COMPARATIVE STANDARDIZATION STUDY OF THREE TRIPHALA CHURNA FORMULATION
HTML Full TextCOMPARATIVE STANDARDIZATION STUDY OF THREE TRIPHALA CHURNA FORMULATION
D. K. Kadam *, P. D. Ahire, J. V. Bhoye, A. R. Patil and D. K. Yadav
NGSPM’s College of Pharmacy, Brahma Valley Educational Campus, Tryambakeshwar Road, Anjaneri, Nashik - 422213, Maharashtra, India.
ABSTRACT: In a few decades, there has been exponential growing in the field of herbal medicines. Most of the traditional system of medicine is effective, but they lack standardization. So, there is a need to develop a standardization technique. Standardization of herbal formulation is essential to assess the quality, purity, safety, and efficacy of the drug based on the concentration of their principles. This articles reports on standardization of Triphala churna. Polyherbal ayurvedic medicines used to treat constipation, gastric disorder. The present research study deal with the comparative Standardization of two reputed marketed Triphala churna formulation, from Patanjali, Shree Ayurveda and laboratory made churna. The standardization of this formulation, organoleptic characteristics, physical properties such as moisture content (LOD), ash value, extractive values, crude fiber content was carried out. The heavy metal content, tannin test, and alkaloid test study also carried out to ascertain the quality, purity, and safety of these herbal formulations.
Keywords: |
Standardization, Triphala churna, Physicochemical parameters
INTRODUCTION: Nature always stands as a golden mark to exemplify the outstanding phenomena of symbiosis. Today about 80% of people in developing countries still rely on traditional medicine based largely on the different species of plants for their primary health care. About 500% of plants with medicinal uses are mentioned in ancient literature, and 800 plants have been used in the indigenous system of medicine. The various indigenous system such as Ayurveda, Siddha, Unani use several plant species to treat different ailments 1, 2, 3. Herbal medicines make up an essential component of the trend toward alternative medicine.
A Harvard study recently found that one in three respondents acknowledged the use of at least one alternative therapy within the past year. Extrapolated, these findings suggest that up to $13.7 billion were spent in 1990 alone for these treatments 4. Tyler defines herbal medicines as "crude drugs of vegetable origin utilized for the treatment of disease states, often of a chronic nature, or to attain or maintain a condition of improved health." 5 current demands for herbal medicines have resulted in an annual market of $1.5 billion and increasingly widespread availability 6.
Potential Benefits of Herbal Drugs: Historically, herbal medicines have played a significant role in the management of both minor and major medical illnesses. One example is foxglove, which contains cardiac glycosides, and serves as a classic treatment for congestive heart failure. Even now, physicians still use many drugs that possess botanical origins. Huxtable notes that one-quarter of the prescriptions currently written in the United States are for plant products, while one quarter is for agents based on botanical compounds. The therapeutic potential of herbal medicines cannot be ignored and is highlighted in the three examples provided next 7.
Advantages of Herbal Medicine:
- They have a large amount of use.
- They have better patient tolerance as well as acceptance.
- The medicinal plants have a renewable source of cheaper medicines.
- Improvements in the quality, efficacy, and safety of herbal medicines with the development of science and technology.
- Prolong and uneventful use of herbal medicines may testify to their safety and efficacy.
- They are cheap.
- They are not harmful.
- They are more effective than any synthetic drug throughout the world herbal medicines have provided many of the most potent medicines to the vast arsenal of drugs available to modern medical science, both in crude form as well as a pure chemical upon which modern medicines are constructed 8, 9.
The Need for Standardization – Producers and Consumers Perspective: 10 In the global perspective, there is a shift towards the use of the medicine of herbal origin, as the dangers and the shortcoming of modern medicine are getting more apparent. It is the cardinal responsibility of the regulatory authorities to ensure that consumers get the medication, which guarantees purity, safety, potency and efficacy. The regulatory authorities rigidly follow various standards of quality prescribed for raw materials and finished products in pharmacopeias, formularies and manufacturing operation through statutory imposed good manufacturing practices. These procedures logically would apply to all types of medication whether included in the modern system of medicine or one of the traditional systems.
