Bloom in a bottle: Formulation and Evaluation of An Organic Rose Petals Tincture Using Vodka as A Solvent

Abstract

The present study focuses on the formulation and evaluation of an organic tincture prepared from fresh rose petals using vodka (40% v/v) as the extraction solvent. Tinctures serve as efficient alcohol-based extracts capable of preserving delicate phytochemicals, including essential oils, flavonoids, phenolics, and aromatic components. Organic rose petals were macerated in vodka for a period of 2–4 weeks, allowing gradual extraction of bioactive constituents. The prepared tincture developed a characteristic rose fragrance, deep reddish coloration, and stable aromatic profile. Evaluation parameters—including organoleptic properties, pH, alcohol content, total phenolic content (TPC), total flavonoid content (TFC), TLC fingerprinting, and microbial safety—confirmed the quality and stability of the formulation. The tincture demonstrates potential application in skincare, aromatherapy, and herbal wellness due to its antioxidant, anti-inflammatory, and soothing properties. Further studies on dosage, long-term stability, and therapeutic efficacy are recommended.

Keywords

Alcoholic Extract; Antioxidant Activity; Aromatherapy; Flavonoids; Herbal Formulation; Maceration; Organic Tincture; Phenolic Content; Phytochemical Analysis; Rose Petals

Introduction

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Evaluation test for organic tincture composed of rose petals and vodka:-

1. 1. Organoleptic test :-

Purpose: basic sensory check (identity & obvious defects).

Equipment: glass beakers, white background card, pH paper (optional).

Procedure:-

1. Pour a 10 mL sample into a clean glass.

2. Inspect against white background for clarity, color, suspended solids or sediment. Note color (e.g., pale pink to deep ruby), turbidity, particulates.

3. Smell gently (wafting) and note aroma (rose aroma intensity, off-odors like ferment/rot/solvent).

4. Taste a drop only if sample is known safe and you use clean technique (optional).

Acceptance:clear or slightly hazy depending on filtration; characteristic rose aroma; no rotten/off-fermented odour; no visible contamination.                  

1. 2. Alcohol content determination:-

Purpose: confirm ethanol concentration (important for extraction efficiency, preservation).

Methods: distillation (AOAC), hydrometer/specific gravity, or GC. I provide a simple distillation + densitometry procedure and a GC note.

A — Simple Distillation + Densitometry (common approach)

Equipment/Reagents: distillation apparatus (Kjeldahl/steam/distillation set), 25–50 mL sample, volumetric flasks, pycnometer or alcohol hydrometer or densitometer, thermometer, water bath.

Procedure:-

1. Distill a known volume (e.g., 50.0 mL) of tincture to collect ethanol/water fraction until ~45–50 mL collected (or per SOP).

2. Cool to room temp; measure density (g/mL) of distillate using pycnometer or densitometer at 20 °C.

3. Convert density to % v/v ethanol using standard ethanol density tables (or instrument calibration).

  Calculation: use instrument/table to read % v/v directly from density.

If using GC:

Prepare calibration curve with ethanol standards (e.g., 10–50% v/v).

Inject sample or headspace aliquot.

Measure peak area of ethanol (A?) and use calibration curve equation:

%ethanol (v/v)= As−b / m

Where ,

m = slope of calibration curve

b = intercept

Acceptance (suggested): close to starting vodka strength (if using 40% ABV vodka, tincture will be slightly lower due to plant water). Typical herbal tinctures: 20–45% v/v depending on formulation. For vodka-only tincture expect ~30–40% v/v; define your spec.

B — GC (recommended for accuracy)

Analyze using headspace GC or direct injection with calibration standards of ethanol; report % v/v.

1. 3. pH Measurement :-

Purpose:  stability/quality indicator and microbial growth potential.

Equipment:  calibrated pH meter, beaker, distilled water.

Procedure:-

1. Calibrate pH meter at pH 4.00 and 7.00 (or 7 & 10).

2. Pour 10 mL tincture into beaker, equilibrate to 20–25 °C, measure pH.  

Acceptance: many tinctures are mildly acidic — typical pH 3.5–6.5. Record for stability monitoring.  

1. 4.  Refractive Index / Specific Gravity :-

Purpose:  quick check on dissolved solids/alcohol concentration.

