Standard Manufacturing Procedure and Quality Control of Karpanpatru Roll-on Prepared with Coconut Oil

Shrestha, Bedarkar, Patgiri, Shukla, and Prajapati: Standard Manufacturing Procedure and Quality Control of Karpanpatru Roll-on Prepared with Coconut Oil

Authors

Introduction

1.

Ayurveda embraces a holistic approach with its gist lying in curing the disease and promoting health of an individual. Ayurveda is unique in its sound foundation of documented theories and operational guidelines. Although Ayurvedic seers have scientifically documented outcomes of pharmaceutico-therapeutic aspects of regular practice and keen observations of medicinal formulations and established theories, still the validation of these theories in terms of modern scientific tools and parameters is needed to assure its credibility and global acceptability. Karpanpatru formulation in powder form has been used effectively since long in the hospitals of Gujarat Ayurved University, Jamnagar. [1] Karpanpatru formulation is derived from a compound name as Karpogi Paste. This is mentioned in Siddha pharmacopeia and has been utilized to treat vitiligo since mid 50’s.[2] Studies were also attempted by modifying into Lepa (medicated paste) and Taila (medicated oil) forms to increase patient compliance and shelf life. [1] The comparative efficacy of the media was assessed by replacing Gomutra (cow urine) with Takra (butter milk) and Kanji (sour gruel).[3] Later studies attempted on converting oil into gel for ease of application, transportation and better acceptability.[4] As the formulation contains Haratala (Arsenic trisulphide); its application using fingertip may pose a risk of dose variation and toxicity. Arsenic toxicity inactivates up to 200 enzymes, most notably those involved in cellular energy pathways and DNA replication and repair, and is substituted for phosphate in high energy compounds such as ATP. Unbound arsenic also exerts its toxicity by generating reactive oxygen intermediates during their redox cycling and metabolic activation processes that cause lipid peroxidation and DNA damage.[5] To minimize this; Karpanpatru formulation was modified into roll-on form that ensures site specific drug application, easy handling and increased stability than liquid dosage forms. Apart from commonly usedTaila (Sarshapa taila) for Sneha Paka of Karpanpatru Taila, in present study, coconut oil was used and converted into roll-on form by the addition of bee’s wax and its in-process standards and quality parameters were established.

Materials and methods

2.

Procurement of raw materials

2.1

Karpanpatru Taila is a herbo-mineral compound used for treating Shvitra (Vitiligo) (Table 1). Bakuchi (Psoralea corylifolia Linn.) and Siktha (bees wax) were procured from Pharmacy, Gujarat Ayurved University, Jamnagar. Tuvaraka (Hydnocarpus laurifolia Dennist.) was obtained from Mangaluru, Karnataka. Krishna Jeeraka (Carum carvi Linn.), Ahiphena (Papaver somniferum Linn.), Karvi (Nigella sativa Linn.), Vatama (Prunus amygdales Linn.), Narikela (Cocos nucifera Linn.) and coconut oil were obtained from local market, Jamnagar. Authentication of raw materials was done at Pharmacognosy Lab of Institute for Post Graduate Teaching and Research in Ayurveda, Jamnagar. Samples of the raw materials were then examined for probable adulterants and for foreign matters, which were removed.[6-7] Shodhita Haratala (processed orpiment) was procured from the Department of Rasa Shastra and Bhaishajya Kalpana of Institute for Post Graduate Teaching and Research in Ayurveda, Jamnagar. Cow urine was procured from Panjarapola Goshala (Cow-shed), Jamnagar. Roll-on glass bottles (10 ml capacity) were purchased from Kerala. The specifications of the vessels are shown in Table 2.

Table 1.

Formulation composition of Karpanpatru Taila for batch size of 1 litre

SNIngredientsBotanical namePart usedRatioQuantity
Kalka Dravya(paste)
1BakuchiPsoralea corylifolia Linn.Dried Seeds1/8th part31.25gm
2TuvarakaHydnocarpus laurifolia Dennist.Dried Seeds1/8th part31.25gm
3Krishna JeerakaCarum carvi Linn.Dried Seeds1/8th part31.25gm
4KarviNigella sativa Linn.Dried Seeds1/8th part31.25gm
5VatamaPrunus amygdales Linn.Dried Seeds1/8th part31.25gm
6AhiphenaPapaver somniferum Linn.Dried Seeds1/8th part31.25gm
7NarikelaCocos nucifera Linn.Endocarp1/8th part31.25gm
8HaratalaYellow arsenic sulphide1/8th part31.25gmOther ingredients
1Coconut oilCocos nucifera Linn.Oil4 parts1 Litre
2GomutraCow’s Urine-16 parts4 Litres
Roll-on preparation
1Karpanpatru Taila--1 part1 litre
2SikthaBees Wax--1/10th of Taila100 gm

Preparation of Karpanpatru Kalka

2.2

The herbal ingredients were powdered and passed through sieve 40# (BSS). Powder of Haratala was mixed with other ingredients, mixed thoroughly to prepare a homogenous blend. This blend was taken in a stainless steel vessel and cow urine (125 ml per 250 gm of Kalka on an average) was added in sufficient quantity to prepare bolus of Kalka.

