INTRODUCTION:

Plants have been used for medicinal purposes since ancient times, and this knowledge has been passed down through generations, forming the basis of traditional medicine in many cultures. The importance of plants as medicine lies in their ability to produce a wide range of bioactive compounds that can be used to treat various ailments.^1,2

In addition to their therapeutic properties, plant-based medicines are often safer and more affordable than synthetic drugs, making them accessible to a wider population. They also offer a sustainable and environmentally friendly option for medicine production, as plants can be grown renewably and without the need for synthetic chemicals. Various plant derived components had shown remarkable pharmacological action and had proven to be medicines for human use^1,3. The theory of using plants as medicine is supported by various scientific researches, which has validated many traditional uses of plant-based medicines and identified new therapeutic compounds. Ongoing research continues to explore the potential of plants as a source of new drugs and treatments.⁴

The World Health Organization (WHO) recognizes the potential of traditional medicine and encourages the use of plant-based medicines as a complementary approach to conventional medicine. In 2014, WHO published a strategy that aims to promote the safe and effective use of traditional medicine, including herbal medicines. The strategy outlines key actions to be taken by member states and WHO in order to integrate traditional medicine into healthcare systems.⁵WHO has developed guidelines for the quality control and standardization of herbal medicines. These guidelines provide a framework for ensuring the safety, efficacy, and quality of herbal medicines, and cover areas such as cultivation, harvesting, processing, and storage. Overall, WHO's policies and guidelines for the use of plants as medicines aim to promote the safe and effective use of traditional medicine, while ensuring the quality and efficacy of herbal medicines.^5,6

Another survey conducted in India in 2019 found that 73% of the participants reported using herbal medicines for various health problems, with a significant proportion using them for chronic conditions such as diabetes and hypertension. The most commonly used herbal remedies in this study were Tulsi (holy basil), ginger, and turmeric.⁷ However, despite the widespread use of herbal medicines, it is important to note that not all herbal remedies are safe or effective. Some can interact with prescription medications or cause adverse effects. Therefore, it is crucial for individuals to consult with a qualified healthcare professional before using herbal medicines.^5,7

In recent years, there has been a growing interest in using herbal remedies as an alternative to conventional medicine. There are many reasons why people choose to use herbal medicines. Some prefer them because they are perceived as more natural and gentler than conventional drugs, which can have side effects. Others may prefer to use herbal remedies because they believe they are more effective than pharmaceuticals or because they want to avoid the high costs associated with prescription drugs.^8,9 Herbal medicine has been an integral part of traditional Indian medicine for centuries, and its use continues to be prevalent among the literate population. According to a survey conducted in India in 2017, 76% of the respondents reported using herbal medicines for various health problems.¹⁰ The awareness of herbal medicine is also high among the literate population in India. According to the same survey, 93% of the respondents reported having knowledge about herbal remedies, with 70% of them having obtained the information from family members and friends. Additionally, 67% of the respondents reported using herbal medicines based on recommendations from healthcare professionals.^10,11 The use of plants in the field of medicines is enormous than ones thinking, the plant resources are not only exploited the way it is useful in medicines but there are various others uses of plants as medicines.

The plants not only show the medicinal and active pharmacological action rather it has its novel emerging use in the field of naturally derived cosmetics. Natural cosmetics are products made from naturally-derived ingredients such as plant extracts, essential oils, and minerals, and are free from synthetic chemicals such as parabens, sulphates, and fragrances. These ingredients are sourced from nature and are minimally processed to retain their natural properties. Natural cosmetics are becoming increasingly popular due to their perceived health benefits and eco-friendly properties. They are thought to be gentler on the skin and less likely to cause allergic reactions or skin irritations compared to synthetic cosmetics.¹² Furthermore, there are some companies in which natural cosmetics are often packaged in biodegradable or recyclable materials, making them more sustainable and environmentally friendly. Some popular natural cosmetic ingredients include aloe vera, lavender oil, chamomile extract, coconut oil, and shea butter etc. These ingredients are majorly known for their moisturizing, nourishing, and soothing properties, and are commonly used in skincare products such as moisturizers, cleansers, and face masks as marketed products. Overall, natural cosmetics offer a safer and more sustainable alternative to traditional cosmetics. However, it is important to note that not all natural ingredients are safe for everyone as it may cause certain skin reactions or allergies varying from person to person, skin type, race and geographical region they belong to. Therefore, it is important to always choose a product wisely and consult with a dermatologist if a person have sensitive skin or are prone to allergies.

Not only medicines but presently plants and natural products are widely accepted in cosmetics and cosmeceuticals. Cosmetic defined by Food, Drug and Cosmetic Act as under section 3(aaa) of the Drugs and Cosmetics Act, 1940 as, any article intended to be rubbed, poured, sprinkled or sprayed on, or introduced into, or otherwise applied to, the human body or any part thereof for cleansing, beautifying, promoting attractiveness or altering the appearance, and includes any article intended for use as a component of cosmetic¹³. Looking forward to the definition the several category of products that can be listed as cosmetics are related to skincare, hair care, nail care, dental care and the fragrance¹². The cosmeceuticals are defined as the cosmetic products used as the pharmaceuticals, which are intended for the use as cosmetics for enhancing the attractiveness and beautifying of body parts, as well as it exhibits its medicinal and pharmacological activity on certain body parts. Cosmetics which are derived from plants and its parts are called as the herbal or natural cosmetics, being naturally derived it has least side effects/harmful impacts on body parts as compared to synthetically manufactured cosmetic products. The beauty products has its own ugly side, the side effects of synthetically derived cosmetics vary from the amount of chemicals present in it to the skin/hair type of the user¹⁴, majority of studies had mentioned the presence of phthalates [Di(2-ethylhexyl) phthalate, Dibutyl phthalate] in the cosmetics to be the proven cause of various reproductive deformities in males as well as causing risk of recurrent miscarriages in females, it is also responsible for raised level of its accumulation in urine of infants too¹⁵.

