INTRODUCTION:

Brucine, a naturally occurring alkaloid extracted from Strychnos nux-vomica, exhibits a numerous of pharmacological action, containing analgesic, anti-inflammatory, and anticancer activities. Despite its therapeutic potential, clinical application of Brucine is significantly restricted due its low water solubility, poor bioavailability, and potential systemic poisoning¹.

To overcome these barriers, advanced drug delivery systems are needed to enhance its solubility and therapeutic effectiveness. SNEDDS acquired attention as successful carriers for improving oral bioavailability of hydrophobic drugs. These systems are made up of oils, co-surfactants, and surfactants in isotropic mixtures that, when disclosed to gastrointestinal fluids, spontaneously emulsify into fine oil-in-water nanoemulsions². This mechanism facilitates drug solubilization, encourages lymphatic uptake, and reduces first-pass hepatic metabolism, thereby improving pharmacokinetic outcomes. The present study focused on developing and optimizing a Brucine-loaded SNEDDS to improve its solubility, physical stability, and bioavailability following oral administration³. Droplet size, polydispersity index (PDI), zeta potential, and formulation's in vitro drug release behavior were among metrics utilized to assess it. Furthermore, in vivo pharmacokinetic analysis had been performed to assess improvements in Brucine absorption. This work introduces a promising formulation strategy that could support the clinical advancement of Brucine for therapeutic use⁴.

MATERIALS AND METHODS:

Materials:

Brucine had been parched Yucca Enterprises, Wadala (E), Mumbai, Tween 80, PEG 400, Arachis oil was parches agrochemical.

Methods:

Brucine nano emulsions were formulated using an ultrasonication technique, adapted with slight modifications from the method described by Kaushal Kumar. Initially, a predetermined amount of Brucine was dissolved in accurately measured volumes of selected oils. Based on solubility screening, Arachis oil had been selected as phase of oil. Surfactant was Tween 80 served as surfactant, while PEG 400 was used as co-surfactant. To prepare oil phase, varying amounts of PEG 400 were added to drug-containing oil and mixed thoroughly under constant stirring. Meanwhile, aqueous phase had been prepared by dissolving Tween 80 in distilled water. This aqueous solution had been stirred on a magnetic stirrer, and oil phase had been added gradually, drop by drop, while maintaining constant stirring at 500rpm for 15 minutes. Resulting mixture was then subjected to ultrasonication for 20 minutes to obtain a stable nano emulsion^5,10.

Construction of Pseudo Ternary Phase Diagram:

Co-surfactants and Surfactants have been mixed in different proportions including 1:1, 2:1, 3:1, 1:2, and1:3, to prepare different surfactant mixtures (Smix). These Smix formulations were gradually incorporated into the oil phase with continuous stirring to ensure proper mixing. The resulting oil-to-Smix ratios were adjusted across a range of 9:1 to 1:9 (i.e., 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8, and 1:9). Every combination was titrated using distilled water, which was added incrementally at 0.5% (w/w) under consistent stirring to ensure uniformity. Following the achievement of equilibrium, the systems were aesthetically evaluated for transparency to identify clear and isotropic mixtures. Pseudo-ternary phase diagrams have been generated for each formulation utilization surfactant, oil, and co-surfactant combinations. These diagrams have been made with help of Chemix Tri-Plot software to determine the optimal nano emulsion regions^11,15.

Figure 1. Pseudo ternary phase diagrams of Brucine with excipients oil-Arachis, Surfactant- Tween 80 and co-surfactant – PEG400, Smix ratio 1:2 Smix indicates surfactant/ co-surfactant. Was selected ratio.

Design of Experiments:

Brucine Loaded-SNEDDS were formulated by the Box-Behnken models. The effect of Arachis oil, Tween 80 and PEG 400 concentration on the responses such as % Drug Release and Particle Size have been statistically analyzed by using design expert software ^16,20. And results were shows in tables 1.

Table 1. Design of experiments

Run	Factor 1	Factor 2	Factor 3	Response 1	Response 2
	A: Arachis oil	B: Tween80	C: PEG 400	Drug release %	Partical size nm
1	0.2	0.25	0.55	93.23	112
2	0.3	0.3	0.45	58.26	200
3	0.3	0.35	0.5	60.56	190
4	0.2	0.3	0.5	88.36	168
5	0.2	0.35	0.55	92.65	100
6	0.3	0.3	0.55	55.98	196
7	0.1	0.25	0.5	65.15	168
8	0.2	0.3	0.5	89.64	150
9	0.2	0.25	0.45	91.56	125
10	0.1	0.35	0.5	64.23	156
11	0.3	0.25	0.5	59.24	189
12	0.1	0.3	0.45	69.24	176
13	0.2	0.35	0.45	92.56	115

Drug Release % model:

Figure 2. Drug Release % model

Particle Size:

Figure 3. Particle Size

In ivitro isolation studies with Dialysis Bag:

Determined by Dialysis bag method in phosphate buffer pH 6.8 It had been discovered that release of brucine from brucine solution was 97.42±0.58 in 2hours and 86.89±0.65percent in 6hours. When compared to pure brucine, brucine exhibits a prolonged release; this could be because the brucine SNEDDS vesicle is encapsulated. Vesicular composition and encapsulation efficiency of SNEDDS may have an impact on slow diffusion of brucine from the SNEDDS vesicles. Due of its sustained release profile, SNEDDS is a promising drug molecule carrier system, particularly when it comes to treating chronic illnesses where long-term drug exposure is necessary for effectiveness³².

