RVS College of Pharmaceutical Sciences, Sulur, Coimbatore, India 641402
India's gomutra tradition is significant. It has several significant medicinal uses in addition to being holy. Eight forms of urine, or Ashta mutra, are discussed along with their characteristics, applications, and formulations in the classic Ayurvedic treatises Charaka, Sushruta, and Vagbhata Samhita. One such example is cow pee. Urine, milk, ghee, curd, and dung are the five ingredients of Panchagavya that are derived from cows. The term Panchagavya Chikitsa (Cowpathy) refers to cow-based therapy.Numerous studies have been conducted that demonstrate the plant's potential for treating a wide range of conditions, including skin disorders, stomach disorders, kidney diseases, heart problems, diabetes, liver problems, jaundice, athlete's foot, cysts, hemorrhoids, and more. These studies also highlight the plant's immune-stimulant, bioenhcer, anticonvulsant, anticancer, wound-healing, antioxidant, and antimicrobial qualities. The physicochemical properties and cytocompatibility of lyophilized cow urine powder from Sahiwal and the pure Indian breed are presented in the current paper. Various analytical techniques have been employed to investigate the physical and chemical properties of cow urine powder obtained through lyophilization. These techniques include Thermogravimetric Analysis (TGA), Fourier Transform Infrared (FTIR) Spectroscopy, Differential Scanning Calorimetry (DSC), Nuclear Magnetic Resonance (NMR) Spectroscopy, and Scanning Electron Microscopy – Energy Dispersive X-ray Spectroscopy (SEM-EDX). A SEM-EDX study of the two samples revealed that 25–30% of the metal ions had a cluster-like shape. The degradation processes revealed by TGA analysis were most likely caused by the first desorption of adsorbed water, urea degradation, and loss of the flavonoid's glycoside moiety. Data from the DSC showed the existence of both irregular (amorphous) andThe lyophilized powder of cow urine contains crystalline molecular structures.
Cow urine serves as a comprehensive solution for various health issues, and cows function as dynamic sources of medicinal properties. According to ancient sacred writings like the Charaka Samhita, Sushruta Samhita, Vridhabhagabhatt, Atharva Veda, Bhavaprakash, Rajni Ghuntu, and Amritasagar, cow urine, a component of "Panchagawya," has the potential to address numerous curable and incurable ailments (1). This has been utilised widely in the preparations of traditional Indian medicine for centuries. This form of therapy is referred to as cow pathy or Panchgavya therapy. In a manner akin to homoeopathy, allopathy, or naturopathy, cowpathy represents a distinct medical system. The cow serves as a cornerstone of Indian culture and the rural economy, fulfilling our needs while representing both cattle wealth and biodiversity. 2 (2). Recognised for its nurturing attributes similar to those of a mother, its significance as the origin of all that contributes to human prosperity, and its ability to preserve medicinal substances, it is referred to as "Kamdhenu" and "Gaumata." The traditional Indian medical system recognised as Ayurveda provides comprehensive insights into the application of cow's milk, curd, ghee, and urine for addressing various health conditions in humans. The Charaka and Sushruta Samhita highlight numerous advantages of cow urine (gomutra), such as its potential to reverse anaemia, address specific cardiac and renal issues, alleviate diarrhoea, reduce oedema, manage haemorrhoids, improve indigestion, combat jaundice, relieve stomach aches, and treat skin conditions like vitiligo. It contributes to weight loss as well. (3) All bodily imbalances can be addressed with gomutra, supporting overall well-being. It has been utilised for oral preparations in treatments and beverages, as well as externally in the form of lotions, ointments, and baths. Cow urine serves multiple functions, such as household cleaning, hygiene, agriculture, and sericulture (including its use as fertiliser, insecticides, and pesticides), as well as acting as a therapeutic agent for various ailments and diseases. Cow urine has potential therapeutic applications. Gomutra has been identified to include a variety of components such as nitrogen, sulphur, phosphate, sodium, manganese, iron, silicon, chlorine, magnesium, citric, succinic, and calcium salts. Additionally, it encompasses vitamins A, B, C, D, and E, along with minerals, lactose, enzymes, creatinine, and hormones. In (4) Thermogravimetric analysis (TGA), Fourier transform infrared (FTIR) spectroscopy, Differential scanning calorimetry (DSC), nuclear magnetic resonance (NMR) spectroscopy, and scanning electron microscopy – energy dispersive X-ray spectroscopy (SEM-EDX) are some of the analytical techniques employed to examine the physical and chemical properties of lyophilised cow urine powder. (5). Gomutra Ark currently holds three US patents that have undergone thorough examination and received approval from the US government. U.S. Patent No. 1 = 6410059, issued