Preparation of Karpanpatru Taila

2.3

Coconut oil was taken in a steel vessel and heated over mild heat maintaining temperature around 90°C for 15 minutes. The vessel was removed from the heating source and allowed to cool slightly. Kalka was added to coconut oil followed by addition of cow urine. Heating was continued for duration of one and half hours per day maintaining temperature in between 100 ± 6°C. The contents were stirred continuously throughout the process to avoid the possibility of settling down of suspended contents and charring. The contents were left undisturbed through the night and heating was continued on the next day. Classical tests of Taila and Kalka for determination of Sneha Paka were conducted. At regular intervals; Kalka was rolled in between fingers to check the consistency. Heating was stopped after observing characteristic features of Taila Paka. It was filtered while hot through clean cotton cloth into sterile stainless steel container. Total five batches of Karpanpatru Taila were prepared [KN], the details of which are presented in Table 3 and 4.

Preparation of roll-on

2.4

The proportion of Siktha required for Taila was determined with 1/6th, 1/7th, 1/8th and 1/10th part of Taila. KN was taken into a stainless steel container and subjected to mild, controlled heating. When the temperature of Taila reached to 60°C, small pieces of Siktha were added and allowed to melt completely with continuous stirring. The contents were filtered while hot through a clean cotton cloth to separate insoluble particles, if any, possibly present in Siktha. After that, it was stirred continuously till the contents reached to room temperature and the blend became homogenous liquid in consistency. Then it was poured and stored in roll-on bottles. The details of the five batches of Karpanpatru roll-on prepared are presented in Table 5.

Analytical study

2.5

Karpanpatru roll-on was subjected to basic analytical tests. The organoleptic characters of Kalka, coconut oil, Karpanpatru Taila and Karpanpatru roll-on were assessed.

Physico-chemical characteristics

2.6

Physico-chemical parameters of cow’s urine, coconut oil and Karpanpatru Taila were assessed.

Table 2.

Equipment Specifications

EquipmentSpecification
Type of VesselStainless Steel Vessel
Depth16 cm
Diameter20 cm
Circumference94.2 cm
Capacity8 Litre
Size of Cotton Cloth2x2 Feet
Length of Spatula32 cm
Measuring Jar2 Litre Plastic Jar
Weighing Machine20 kg sensitivity ±0.2gm
Heating Device ThermometerLPG Cylinder
Mercury thermometer (0°C to 360°C)
Table 3.

Observations and results of Karpanpatru Taila

ParametersBatchAverage
12345
Weight of powder of Kalka Dravya (g)250250250250250250
Quantity of Cow urine required for preparation of bolus (ml)115120130130125124
Weight of bolus (g)349353368372363361
Initial quantity of Taila (ml)100010001000100010001000
Final quantity of Taila (ml)910900890900880896
Residual Bolus after Sneha Paka (g)574578584585582580.6
Loss of Taila (ml)90100110100120104
Percent loss of Taila91011101210.4
Table 4.

Time taken for preparation of Karpanpatru Taila

SNObservations and temperature of mixture (at stage of observation and duration)Duration required (Batches 1-5)Average
12345
On 1st day
1Duration of heating coconut oil to de- moisturize at 90°C (min)171515161716
2Addition of Bolus at 80°C (min)282427283027.4
3Addition of Cow urine at 80°C (min)322931323431.6
4Boiling started at 100°C (min)444042444843.6
On last day (Duration required to get Sneha Siddhi Lakshana)
5Phenodgama at 104°C(hr)7:557:497:528:008:077:56
6Mridupaka at 104°C (hr)8:208:158:178:248:358:22
7Madhyama Paka at 106°C (hr)9:008:559:009:049:159:02
8Total Time (hr)9:008:559:009:049:159:02
Table 5.

