in a
dose-dependent manner and the concentration of 1000 µg mL_,
presented an activity similar to that of 5 mM aminoguanidine as
a positive control. The incubation of Human Umbilical Vein
Endothelial Cells (HUVEC) with 100 µg mL_, of advanced glycation
end products (AGEs) caused a considerable increase in THP-1
monocytic cell adhesion, but this adhesion was reduced by the
treatment of TCE. This showed that TCE is a potential agent
Page 13
➢ Hypolipidaemic activity: Terminalia chebula was found to possess
significant hypolipidaemic activity. In atherogenic diet induced
hyperlipidemic model, the rats receiving treatment with Terminalia chebula showed significant reduction in total cholesterol, triglycerides, total protein and elevation of high density lipoprotein
cholesterol. The results also suggested that Terminalia chebula at 1.05
and 2.10 mg kg concentrations are an excellent lipid-lowering
agent70,71.
➢ Anti-arthritic effect: Terminalia chebula hydroalcoholic Extract
(TCHE) has the potential to be used as a disease-modifying
agent in treatment of rheumatoid arthritis72,73. TCHE produced
a significant inhibition of joint swelling as compared with
control in both formaldehyde-induced and Complete Freund's
adjuvant CFA-induced arthritis. The TCHE treatment also reduced
serum TNF-a level and synovial expression of TNF-Rl, IL-6 and
IL-1 ~- Results of acute toxicity study showed that the oral LD50
of TCHE was >2000 mg kg ". Chronic administration also did
not produce any significant physiological changes as compared
with normal rats.
➢ Antimutagenic/anticarcinogenic activity: The inhibitory action on
cancer cell growth by the phenolics of Terminalia chebula fruit
and found that chebulinic acid, tannic acid and ellagic acid were the
growth inhibitory phenolics74. Acetone extract of bark and fruit
powder of Terminalia chebula harbors constituents with promising
antimutagenic/anticarcinogenic activity75.
➢ Molluscicidal activity: Terminalia chebula fruit is a potential
source of biomolluscicides against Lymnaea acuminata76. These
snails are the intermediate host of liver fluke Fasciola gigantica which
causes 94% fascioliasis in the buffalo population of northern India 77.
The active molluscicidal component of Terminalia chebula fruit is soluble in carbon tetrachloride, chloroform, ether, acetone and ethanol.
Page 14
component present is more soluble in ethanol than other organic
solvents. The tannic acid is the active component present in
Terminalia chebula fruit by High Performance Liquid
Chromatography76. Further it was evaluated that in vivo and in
vitro exposure of tannic acid significantly inhibited the
acetylcholinesterase (AChE), acid phosphatase (ACP) and alkaline
phosphatase (ALP) activity in the nervous tissue of Lymnaea
acuminata78.
➢ Immunomodulatory effect: The immunosuppressive effects of gallic
acid and chebulagic acid79, the active phytoconstituents of Terminalia
chebula extract., on cytotoxic T - lymphocyte (CTL) mediated cytotoxicity. It has been noted that gallic acid and chebulagic acid
blocked the CTL- mediated cytotoxicity. Moreover, gallic acid and
chebulagic acid has been shown to inhibit the killing activity of CD8 +
CTL clone at IC 50 values of 30 and 50 µM respectively.
Additionally, the granular exocytosis in response to anti-CD3
stimulation was also blocked by gallic acid and chebulagic acid that
further evidenced its immunosuppressive effect.
➢ Anaphylactic effect: Inhibitory action of water soluble fraction of
Terminalia chebula on systemic and local anaphylaxis has also been
evaluated. The effects of the water soluble fraction of Terminalia
chebula showed the reduction and frequency of anaphylactic shock that further confirmed the fact that it may possess a strong anti-
anaphylactic action80.
➢ Anti-caries effect: The potential of the aqueous extract of Terminalia
chebula as an anticaries agent have also been evaluated. The extract strongly inhibited the growth, sucrose-induced adherence and
glucan-induced aggregation of Streptococcus mutans81.
➢ Wound healing: The herbal paste preparation obtained from T.