on June 25, 2002. U.S. Patent No. 2 = 6896907, issued on May 24, 2005. Dated June 26, 2007, U.S. Patent No. 3,723,5262. No. (6) It possesses several notable qualities, including bioenhancing capabilities. Milk urine does not demonstrate the same efficacy as cow urine when it comes to enhancing biological activity; rather, cow urine distillate significantly improves the effectiveness of antibacterial, antifungal, and anticancer treatments. In addition to zinc and bovine serum albumin (GnRH–BSA), it further amplifies the effects of the gonadotropin-releasing hormone conjugate. The combination of medications with bioenhancers—chemical entities that do not exhibit synergistic effects with the drug—results in improved and promoted bioavailability of the combined drugs. The origins of herbal bioenhancers can be traced back to the ancient practices of Ayurvedic medicine, which has a rich history spanning thousands of years. The concept known as "yogvahi" in Ayurveda serves to reduce dosage and side effects, enhance oral bioavailability, optimise tissue distribution—especially for medications with low oral bioavailability—and eliminate the need for parenteral drug administration. The Sanskrit term "Trikatu," translating to "three acrids," refers to an ayurvedic medicine with origins tracing back to the 7th and 6th centuries A.D.
Optimal Supplementary Energy
The most crucial characteristic of the perfect bioenhancers is that they should improve the drug molecules' uptake/absorption and activity. Other unique qualities include: being harmless to humans or animals; being effective at very low concentrations when combined; and being simple to mix. (7)
Bioenhancers Are Necessary
Aqueous solubility, drug permeability, dissolving rate, first pass metabolism, pre-systemic metabolism, and sensitivity to efflux mechanisms are some of the variables that affect oral bioavailability. Drug dosage, toxicity, and cost can all be decreased by using bioenhancers, which alter one or more of these variables to improve absorption and slow down metabolism. Drugs that are poisonous, costly, poorly obtainable, and administered over extended periods of time necessitate the use of bioenhancers (7).
Drug interactions with bioenhancers
Drug resistance is kept to a minimum and the dosage of the medication is lowered. In particular, anti-tubercular medications will have lower costs and less dose-dependent toxicity. Research has demonstrated that the use of bioenhancers at modest dosages has improved the bioavailability and bioefficacy of various therapeutic classes, including anti tubercular, antibiotic, antiviral, antifungal, and anticancer medications. (7)
GOMUTRA'S ATTRIBUTES[8]
Synonyms:
Rasapanchak [8]
HOW COW URINE WORKS [6]
Due to the presence of many volatile and non-volatile components such as urea, creatinine, arum hydroxide, phenol, carboxylic acid, and calcium and manganese salts, cow urine has strong antibacterial, germicidal, and antifungal properties. Numerous amino acids and urine peptides are present, which contribute to the bacterial cell wall and surface hydrophobicity, enhancing this germicidal activity even further. Furthermore, because fresh cow urine has a higher phenol concentration than its distillate, it has been shown to be more effective as an antibacterial agent. Cow pee that has been void in the morning has the highest concentration of antioxidants, uric acid, and allantois, which can be used to treat cancer. This pee that has been voiding in the morning has been found to be more effective since it contains more micro and macronutrients, enzymes, and other components, making it more sterile. It has been discovered that cow urine increases lymphocyte survival by reducing apoptosis. This is explained by the components' ability to scavenge free radicals, which in the end helps to slow down the aging process. The best urine for medicinal purposes is that from purebred Indian cows. It possesses immune-modulatory properties that protect the host body from a range of illnesses. Mice's immune systems have been found to be modulated by distillate of cow urine. It has been discovered that it raises IgG levels and promotes T- and B-cell development. The distillate of cow urine exhibits synergistic effects when used with antifungal, anticancer, and antibiotic medications. CUD therefore serves as a bio-enhancer. CUD has been shown to enhance T and B cell blast genesis. The research of CUD's impact on humoral and cell-mediated immune responses to NDV vaccination was made possible by this work. Analogous studies have been conducted using cow urine to increase the production of lymphoblast. In order to assess their capacity to foster a protective immune response against disease, the study was designed to look at the blast genic activity of lymphocytes and how in-vivo cow urine therapy affected it. With no possibility of contamination or the release of hazardous chemicals into the environment, this therapy not only strengthens the immune systems of living things but is also environmentally beneficial.