Particle size:

The size of particles is one of the key factors that determines the effectiveness of various drug delivery systems, absorption, distribution and also bioavailability. Usually, the particle size determines the amount of surface area where the drug will be absorbed, therefore increasing the bioavailability. The values of F1 at 112± 0.29nm confirm that they have excellent properties that might enhance absorption of drug. Lowered size of droplets ensures that Brucine dissolves and is absorbed more quickly²¹

Thermodynamic Stability Studies:

Developed SNEDDS formulations have been assessed for their physical stability through a series of stress tests, including “heat-cool cycles, centrifugation, and freeze thaw cycles. During each phase, physical characteristics of formulations have been visually” inspected²². For heat-cool stability test, the formulations underwent six alternating temperature cycles between 4°Celcius and 45°Celcius, with each temperature maintained for 48hours. Formulations that remained stable—without any signs of “phase separation, creaming, or cracking—were then subjected to centrifugation at 3500rpm for” 30minutes²³. Only formulations that successfully passed both the heat-cool and centrifugation tests were further evaluated using freeze-thaw cycles. In this step, the samples have been alternately stored at −21°Celcius and 25°Celcius, holding each temperature for 48 hours across 3 complete cycles²⁴.

Robustness to Dilution:

To mimic the dilution behaviour of the formulations under physiological conditions, the impact of dilution on emulsion properties was “examined. Distilled water, 0.1N hydrochloric acid, and phosphate buffer with a pH02 of 6.8 were 3 media employed to dilute 1mL of each formulation at ratios of 1:10, 1:100, and 1:1000 for this investigation. Diluted mixtures have been stirred employing “a magnetic stirrer at 100rpm and maintained at 37°Celcius to replicate body temperature, ensuring complete mixing and homogeneity. After mixing, samples have been kept at ambident temperature for 24hours, following which they were visually assessed for any signs of instability”, like phase separation²⁵.

Self-Emulsification Time:

For “this evaluation, a fixed volume (1mL) of each formulation have been mixed to 300mL of distilled water managed at 37±0.5° in a glass beaker. Gentle mixing was carried out employing a magnetic stirrer set at a constant speed of 100rpm. Emulsification time—defined” as duration needed for formulation to completely disperse and make a uniform nano emulsion was determined by visually tracking the point at which the SNEDDS fully dissolved and a clear or milky emulsion appeared²⁵.

Droplet Size Analysis and Polydispersibility Index (PDI) Determination:

Droplet size plays a crucial role in the self-emulsification process, as it directly influences drug's release rate and absorption efficiency. To evaluate this parameter, 1mL of each SNEDDS formulation had been diluted tenfold with distilled water prior to analysis. Resulting nano emulsions have been examined for droplet size and PDI using DLS (dynamic light scattering), employing “a photon correlation spectrometer (Zetasizer, Malvern Instruments Ltd., Malvern, UK). This approach measures variations in scattering of light caused by particles’ Brownian motion in suspension. Measurements were conducted at 25°C with a fixed scattering angle of 90°. Each sample had been analysed in triplicate, and outcome were expressed as the mean ± standard deviation” (SD)²⁶.

Zeta Potential Determination:

Zeta “potential of diluted SNEDDS formulations had been measured utilizing a Zetasizer (Malvern Instruments, UK). Each sample had been transferred into a clear disposable cuvette for analysis. Surface charge of emulsion droplets, along with their corresponding zeta potential” values, had been recorded to assess stability of nano emulsions²⁷.

Drug Loading Efficiency:

To quantify chlorzoxazone content, 1mL of each SNEDDS formulation (equivalent to 20mg of Brucine) had been transferred into a volumetric flask and diluted with methanol. Mixture had been thoroughly homogenized by “gently shaking or inverting flask 2-3 times. All samples have been prepared in triplicate. After appropriate dilution, absorbance was measured at 256nm utilizing a UV–Visible Spectrophotometer” (Hitachi U-2900, Tokyo, Japan). The Brucine concentration in each formulation had been determined utilizing a previously established calibration curve²⁸.

RESULTS AND DISCUSSION:

Melting Point:

Melting point observed in the range of 190-19°C by using capillary method²⁸.