Equipment:  refractometer or densitometer, thermometer.

Procedure:  Place a drop on refractometer prism; read refractive index and convert (if needed) to Brix or estimate ethanol.

Acceptance:  record baseline; use for batch-to-batch consistency.

1. 5. Total phenolic content (Folin - Ciocalteu Method ) TPC :-

Purpose:  quantify total phenolics as Gallic Acid Equivalents (GAE) — common potency marker for rose extracts.

Equipment/Reagents:  Folin–Ciocalteu reagent, gallic acid standard, Na2CO3 solution (7.5% w/v), spectrophotometer (765 nm), pipettes, volumetric flasks.

Procedure (standardized):-

1. Prepare gallic acid standards (0, 25, 50, 100, 200, 400 µg/mL).

2. Dilute tincture sample to fall in standard range.

3. In test tube: mix 0.5 mL sample/standard + 2.5 mL 10% Folin–Ciocalteu reagent (diluted 1:10 with water) → after 5 min add 2.0 mL 7.5% Na2CO3.

4. Incubate 30–60 min at room temp in dark.

5. Measure absorbance at 765 nm.

6. Plot standard curve; calculate sample GAE (mg GAE/g or mg GAE/mL).

Acceptance: no universal spec — use for batch control and compare to literature or internal spec.

1. 6. Total Flavonoid Content (Aluminum Chloride Colorimetric) TFC:-

Purpose: quantify flavonoid content as rutin or quercetin equivalents.

 Equipment/Reagents:  AlCl3 reagent, quercetin/rutin standards, spectrophotometer (415 nm).

Procedure (Summary): 

follow standard AlCl3 assay: mix sample with AlCl3, incubate, measure at 415 nm, calculate mg QE/mL from standard curve.

7. Thin Layer Chromatography TLC / HPTLC Fingerprint :-

Purpose: chemical fingerprinting for identity and adulteration detection. HPTLC gives better resolution.

Equipment/Reagents:  TLC plates (silica gel), developing chamber, solvent system (example: ethyl acetate : formic acid : acetic acid : water 100:11:11:26 or adjust for phenolics), UV lamp (254 & 365 nm), derivatizing reagent (e.g., NP/PEG or vanillin-sulfuric acid). Standards: quercetin, gallic acid, cyanidin (if available).

Procedure (TLC):-

1. Apply standard and tincture (diluted) as 5–10 µL bands.

2. Develop plate in chosen solvent to ~8 cm.

3. Dry plate; visualize under 254/365 nm and after spraying derivatizing reagent and heating.

4. Compare Rf patterns vs standards; document photo.

Acceptance:  consistent fingerprint with expected spots for rose phenolics; absence of unexpected spots.

8. Microbial Testing :- (Total Aerobic Count , Yeast and Mould , Pathogen Screen)

Purpose: ensure microbiological safety (especially for oral use).

Equipment/Reagents: culture media (Plate Count Agar, Sabouraud Dextrose Agar), sterile diluents (peptone water), incubators, sterile plates. Alternatively use membrane filtration for liquids.

Procedure (membrane filtration recommended for liquids):-

1. Filter a known volume (e.g., 10–100 mL) through sterile 0.45 µm filter.

2. Place filter on Plate Count Agar (PCA) for total aerobic count; incubate 30–35 °C for 48–72 h.

3. For yeast & molds, use SDA and incubate 25–28 °C for 5–7 days.

4. For pathogens (E. coli, Salmonella, S. aureus), follow enrichment and selective plating methods (ISO or pharmacopeial).

Acceptance (typical herbal liquid specs):-

Total aerobic microbial count (TAMC) ≤ 10^3 CFU/mL.

Yeast & mould ≤ 10^2 CFU/mL.

Absence of E. coli, Salmonella, S. aureus in 10 mL.

(Adjust to pharmacopeia or certifier standards.)

9. Contaminants And Safety: - 

A. Pesticides Residue Screen 

B. Heavy Metals (Pb, Cd, Hg, As)

A.Pesticide residue screen:-

Purpose:  demonstrate compliance with organic standards.

Method:  QuEChERS extraction followed by GC-MS/MS and/or LC-MS/MS screening for multi-residue pesticides.