HOW BIOENHANCER ACTS MECHANISTICALLY[7]
The primary mechanisms by which various bio enhancers influence drug molecules to enhance their bioavailability are outlined here. Enhancing bioavailability can be accomplished through various strategies, such as optimising drug absorption via the gastrointestinal tract, modifying the pharmacokinetic and pharmacodynamic characteristics of various medications, employing immunomodulation to improve interactions among drugs, pathogens, and the host, facilitating better drug penetration into pathogens that persist within macrophages (e.g., Mycobacterium tuberculosis) to promote effective eradication of these organisms, and augmenting drug penetration into cerebrospinal fluid. Enhancing blood circulation facilitates the body's capacity to effectively absorb orally administered medications. They modify various active transporters, such as P-glycoprotein (Pgp), an efflux pump that expels medications and prevents them from arriving at their designated target. In these conditions, bio enhancers operate by inhibiting the P-glycoprotein. By inhibiting the activity of drug metabolising enzymes like CYP 3A4, CYP1A1, CYP1B2, and CYP2E1 in the liver, gut, lungs, and various other locations, the elimination process is also shortened, resulting in an extended duration of the medication's presence in the body. This will also aid in mitigating the first pass effect associated with medication administration. Through the inhibition of glomerular filtration and the reduction of P-glycoprotein activity, bio enhancers effectively diminish renal clearance, simultaneously facilitating passive tubular reabsorption and active tubular secretion. The inhibition of the enzyme uridine diphosphate (UDP) glucuronyl transferase, responsible for conjugating and inactivating the medication, may also influence biliary clearance. In addition to the previously discussed mechanisms, several other theories concerning herbal bio enhancers can be identified: reduced secretion of hydrochloric acid and enhanced blood flow to the gastrointestinal tract; inhibition of intestinal motility, gastric emptying time, and gastrointestinal transit; alterations in the permeability of the gastrointestinal epithelial cell membrane; cholagogous effects; properties related to bioenergetics and thermogenesis; suppression of first-pass metabolism; and the stimulation of gamma-glutamyl transpeptidase (GGT) activity, which facilitates improved absorption of amino acids. Recent advancements in the pharmaceutical industry include salt preparation, prodrug creation, chemical change, micronization, prolonged drug administration, and liposomal microencapsulation, all aimed at enhancing bioavailability.
EQUIPMENT AND TECHNIQUES[9]
Collecting and preparing the samples The selected healthy Badri cow had its midstream urine sample obtained using aseptic technique and sent to the lab in a sterile, transparent container with a screw lid. Whatman No. 1 filter paper was used to filter the cow urine after the precipitates and debris were removed.
Analysing cow Urine Chemically Qualitatively
Molisch’s test: In a test tube, 2 ml of cow urine was measured, followed by the addition of two drops of α-napthol, which were then thoroughly mixed. Concentrated sulphuric acid (H2SO4) was carefully introduced along the edge of the test tube, and observations were made for the emergence of a purple colour ring.
Test for reducing sugar
Benedict test: A clean test tube was prepared with 5 ml of Benedict reagent. Eight drops of cow urine were introduced and thoroughly mixed. The solution was subsequently heated to a boil for a duration of 2 minutes, during which a careful observation for any colour change was conducted.