Page 15
and deposition of collagen. Thus, it offers a distinct advantage to
wound healing82.
➢ Prokinetic effect: The oral administration of Terminalia chebula
on gastric emptying has been investigated to confirm its potent
prokinetic effect. Metoclopramide significantly increased the
gastric emptying (76.33±12.37%; p<0.01) and atropine inhibited the
motility percent gastric emptying (7.26±19.76%; p<0.01 ).
Terminalia chebula extract was found to increase the percent gastric
emptying (86.57±6.65%; p<0.01) which showed that Terminalia
chebula extract may serve as a useful alternative to prokinetic drugs available benefits.
TOXICITY STUDIES
➢ To support the safety-in-use of the ethyl acetate (EtOAc)-soluble
portion of a T. chebula ethanol extract containing 29.4% chebulic acid
content, the prepared portion was tested in an in vitro mutagenicity
assay, and a single and 14-day repeated dose oral toxicity study. In the
bacterial mutation assay, up to 5000 µg/mL concentration of the
EtOAc-soluble portion, the numbers of colonies did not increase
whether with or without metabolic activation. In the oral toxicity study,
the single oral dose of the extract at 2000 mg/kg did not produce
mortality or abnormal lesions in the internal organs of rats. The results
of a 14-day orally repeated dose showed that the EtOAc-soluble
portion of T. chebula ethanol extracts gave no adverse effects at dosages of 2000 mg/kg in rats83.
ANTI MICROBIAL ACTIVITY
➢ Anti-bacterial activity: The extract of Terminalia chebula shows
broad spectrum activitiy84. The ethanol extract at a concentration of
1mg per disc showed maximum inhibition against Salmonella
epidermidis followed by Bacillus subtilis85. The methanolic and
Page 16
against gram-negative bacteria particularly to the agents causing
gastroenteritis86. Terminalia chebula exhibited antibacterial activity
against a number of bacterial species87. It is effective in inhibiting
the urease activity of Helicobactor pylori88, an ubiquitous bacterium implicated in the development of gastritis, ulcers and stomach
cancers. Gallic acid and its ethyl ester isolated from ethanolic extract
of Terminalia chebula showed antimicrobial activity against methicillin-resistant Staphylococcus aureus89. It has also growth inhibitory action against Salmonella typhi90 and intestinal bacteria91.
The extracts of Terminalia chebula proves to be an effective
anti-bacterial agent by forming the inhibitory zone against
Pseudomonas aeruginosa, P. fluorescens, Bacillus bronchiseptica, Staphylococcus aureus, Salmonella epidermidis, B. cereus, B. pumilis, Shigella boydii and Escherichia coli92.
➢ Antifungal activity: An aqueous extract of Terminalia chebula
exhibits antifungal activity against a number of dermatophytes and
yeasts93,94. It is effective against the pathogenic yeast Candida
albicans and dermatophytes Epidermophyton, Floccosum,
Microsporum gypseum and Trichophyton rubrum95.
➢ Antiviral activity: Terminalia chebula fruits afforded four
immunodeficiency virus type 1 (HIV-1) integrate inhibitors, gallic
acid and three galloy glucoses. Their galloyl moiety plays a major role
for inhibition against the 3'-processing of HIV-1 integrase of the
compounds (Ahn et al., 2002). It protects epithelial cells against
Influenza A virus, supporting its traditional use for aiding in
recovery from acute respiratory infections96. The therapeutic
activity against Herpes simplex virus (HSV) both in in vitro and in
vivo tests97. Terminalia chebula's effect on human cytomagalovirus
(CMV). They found that Terminalia chebula was effective in
inhibiting the replication of human cytomagalovirus in vitro and in
anAIDS model with immunosuppressed mice and concluded that it
Page 17
immunocompromised patients. It is also helpful in sexually transmitted
diseases and AIDS98.
CLINICAL STUDIES
➢ Oral rinsing with extract of T. chebula was found to significantly
reduce both total bacterial counts and streptococcal counts in saliva samples. The protective effect lasted for about 3 h after rinsing,
demonstrating a potential role of T. chebula in the prevention of dental
caries99.
