Preparation, Characterization, and Pharmacokinetic Evaluation of BerbiQ: An Advanced Bioavailable Berberine Formulation Using OMICS Technology by Synergistic Complexation

Abstract

A novel bioavailable berberine formulation, BerbiQ, was developed using OMICS technology by complexing berberine hydrochloride with synergistic molecules, particularly silymarin, to enhance its therapeutic efficacy. The formulation incorporated coconut milk containing proteins and lipids through an advanced bionanotechnology approach. Morphological analysis via scanning electron microscope (SEM) and transmission electron microscopy (TEM) confirmed that BerbiQ consists of spherical, well-dispersed particles with smooth surfaces and no aggregation, indicative of successful berberine complexation. Additional characterization using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and differential scanning calorimetry (DSC) analyses, along with stability studies, validated the effective complexation and stability of berberine hydrochloride within the BerbiQ formulation. Pharmacokinetic studies revealed that BerbiQ significantly enhances the oral bioavailability of berberine compared to conventional formulations, demonstrating a 4.26-fold increase in area under the curve (AUC) and a 4.10-fold increase in maximum plasma concentration (C _max). Moreover, BerbiQ exhibited a shorter time to reach the maximum plasma concentration (T _max), a lower elimination rate constant (K _el), and an extended terminal half-life (t _1/2), indicating sustained release and prolonged systemic availability of berberine. These improvements are attributed to the advanced complexation and sustained-release properties enabled by OMICS technology. The enhanced bioavailability and pharmacokinetic profile of BerbiQ suggest it as an efficient delivery system for berberine, potentially reducing the required dosage and minimizing side effects. These findings position BerbiQ as a promising candidate for various therapeutic applications, offering improved efficacy and therapeutic potential.

1. Introduction

Herbal medicines are widely used in traditional systems due to their rich phytochemical diversity and therapeutic potential. Natural products, such as plant-derived alkaloids and flavonoids, offer broad pharmacological effects with fewer side effects, making them attractive candidates for drug development. − Berberine is a bioactive alkaloid present in several medicinal plants, notably Berberis aristata, Berberis aquifolium, Coptis rhizome, Coptis chinensis, and Coptis japonica, among others. It is primarily localized in the roots, rhizomes, stems, barks, fruits, and to some extent in the leaves of these plants. Widely used in Chinese, Unani, and Indian Ayurvedic medicine, berberine is also employed in traditional medicine across South Asia, Europe, and America. It is characterized as a yellow, odorless, and bitter-tasting crystalline powder. Berberine has been indexed in the Medical Subject Headings (MeSH) since 1975 and has recently attracted significant interest from the Western scientific community. , It exhibits a broad spectrum of pharmacological activities, including antitumor, antibiotic, antioxidant, anti-inflammatory, anticancer, antimicrobial, antiviral, antidepressant, antiplasmodial, antidiarrheal, cardiovascular, antidiabetic, hepatoprotective, neuroprotective, memory-enhancing, and immune-modulating properties. −

Despite its potent bioactivity and low toxicity, berberine suffers from extremely poor oral bioavailability (<1%) and gastrointestinal side effects. This is mainly due to its low aqueous solubility, poor intestinal absorption, first-pass metabolism, and efflux by P-glycoprotein (P-gp). − To address the challenges associated with oral administration of berberine and enhance its bioavailability, various dosage forms have been developed. Advanced drug delivery systems have been explored, including polymeric nanoparticles, silica-based nanoparticles, dendrimers, micelles, liposomes, graphene-based carriers, and lipid-based nanostructures, as well as metal-based carbon nanoparticles. − These systems utilize diverse technologies such as nanoprecipitation, ionic complexation, emulsion solvent evaporation, salting-out, and nanoencapsulation. , However, many of these systems face challenges such as low drug loading, instability in gastric conditions, or limited food/nutraceutical applicability. Therefore, sustainable and food-compatible delivery systems are critically needed. −

Biodegradable carriers, particularly protein-based systems, have shown promise in enhancing the solubility and stability of poorly soluble drugs. , Coconut milk-derived protein is an underutilized byproduct rich in essential amino acids (71–77%), lipids (3–4.8%), and emulsifying properties, making it suitable for encapsulation. − The amphiphilic nature of coconut protein enables complexation with bioactives, while the medium-chain fatty acids in coconut lipids facilitate micelle formation and absorption in the gastrointestinal tract. −

Silymarin, a flavonoid complex composed of silybin, silydianin, and silychristin extracted from the seeds of Silybum marianum (milk thistle), is well-known for its hepatoprotective properties but is correspondingly notable intended for its P-gp inhibitory activity. The coadministration of silymarin with berberine has been shown to enhance beneficial effects of berberine on lipid and glucose metabolism in humans. This combination allows for lower doses of berberine to be used, thereby reducing the risk of gastrointestinal discomfort. , It is hypothesized that silymarin primarily improves the oral bioavailability of berberine through direct interaction with P-gp, rather than exerting a significant independent effect on glucose and lipid metabolism.