COSMETICS AS THE SKIN CARE PRODUCTS:

Cosmetics are products that are used to enhance or improve a person's appearance. One of the most common uses of cosmetics is for skin care. Skin is the largest organ of our body, it functions widely in providing a barrier to water and several pathogens as well as it protects the body from various types of trauma such as thermal, chemical traumas and the radiations. The skin helps in regulating the body temperature by maintaining the homeostasis of the body and it also enhances the metabolic functions, the human skin is also responsible for the synthesis of vitamin D in the body.¹⁷ The anatomy of skin depicts that the skin comprises of the three layers:

1. The Epidermis:- This is the outermost layer of the skin which acts as the water barrier and contributes in providing the skin colour/tone.

2. The Dermis:- This layer is present beneath the epidermis layer and it comprises of the connective tissues, hair follicles, blood vessels, lymphatic vessels and the sweat glands.

The Subcutaneous layer/Hypodermis:-

This layer of skin comprises of fat and connective tissues.¹⁸

COSMETICS AS THE LIP CARE PRODUCTS:

Lips are the outermost muscular folds which are projected out from the one end i.e. mouth of the alimentary canal. The skin of lips differ in various ways from the overall regular skin of the body. Generally the regular skin of the body comprises of 14–16 layers for the protection whereas the skin of the lips in case of facial skin too is comparatively thin consisting of the 4 -5 layers. The concentration of melanin cells is less in the skin of lips as compared to regular skin due to which the blood vessels beneath the outer layers are more visible through those layers providing lips there pinkish to peach colour in most of the people. The skin of lips does not contain any hair follicle, sebum and oil glands, therefore they cannot produce any sweat or natural body oil in order for there protection.^19,20

The marketed product of petroleum jelly which has its general purpose for the use on the skin and lips, the primary purpose of those products is to keep the skin moisturised and hydrated as well as prevent the drying, chapping, wearing and cracking of the skin. The marketed product of the plain petroleum jelly does not contain any of the active ingredient which could show its therapeutic or remedial effect. On the contrary instead of marketed plain petroleum jelly the herbal petroleum jelly containing the various plant extracts has numerous advantages over it. There are various therapeutic effects of plant extracts such as antioxidant and antimicrobial activities which enhances the health of the skin or the lips that produces the remedial effects.²¹

Various studies have found that many plants possess such plant pigments and the phenolic compounds have the free radical and ROS (reactive oxygen species , ex- superoxide radical ion, hydrogen peroxide, hydroxyl radical) scavenging power and thus act as the natural antioxidant, these entities work either by the termination of free radical chain reaction or by reduction of ROS and other free radical species in the biological system, these are generated during the oxidative stress conditions which are further responsible for the damages to the macromolecules such as proteins, lipids and DNA.^22,23,24

Antimicrobial studies of a formulation aim to determine whether the formulation has any antimicrobial properties and how effective it is at killing or inhibiting the growth of microorganisms such as bacteria, viruses, fungi, and other pathogens. These studies are important in the development of various products, such as pharmaceuticals, cosmetics, and disinfectants, to ensure that they are safe and effective for use. The antimicrobial studies typically involve testing the formulation against a panel of microorganisms under specific conditions, such as time, temperature, and concentration. The results of these studies can provide information about the formulation's potency, spectrum of activity, and potential for resistance development.^25,26

Amaranthus cruentus L., commonly known as red amaranth or Lal Sag, is a species of flowering plant in the family Amaranthaceae. Beyond its various culinary importance as ornamental beauty and its producing edible leaves and seeds , Amaranthus cruentus L. has also been employed in traditional medicine for its potential health benefits. Introduction Medicinal plants are used locally in the treatment of infections caused by fungi, bacteria, viruses and parasites. And over 60% of people in Nigeria rural areas depend on the traditional medicine for the treatment of their ailments. Plant derived medicines are widely used because they are relatively safer than the synthetic alternatives, as they are easily available and cheaper. Amaranthus belong to Amaranthaceae family. Amaranthus, communally known as Green amaranth or locally as “Karund”, is a multinational genus of herbs. Amaranthus or Amaranth is defined as “never-fading flower” in Greek. Several species of Amaranthus are often considered as weeds, people around the world worth amaranths as leaf vegetables, cereals and ornamentals.²⁷

Antioxidants protect cells against damage caused by molecules known as free radicals. The antioxidant effects in plants are mainly due to the presence of phenolic compounds such as flavonoids, phenolic acids, tannins and phenolic diterpenes. Oxidative damage is implicated in most disease processes such as cardiovascular disease, cancer, inflammatory conditions, asthma, liver disease and muscular degeneration. Epidemiological, clinical and laboratory research on flavonoids and other antioxidants suggest their use in the prevention and treatment of a number of these disorders. It is recently found that Amaranth seeds and leaves are rich in quite a few phytonutrients that take part in a significant role in inhibition of both oxidative chain reactions and free radicals reactions with tissue and membranes. Existence of microorganisms causes spoilage, and results in reduction of the quality and quantity of processed foods. Some biologically active compounds isolated from herbs have been in use for the inhibition of growth pathogenic microbes because of the resistance against antibiotics. The medicinal values of plants and vegetables are dictated by their phytochemical and other chemical constituents.²⁸

The pharmacological properties of amaranth products are considered of vital importance. For reducing tissue swelling the leaves are well thought-out to be constructive, and they have a cleansing effect too. The plant has also been used curatively for diarrhoea, dysentery, excessive menstrual flow, ulcers and intestinal haemorrhaging^.27 For the treatment of intestinal bleeding, excessive menstruation, diarrhoea and other related problems, a tea made from its leaves is used. Aim of the present study was to evaluate the phytochemicals, antioxidant and to detect the presence of natural therapeutic agents, especially those related to control the microbes that cause diseases in human beings from Amaranthus leaf extract with ethanolic solvent system. The antioxidant molecules that reduce effect of radicals important for protection against cancer and degenerative disorder are in a abundance in Amaranthus. Antioxidant potential has been attributed to the presence of appreciable levels of phenolic and flavonoids. Leaves and flowers of Amaranthus.