➢ A short term clinical trials have been carried out on patients with
simple constipation. T. chebula increases the stools and has got
property of evacuating the bowel completely100.
➢ Besides, some Ayurvedic drugs, consisting of T. chebula as one of the
constituents have been subjected to clinical trials regarding their
effects on constipation, mental and physical disability, allergic rhinitis
and mental stress. In all the cases T. chebula containing drugs showed
good effects in the treated groups when compared to their normal
Page 18
3.1.2 PLANT PROFILE (Phyllanthus emblica L.)
Botanical Source : Phyllanthus emblica L.
Family : Phyllanthaceae
Synonyms : Emblica officinalis Gaertn, Cicca emblica (L.) Kurz, Diasperu
emblica (L.) Kuntze .
Parts Used : Dried fruit
Taxonomic classification
Kingdom : Plantae
Division : Tracheophyta
Class : Magnoliopsida
Order : Malphigiales
Family : Phllanthaceae
Genus : Phyllanthus L.
Species : Phllanthus emblica
Vernacular Names
Sanskrit : Amalaki
Hindi : Amla, Amlika
Kannda : Nellikayi, Amalaka
Malayalam : Nellikai, Nelli
English : Emblic myrobalan
Tamil : Nellikai, Nelli
Telugu : Amalakamu, Nelli
Distribution
The tree is found in the mixed deciduous forests of India ascending to 1280 m in
the hills. It is often cultivated in gardens and homeyards103
Description of drug
Drug consists of curled pieces of pericarp of dried fruit occurring as separated
segments; 1 to 2 cm, long or united with 3 or 4 segments; bulk colour grey to black,
pieces showing a broad, highly shriveled and wrinkled external convex surface to
somewhat concave, transversely wrinkled lateral surface, external surface shows a few
whitish specks, occasionally, some pieces show a portion of stony testa; texture rough
Page 19
Habit
Small or medium sized deciduous tree103
Chemical Constituents
Fruit
The fruits are reported to possess most stable ascorbic acid content and tannins,
and can be retained even when dried, powdered, or prepared candies or tablets105,106.
Some of the tannins which were found in all tissues except seeds in all stages of growth
were glucogallin, corilagin, gallic acid, ellagic acid. Immature fruits consisted of
chebulagic acid, chebulinic acid, 3, 6-dialloylglucose and five unidentified tannin
components107. Pericarp consisted emblicanins A and B, punigluconin and
pedunculagin108. The juice of the fruit contains tannic acid, gallic acid, phoroglucinol,
pyrogallol and catechol109. The major amino acids found in the fruits were glutamic
acid, proline, aspartic acid, alanine and lysine110. The free amino acids found in the
fruits were serine and traces of alanine and beta-alanine along with fructose, glucose
and sucrose as sugars111. The organic acids in the fruits were citric, malic, oxalic and
tartaric acid112. The macro and micro contents of the fruits showed the presence of
potassium, phosphorus, calcium, magnesium, sulphur, iron, manganese, zinc, boron,
copper, sodium, chloride, selenium and silica110.
Traditional Medicinal Uses
The fruits are used as carminative113, appetizer114, laxative, astringent115,
purgative, refrigerant116, nausea 117, vomiting115, stomach troubles118, constipation119,
indigestion120, diarrhea, dysentery, for gastritis, jaundice116, stomach ache121,
flatulence122, liver tonic123, liver disorders124, hepatitis125, cholera126, burning sensation,
emaciation, peptic ulcer, anorexia, dyspepsia115, to promote salivary secretion127, skin
diseases128, leucoderma129, scurvy130, hair tonic131, alopecia132, for dandruff133,
bronchitis134, aphrodisiac115, abortifacient135, menorrhagia136, leucorrhoea137,
spermatorrhoea138, anaemia116, high blood pressure139, urinary ailments140,
hemorrhage116, cardiac disorders141, painful micturition during pregnancy142, alexiteric,
Page 20
PHYTOCHEMICAL INVESTIGATIONS
➢ Emblica officinalis Gaertn contains most stable ascorbic acid content
and tannins, and can be retained even when dried, powdered, or
prepared candies or tablets105,106. Some of the tannins which were
found in all tissues except seeds in all stages of growth were
glucogallin, corilagin, gallic acid, ellagic acid. Immature fruits
consisted of chebulagic acid, chebulinic acid, 3, 6-dialloylglucose and
five unidentified tannin components107. Pericarp consisted emblicanins
A and B, punigluconin and pedunculagin108. The juice of the fruit
contains tannic acid, gallic acid, phoroglucinol, pyrogallol and
catechol109. The major amino acids found in the fruits were glutamic
acid, proline, aspartic acid, alanine and lysine110. The free amino acids
found in the fruits were serine and traces of alanine and beta-alanine
along with fructose, glucose and sucrose as sugars111. The organic
acids in the fruits were citric, malic, oxalic and tartaric acid112. The
macro and micro contents of the fruits showed the presence of
potassium, phosphorus, calcium, magnesium, sulphur, iron,
manganese, zinc, boron, copper, sodium, chloride, selenium and
silica110.