In the present study, Molecules Biolabs designed and developed a bioavailable berberine formulation, commercially known as BerbiQ based on the complexation of berberine hydrochloride with synergistic molecules particularly silymarin through a fascinating concept, OMICS technology to enhance the efficacy of the berberine and process with coconut milk containing protein and lipids using advanced bionanotechnology process. OMICS technologies can be employed to preserve the functional properties of bioactive compounds, enhance their stability, improve bioavailability, optimize health benefits, and enable controlled release at specific targets and time points. OMICS technologies are regarded as among the most promising approaches for improving the dissolution of poorly soluble berberine. Its simplicity, cost-effectiveness, and commercial viability make it an attractive option for industrial-scale production. The characterization of BerbiQ was further analyzed using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscope (SEM), transmission electron microscopy (TEM), differential scanning calorimetry (DSC), stability tests, and various physicochemical parameters to assess its suitability as a nutraceutical and dietary supplement. Additionally, the study evaluated the oral bioavailability of berberine by comparing the administration of BerbiQ and conventional berberine in a randomized, double-blind, single-dose, two-period, two-way crossover design involving healthy adult human subjects under fasting conditions. This research aims to provide an efficient and scalable delivery system for berberine as a nutraceutical or dietary supplement.

2. Materials and Methods

2.1. Materials

Roots of B. aristata were procured from Natura Herbal Ashram LLP, Jaipur, Rajesthan, India, and seeds of S. marianum (milk thistle) were procured from The Hill Herbs India, Uttarakhand, India, which were authenticated by a local botanist; however, no voucher specimens were retained. Coconut milk powder (containing ∼56% fat and 8% protein) was purchased form Kaira Organic World, Kanniyakumari, Tamil Nadu. Organic solvents used in the study were of HPLC and LC-MS grade and were procured from Merck India. Millipore Milli-Q distilled water was employed throughout the entire experimental process.

2.2. Method of Preparation of BerbiQ Using OMICS Technology

2.2.1. Extraction and Purification of Berberine and Preparation of Berberine Hydrochloride

Roots of B. aristata were thoroughly shade-dried, which were coarsely powdered using a mechanical grinder and sieved to obtain a uniform particle size suitable for solvent extraction. The powdered root material was subjected to maceration using 80% ethanol as the extraction solvent in a plant material-to solvent ratio of 1:5 (w/v). The extraction was carried out at 65 °C for 8 h per cycle, and repeated for three successive cycles to ensure exhaustive extraction. All ethanolic extracts were pooled and concentrated under reduced pressure using a rotary evaporator (Rotary Evap, Rotary Vacuum Film Evaporator) at 45 °C to remove the solvent and yield a thick, crude extract containing berberine alkaloid.

The resulting crude berberine was weighted and transferred to a reaction flask, which was solubilized in methanol in a 1:10 (w/v) ratio, followed by continuous stirring at 5 °C for 30 min. The mixture was then filtered to remove insoluble impurities. The filtrate was concentrated to dryness and the residue was dissolved in distilled water. To this aqueous solution 10% hydrochloric acid was added dropwise at 5 °C, in a quantity equivalent to twice the weight of the initial crude berberine. The mixture was stirred continuously until yellow crystalline berberine hydrocholoride began to precipitate. The precipitated crystals were collected by filtration, washed thoroughly with chilled ethanol to remove unreacted components and impurities, and dried in a hot-air oven at 45 °C. The dried berberine hydrochloride crystals were stored in a desiccator.

2.2.2. Water Extract of B. aristata Roots

The uniformly powdered B. aristata root was soaked in 1 L of distilled water (1:10 w/v) and heated at 100 °C for 2 h with occasional stirring to facilitate extraction of water-soluble phytoconstituents. After heating, the decoction was allowed to cool to room temperature, and subsequently filtered to remove coarse plant debris. The filtrate was subsequently centrifuged at 5000 rpm for 10 min to remove any remaining fine particulate matter. The clear supernatant was collected and concentrated under reduced pressure using a rotary evaporator at 40 °C. The semisolid residue was then lyophilized to obtain a dry powdered aqueous extract of B. aristata roots, which was stored in an airtight container at 4 °C.