Observations of pharmaceutical preparation of Roll-on from Karpanpatru Taila prepared with Coconut oil

ParametersBatch 1Batch 2Batch 3Batch 4Batch 5Average
Quantity of Coconut oil (ml)910900890900880896
Quantity of Bee’s wax (gm)919089908889.6
Duration of Stirring (min)211920182220
Yield (gm)996986975985962980.8
Percent yield (%)99.599.5999.5999.4999.3899.51

Results and discussion

3.

Standardization is an essential factor for medicinal formulations in order to assess their quality based on the concentration of chemical or bioactive marker.[8] The use of fine powder in Kalka reduces the final yield of the Taila[2] and also increases the chances of passing through the sieve during filtration, increasing turbidity of Siddha Taila leading to reduction of its shelf life. Hence ingredients were powdered and passed through sieve 40# BSS. Mild fragrance of coconut oil was perceived when it was heated to attain moisture free state. After being free from moisture, colour of Taila became darker. During this period, popping sound was perceived along with slight bubbling. The popping noise denotes the water leaving the Sneha, which quickly got subsided. Frothing was noted in initial hours of boiling after addition of cow urine in oil which may be due to partial emulsification and sudden release of ammonia from cow’s urine. The bubbling in Sneha, on the verge of Sneha Siddhi was because of the moisture inside the Kalka. Cow urine is heavier than coconut oil so, it sank to the bottom and then boiled. The steam thus produced due to more quantity of hot oil and very little moisture fragmented into smaller compartments throughout the bolus eventually make bubbles rise suddenly to the surface. The contents were stirred well to allow equal dissipation of heat. Sneha was heated intermittently maintaining temperature in between 100±6°C for approximately 1.5 h/day for four days. On the fifth day, Sneha was heated till the Paka lakshana (completion test for chief desired characteristics) were obtained like Gandha-Varna-Rasotpatti (desired smell, color and taste), Shabdahinata (no cracking sound on fire), Phenodgama (appearance of froth) and Vartivat Kalka (stickiness in bolus to get rolled in to a wick). An average duration of heating per day was decided from pilot batch of the same batch size so as to get Sneha Siddhi Lakshana in five days.[9] The longer duration of interaction of the ingredients and the media was provided with a view to extract more chemical moieties. Repeated boiling causes several oxidative and thermal reactions bringing about a change in the physico-chemical and therapeutic properties in Siddha Sneha. Phenodgama occurred due to presence of unsaturated fatty acids in Taila.

Madhyama Paka of the Siddha Sneha was done so as to extract maximum active chemical constituents. The temperature during the Paka Kala (preparation time) was maintained near about 106°C. By this, reaction between the liquid and fat molecules occurs in a consistent manner over a specific time. This temperature facilitates easy evaporation of liquid molecules with the retention of water soluble extractives, which is slowly imbibed into the Sneha by loosening the bondage in between the fat molecules. The weight of the residual Kalka was comparatively more i.e. an average of 580.6 g than that of initial weight of bolus before heating. Gain in weight may be due to retention of oil by particles of bolus.

Table 6.

Organoleptic characters

CharacterKalkaNTKNKN Roll-On
ColorBrownish YellowWhitish YellowLight Brownish yellowBrownish Yellow
SmellGomutraMildCharacteristicCharacteristic
TouchSlightly RoughCreamyCreamyCreamy
FormBolusLiquidSmooth - oilySemi-solid

NT = Coconut oil; KN = Karpanpatru Taila prepared with Coconut oil; KN Roll-On = Karpanpatru Roll-on prepared with coconut oil

Table 7.

Physico-chemical parameters of Cow’s urine and Coconut oil

ParametersCow’s urineCoconut oil
pH7.46-
Total Solid Content6.51-
Specific Gravity1.0290.931
Refractive Index-1.44
Saponification Value-248
Acid Value-1.76
Table 8.

Physico-chemical parameters of Karpanpatru Taila

ParametersKarpanpatru Taila
Moisture Content0.184 % w/w
Refractive index1.457
Specific gravity0.9233
Viscosity24.652
Cp Acid value8.477
Saponification value255.53
Peroxide value0.796
Iodine Value155.84
Lead (Pb)ND
Cadmium (Cd)ND
Arsenic (As)33.44ppm
Mercury (Hg)ND
Total microbial plate count02 cfu/mL
Total yeast & mouldAbsent
Staphylococcus aureusAbsent
Escherichia coliAbsent
Salmonella Spp.Absent
Pseudomonas aeruginosaAbsent

ppm - parts per million; ND - Not Detected; cfu/mL - Colony forming unit per mililitre; cP - Centipoise

The roll-on was prepared with the addition of Siktha in 1/10th parts of Karpanpatru Taila as it was found to be the most appropriate ratio to get the desired consistency from the results of the pilot batch made by incorporating different proportions of Siktha. The homogenous liquid blend was poured into the roll-on bottles and the cap was properly fitted. The organoleptic characters were assessed as shown in Table 6.