Allium cepa L., have been an important dietary resource and have also been of interest of medical purposes. Traditionally onions and plants belonging to allium genus have been used as herbal remedy for the wide range of ailments, due to their association with many pharmacological effects. Biological effects attributed to onions have been commonly described to the volatile sulphur containing compounds, such as thiosulfates, mainly responsible for the characteristics, taste, aroma and lachrymatory effects. Onion is known for being a good natural source of flavonoids mainly represented by the flavonols quercetin and kaempferol which are present as their glycosides. Onion can be considered as the good source of the natural additives to retard food deterioration. Onion belongs to the genus Allium chives, shallots, and leeks. Onion has been characterized for its flavonols, quercetin, and quercetin derivatives with red onion containing a higher amount of antioxidant compound.²⁴

The aim of the study was to test the efficacy of peels of Allium cepa L. against lip disorders and its antioxidant activity. Allium cepa L. may be a new potential source as natural antimicrobial and antioxidants agents applied in food systems. Antioxidant that retard the oxidation process may additionally exhibit antimicrobial activity.

Tagetes erecta L. known as marigold (common ornamental plant) which bears bright yellow and orange flowers, is available in many parts of the world. The species Tagetes minuta erecta, patula and tenuifolia are most common, other species referred to are often specific to a region. Marigold is also widely used as the medicinal herb for its anti-inflammatory, analgesic, anti-oedematous properties which are important to phytotherapy as well as to dermatology and cosmetology. The pharmacological activity is related to the content of several secondary metabolites, and the most important compound such as terpenes, essential oils, flavonoids and carotenoids. Many reports and clinical studies describe this plant as having a numerous biological activity such as wound healing, anti-inflammatory effects. Phytochemical investigation of these flower and leaves extract reported the presence of the highest flavonoids, flavonols glycosides, coumarins, saponins, triterpenes, fatty acids, esters, essential oil, and hydrocarbons.²⁹The aim of this study was to validate the antioxidant activity and anti-inflammatory effects of alcoholic extracts marigold petals for the cure of lip disorders.

The utilization of antioxidants and antimicrobial properties in the formulation of herbal lip petroleum jelly using amaranth, onion, and marigold brings about significant benefits. These natural ingredients possess unique properties that contribute to the overall effectiveness and appeal of the product.

(a) (b)

(c)

Fig. 1: (a) Amaranthus cruentus L. (red amaranth) (b) Allium cepa L. (red onion) (c) Tagetes erecta L. (marigold)

The combination of these three ingredients in the herbal lip petroleum jelly results in a comprehensive and effective lip care product. The antioxidants derived from amaranth, onion and marigold work synergistically to provide robust protection against oxidative damage, promoting lip health and vitality. Simultaneously, the antimicrobial properties from onion and marigold extracts contribute to maintaining the lips cleanliness and preventing microbial infections. Moreover, the use of natural ingredients like amaranth, onion, and marigold aligns with the increasing consumer demand for sustainable and eco-friendly products. By opting for herbal alternatives instead of synthetic additives, the formulation of this lip petroleum jelly offers a more environmentally conscious choice. These natural ingredients contribute to the overall efficacy of the product by protecting the lips from oxidative damage, combating microbial growth, and promoting lip health. Additionally, the use of sustainable and eco-friendly ingredients aligns with the growing consumer preference for natural and environmentally conscious products.

PLANT PROFILE:

1) RED AMARANTH LEAVES:

Synonym: Indian spinach, Pigweed, Amaranthus albus, Amaranthus caudates, Genus Amaranthus

Scientific name: Amaranthus cruentus L.

Biological source: Obtained from the leaves of Amaranthus cruentus L., native to Mexico and Guatemala

Family: Amaranthaceae

Chemical constituents: Dietary fiber, protein with essential amino acids, such as lysine and methionine, carotenoids, vitamin C, and minerals, such as calcium, magnesium, potassium, phosphorus, iron

Uses:

1. Amaranth's use as an herbal medicine is derived mainly from its high content of vitamins and minerals. Because of its high protein and low calorie value, it is an effective weight loss tool, additionally preventing excess buildup of LDL, or "bad" cholesterol.

2. Amaranth is an excellent source of manganese, a mineral that plays an important role in the metabolism of carbohydrates, promotes calcium absorption, and helps regulate blood sugar. Amaranth have been used mainly as a food for its amazing nutritional content.

3. Amaranth has been consumed for supplementing calcium intake, since rich in this amaranth can decrease the anemia prevalence and increase hemoglobin levels in children.³⁵

4. Amaranth is tricolor whole plants are used by folk medicinal practitioners for treatment of pain, anemia, dysentery, skin diseases and as a blood purifier.

5. It has the significant oral hypoglycemic activity. ³⁶

Purpose of choosing Amaranth:

1) The leaves of amaranth are obtained from natural sources hence it does not produce any side effects.

2) It get easily available in Vidarbha region.

3) Various studies had reported potential effect of amaranth as antioxidant and antimicrobial plant.

4) The pharmacognostical investigation of plant was done sufficiently in previous studies.

5) It gives coloration to the formulation

2) RED ONION PEELS:

Synonym: Onion plant, Allium cepa L., Allium ascalonicum

Scientific name: Allium cepa L.

Biological source: -It is derived from the plant Allium ascalonicum

Family: Amaryllidaceae

Chemical constituents: The principal constituents of onion is containing various sulfur compound such as mono-, di-,tri-,tetra sulfide, thiols and thiophene derivatives, carbohydrates, proteins, amino acids, polyphenols, vitamin B and C, saponins, sterols, phenolic acids and phenols.

Uses:

1. The antibacterial activity of onion can be attributed to the presence of flavonoids and polyphenols which has been reported to the broad-spectrum antimicrobial activity.

2. Onion is the used for its characteristics in the management of various ailments. It is also used as blood purifier.³⁷

3. Onions are found to be diuretic and wound healer.³⁸

4. Onions are a major source of the dietary flavonoid, however there may exist varietal difference in composition and beneficial activities of onion. Onion is also used in skin problems. Onions are rich in vitamin C which helps boost the production of a collagen and thus improve skin elasticity. ³⁰

5. Onion (Allium cepa L. L.) among the oldest cultivated plant also used in the treatment and prevention of several diseases, including cancer, coronary heart disease, obesity, hypercholesterolemia, diabetes type 2, hypertension, cataract, and disturbances of the gastrointestinal tract. Onion is the potential antioxidant.