Though, herbal products have become increasingly popular throughout the world, one of the impediments in its acceptance is the lack of standard quality control profile. The quality of herbal medicine that is, the profile of the constituents in the final product has implications in efficacy and safety.
However, due to the complex nature and inherent variability of the constituents of plant-based drugs, it is difficult to establish a quality control parameter through the modern analytical technique is expected to help in circumventing this problem. Furthermore, the constituents responsible for the claimed therapeutic effects are frequently unknown or only partly explained. This is further complicated by the use of a combination of herbal ingredients as being used in traditional practice. It is common to have as many as five different herbal ingredients in one product. Thus, batch to batch variation starts from the collection of the raw material itself in the absence of any reference standard for identification.
These variations multiply during storage and further processing. Hence, for herbal drugs and products, standardization should encompass the entire field of study from the cultivation of medicinal plant to its clinical application. Plant materials and herbal remedies derived from them represent a substantial portion of the global market, and in this respect internationally recognized guidelines for their quality assessment and quality control are necessary.
Plan of Work: Comparative standardization of Triphala Churna formulated by Patanjali, Shree Shree Ayurveda and laboratory made Triphala churna was planned to carry out the development of quality standards for the finished marketed formulation. The method used for the comparative standardization was planned to be carried out as follows:
Development of standardization parameters for Triphala Churna:
Study of Organoleptic Characters:
- Colour
- Odor
- Taste
Determination of Physicochemical Parameters:
- Total ash
- Acid-insoluble ash
- Water soluble ash
- Moisture content/ Loss on drying
- Water-soluble extractive
- Alcohol soluble extractive
- Crude fiber contents
Evaluation of Churna:
- Powder fineness
- Bulk density
- Tap density
- Angle of repose
- Compressibility
- Hausner’s ratio
Determination of pH:
Establishing the Safety about Heavy Metals & Microbial Load:
Fluorescence Analyses:
MATERIALS AND METHODS:
Samples Preparation: Triphala churn contains mainly three ingredients as Harad or Haritaki (Chebulic Myrobalans or Terminalia chebula), Baheda or Bibhitaki (Terminalia Bellirica) and Amla or Amalaki (Indian gooseberry or Emblica officinalis).
Sample No. 1: Patanjali Triphala Churna
FIG. 1: SAMPLE NO. 1 PATANJALI TRIPHALA CHURNA
Sample No. 2: Shree Shree Ayurveda Triphala Churna
FIG. 2: SAMPLE NO. 2 SHREE SHREE AYURVEDA TRIPHALA CHURNA
Sample No. 3: Lab made Triphala churna
FIG. 3: SAMPLE NO. 3 LAB MADE TRIPHALA CHURNA
Method to Prepare the Triphala Churna: Harad or Haritaki (Chebulic Myrobalans or Terminalia chebula), Baheda or Bibhitaki (Terminalia Bellirica) and Amla or Amalaki (Indian gooseberry or Emblica officinalis) collected from the local market. The fine powder was made both by grinding and filtering them.
All the powders were mixed properly in a ratio 1:2:4. The Triphala Churna is prepared and ready to use. For future use, it can keep into a plastic box.
Developments of Standardization Parameters for Triphala Churna:
Study of Organoleptic Characters: The polyhedral formulation is studied for organoleptic characters like color, odor and taste using the sensory organs of our body.
Physicochemical Analysis: 11
A) Ash Value:
Determination of Total Ash: About 2 to 3 g of sample was accurately weighed in a tarred silica dish at a temperature not exceeding 45 °C until it was free from carbon. Then it was cooled and weighed. The percentage of total ash was calculated concerning the air-dried drug.
Determination of Acid Insoluble Ash: The total ash obtained was boiled for 5 minutes with 25 ml of dilute hydrochloric acid; the insoluble matter obtained was collected on an ashless filter paper, washed with hot water and ignited to constant weight. The percentage of acid insoluble ash was calculated concerning the air-dried drug.