Table 2: Determination of Meting point

Parameter	Observed value	Reference value	Inference
Melting Point	177°iC	175-178° C	Complies with the standard

UV Visible Spectrum of Brucine

Linearity curve was constructed in phosphate buffer at pH 7.4 at concentration rang of 2, 4, 5, 6, 8, 10 and it shows linear relationship with R² = 0.9948.

Figure 4. UV Visible Spectrum of Brucine at λ max 265nm

UV visible Spectrum:

Table 3. UV visible Spectrum of Brucine

	Observed value	Standard value
Spectrum (λ max.)	265 nm	265nm

Calibration Curve of Brucine:

a) Preparation of Standard Stock Solution:

10mg of Brucine precisely weighed, had been mixed to “a 100ml volumetric flask, and volume” had been increased Linearity curve had been constructed in phosphate buffer at pH7.4 at concentration rang of 2, 4, 5, 6, 8, 10 and it demonstrates linear relationship with R² =0.9948.

Figure 5. Calibration Curve of Brucine

B) Infrared Spectroscopy:

The FT-IR spectrophotometer (S) was used to conduct an IR spectroscopy analysis of Brucine, physical mixture of SNEDDS component and optimized Brucine loaded SNEDDS.iAi400-4000nm range using the KBr disc method. icm-1 wavelength range was covered by the spectra scan. FTIR analysis had been employed to examine structural properties of generated SNEDDS and compatibility of medicine with excipient²⁹.

A) pure Brucine

B) Physical mixture

C) Formulation of SNEDDS

Figure 6. A) pure Brucine B) Physical mixture C) Formulation of SNEDDS

C) Differential Scanning Calorimetry:

DSC “thermogram of pure Brucine shows a prominent endothermic peak at 181.99℃, confirming its purity. Melting peaks of brucine and excipients have been visible in physical mixture, however they somewhat shifted due to mixing. This indicated that there is no chemical interaction between” excipients and brucine³⁰.

A) pure Brucine

B) Physical mixture

C) SNEDDS Formulation

Figure 7. A) pure Brucine B) Physical mixture C) SNEDDS Formulation

Particle size and PDI:

For optimized batch particle size is 112.0nm and PDI is 0.162 Smaller particle size of SNEDDS improves bioavailability of drug by increasing penetration and cellular uptake of drug.PDI is heterogenicity index. PDI of 0.3 and below is considered to be acceptable in different vesicular drug delivery system³¹.

Figure 8. Praticle size and PDI

Zeta-Potential:

Increased negative charge may be decreasing propensity of SNEDDS to aggregate, increasing SNEDDS stability under storage conditions³². Optimized batch of Brucine loaded SNEDDS shows zeta potential -34.1

Figure 9. Zeta Potential

Thermodynamic Stability Studies, Dilution Study, Self-Emulsification time, Precipitation, Clarity:

F1 batch is considered as optimized batch based on Thermodynamic Stability Studies, Dilution Study, Self-Emulsification time, Precipitation, Clarity.

Table 4: Formulation and optimization results of Brucine SNEDDS

F. code	Brucine: oil: Suf: co- “s (%w/w/w)	Centrifuga-tion	Freeze thaw cycle	Distilled water	0.1 N HCL”	Phosphate Buffer pH	S.E.T.	Precipitation	Clarity
F1	10:0.2:0.25:0.55	pass	pass	Stable	Stable	Stable	25	Stable	Clear
F2	10:0.3:0.3:0.45	pass	pass	UnStable	UnStable	UnStable	35	UnStable	Bluish
F3	10:0.3:0.35:0.5	pass	pass	UnStable	UnStable	UnStable	41	UnStable	Turbid
F4	10:0.2:0.3:0.5	pass	pass	Stable	Stable	Stable	30	Stable	Clear
F5	10:0.2:0.35:0.55	pass	pass	UnStable	UnStable	UnStable	52	UnStable	Turbid
F6	10:0.3:0.3:0.55	pass	pass	UnStable	UnStable	UnStable	40	UnStable	Turbid
F7	10:0.1:0.25:0.5	pass	pass	Stable	Stable	Stable	37	Stable	Bluish
F8	10:1.0.2:0.3:0.5	pass	pass	UnStable	UnStable	UnStable	80	UnStable	Turbid
F9	10:0.2:0.25:0.45	pass	pass	UnStable	UnStable	UnStable	45	UnStable	Turbid
F10	10:0.1:0.35:0.5	pass	pass	UnStable	Stable	Stable	35	UnStable	Bluish
F11	10:0.3:0.25:0.5	pass	pass	unStable	Stable	Stable	41	UnStable	Clear
F12	10:0.1:0.3:0.45	pass	pass	Stable	Stable	Stable	30	Stable	Clear
F13	10:0.2:0.35:0.45	pass	pass	UnStable	UnStable	UnStable	52	UnStable	Turbid
F14	10:0.1:0.3:0.55	pass	pass	UnStable	UnStable	UnStable	40	UnStable	Turbid

The stability study of Brucine-loaded SNEDDS showed that SNEDDS stored at 4°C preserved their physical properties, Phase Separation, pH and entrapment efficiency with minimal changes. In contrast, SNEDDS stored at room temperature caused particle aggregation, slightly phase separation and entrapment efficiency, emphasizing the need for refrigeration to maintain stability.