Biuret test: 1 ml of bovine urine was collected in a test tube. Three drops of biuret reagent were added, thoroughly mixed, and then observed for any colour change.
Ninhydrin test: 1 ml of bovine urine was collected in a test tube. Two drops of a 2% ninhydrin solution were added and thoroughly mixed. The test tube was placed in a water bath for a duration of 5 minutes, after which it was examined for the emergence of a purple colour.
Test for Bile salt
Hay’s Sulphur test: 5 ml of cow urine were placed in a test tube, followed by the addition of sulphur powder to the cow urine.
Test for Urea
Sodium Hypobromite test: In a test tube, two millilitres of cow urine was collected. After that, three drops of sodium hypobromite were added.
ENZYMES PRESENT AND ITS UNITS [4]
Benefits of Gomutra for Healing [8]
ANTIMICROBIAL AND ANTHELMINTIC ACTIVITY. [8]
Pathogenic bacteria, opportunistic fungi, and parasitic helminthes can all cause ailments that can be treated with the Gomutra. Cow urine concentrate kills gram positive bacteria more effectively than gram negative bacteria, according to several analyses. Strong anthelmintic activity found in cow urine causes dose-dependent paralysis and death of worms. Cow urine was found to boost antibiotic action against a range of illnesses because of its bio-enhancing property. Khadiradi Kashaya and Gomutra have shown remarkable effectiveness in treating Purishaj Krimi in this study, with appreciable improvements in the condition's main symptoms.
Analgesic activity:
In a study of cow urine and its distillate to test analgesic efficacy using the rat tail immersion method, the rats administered cow pee demonstrated good analgesic effects that were comparable to those of the standard treatment group, which received Diclofenac Sodium. Cow urine has an analgesic effect because it contains certain volatile fatty acids and steroidal moieties.
Hemorrhoid:
Cow urine extract has been clinically evaluated in individuals with hemorrhoids. According to the findings, patients with hemorrhoids have not experienced the uncomfortable, expensive, or time-consuming side effects of the ailment since they were given cow urine supplements orally.
Wound healing activity:
A study was conducted utilizing an excision wound model on wistar albino rats to examine the healing potential of cow urine. The criterion that was being studied was the rate of wound contraction. A decrease in the wound zone was found in studies on the healing of excision wounds. The wound healing of the male and female rats treated with external urine treatment on Day 4 was significantly better than that of the other groups. By the conclusion of the fourteenth day, however, the animals continued to show that only 0% and 0.40.4 % of healing remained. This could be the case even if animals treated with the standard medication, nitrofurazone, showed healing rates of 0% and 0.50.3 %. The animals nevertheless had normal immunity. The study's findings demonstrated that external application of cow urine to a lesion speed up the healing process.
Immunomodulatory Activity:
Cow dung can be used to increase the immune systems of plants and animals. It can also be used to strengthen the body's general defenses against social ailments by applying minerals and herbs (such chyavanprash and panchgavya). Rulings indicate that the immune system was shown to benefit from the presence of methanol, ethyl acetate, acetone, and watery components in dried cow urine. Ancient Ayurvedic teachings state that humoral and cell-mediated immunity with consistent use of CU. The origin of the distillate of cow urine used to increase IgG levels, T and B cell proliferation, and blastogenesis in mice and chicks (avian species)
Hepatoprotective Activity:
For the treatment of liver cirrhosis, 1-2 ounces of warm cow pee are taken daily. Urine taken either on its own or combined with salt or honey is also recommended for liver and other issues. Internal administration of antimony sulfide and cow urine is used to treat malignant jaundice.
Antidiabetic activity:
Rats with experimental alloxan-induced diabetes were examined to see if cow urine formulation (Gomutra ark, GoA) affected the condition. We utilized 200–250 g Wistar albino rats of both sexes. Blood sugar, vitamin C, and the release of malondialdehyde (MDA) were the biochemical characteristics that were noted. Although the observed effect was found to be less than that of the standard antidiabetic glibenclamide, GoA symptomatically lowers blood glucose in diabetic rats. The distillate of cow urine also reduces serum glucose levels parallel to the levels of serum triglycerides and cholesterol. Volatile fatty acids that act as antioxidants and contribute to the antihyperglycemic effect include quinoline, butanoic acid-3methyl, propyl ester, acetic acid 2 propenyl ester, acetic acid methyl ester, 2,2,3-trichloro propionic acid, and 1H indol-3-acetate.