6. The phytochemicals present in Onion are polyphenols, anthocyanins, flavonoids, quercetin, and their glycosides. All phytochemicals have health benefits, the high qualities of flavonoids, organosulfur compounds, phenolic acids are important for the antioxidant activity. ³⁰

7. Onions are helps to reduce the severity of symptoms associated with conditions such as the pain and swelling of the osteo and rheumatoid arthritis, the allergic inflammatory response of asthma, and the respiratory congestion associated with common cold.

Purpose of choosing Onion:

1) Onion peels are the major vegetable wastes from the common households and restaurants; hence it has proven antioxidant and antimicrobial activity, the onion peels can be used sustainably for the formulation.

2) The A. Cepa is selected for the study as it is available throughout the year.

3) There are very fewer skin cosmetics containing the onion are available in market.

4) Various studies have shown the potential antioxidant and antimicrobial activities of the onion hence it becomes useful for the lip disorders.

3) MARIGOLD FLOWER PETALS:

Synonym:Cornflower, Gold bloom, geranium, meadowsweet

Scientific name: Tagetes erecta L.

Biological Source: Calendula officinalis, the pot marigold, common marigold, ruddles, Mary's gold or Scotch marigold

Family: Asteraceae

Chemical constituents: -Bicyclogermacrene, Piperidones, Terthiophene, carotenoids such as lutein, neoxantin, zeaxantin, β-carotene

Uses: Pot marigold petals have been keyed into ointments, extracts, infusions and used for the variety of ailments including fever, jaundice, stomach ulcers, burns and wounds

1. Marigold is may play a role in improving skin appearance. Plant compound of calendula, like flavonoids and saponins, may help prevent the body for releasing enzymes that cause inflammation and sensitivity. ³⁹

2. Different parts of these plant are useful in fevers, astringent, carminative, stomachic, scabies and liver complaints, diseases of the eyes, purify the blood, bleeding piles, rheumatism, colds and bronchitis.

3. The plant Tagetes erecta L. contains various important phytochemical constituents from the different part of the plant. It shows different pharmacological activities like anti-nociceptive, anti-inflammatory, antioxidant, insecticidal, larvicidal, hepatoprotective, antipyretic, wound healing, antibacterial, antimicrobial, antiepileptic and antifungal. ⁴⁰

4. Marigold flower has long been used as a food colorant and ingredient in human food and animal feed. These phytochemicals are precursors to the production of antioxidant vitamin A.

5. Marigold was an important medicine in Ancient Greece, Rome, and Arabia. It was most used in skin treatment preparation for treating minor wounds, callouses, insect bites and stings, eczema, itches, and burns.

6. Marigold has high content of antioxidants. These antioxidants are known to be effective in combating the damage caused by free radicals which are byproduct of environmental factor such as pollution or cigarette smoke or even normal body functions.

7. Marigold flower petals are excellent and important source of carotenoids.

Purpose of choosing Marigold:

1) The petals of marigold are obtained naturally from the flowers of the T. erecta, hence it does not cause any side effect to the humans.

2) Various studies had shown the antioxidant and antimicrobial activities of the marigold flower petals which makes it useful in formulation of the various cosmetic products.

3) There has been not any such formulation containing the extract of marigold flowers.

MATERIAL AND METHODS:

COLLECTION OF PLANT MATERIALS:

· The Red amaranth (Amaranthus cruentus L.) was purchased from the vendors of the local market of Hingana, the whole plant was washed off and air dried at room temperature, the leaves were separated from the whole plant and shed dried for 5-6 days. Completely dried leaves of red amaranth were grinded to form fine powder in a laboratory grinder of about 250grams. The powdered crude drug was stored in an air tight container for further investigation and about 20grams of powder was taken for the extraction process.

· The Red onion peels (Allium cepa L.) was collected from the daily household vegetable waste and from the local vendors of the Hingana market. The collected peels were dusted off and desirable peels were shed dried for about 2-3 days. Further the peels were grinded to form the fine powder in a laboratory grinder of about 250 grams. The powdered crude drug was stored in an air tight container for further investigation and about 20 grams of powder was taken for the extraction process.

· The Marigold flowers (Tagetes erecta L.) were collected from the cultural head of Nagpur college of Pharmacy, which were used in the traditional programs held in the premises. The flowers were collected washed and strained for few hours, after which the petals were separated and shed dried for 5-7 days. Completely dried petals of marigold flowers were grinded to form the fine powder in a laboratory grinder of about 250 grams. The powdered crude drug was stored in an air tight container for further investigation and about 20 grams of powder was taken for the extraction process.

EXTRACTION PROCESS:

The extraction process was carried out by following techniques for different plant materials and the extractive process:

1. Red amaranth leaves (Amaranthus cruentus L.):

The alcoholic extract of dried powdered leaves of red amaranth was prepared by the process of Soxhlet extraction. The 60ml extractor of the Soxhlet apparatus was packed with about 20 – 25grams of dried powder of red amaranth leaves, the condenser was attached on the opening of the extracting thimble which was provided with the continuous cold water supply, the bottom opening pf the extracting thimble was attached with the round bottom flask of about 250ml which was placed on an electric heating mantle. About 200-250ml of 90% ethanol was filled into the round bottom flask which after switching on the heating mantle ( 50^oC ) gets evaporated through the solvent evaporating tube, the ethanolic vapours gets condensed into the droplets into the condenser with continuous water supply wetting the powdered crude drug continuously. The alcohol soluble contents of the crude drug move down along with the solvent (ethanol) through the siphon tube and gets collected into the round bottom flask below. This process was continued for 25hours, after which the solvent was allowed to get concentrated into the same round bottom flask and the recovery solvent was collected into the extracting thimble. The further concentration of the extract was obtained by putting the concentrate into a china dish which was heated on the water bath for about 75mins.