Determination of Water-soluble Ash: The ash obtained in the determination of total ash was boiled for 5 min with 25 ml of water. The insoluble matter was collected on an ashless filter paper and washed with hot water. The insoluble ash was transferred into a tarred silica crucible and ignited for 15 min at a temperature not exceeding 45 °C. The weight of the insoluble matter was subtracted from the weight of the total ash. The difference in weight was considered as the water-soluble ash was calculated concerning the air-dried drug.
B) Determination of Loss and Drying: 10 g of the sample (without preliminary drying) was weighed and placed in a tared evaporating dish. It was dried at 105 °C for 5 h, and at 1-h interval until difference two successive weighing corresponded to not more than 0.25%.
C) Determination of Extractive Values:
Determination of Water-Soluble Extractive: 5 g of the test sample was weighed and macerated with 100 ml of chloroform water in a closed flask for twenty-four hours, frequently shaking during six hours and allowing standing for eighteen hours. It was filtered rapidly, taking precautions against the loss of solvent. 25 ml of the filtrate was taken and evaporated to dryness in a tarred flat bottomed shallow dish at 105 °C, to constant weight and weighed the percentage of water-soluble extractive was calculated concerning the air-dried sample.
Determination of Alcohol-Soluble Extractive: Procedure for water-soluble extractive was followed for the determination of alcohol-soluble extractive, but 90% ethanol was used instead of chloroform water.
D) Determination of Crude Fiber Content: Mix about 2g of the powdered drug in no.60 with 50 ml of 10% nitric acid. Bring to boil and maintain at the boiling point for 30 sec. Dilute with water and strain through a fine filter cloth held over the mouth of filter funnel. Transfer the washed residue to the beaker and boil further 30 seconds with 50 ml of a 2.5% solution of sodium hydroxide. Collect and wash residue as before, mount and examine.
Qualitative Phytochemical Screening: 12, 13, 14
A) Detection of Tannins: 2-3 ml of aqueous or alcoholic extract of powders were tested carefully with various tannins test reagents as:
5% FeCl3 Solution: A deep blue-black color indicates the test is positive.
Lead Acetate Solution: A white precipitate indicates the test is positive.
Bromine Water: Deceleration of bromine water indicates the test is positive.
Dilute Iodine Solution: Transient red color indicates the test is positive.
B) Detection of Alkaloids: 50 mg of solvent-free extract was hydrolyzed with dil. HCl and filtered. The filtrates were tested carefully with various alkaloid test reagents as follows
Dragendroff’s Test: To a few ml of filtrates, 1 to 2 ml of Dragendroff’s reagent was added. A prominent yellow precipitate indicates the test is positive.
Wagner’s Test: To a few ml of filtrates, few drops of Wagner’s reagent were added by the side of the test tube. A reddish-brown precipitate confirms the test as positive.
Mayer’s Test: To a few ml of filtrates, few drops of Mayer’s reagent were added by the side of the test tube. A white or creamy precipitate if obtained indicates the presence of alkaloids.
Determination of Physical Characteristics: 15
A) Bulk Density: It is the ratio of the given mass of powder and its bulk volume. It is determined by transferring an accurately weighed amount of powder sample to the graduated cylinder with the aid of a funnel. The initial volume was noted. The ratio of the weight of the volume it occupied was calculated.
Bulk density= w / v0 g/ml
Where, w = mass of the powder, v0 = untapped volume.
B) Tapped Density: It is measured by transferring a known quantity (25g) of powder into a graduated cylinder and tapping it for a specific number of times. The initial volume was noted. The graduated cylinder was tapped continuously for 10-15 min. The density can be determined as the ratio of the mass of the powder to the tapped volume.
Tapped volume = w/vf g/ml
Where, w = mass of the powder vf = tapped volume.
C) Compressibility Index/ Carr’s Index: It is the propensity of the powder to be compressed. Based on the apparent bulk density and tapped density the percentage compressibility of the powder can be determined using the following formula.
Compressibility index/ Carr’s index = [(V0-Vf)/V0] × 100
Or
% Compressibility/ Carr’s Index = [(Tapped density – Bulk density)]/ Tapped density] × 100
D) Hausner’s Ratio: It indicates the flow properties of the powder. The ratio of tapped density to the bulk density of the powder is called Hausner’s ratio.