Stomach, kidney and heart diseases:
The best remedies for renal problems, heart problems, stomach disorders, and tuberculosis are cow dung and pee. Cow pee is used as an ingredient in a variety of dressings and mixes (often combined with sour wine) or as a stand-alone medicine for a variety of ailments, including obesity, intestinal dropsy, Vayu problems, ulcer insect removal, and many more. When it comes to tuberculosis and malaria, cow dung has antibacterial properties.
Stones:
It works for stones in the kidneys, gallbladder, etc. For 21 days, the first item that should be consumed in the morning is a glass of freshly drawn cow pee. These stones dissolve into a manageable size thanks to the uric acid in cow urine.
Skin diseases:
In terms of all skin conditions, such as eczema, psoriasis, acne, sunburns, and itching, it is quite helpful. The hair is also left clean and silky by it. After secretion, cow pee has an antibacterial effect and is not totally sterile.
Anticonvulsant action
When it comes to treating convulsive illnesses in children, gomutra mixture is the traditional herbal remedy of choice for Yoruba speakers in Nigeria, regardless of the underlying cause.
Good for liver
Gomutra corrects the liver's function. Therefore, liver produces pure, healthy blood. It provides the body with the ability to resist sickness. Liver cirrhosis is treated with 1-2 ounces of warm cow pee given daily.
Good antiseptic
Research has shown that cow urine has antiseptic qualities that aid in wound healing, and that these qualities cause wounds to heal more quickly than wounds treated with antiseptic cream.
Protection against parasites
Cow urine has the ability to shield the body from inside parasites. Numerous health issues, including diarrhea, stomach pain, and nutritional loss in the body, can be brought on by intestinal parasites. A study demonstrates that utilizing a large amount of cow urine is an effective way to combat these disease-causing parasites.
Protects heart and brain
Nervous system damage is caused by mental strain. It is known as medhya and hradya, which implies that it strengthens the heart and brain. Hence, gomutra guards the brain and heart against illnesses and disorders as well as against the harm that mental stress may do to these vital organs. The heart and mind become limited in their thinking when they act immorally or sinfully, which goes against the voice of the soul. This affects how the body functions and leads to diseases. A happy mood is produced by cow urine. Consequently, it facilitates our ability to carry out the right actions mentally. therefore, guards against illnesses
Anticancer properties
Gomutra possesses potent antioxidant qualities that aid in scavenging free radicals and mitigating oxidative stress. These characteristics suggest that cow pee may have anticancer effects. This is not to imply that it can treat cancer; rather, it can lower the chance of developing the disease. Gomutra treats kidney problems. For stones, it can be applied. For 21 days, the first thing you should do when you wake up is drink a glass of fresh cow urine. The uric acid included in cow pee dissolves the stones into smaller, more manageable pieces.
Antidiabetic effect
A study was carried out to validate and determine the antidiabetic effects of gomtra and its concentrate, which remains after the distillation process. Rats with alloxan-induced diabetes were utilised to evaluate the antidiabetic effects of cow urine and glibenclamide treatment as per standard procedures. The GOD-POD method (glucose oxidase–peroxidase) was employed to assess their blood glucose levels. The results were analysed in relation to those achieved in rats undergoing standard glibenclamide treatment. In comparison to the diabetic control group, the distillate of cow urine demonstrated a significant reduction in elevated levels of blood glucose, serum cholesterol, and serum triglycerides. It additionally facilitates the management of diabetes in individuals through the use of commercially available formulations, such as cow urine.
CHEMICAL COMPOSITION OF COW URINE [10].