2. Red onion peels (Allium cepa L.):

The alcoholic extract of dried powdered peels of red onion was prepared by the process maceration. About 20 grams of the powdered red onion peels was taken into the 250 ml iodine flask, 200 ml of 90% ethanol was added to the same iodine flask. The crude drug was allowed to soak into the ethanol for about 26-28 hours, the extract was filtered through the wattman filter paper and concentrated by heating on the water bath by evaporating it to dryness.

3. Marigold flowers petals (Tagetes erecta L.):

The alcoholic extract of dried powdered petals of marigold flower was prepared by the process of Soxhlet extraction. The 60ml extractor of the Soxhlet apparatus was packed with about 20 – 25 grams of dried powder of petals of marigold flower, the condenser was attached on the opening of the extracting thimble which was provided with the continuous cold water supply, the bottom opening pf the extracting thimble was attached with the round bottom flask of about 250ml which was placed on an electric heating mantle. About 200-250 ml of 90% ethanol was filled into the round bottom flask which after switching on the heating mantle ( 50^oC ) gets evaporated through the solvent evaporating tube, the ethanolic vapours gets condensed into the droplets into the condenser with continuous water supply wetting the powdered crude drug continuously. The alcohol soluble contents of the crude drug move down along with the solvent (ethanol) through the siphon tube and gets collected into the round bottom flask below. This process was continued for 28 hours, after which the solvent was allowed to get concentrated into the same round bottom flask and the recovery solvent was collected into the extracting thimble. The further concentration of the extract was obtained by putting the concentrate into a china dish which was heated on the water bath for about 90 mins.

MORPHOLOGICAL STUDY OF PLANTS:

1. Red amaranth (Amaranthus cruentus L.)^{41, 42}

a) Plant Type: Amaranthus cruentus L. is an annual plant species that belongs to the Amaranthaceae family. It is commonly known as red amaranth or blood amaranth.

b) Stem: The plant has an erect, branched stem that can grow up to 2-8 feet in height. The stem is typically green or reddish in colour, depending on the variety.

c) Leaves: The leaves of Amaranthus cruentus L. are simple, alternate, and ovate to lanceolate in shape. They are typically dark green or purplish-red in colour. The leaf margins may be smooth or slightly serrated.

d) Flowers: The flowers of Amaranthus cruentus L. are small and inconspicuous. They are usually clustered together in dense clusters along the flower spikes. Each flower has a short pedicel and consists of a perianth with five tepals.

e) Fruit and Seeds: After successful pollination, the flowers develop into small, oval-shaped fruits called utricles. These utricles contain a single seed each, which is tiny, shiny, and black in colour.

f) Root System: The plant has a fibrous root system that spreads wide rather than growing deep into the soil. The roots help anchor the plant and absorb nutrients and water from the surrounding soil.

g) Growth Habit: Amaranthus cruentus L. exhibits a vigorous growth habit and can quickly colonize an area under favourable conditions. It is a sun-loving plant that thrives in warm climates.

h) Varieties: There are several cultivated varieties of Amaranthus cruentus L., each with its unique characteristics, including variations in leaf colour, stem colour, and inflorescence shape.

2. Red onion peels (Allium cepa L.)^31,43

a) Plant Type: Allium cepa L. is a biennial plant species that belongs to the Amaryllidaceae family. It is widely cultivated as a vegetable crop.

b) Bulb: The most distinctive feature of Allium cepa L. is its bulbous underground storage organ. The bulb is composed of fleshy, concentric layers called scales. It serves as a food reserve for the plant.

c) Leaves: The plant has long, cylindrical leaves that arise from the base of the bulb. The leaves are hollow and tubular, with a waxy coating. They are typically green in colour.

d) Flowers: Each flower of Allium cepa L. is composed of a perianth with six tepals, which are usually white or pale pink in color. The flowers also have a prominent, spherical-shaped structure called the ovary, which develops into the onion bulb after fertilization.

e) Roots: Allium cepa L. has a fibrous root system that originates from the base of the bulb. The roots help anchor the plant in the soil and absorb water and nutrients.

f) Stem: The flowering stalk of Allium cepa L. is tall and slender. It emerges from the center of the bulb and supports the umbels of flowers. The stem is typically hollow and leafless.

g) Reproduction: Allium cepa L. reproduces sexually through the production of flowers and subsequent seed formation. However, it is primarily propagated vegetatively by planting bulbs or bulbils (small bulbs) rather than relying on seeds.

h) Varieties: There are numerous cultivated varieties of Allium cepa L., each with different characteristics such as bulb size, shape, color, and pungency. Varieties range from mild, sweet onions to pungent and spicy ones.

3. Marigold flowers (Tagetes erecta L.)⁴⁴

a) Plant Type: Tagetes erecta L. is an annual plant species that belongs to the Asteraceae family. It is native to Mexico and widely cultivated as an ornamental plant.

b) Height: The plant can grow to a height of 1 to 3 feet, with a compact and bushy habit.

c) Leaves: The leaves of Tagetes erecta L. are alternate and deeply divided into lanceolate or oblong leaflets. The leaf color ranges from dark green to medium green, and the leaflets may have toothed or serrated edges.

d) Flowers: The plant produces large, showy flowers on sturdy stems. The flowers are composed of densely packed, overlapping petals. The typical color of the flowers is bright orange or yellow, but cultivars with red, maroon, and bi-coloured flowers are also available.

e) Inflorescence: Tagetes erecta L. has solitary flower heads that are terminal or axillary in position. Each flower head consists of a central disc of tiny tubular florets surrounded by a ring of ray florets.

f) Ray Florets: The ray florets are the outermost part of the flower head and are characterized by long, narrow petals that radiate outward. They are responsible for the vibrant colours and ornamental appeal of the plant.

g) Disc Florets: The disc florets are small and tubular, forming a dense, rounded center within the flower head. They are typically yellow or orange in color.

h) Scent: Tagetes erecta L. is known for its strong, distinctive fragrance, which can vary in intensity depending on the cultivar and individual plant.

i) Fruit and Seeds: After pollination, the flowers develop into small, dry fruits called achenes. Each achene contains a single seed and is equipped with bristles or barbs that aid in seed dispersal.

j) Growth Habit: African marigold has a compact and upright growth habit. It is known for its vigorous growth, rapid development, and ability to tolerate hot and dry conditions.