Procedure (summary):-

1. Perform QuEChERS extraction adapted for liquid extracts (acetonitrile partition, salts).

2. Clean up extract (dispersive SPE).

3. Analyze by GC-MS/MS and LC-MS/MS using a multi-residue method and compare to regulatory MRLs (or LOQs of method).

Acceptance:  non-detect or below applicable MRLs / certifier limits. For organic certification, many certifiers require residues below detection limits or within allowable traces — check certifier rules.

B . Heavy metals (Pb, Cd, Hg, As):-

Purpose:  ensure safety regarding heavy metal contamination.

Method:  acid digestion of sample and analysis by ICP-MS or AAS.

Procedure (summary):-

1. Digest measured aliquot (e.g., 5 mL) with concentrated HNO3 (and H2O2) in microwave digester.

2. Dilute to volume; analyze by ICP-MS or atomic absorption for Pb, Cd, As, Hg.

Acceptance (typical guidance):-

 keep as low as reasonably achievable; many herbal product guidelines suggest Pb ≤ 10 ppm, Cd ≤ 3 ppm, As ≤ 3 ppm, Hg ≤ 0.1–0.5 ppm — check local regulation and stricter organic certifier limits.

10. Stability Testing: -

Stability / accelerated testing (basic)

Purpose:  check how tincture holds up over time (color, aroma, potency, microbial).

Procedure:-

store aliquots at :- Room temp (25 °C),

Accelerated (40 °C ± 2 °C, 75% RH) for 1–3 months,

Refrigerated (4 °C).

At timepoints (0, 1, 3 months) test: appearance, pH, alcohol content, total phenolics, microbial. Monitor for unacceptable changes.

CONCLUSION: -

The prepared organic rose petal tincture using vodka as a solvent proved to be a simple, effective, and efficient extraction method for obtaining key phytoconstituents of rose petals. The maceration process enabled the transfer of essential oils, phenolics, flavonoids, and aromatic compounds into the alcohol medium, resulting in a potent and stable extract. Evaluation studies demonstrated favorable organoleptic, chemical, and microbiological profiles, supporting the tincture’s suitability for use in natural skincare, aromatherapy, and herbal wellness applications. The formulation also shows promise for incorporation into pharmaceutical preparations such as tonics, elixirs, and dermatological products. Future research should focus on optimizing extraction parameters, determining clinical efficacy, and establishing long-term stability and safety data.

Applications: -

1. Skincare & beauty as well as in acne treatment.
2. Herbal medicine as well as work as a digestive aid.

• Pharmaceutical uses: -

• Skincare (topical use):- Work as Astringents, Anti-inflammatory & Anti-microbial.
• Internal use (Herbal medicine):- Work as Digestive aid and mood support.(Note - For internal use only when it is prescribed by healthcare prescriber)
• Pharmaceutical preparations:- It is ingredients in elixirs & tonics.In compound preparations, In dermatologically basis & aromatherapeutic products.
• Preservations and solvent uses:- Solvent for active constituents and as a preservatives.

RESULT: -

1. Appearance:

The tincture exhibited a deep pink to reddish-amber color, depending on the rose variety. No suspended particulate matter was observed after filtration.

2. Aroma:

A strong floral, characteristic rose fragrance was recorded, indicating successful extraction of volatile oils.

3. Alcohol Content:

The final alcohol strength decreased slightly from the initial 40% v/v due to moisture in the petals, yielding ~32–38% v/v, which is suitable for preservation.

4. pH:

The tincture showed mild acidity (pH 4.2–5.1), aligning with typical herbal extracts and favoring microbial stability.

5. TPC (Total Phenolic Content):

The tincture demonstrated measurable phenolic content expressed as mg GAE/mL, confirming antioxidant potential.

6. TFC (Total Flavonoid Content):

The tincture showed significant flavonoid levels expressed as mg QE/mL, supporting anti-inflammatory activity.

7. TLC Fingerprinting:

TLC plates revealed multiple spots corresponding to rose phenolics and flavonoids—consistent with standard markers such as quercetin and gallic acid.

8. Microbial Safety:

Total microbial load remained within acceptable pharmacopeial limits, with no pathogenic organisms detected.

Overall Result:

A stable, aromatic, antioxidant-rich organic tincture was successfully formulated and met quality evaluation parameters.

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