PHARMACOLOGICAL ACTION
➢ Hepatoprotective Activity: Hepatoprotective activity of Emblica
officinalis (EO) and chyawanprash (CHY) extracts was studied using
Carbon tetrachloride induced liver injury model in rats. Emblica
officinalis and CHY extracts were found to inhibit the hepatotoxicity produced by acute and chronic administration as seen from the
decreased levels of serum and liver lipid peroxides (LPO),
glutamate-pyruvate transaminase (GPT), and alkaline phosphatase (ALP). Chronic
CCI (4) administration was also found to produce liver fibrosis as seen
from the increased levels of collagen hydroxyl proline and pathological
analysis. Emblica officinalis and CHY extracts were found to reduce the
elevated levels significantly, indicating that the extract could inhibit the
Page 21
➢ Anti-Inflammatory Activity: Leaves and fruits of Phyllanthus emblica
L. have been used for the anti-inflammatory and antipyretic treatment of
rural populations in its growing areas in subtropical and tropical parts of
China, India, Indonesia, and the Malay Peninsula. In the present study,
leaves of Phyllanthus emblica were extracted with ten different solvents
(n-hexane, diethyl ether, and methanol, tetrahydrofuran, acetic acid,
dichloromethane, 1, 4 - dioxane, toluene, chloroform and water).
➢ The inhibitory activity of the extracts against human poly morphs
nuclear leukocyte (PMN) and platelet functions was studied. Methanol,
tetrahydrofuran, and 1, 4-dioxane extracts (50 micrograms/ml) inhibited
leukotriene B4-induced migration of human PMNs by 90% and
N-formyl-L-methionyl- L-phenylalanine (FMLP)-induced degranulation
by 25-35%. Diethyl either extract (50 micrograms/ml) inhibited calcium
ionosphere A23187-induced leukotrienes Release form human PMNs by
40% thromboxane B2 production in platelet during blood clotting by
40% and adrenaline induced platelet aggregation by 36%. Ellagic acid,
garlic acid and rutin all compounds isolated earlier from P. emblica.
Anti-inflammatory activity was found in the water fraction of methanol
extract of the plant leaves. The water fraction of the methanol extract
inhibited migration of human PMNs in human platelets during clotting;
suggesting that the mechanism of the anti-inflammatory action found in
the rat paw model does not involve inhibition of the synthesis of the
measured lipid mediators144.
➢ Cardio-protective Activity: The effects of chronic oral administration
of fresh fruit homogenate of Amla on myocardial antioxidant system
and oxidative stress induced by ischemic-reperfusion injury (IRI) were
investigated on heart in rats. Chronic Emblica officinalis administration
produces myocardial adaptation by augmenting endogenous antioxidants
and protects rat hearts from oxidative stress associated with IRI145.
➢ Anti-cancer activity: The important advantages claimed for therapeutic
use of medicinal plants in chemoprevention. Chemoprevention is a
rapidly growing area of oncology which focuses towards the cancer
preventive strategy of natural or synthetic interventions, nowadays
Page 22
use of synthetic agents which shows toxic and harmful side effects.
Amla is one of the foremost plants utilized from antiquity till to date.