Hausner’s ratio= Tapped density/bulk density
E) Angle of Repose: The internal angle between the surface of the pile of powder and the horizontal surface is known as the angle of repose. The powder is passed through funnel fixed to a burette at s height of 4 cm. A graph paper is placed below the funnel on the table. The height and radius of the pile were measured. The angle of repose of the powder was calculated using the formula
Angle of repose= tan-1(h/r)
Where, h=height of the pile r = radius of the pile.
Determination of pH Range: The powder sample of Triphala churna was weighed to about 5g and immersed in 100 ml of water in a beaker. The beaker was closed with aluminium foil and left behind for 24-h s in room temperature. Later the supernatant solution was decanted into another beaker, and the pH of the formulation was determined using a calibrated pH meter.
Heavy Metals Test: 16
TABLE 1: FOR CADMIUM
Test | Observation | Inference |
NH4OH added in the sample solution | White ppt. of cadmium hydroxide soluble in excess NH4OH | Presence of cadmium |
Potassium ferrocyanide added | White ppt. of cadmium ferrocyanide | Presence of cadmium |
TABLE 2: FOR BISMUTH
Test | Observation | Inference |
H2S gas added in the sample solution | Dark brown ppt. soluble in hot dil. HNO3 but insoluble in NH4S | Presence of bismuth |
NH4OH | White ppt. insoluble in excess NH4OH dissolved in dil. HCl | Presence of bismuth |
TABLE 3: FOR LEAD
Test | Observation | Inference |
Dil. HCl added in the sample solution | White ppt. of CaCl2 soluble in boiled water & conc. HCl | Presence of lead |
KI is added in the sample solution | Yellow ppt. soluble in boiling water | Presence of lead |
Fluorescence Analysis: 14, 17 A little amount of churna was macerated with a small quantity of solvents like 1N sulphuric acid, 1N nitric acid, 1N hydrochloric acid, iodine, potassium hydroxide, ammonia, 1N sodium hydroxide for an hour and then filtered. The filtrate was then analyzed under daylight and UV light for color and fluorescence.
RESULTS AND DISCUSSION:
TABLE 4: DETERMINATION OF ORGANOLEPTIC CHARACTERS
Characteristics | Sample 1 | Sample 2 | Sample 3 | |
Colour | Light yellow | Yellowish | Yellowish | |
Odor | Characteristics | Characteristics | Characteristics | |
Taste | Very bitter | Astringent | Astringent | |
TABLE 5: ASH VALUES
Type of ash | Sample 1 | Sample 2 | Sample 3 |
Total ash | 6.65 | 7.45 | 6.35 |
Acid insoluble ash | 2.55 | 3.4 | 3.4 |
Water soluble ash | 2.20 | 4.55 | 3.5 |
TABLE 6: MOISTURE CONTENT/ LOSS ON DRYING
Characteristics | Sample 1 | Sample 2 | Sample 3 |
Moisture Content/ Loss On Drying | 0.779 | 1.25 | 1.8 |
TABLE 7: EXTRACTIVE VALUES
Characteristics | Sample 1 | Sample 2 | Sample 3 |
Water | 3.5 | 3.3 | 3.24 |
Alcohol | 1.24 | 2.06 | 1.52 |
TABLE 8: QUANTITATIVE ESTIMATION
Test | Sample 1 | Sample 2 | Sample 3 | |
Test of Tannin | ||||
5% FeCl3 solution | Positive | Positive | Positive | |
Lead acetate solution | Positive | Positive | Positive | |
Bromine water | Positive | Positive | Positive | |
Dilute iodine solution | Positive | Positive | Positive | |
Test for Alkaloids | ||||
Dragendroff’s test | Positive | Positive | Positive | |
Wagner’s test | Positive | Positive | Positive | |
Mayer’ test | Positive | Positive | Positive | |
TABLE 9: BULK DENSITY & TAP DENSITY
Characteristics | Sample 1 | Sample 2 | Sample 3 |
Bulk Density | 0.666 | 0.476 | 0.555 |
Tap Density | 0.909 | 0.625 | 0.80 |
TABLE 10: CARR’S INDEX & HAUSNER’S RATIO
Characteristics | Sample 1 | Sample 2 | Sample 3 |
Carr’s index | 26.73 | 23.84 | 30.625 |
Hausner’s ratio | 1.36 | 1.31 | 1.44 |
TABLE 11: ANGLE OF REPOSE
Characteristics | Sample 1 | Sample 2 | Sample 3 |
Angle of repose | 36.50 | 39.69 | 35.75 |
TABLE 12: DETERMINATION OF PH SAMPLE
Characteristics | Sample 1 | Sample 2 | Sample 3 |
pH | 5 (acidic) | 6 (acidic) | 6 (acidic) |
TABLE 13: ESTIMATION OF CRUDE FIBER
Characteristics | Sample 1 | Sample 2 | Sample 3 |
Crude Fiber | 4.7 | 4.15 | 4.2 |
Heavy Metal Test: Triphala Churna of Patanjali, Shree Shree Ayurveda, and Laboratory made.