MICROSCOPICAL CHARACTERISTICS OF POWDERED CRUDE DRUGS:

The microscopical study of the crude drugs is essential to determine the several microscopical features of the plant such as crystals, starch grains and lignified cells etc. which determines the quality of the drug. The powder of Red amaranth (Amaranthus cruentus L.), Red onion peels (Allium cepa L.) and Marigold flower petals (Tagetes erecta L.) was passed through the sieve no. #80 and were subjected for further microscopic evaluation. The slides were mounted of each crude drug powder and was treated with following reagents for microscopic evaluation and the mounted slides were observed under the light microscope and examined for the following characteristics along with it, the results were recorded in Table no. 1³⁴:

Table no. 1: Staining / Diagnosis / Microscopical studies

S.N.	Reagents	Observations
1	Phloroglucinol + Conc. HCl (1:1)	Red or pink colour indicates the presence of lignified xylem, lignified pericyclic fibres, vessels (V.B.) and xylem of V.B.
2	HCl / dil. HCl	Calcium oxalate crystals gets dissolved
3	Sulphuric acid (60% w/w)	Formation of needle shaped crystals of calcium oxalate
4	Dil. Acetic acid	Calcium oxalate crystals are insoluble
5	Dil. Iodine solution	Blue colour indicates the presence of starch in endodermis
6	Baljet reagent	Yellow or orange colour formation indicates the presence of the glycosidic cells
7	Ruthenium red	Formation of red or pink colour indicates the presence of mucilaginous cells
8	Alc. Picric acid	Yellow colour indicates the presence of aleurone grains in the cells of endosperm and embryo
9	Dil. Iodine solution + Conc. H₂SO₄	Blue colour indicates the presence of hemicellulose in the endospermic walls
10	Strong KOH solution	Formation of the needle shaped potassium eugenate crystals indicates the presence of eugenol of volatile oil
11	Conc. H₂SO₄	Green colour indicates the presence of the stone cells

PHYSICOCHEMICAL INVESTIGATION OF POWDERED CRUDE DRUGS:

Physicochemical investigation of crude drugs before extraction is vital for quality control, authentication, standardization, determination of extraction conditions, shelf-life determination, and compliance with pharmacopeial standards. The various physicochemical parameters that were determined for powdered Red amaranth (Amaranthus cruentus L.), Red onion peels (Allium cepa L.) and Marigold flower petals (Tagetes erecta L.) as per Indian Pharmacopoeia Vol., and the results were recorded as per Table no. 4 are as follows ^45,
46

PRELMIINARY PHYTOCHEMICAL SCREENING OF CRUDE DRUG EXTRACT³²

Preliminary phytochemical screening of plant materials is an essential step in the study of medicinal plants and natural products. It involves the qualitative analysis of various chemical constituents present in plant extracts. The phytochemical tests were performed for detection of chemical constituents of the crude drugs and the results were recorded in Table no. 3:

QUALITATIVE ANALYSIS FOR THE INORGANIC ELEMENTS ³²

Qualitative analysis for the inorganic elements provides valuable information about the elemental composition of plants. Prepare ash of drug material. Add 50% v/v HCl to ash. Keep for 1 hour or longer. Filter. With filtrate perform the following tests, and the results were recorded as in table 6.

FORMULATION OF POLYHERBAL LIP PETROLEUM JELLY¹

The herbal lip jelly was prepared from the ethanolic extracts of Red amaranth (Amaranthus cruentus L.), Red onion peels (Allium cepa L.) and Marigold flower petals (Tagetes erecta L.), petroleum jelly, lanolin, pineapple essence and vanillin were added as per the formulation in Table no. 2. Petroleum jelly and lanolin was melted and required quantity of Red amaranth (Amaranthus cruentus L.), Red onion peels (Allium cepa L.) and Marigold flower petals (Tagetes erecta L.) was incorporated into the base mixture later pineapple essence and vanillin were added and it was allowed to get a consistency at a room temperature only. Prepared formulations are shown in following Table no.

Fig. 2: Formulated polyherbal lip petroleum jelly

ANTIMICROBIAL ACTIVITY OF CRUDE DRUG EXTRACT:

Various studies had determined the promising antibacterial activity of red amaranth against E. coli strains, by demonstrating the inhibitory effects of red Amaranth extracts or its bioactive compounds against E. coli growth. The presence of natural compounds, such as phenols, flavonoids, and saponins, in red Amaranth contributes to its antimicrobial properties. These compounds disrupt the bacterial cell membrane, inhibit enzyme activity, and interfere with essential cellular processes of E. coli. The antibacterial activity of red Amaranth suggests its potential as a natural alternative for combating E. coli infections, supporting its use in traditional medicine and as a functional food ingredient and its use in cosmetics.^28,47,48

Similarly, Red onion exhibits potent antibacterial activity against E. coli strains. This can be attributed to the presence of bioactive compounds like organosulfur compounds, flavonoids, and quercetin. These compounds possess antimicrobial properties that disrupt the integrity of the bacterial cell membrane, inhibit crucial enzymes, and interfere with essential cellular processes of E. coli. Red onion extracts or their bioactive components have been shown to effectively inhibit the growth and proliferation of E. coli, making red onion a potential natural remedy for E. coli infections.^49,50,51

The antimicrobial properties of marigold can be attributed to its diverse range of bioactive compounds, including flavonoids, terpenoids, and essential oils. Several studies have explored the antibacterial potential of marigold extracts or essential oils against E. coli. These studies have revealed that marigold exhibits inhibitory effects on the growth and proliferation of E. coli, making it a promising natural remedy for E. coli infections. One of the key mechanisms through which marigold exerts its antibacterial activity is by disrupting the integrity of the bacterial cell membrane. The bioactive compounds present in marigold interact with the cell membrane, leading to membrane damage and leakage of cellular contents. This disruption impairs the normal functioning of the bacteria, ultimately inhibiting their growth.^18,52