The physico-chemical parameters of cow’s urine, coconut oil and Karpanpatru Taila were assessed as shown in Table 7 and 8. An acid value of coconut oil has been significantly increased after Sneha Paka suggesting chemical breakdown of glycerides of Sneha during process of Sneha Paka. The viscosity of prepared oil was found 36800 cP which may also influence rate of absorption of topical applicant. Free fatty acids are nascent and hence susceptible for formation of newer compounds in an attempt to get stabilized which has been seen in the form of changes in saponification value after Sneha Paka. Saponification value of Karpanpatru Taila was 255.53. The long chain fatty acids found in fats have a low saponification value because they have a relatively fewer number of carboxylic functional groups per unit mass of the fat as compared to short chain fatty acids.[10] High saponification values of fats and oils are due to predominantly high proportion of shorter carbon chain lengths of the fatty acids and more number of low molecular weight fatty acids.[11-12] Shorter chain fatty acids (high saponification value) have faster rate of absorption than longer chain fatty acids. Comparatively higher saponification value of KN points out that the absorption of KN may be quicker than coconut oil itself. Increase in saponification value of KN than coconut oil may be due to different reactions during Sneha Paka including oxidation, hydrolysis, polymerization due to repetitive heating of fats[13] saturation due to heating with aqueous media etc.

All these physico-chemical parameters such as viscosity, specific gravity, peroxide value, iodine value and saponification values are qualitative properties of oils and do not indicate the position of the double bonds or the amount of olefinic carbon but rather it provides an overall status of unsaturation of the oils so it is not possible to point out the position of double bond(s) which are more susceptible to oxidation.[14]

Table 9.

Physico-chemical parameters of Karpanpatru Roll on

ParametersResult
Moisture content (%w/w)0.148
Viscosity (cP)36800 cP
Spreadability (mm/10g)22

cP - Centipoise

Table 10.

HPTLC profile of samples of Roll-on at 254nm, 366nm and 540nm

Spot No. (10uL)Short UVLong UVWhite light
10.110.140.05
20.290.240.23
30.520.350.33
40.600.430.54
50.670.520.79
60.920.750.91

Solvent System- Ethyl Acetate: Methanol: Ammonia (8:2:0.2)

The spreadability of the roll-on form was found to be 22mm/10g as shown in Table 9. This indicates that the formulation has a good flow on skin. Ultimate acceptability and clinical efficacy of topical preparations require them to possess optimal mechanical properties (ease of removal from the container, spreadability on the substrate), rheological properties (viscosity, elasticity, thixotropy, flowability) and other desired properties such as bio-adhesion, desired drug release, and absorption.[15] KN was found to be free from fungal and bacterial growth suggesting that KN is free from microbial contamination and contain moisture in negligible limits. Arsenic content was found to be 33.44 ppm. HPTLC study showed same number of spots in the visualizing conditions as shown in Table 10 and Figure 1.

Karpanpatru roll-on prepared with coconut oil solidifies in winter. Coconut oil melts at 24°C (75oF)[16] and is solidified below that temperature. The roll-on had Siktha added to it, which further helped it to solidify in winter. Hence the flow of the roll-on prepared with coconut oil was not identical in all seasons. The rollon needed to be placed in hot water bath to attain a good flow during winter season. Appropriate excipient may be added to maintain the identical consistency of roll-on.

Conclusion

4.

Karpanpatru Taila prepared with coconut oil was light brownish yellow with a characteristic smell and had an average loss of 10.4% on an average and required 9:02 h for preparation. It took 20 min for the preparation of roll-on from Karpanpatru Taila and the yield was 99.51% with addition of 1/10th part of Siktha. The evaluated parameters i.e. moisture content, viscosity and spreadability were 0.148% w/w, 36800cP and 22 respectively of roll-on (Table 9) and 6 spots of HPTLC of KN Roll-on were at 254 nm, 366 nm and 540 nm.

1.

HPTLC of Karpanpatru Roll-on with coconut oil

https://s3-us-west-2.amazonaws.com/jourdata/jams/JAyuMedSci-2-225-g001.jpg

The standard operating procedures for preparation along with quality control parameters of Karpanpatru roll-on with coconut oil observed in the current study may be considered as standards for future studies.