TABLE 14: TEST FOR CADMIUM
Test | Observation | Result |
NH4OH added in the sample solution | White ppt. is absent | Absence of cadmium |
Potassium ferrocyanide added | White ppt. is absent | Absence of cadmium |
TABLE 15: TEST FOR BISMUTH
Test | Observation | Result |
H2S gas added in the sample solution | Dark brown ppt. is absent | Absence of bismuth |
NH4OH | White ppt. is absent | Absence of bismuth |
TABLE 16: TEST FOR LEAD
Test | Observation | Result |
Dil HCl added in the sample solution | White ppt. of CaCl2 is absent | Absence of lead. |
KI is added in the sample solution | Yellow ppt. is absent | Absence of lead. |
TABLE 17: FLUORESCENCE ANALYSIS FOR SAMPLE 1
Solvent
added |
Colour observed under | ||
Daylight | Short UV wavelength (256 nm) | Long UV wavelength (365 nm) | |
1N Sulphuric acid | Light brown | Light green | Dark green |
1N Nitric acid | Light brown | Light green | Dark green |
1N Hydrochloric acid | Light brown | Light green | Dark green |
Iodine | Greenish brown | Dark green | Dark blue |
Potassium hydroxide | Brown | Green | Dark blue |
Ammonia | Brown | Green | Dark blue |
1N Sodium hydroxide | Dark brown | Dark green | Dark blue |
TABLE 18: FLUORESCENCE ANALYSIS FOR SAMPLE 2
Solvent added | Colour observed under | ||
Daylight | Short UV wavelength (256 nm) | Long UV wavelength (365 nm) | |
1N Sulphuric acid | Light brown | Light green | Green |
1N Nitric acid | Light brown | Light green | Green |
1N Hydrochloric acid | Light brown | Light green | Green |
Iodine | Greenish brown | Dark green | Dark blue |
Potassium hydroxide | Brown | Green | Dark blue |
Ammonia | Brown | Green | Dark blue |
1N Sodium hydroxide | Dark brown | Dark green | Dark blue |
TABLE 19: FLUORESCENCE ANALYSIS FOR SAMPLE 3
Solvent
added |
Colour observed under | ||
Daylight | Short UV wavelength (256 nm) | Long UV wavelength (365 nm) | |
1N Sulphuric acid | Light brown | Light green | Dark Green |
1N Nitric acid | Light brown | Light green | Dark Green |
1N Hydrochloric acid | Light brown | Light green | Dark Green |
Iodine | Greenish brown | Dark green | Dark bluish |
Potassium hydroxide | Light Brown | Green | Light bluish |
Ammonia | Light Brown | Green | Light bluish |
1N Sodium hydroxide | Brown | Dark green | Dark bluish |
DISCUSSION: From the heavy metal test it is concluded that Triphala Churna of Patanjali, Shree shree Ayurveda and Lab made formulation are free from heavy metals.
CONCLUSION: From the present investigation various standardization parameters such as physicochemical standards like total ash, acid insoluble ash, water & alcohol-soluble extractive values, loss on drying, phytochemical analysis, flow properties, and safety evaluation were carried out, it can be concluded that the formulation of Triphala churna contains all good characters of an ideal churna and it was found to be harmless, more effective, and economic.