The antimicrobial study for the extracts of Red amaranth (Amaranthus cruentus L.), Red onion peels (Allium cepa L.) and Marigold flower petals (Tagetes erecta L.) was done by antibacterial testing for the strains of E.coli and the zone of inhibition was observed. The platform where all the activities was carried out was cleaned and sterilized with 90% absolute ethanol and the surface was exposed to the flame of the burner, four burners were ignited and the further activities was performed within the sterile platform. The prepared bacterial culture was used for the study, the sterilized petri plates were prepared by pouring the agar (prepared and autoclaved for 15min at 150 Pa) into it, the petri dish were allowed to solidify on a sterilized platformed. After the solidification of agar media, the cork borer was used to create the cavities into the solidified agar media. Further the wells were loaded with crude drug extract and plates were allowed to stand for some time at room temperature to allow the extract to diffuse into the agar. Then, the plates were incubated at 40^oC temperature for 24hours. After that the zone of inhibition was observed.

EVALUATION OF POLYHERBAL LIP PETROLEUM JELLY^33,1,19

For the evaluation of the herbal petroleum jelly, several parameters should be considered to assess its quality and effectiveness. These are:

1. Organoleptic characteristics (colour and appearance):

Colour and appearance of the polyherbal lip petroleum jelly was characterized by visualizing the samples in 10x magnification under microscope, while the odour of the formulation was compared by the group of evaluators (two groups of evaluators having 3 individuals in each, they have scored the perfume stability of formulation in the ‘good/intermediate’ mark as per result).

2. Test for spreadability:

The test for spread ability was performed by applying the prepared formulation on a glass slide at room temperature repeatedly for each formulation to observe uniformity in the formulation of protective layer, the spreading length was measured in centimetres.

3. Test for pH:

The pH of formulations was determined by using digital pH meter. 0.5 gm of jelly was accurately weighed and dispersed in 20ml of distilled water and stored for 90 mins. The measurement of pH of each formulation was carried out three times and the average values are represented in the table no. 8.

4. Skin irritation test:

The test was carried out by applying the product on the human skin for 10 mins.

5. Viscosity:

The viscosity of the formulation was measured using Brookfield viscometer using rotor number #4 at 6,12 and 30 rpm each for every sample. Samples of the lip jelly were allowed to settle over 30 min at the temperature of test (25±10 c) before the measurement were taken. All readings were taken in triplicate for each formulation respectively and recorded in Table no. 9.

RESULTS AND DISCUSSION:

MICROSCOPICAL CHARACTERISTICS OF POWDERED CRUDE DRUGS³²

The results of the microscopical investigation of Red amaranth (Amaranthus cruentus L.), Red onion peels (Allium cepa L.) and Marigold flower petals (Tagetes erecta L.) is formulated in the following table

Table no. 3 Microscopical characteristics of powdered crude drugs

S. No	Reagents	Observations	Inference
S. No	Reagents	Observations	Amaranth	Onion	Marigold
1.	Phloroglucinol + Conc. HCl (1:1)	Red, Pink	-	+	+
2.	HCL/dil.HCL	Crystal get dissolved	-	-	-
3.	Sulphuric acid(60%w/w	Formation of needle shaped crystals of calcium sulphate	+	+	+
4.	Dil. Acetic acid	Insoluble crystals	-	-	-
5.	Dil. Iodine solution	Blue	+	+	+
6.	Baljet reagent	Yellow or orange colour	+	+	+
7.	Ruthenium red	Red pink	+	+	+
8.	Alc. Picric acid	Yellow	+	+	+
9.	Dil. Iodine solution + Conc. H₂SO₄	Blue	+	+	_
10.	Strong KOH solution	Needle shaped potassium eugenate crystals	+	_	_
11.	Conc. H₂SO₄	Green	_	_	_

Table no. 5: Preliminary phytochemical screening of crude drug extract

Sr No	Test for	Test	Observation	Inference
Sr No	Test for	Test	Observation	Amaranth (A)	Onion (O)	Marigold (M)
1.	Carbohydrate	Molisch test	Violet ring is formed at the junction of two liquids	_	+	+
		Fehling	Brick red ppt is observed	+	_	+
		Benedict’s test	Solution appears green, yellow or red	+ (Green)	+ (Red)	+ (Yellow)
		Barfoed’s	Red ppt is observed	+	+	+
		Iodine polysaccharides	Blue colour appears, it disappears on boiling and reappears on cooling	_	_	_
2	Proteins	Biuret	Violet or pink colour appears	_	_	_
		Million’s reagents	Brick red colour ppt form a red colour solution	_	_	_
		Precipitation test	White colloidal ppt	_	_	_
3	Amino acids	Ninhydrin test	Purple or bluish colour appears	+	+	+
3	Amino acids	Cysteine	Black ppt of lead sulphate	+	_	_
4.	Volatile oils	Characteristics odour	Characteristics odour aroma	_	_	_
5.	Steroid	Salkowski	Red and acid layer shows greenish yellow fluorescence	+	+	+
6.	Glycosides	Keller-Killani	Reddish brown colour appears and upper layer bluish green	+	+	_
6.	Glycosides	Saponins (Foam test)	Stable foam observed	+	_	+
7.	Flavonoids	Shinoda test	Orange, Pink, Red to Purple colour appears	+	+	_
7.	Flavonoids	Sulphuric acid test	Flavanes gives orange to red colour	_	_	_
8.	Tannins andPhenolic compound	Lead acetate	White ppt formed	+	+	+
		Acetic acid solu.	Red colour solution	+	+	_
		5%FeCl3	Deep blue- black colour	+	+	+
9.	Alkaloids	Dragendorff’s	Orange brown ppt is formed	+	_	_
		Mayer’s	White ppt formed	_	_	_
		Hager’s	Gives yellow ppt	-	-	-
		Wagner’s	Gives reddish brown ppt	-	-	-

Table no. 6: Qualitative analysis of inorganic elements present in the powdered crude drugs.