➢ Emblica officinalis is valued for its unique tannins and flavanoids, which
exhibit very powerful antioxidant properties. The inhibition of tumor
incidences by fruit extract of this plant has been evaluated on two-stage
process of skin carcinogenesis in Swiss albino mice. Chemopreventive
potential of Emblica officinalis fruit extract on
7,12-dimethylbenz(a)anthracene (DMBA) induced skin tumourogenesis in
Swiss albino mice have been found146.
➢ Experiment using Emblica officinalis powder (EOP) to understand the
activity in Alzheimer’s disease147, total serum cholesterol level and
brain cholinesterase activity in mice. EOP was administered orally in
three different doses for 15 days to different groups of young and aged
mice. Study was performed using different models to test the cognition
activity. Diazepam, Scopolamine and ageing induced amnesia served
as the interceptive behavioral model, different blood biochemical
parameters were also estimated. EOP showed a dose dependent growth
in learning behavior of young and elderly mice. It also reversed the
amnesia induced by scopolamine and diazepam. EOP helped to reduce
brain cholinesterase and total cholesterol levels. EOP may prove to be
a useful medication for Alzheimer’s disease.
➢ The antiaging effect148 of Embelica officinalis fruit (EO).
Mitochondrial activity on human skin fibroblasts was established using
MTT assay. Valuation of pro-collagen-1 and matrix
metalloproteinase-1 (MMP-metalloproteinase-1) release was performed. EO demonstrated inhibitory
activity of hyaluronidase. EO also banned UVB disturbed cell cycle to
normal phase. EO helps to control UVB induced photoaging in human
skin fibroblasts via its strong ROS scavenging ability.
➢ Photoprotection efficacy149 of Amla (Emblica officinalis) fruit extract
through various in vitro models. It was found that
1-O-Galloyl-β-D-glucose (β-glucogallin) active in Amla extract contributes for photoprotection activity in cell cultures. β-glucogallin efficacy was a at range between 10 to 80μg/mL in B16F1 mouse melanoma cells which
Page 23
melanogenesis. It showed very high oxygen radical absorbance
capacity (ORAC) value of 4200μmol Trolox equivalents/g.
β-glucogallin from amla extract has potential photo protection efficacy.
➢ The efficacy of Emblica officinalis (EO) against radiation and
cadmium induced biochemical changes150. Total proteins, glycogen,
cholesterol, acid phosphatase, alkaline phosphatase activity, DNA and
RNA was examined. The value of cholesterol, glycogen, RNA, acid
phosphatase and alkaline phosphatase activity increased till day 14 in
non-drug treated groups and in day-7 (EO) treated groups and
thereafter decreased up to the last autopsy interval studied.
➢ An experiment to evaluate the antitumor efficacy151 of the ethanolic
extract of Amalakyadi churna (AC) against Ehrlich Ascites Carcinoma
(EAC) tumor model. The ethanolic extract showed significant
increased survival time, life span and increases body weight of treated
tumorous mice was almost normal and reducing the rate of tumor
incidence. This experiment informs that AC possesses significant
antitumor activity when tumor is at preliminary stage. The chemical
compounds present in the extract might synergistically exhibit the
antitumor activity and provided compounds responsible for the folk
anticancer uses of this plant.
➢ Anti-oxidant properties152 of Emblica officinalis (Amla) fruits on
Streptozotocin induced type-I diabetes rats. Diabetic induced animals
showed significant increase in BSL, malonadialdehyde and reduced
superoxide dismutase, catalase and decreased glutathione levels in
liver tissue compared to control groups. This infers potent antioxidant
and lipid peroxidation inhibiting activity of fresh juice and
hydroalcoholic extract of fruits of E.officinalis in diabetic rats. The
study indicates link between oxidative stress and diabetes.
➢ Anti-diabetic activity153 of fresh and hydroalcoholic extract of fruits
of E.officinalis on Streptozotocin (STZ) induced type-I diabetic rats. Treatment with freshly extracted juice and hydroalcoholic extract
significantly reduced elevated fasting glucose levels in type-I diabetic
rats. Treatment with freshly extracted juice and hydroalcoholic extract