Table 1.1 lists the components and their respective percentages found in cow pee. According to numerous studies, the contents of cow urine include nitrogen, sulfur, phosphate, sodium, manganese, carbolic acid, iron, silicon, chlorine, magnesium, citric, titric, succinic, and calcium salts; vitamins A, B, C, D, and E; minerals; lactose; enzymes; creatinine; hormones; urea; and gold acids. The main ingredient in urine is urea, which explains why cow poop has antibacterial and fungicidal properties. Interleukin-1 and interleukin-2 release is increased by cow urine. Because it contains more macro and micronutrients as well as other enzymes, early morning first-void cow urine is more sanitary and effective. Important substances including pheromones, urine proteins, calcium, and estrogen are found in cow pee.
TABLE 1.1
|
Name of the ingredients |
Percentage |
|
Water |
95 % |
|
Urea |
2.5 % |
|
Minerals, Salts, Hormones and enzymes |
2.5 % |
|
Ammonium nitrogen |
1-1.7 ml/kg/day |
|
Calcium |
0.1-1.4 ml/kg/day |
|
Chloride |
0.1-1.1 mmol/kg/day |
|
Creatinine |
15-20 mg/kg/day |
|
Potassium |
0.08-0.15 mmol/kg/day |
|
Uric acid |
1-4 mg/kg/day |
|
Allantoin |
20- 6 0ml/kg/day |
COSMETIC PRODUCTS MADE OF COW URINE[10]
Cow urine cures all skin illnesses and purifies the blood. Toxic substances secreted by skin microorganisms cause microbial skin illnesses. Cow urine is used to skin care products to increase their efficacy in protecting the skin. It is one of the main elements in a number of cosmetic items.the inventory of makeup items containing cow urine. Additionally added to cosmetics is cow urine.
ANALYTICAL TECHNIQUES[5]
Scanning Electron Microscopy (SEM) Analysis with Energy Dispersive X-Ray Spectroscopy (EDX) Measurement of Freeze-Dried Cow Urine Powder
All prepared samples were analysed for their external appearance and particle size using Scanning Electron Microscopy (SEM). The lyophilised samples underwent a coating process with gold at 20 kV in preparation for SEM examination using the Zeiss EVO 18 SEM from Germany. Furthermore, Energy-Dispersive X-ray Spectroscopy (EDX) mapping was conducted to examine the elemental composition of freeze-dried cow urine powder.
Thermogravimetric Analysis (TGA) Analysis of Freeze-Dried Cow Urine Powder
A TGA50 thermogravimetric analytical instrument from Shimadzu (Asia Pacific) Pte Ltd. was utilised to conduct the Thermogravimetric Analysis (TGA). The temperature range, spanning from 20°C to 500°C, was optimised for experimentation with a heating rate of 10°C/min. TGA facilitates the examination of materials regarding their thermal stability by evaluating weight changes in relation to temperature, time, and atmospheric conditions.
Differential Scanning Calorimetry (DSC) Analysis of Freeze-Dried Cow Urine Samples
The thermal analysis and behaviour of cow urine samples were evaluated through differential scanning calorimetry (DSC) using the DSC-60 Plus instrument from Shimadzu [Asia Pacific] Pte Ltd. Measurements were conducted in a nitrogen atmosphere with a flow rate of 100 mL min-1 and a heating rate of 10°C min-1, starting from room temperature and extending to 300°C.
Nuclear Magnetic Resonance (NMR) Spectroscopy of Freeze-Dried Cow Urine Samples
The chemical structures of the primary components of lyophilised cow urine powder were elucidated through 1 H and 13C NMR spectroscopic analyses. The 1 H and 13C NMR spectra were recorded using a JEOL AL300 FTNMR (500 MHz) spectrometer at room temperature, utilising either DMSO-d6 or CDCl3 as the solvent.
Fourier-Transform Infrared (FT-IR) Spectroscopic Analysis of Freeze-Dried Cow Urine Powder
Potassium bromide (KBr) pellets within the 400–4000 cm-1 range were utilised to examine FT-IR spectra with a Thermo Scientific Nicolet iS5 spectrometer. The influence of the lyophilization process on cow urine samples was investigated through the analysis of FTIR spectra.