The comparison between the two marketed samples and lab-made churna have been done on the basis of the above mentioned parameters which shows satisfactory results, but the efficacy of the products can only be judged by doing the pharmacology of which is suggested as the future scope of R & D.
ACKNOWLEDGEMENT: Authors are very much grateful to NGSPM’s College of Pharmacy for providing necessary facilities for completion of the research work.
CONFLICT OF INTEREST: Nil
REFERENCES:
- Sane RT: Standardization, quality control and GMP for the herbal drug. Indian drugs 2002; 39(3): 184-190.
- Farnsworth NR, Akerele O, Bingle AS, Sojarto DD and Guo Z: Medicinal plant in therapy. Bulletin of the World Health Organization 1985; 63: 965-981.
- http:/www.umm.edu/altmed/articles/herbal medicines-000 351.htm, University of Maryland Medical Center, [complementary medicine] 9-01-09.
- Eisenberg DM, Kessler RC and Foster C: Unconventional Medicine in the United States. N Engl J Med 1993; 328; 246-252. [Medline]
- Tyler VE: Herbs of Choice: The Therapeutic Use of Phytomedicinals. Binghampton. Pharmaceutical Products Press, NY, 1994.
- Marwick C: Growing use of medicinal botanicals forces assessment by drug regulators. JAMA 1995; 273: 607-609.
- [Abstract/Free Full Text] Huxtable RJ: “The harmful potential of herbal and other plant products. Drug Safety” 1990; 5(S-1); 126-136.
- Zhang X: 2004, traditional medicine: its importance and production, In Twarog S., Kapoor P., (Eds), 2002. Protecting and promoting traditional knowledge: system, National Experience and International Dimensions, Part 1. The role of traditional knowledge in Helth Care and Agriculture, United nation. New York document UNCTAD/DITC/TED/10. 3-6.
- Gogtay NJ, Bhatt HA, Dalvi SS and Kshirsagar NA: The use and safety of non-allopathic Indian medicines. Drug Safety 2002; 25(14): 1005- 1019, 498-499.
- Kunle, Folashade O, Egharevba, Omoregie H and Ahmadu: Standardization of herbal medicines - A review. International Journal of Biodiversity and Conservation; 2012; 4(3): 101-112.
- Anonymous: Indian Pharmacopoeia. Government of India, Ministry of Health, Controller of Publication, Delhi, India, 1996.
- Khandelwal KR: Practical Pharmacognosy, Techniques and Experiments. Nirali Prakashan, Edition 20th, 25.6, 23.8-23.10.
- Harborne JB: Phytochemical methods- A Guide to Modern Techniques of Plant Analysis, Edition 3rd, 3-31.
- Trease and Evans: Pharmacognosy, (International edition); Harcourt Brace and company Asia Pvt. Ltd. Singapore, Edition 16th, 131, 228.
- “The United State Pharmacopeia (USP 31): The National Formulary (NF 26)”, Asian edition, by authority of The United States Pharmacopeial Convention, Vol. I, 2008: 188, 189, 231, 639, 640.
- Chatwal GR: Pharmaceutical chemistry. Inorganic, 419-422.
- WHO: Quality control methods for medical plants materials. ATTBS Publisher, Delhi, 2002b: 65-67.
- Aulton ME: Pharmaceutics, the design and manufacture of medicine. Churchill Livingstone Elsevier; Edition 3rd, 175-177.
How to cite this article:
Kadam DK, Ahire PD, Bhoye JV, Patil AR and Yadav DK: Comparative standardization study of three Triphala churna formulation. Int J Pharmacognosy 2016; 3(11): 482-90. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.3(11).482-90.
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Article Information
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482-490
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English
IJP
D. K. Kadam *, P. D. Ahire, J. V. Bhoye, A. R. Patil and D. K. Yadav
NGSPM’s College of Pharmacy, Brahma Valley Educational Campus, Tryambakeshwar Road, Anjaneri, Nashik, Maharashtra, India.
deepalikadam777@gmail.com
28 September 2016
26 October 2016
07 November 2016
10.13040/IJPSR.0975-8232.IJP.3(11).482-90
30 November 2016