DISCUSSION:

The formulation of a SNEDDS for Brucine was undertaken to address “its poor water solubility and limited oral bioavailability, which are further hindered by potential degradation within gastrointestinal tract. SNEDDS are composed of a surfactant, an oil phase, and a co-surfactant, which upon gentle agitation in gastrointestinal fluids, spontaneously produce fine oil-in-water nano emulsions. Chosen as lipid” component due to its high solubility potential for Brucine, Arachis oil is a long-chain triglyceride known for supporting substantial drug loading. It also promotes lymphatic absorption, which may help avoid hepatic first-pass metabolism. Its proven biocompatibility makes it a common choice in lipid-based oral delivery systems. Selected as the surfactant, “Tween 80 has a high hydrophilic-lipophilic balance (HLB ~15), which Favors creation of oil-in-water emulsions”. It effectively lowers interfacial tension, thereby enhancing the dispersion of oil droplets in aqueous environments and facilitating the formation of nano-sized emulsions. Serving as the co-surfactant, PEG 400 aids in improving the elasticity of interfacial film and contributes further to reduction of interfacial tension. Its presence enhances emulsification efficiency and aids in development of stable, transparent nano emulsion with reduced droplet size. Finalized SNEDDS formulations comprising Arachis oil, Tween 80, and PEG 400 rapidly emulsified upon dilution in aqueous media, producing stable nano emulsions with droplet sizes generally below 100nm. These systems exhibited clear to slightly bluish appearances, reflecting efficient self-emulsification and fine dispersion at the nanoscale.

This delivery strategy markedly enhanced the dissolution rate of Brucine—a crucial parameter for effective absorption. Small droplet size offers a large surface area for drug release, while lipid-based matrix may shield drug from enzymatic degradation. Furthermore, system may facilitate lymphatic uptake, potentially leading to improved systemic bioavailability.

CONCLUSION:

Prepared Brucine SNEDDS successfully demonstrated their potential as an effective drug delivery system for Brucine, offering improved stability and improved therapeutic properties. Favorable physicochemical properties of SNEDDS formulation, such as ideal particle size and high encapsulation effectiveness, contributed to enhanced bioavailability and delayed release. Stability studies revealed that Brucine SNEDDS maintained their integrity and efficacy over time at 4⁰C.This research highlights potential of brucine SNEDDS as an advanced delivery system that significantly enhances therapeutic properties of Brucine.

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Received on 24.03.2026 Revised on 21.04.2026

Accepted on 15.05.2026 Published on 07.07.2026

Available online from July 10, 2026

Res. J. Pharma. Dosage Forms and Tech.2026; 18(3):185-192.

DOI: 10.52711/0975-4377.2026.00028

This work is licensed under a Creative Commons Attribution-Non Commercial-Share Alike 4.0 International License. Creative Commons License.

Sr. No.	F. code	Brucine: oil: Suf:co-s (%w/w/w	% Drug Content	Particle Size (nm)	PDI	Zeta potential (mv)	EE (%)
1.	F1	0.2:0.25:0.55	88.86 ± 0.39	112	0.162	-34.1	88.44 ± 2.16
2.	F2	0.2:0.35:0.55	79.74 ± 0.17	141.1	0.098	-26.10	64.18± 1.29
3.	F3	0.2:0.35:0.45	62.26 ± 0.47	137.2	0.078	-24.8	52.37 ± 2.87
4.	F4	0.2:0.3:0.5	43.06 ± 0.58	125.8	0.380	-20.0	49.2 ± 1.74
5.	F5	0.2:0.3:0.5	53.86 ± 0.78	115.45	0.260	-27.0	63.2 ± 1.74

Parameter	4^oC			RT
Parameter	0day	30day	90day	0day	30day	90day
Physical Appearance	Good	Good	Good	Good	Redispersible flocculation	Redispersible flocculation

Parameter	4^oC			RT
Parameter	0day	30day	90day	0day	30day	90day
Physical Appearance	Good	Good	Good	Good	Redispersal flocculation	Redispersal flocculation
Phase Separation	No	No	No	No	Slightly phase Separation	slightly phase separation
pH	5.9	5.9	5.8	5.9	6.0	6.5
%EE	97.33	96.12	96.00	97.33	90.14	88.23