Cytocompatibility Study In this study,
The objective of our study was to assess the cytocompatibility of lyophilised cow urine powders utilising mouse embryonic fibroblast NIH-3T3 cells. To culture the cells in a CO2 incubator (Galaxy® 170 S, Eppendorf, Germany), a complete culture medium was prepared by reconstituting Dulbecco's Modified Eagle's Medium (DMEM) with 10% Foetal Bovine Serum (FBS) and 1% Penicillin (10,000 U/ml)/Streptomycin (10 mg/ml) (PS).The incubator was kept at 37°C with 5% CO2 and fed at 95% relative humidity. A 96-well plate was used to cultivate cells in order to examine the effects of powdered cow urine. To treat the cells, lyophilized cow urine powders were combined in a full culture media at different concentrations (e.g., 0.5 mg/ml and 1.0 mg/mL). Following the application of cow urine powders for a full day, cells were observed under a bright-field microscope (Nikon Ti-U Eclipse, Japan) to evaluate the qualitative evaluation of cellular morphology.
Assessment of Cellular Viability Using MTT Assay
The cellular viability was assessed using the MTT (3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyltetrazolium bromide) reduction test. For this, fibroblast cells from NIH-3T3 were utilized. Using cells cultivated in the 96-well plate, the standard assay protocol was optimized to perform the MTT experiment. The cells were cultured in a CO2 incubator for 24 hours at a density of 104 per scaffold. Every experiment had a positive control, which consisted of cells cultivated without any treatment, and a negative control, which consisted of treating the growth media alone. A solution containing MTT dye at a concentration of 5 mg/mL in PBS was made using 90 μL of complete growth medium and 10 μL of MTT solution. The treated samples were then incubated with the solution for four hours after the cells were incubated with freeze-dried cow pee powders. Following at least 15 minutes of mixing, the cells' produced formazan crystals were completely dissolved in 100 μL of dimethyl sulfoxide (DMSO)18. Gentle mixing was followed by the addition of the mixed solution to a brand-new multi-well plate. A multimode reader (Synergy H1 hybrid, Biotek, USA) was employed to document the absorbance values at a wavelength of 570 nm. In all cytocompatibility investigations, MTT tests were carried out in triplicate.
Statistical Analysis
The findings of each experimental investigation were presented as mean ± SD (standard deviation), and each study was conducted in triplicate. To determine the statistical significance of the data, a one-way Analysis of Variance (ANOVA) approach was used in combination with Tukey's multiple comparison tests. All of the data were compared and found to be statistically different when the transition temperature and melting temperature were investigated using the obtained DSC thermograms.
CONCLUSION:
It is concluded that cow urine is a truly multifaceted medicine after examining diverse findings on the subject in numerous study articles. Fresh cow pee from native cows is considered the best, according to Indian medicine. Not only does it boost human and animal immunity, but it also has anti-cancer, antibacterial, anti-diabetic, anti-urolithiatic, antipsychotic, anti-hemorrhoidal, anti-aging, and skin-related properties. Several analytical techniques were used to examine cow urine, including nuclear magnetic resonance (NMR) spectroscopy, Fourier Transform Infrared (FTIR) spectroscopy, Thermogravimetric Analysis (TGA), Differential Scanning Calorimetry (DSC), and Scanning Electron Microscopy – Energy Dispersive X-ray Spectroscopy (SEM-EDX). The lyophilized powder of cow urine had both ordered and irregular (amorphous) molecular structures, had three stages of disintegration, and had a cluster-like appearance. Fresh cow pee from native cows is considered the best, according to Ayurveda. Cow urine may be thoroughly examined in the fields of research and medicine, and this knowledge can also be applied to the field of analytical techniques.
ACKNOWLEDGEMENT:
I would like to express my sincere gratitude to the Management, Principal of RVS college of pharmaceutical sciences for providing the necessary resources and which greatly contributed to the successful completion of this Research.
CONFLICT OF INTEREST:
The authors declare that there is no conflict of interest.
REFERENCES
Supriya. K, Angala Parameshwari. S, A Comprehensive Review on the Medicinal Potential of Cow Urine (Bos indicus), Int. J. of Pharm. Sci., 2025, Vol 3, Issue 10, 516-527. https://doi.org/10.5281/zenodo.17278346
10.5281/zenodo.17278346