S. No.	Inorganic Elements		Observations	INFERENCE
S. No.	Inorganic Elements		Observations	Amaranth (A)	Onion (O)	Marigold (M)
1.	Potassium		Flame test : Gives violet colour to flame	-	-	+
2.	Iron		Blue colouration with 2% K₄[Fe(CN)₆].3H₂O	-	-	+
3.	Sulphate		White ppt of lead acetate solution which is soluble in NaOH	+	+	-
4.	Phosphate		Yellow crystalline ppt of ammonium phosphomolybdate is formed	-	-	-
5.	Chloride		White ppt of lead acetate solution which is soluble in hot water	+	+	+
6.	Carbonate	Test with mercuric chloride	Brownish red ppt	-	-	-
6.	Carbonate	Test with magnesium sulphate	White ppt is formed	-	-	-

QUALITATIVE ANALYSIS FOR THE INORGANIC ELEMENTS³²

The results of the qualitative analysis for the inorganic elements of Red amaranth (Amaranthus cruentus L.), Red onion peels (Allium cepa L.) and Marigold flower petals (Tagetes erecta L.) is formulated in the following table:

EVALUATION OF POLYHERBAL LIP PETROLEUM JELLY^{33, 1}

The evaluation for the four formulations of polyherbal lip petroleum jelly was carried out for various parameters and the results were recorded in the following table:

Table No. 7 Colour, odour, spreadability, pH and skin irritation test.

Sr No.	Parameter	Control	F1	F2	F3
1.	Colour	Off white	Reddish marron	Pinkish red	Red orange
2.	Odour	++	+++	+++	++
3.	Spreadability test	Good	Good	Good	Intermediate
4.	pH	6.5	6.7	6.8	6.7
5.	Skin irritation	No	No	No	No

+++: Very good ++: Good

Table No. 8: Viscosity measurement

Sr No.	Batch	RPM (mPa’s)			Percent
Sr No.	Batch	6	12	30	Percent
1.	CONTROL	13000	7150	3780	17.3%
2.	F1	22000	7150	2840	42.2%
3.	F2	24400	7190	2140	43.8%
4.	F3	25200	7350	1770	41.5%

SUMMARY:

The current study briefs about the benefits of herbal cosmetics over the synthetic ones as well as it highlights about the pharmacognostical characterization of the Red amaranth (Amaranthus cruentus L.), Red onion peels (Allium cepa L.) and Marigold flower petals (Tagetes erecta L.) and the development of polyherbal lip petroleum jelly from the crude drug extracts for its use in curing various lip diseases. The petroleum jelly acts as the base for the formulation which helps in providing various positive effects on the lips such as moisturization, and the extracts of the crude drugs incorporated into the base gives various therapeutic effects on the skin of lips. The crude drug extracts of all the three drugs were studied thoroughly for the presence of various chemical constituents and physicochemical characteristics in it, and also its antioxidant and antimicrobial properties. From the above observation it has been concluded that use of natural ingredients over synthetic gives minimal side effect and rapid healing of the skin of the lips without irritation. The formulation was developed at room temperature and was studied for various evaluation parameters. At room temperature the formulation was observed for changes in its colour, order, texture, and appearance and there was no change found in it, hence it was found to be stable. It can be easily stored and used by any person not only for the prevention of the diseases but also as the daily lip care product. There are very few such formulations available in the market at present time, hence the developed formulation can also be subjected for the pilot scale upgradation and further commercialization.

CONCLUSION:

The use of natural cosmetics over synthetic ones offers several advantages for individuals and the environment. Natural cosmetics are derived from plant-based ingredients and do not contain harmful chemicals or toxins that can potentially damage the skin or cause health issues. They are often hypoallergenic and suitable for sensitive skin types, reducing the risk of irritation or allergic reactions. Moreover, natural cosmetics are typically produced through sustainable and ethical practices, minimizing the negative impact on the environment. The present study evaluates the pharmacognostical studies and the formulation of the polyherbal lip petroleum jelly from the alcoholic extract of Red amaranth (Amaranthus cruentus L.), Red onion peels (Allium cepa L.) and Marigold flower petals (Tagetes erecta L.). In this research, we found effective properties of the polyherbal lip petroleum jelly and further studies are needed to be performed to explore more useful benefits of such herbal lip petroleum jelly, as there are very few such formulations available in the market. So, with this we achieved our aim and objectives.

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Received on 13.11.2025 Revised on 20.02.2026

Accepted on 23.04.2026 Published on 20.05.2026

Available online from May 25, 2026

Research J. Pharmacy and Technology. 2026;19(5):2283-2296.

DOI: 10.52711/0974-360X.2026.00329

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

S.No.	Ingredients	Control	F1	F2	F3	USES
1.	Petroleum jelly	7 gms	7gms	8gms	7gms	Base
2.	Lanolin	2.5gms	2.5gms	1.5gms	2.5gms	Glossy and softening agent
3.	Pineapple essence	0.1ml	0.1ml	0.1ml	0.1ml	Flavouring agent
4.	Vanillin	0.1gm	0.1gm	0.1gm	0.1gm	Preservative
5.	Red amaranth (Amaranthus cruentus L.) extract	-	0.3gm	0.2gm	0.1gm	Antimicrobial and antioxidant agent
6.	Red onion peels (Allium cepa L.) extract	-	0.3gm	0.2gm	0.1gm	Antimicrobial and antioxidant agent
7.	Marigold flower petals (Tagetes erecta L.) extract	-	0.3gm	0.1gm	0.2gm	Antimicrobial and antioxidant agent
8.	Honey	q.s.	q.s.	q.s.	q.s.	Sweetening agent, Humectant

S No.	Test Parameters		Observations (% w/w)
S No.	Test Parameters		Amaranth	Onion	Marigold
1.	Ash value	Total ash	26.50	28.50	39.25
		Acid insoluble ash value	9.45	9.10	9.45
		Water soluble ash value	15.35	15.85	9.55
2.	Extractive value	Alcohol soluble extractive value	6.16	3.76	23.6
2.	Extractive value	Water soluble extractive value	23.44	9.28	26.24
3.	Moisture content / Loss on drying		8.80	9.20	7.20
4.	Swelling index		Nil	Nil	Nil