ACKNOWLEDGEMENT

The authors would like to thank Pharmacognosy Laboratory, IPGT&RA, Jamnagar; Dr Galib, Associate professor, AIIA, New Delhi; IIT Powai and VASU research Centre, Vadodara for support.

Notes

[1] Supported by SOURCE OF SUPPORT

IPGT&RA, Gujarat Ayurved University, Jamnagar, India.

Notes

[2] Conflicts of interest CONFLICT OF INTEREST

None declared

CONTRIBUTORS

Dr Shrestha contributed to design, literature study and data acquisition. Dr Bedarkar contributed to the conceptualization of the topic, data analysis and manuscript editing. Prof. Patgiri contributed to the manuscript review and analysis. Dr Shukla contributed to the pharmaceutical experiments and data analysis. Prof Prajapati contributed to the intellectual content, design and literature study.

REFERENCES

1. 

Tank ZG , author. A Pharmaceutico-clinical study of VIT 8 lepa & VIT 8 oil (modified Karpanpatru- A herbomineral compound) in the management of Shvitra (Vitiligo). PhD Thesis, Department of Rasa Shastra & Bhaishajya Kalpana, IPGT & RA, Gujarat Ayurved University, Jamnagar. 2006

2. 

Goyal M, Patgiri BJ, Ravishankar B, Prajapati PK , authors. Role of different media in Karpanpatru Taila preparation. AYU. 2010;31(1):15–8

3. 

Goyal M , author. A comparative pharmaceutico clinical study of Karpanpatru taila prepared with different medias and its effect on Switra (vitiligo). MD Dissertation, Department of Rasa Shastra & Bhaishajya Kalpana, Institute of Post Graduate Teaching & Research in Ayurveda, Gujarat Ayurved University, Jamnagar. 2008

4. 

Jadhav HR , author. Pharmaceutico-therapeutic evaluation of gel prepared from Karpanpatru Taila and Shvitrahara Yoga on Shvitra (vitiligo). Ph.D. Thesis, Department of Rasa Shastra & Bhaisajya Kalpana, IPGT & RA, Gujarat Ayurved University, Jamnagar. 2017

5. 

Ratnaike RK , author. Acute and chronic arsenic toxicity. Postgrad Med J. 2003;79:391–6. doi: 10.1136/pmj.79.933.391.

6. 

European Agency for the Evaluation of Medicinal Products, European Community. EMEA/HMPWG/ 25/99. 1999. p. 56–60

7. 

Quality Control Methods for Medicinal Plant Materials. World Health Organization (WHO); Geneva: 1998. p. 8–9. 22-5,61-3.

8. 

Shailajan S, Singh A, Tiwari B , authors. Quality control and standardization of an Ayurvedic Taila formulation. International Journal of Biomedical Research and Analysis. 2010;1(2):78–81

9. 

Reddy RC, Suresh P , authors. Translators. Vaidyaka Paribhasa Pradipa of Govindasena, 1st Ed. Tritiya Khanda: Chapter 3, Verse 26. Varanasi: Chaukhambha Sanskrit Bhawan; 2003. p. 45

10. 

Wikipedia.org [homepage on the Internet]. Saponification value. Available on https://en.wikipedia.org/wiki/Saponification_value. cited on 31.05.2017.

11. 

Kirk R, Sawyer R , authors. Pearson’s composition and analysis of foods. 9th edition. England: Addison Wesleylongman ltd; 1991. p. 608–40

12. 

Manual of methods of analysis of foods- oils and fat. New Delhi: Food safety and standards authority of India, Ministry of health and family welfare government of India. 2016

13. 

Kamsiah J, Kamisah Y , authors. Repeatedly Heated Vegetable Oils and Lipid Peroxidation. Lipid Perioxidation. Catala Angel , editor. Role: EditorCroatia: InTech Prepress; 2012. p. 211–28

14. 

Knothe G, Dunn R , authors. Dependence of oil stability index of fatty compounds on their structure and concentration and presence of metals. J Am Oil Chem Soc. 2003;80:1021–6. Accessed on 20/5/2017 at 10:50 PM. Available from: http://biodiesel.org/reports/20030101_gen-343.pdf.

15. 

Jones DS, Woolfson AD, Brown AF , authors. Textural, viscoelastic and mucoadhesive properties of pharmaceutical gels composed of cellulose polymers. Int J Pharm. 1997;151(2):223–33. Accessed on 3/5/2017 at 8:45 PM. Available from: https://doi.org/10.1016/S0378-5173(97)04904-1.

16. 

Wikipedia.org [homepage on the Internet]. Last edited on 23 June 2017 at 23:06. Available from: https://en.wikipedia.org/wiki/Coconut_oil.