2.2.3. Extraction and Purification of Silymarin

The milk thistle seeds were cleaned to remove extraneous material and then dried in a hot-air oven at 40 ± 2 °C for 24 h to reduce moisture content. The dried seeds were then milled to a coarse powder using a laboratory grinder and passed through a 40-mesh sieve to ensure unform particle size. The powdered seed material was extracted with 70% (v/v) ethanol (solvent-to-solvent ratio 1:20 w/v). The mixture was subjected to maceration on an orbital shaker at 150 rpm and maintained at 25 ± 2 °C for 24 h to facilitate the extraction of flavonolignans. Then, the mixture was filtered, and residual plant materials was subjected to a second round of extraction under identical conditions to ensure maximal recovery of silymarin. The filtrates from both extractions were combined. The pooled ethanolic extracts were concentrated using a rotary evaporator at 40 °C under reduced pressure to remove ethanol. This resulted in a thick, silymarin-rich semisolid extract, which was further dried under vacuum and stored at 4 °C.

2.2.4. Preparation of BerbiQ Using OMICS Technology

The development of BerbiQ, a bioavailable berberine formulation, was achieved through the application of OMICS technology, an integrated bioprocessing approach designed to enhance the solubility, stability, and intestinal absorption of plant-based actives. This advanced bionanotechnology platform, supported by OMICS data sets, allowed for precise modulation of particle size, encapsulation efficiency, and surface characteristics to yield a stable, bioavailable BerbiQ complex suitable for oral administration. This approach combines nanomilling, synergistic bioactive complexation, and lipid–protein encapsulation to address the core pharmacokinetic challenges associated with berberine, such as poor water solubility, limited intestinal permeability, and high first-pass metabolism.

To begin with, berberine hydrochloride (85 g) was dissolved in 1 L of ultrapure Millipore water and subjected to nanomilling using a Dyno-agitator bead mill. This process reduced the particle size of berberine hydrochloride to below 200 nm, dramatically improving its aqueous dispersibility and surface area, thus setting the stage for betting intestinal interaction. Following this, 5 g of water extract of B. aristata root containing a natural matrix of alkaloids and supportive phytochemicals was incorporated into the berberine hydrochloride solution under constant stirring. This combination was again nanomilled to promote intermolecular interaction, the resulting dispersion, referred to as the berberine whole matrix (BWM).

In parallel, 5 g of purified silymarin extract (a flavonolignan complex known for its hepatoprotective and bioavailability-enhancing effects) was dispersed in 1 L of Millipore water and nanomilled to achieve a uniform submicron suspension. To this, 5 g of coconut milk powder was added, which serves a dual function; the lipid fraction acts as a natural encapsulating agent that facilitates lipophilic transport across intestinal membranes, while milk proteins aid in the stabilization of the bioactive complex through hydrogen bonding and hydrophobic interactions. The coconut-silymarin mixture was again nanomilled to form a stable dispersion.

The two dispersions, BWM and the silymarin-coconut milk complex were then combined and stirred gently for 30 min to allow proper integration of all molecular components. The final mixture underwent a third and final nanomilling process to ensure a homogeneous nanosuspension with a well-integrated phytochemical profile. This nanosuspension was subsequently spray-dried using a Spray Tech Systems Mini Laboratory Spray Drier (STS001-SS) under controlled conditions: an inlet temperature of 175–185 °C, an outlet temperature of 75–85 °C, and a during air pressure of 10 kg/cm². The process yielded a dry, flowable powder rich in bioavailable berberine complexes. The resulting formulation, BerbiQ was collected and stored in amber-colored, airtight glass containers at room temperature to preserve chemical stability and photoprotection. The innovative application of OMICS technology allowed for the preservation, protection, and potentiation of pharmacological activities of berberine by improving its molecular compatibility with the gastrointestinal environment, significantly enhancing its potential for absorption and therapeutic efficacy. A schematic representation of the preparation steps is presented in Figure .

1.

Schematic representation of preparation of BerbiQ by OMICS technology.

2.3. Physical Parameters

2.3.1. Water Solubility Index (WSI) and Bulk Density

The water solubility index (WSI) of berberine hydrochloride and BerbiQ was determined following the method described by Amalraj et al. Briefly, 2.5 g of each sample was mixed with 30 mL of distilled water in a 100 mL centrifuge tube and vigorously shaken. The mixture was then incubated in a water bath at 37 °C for 30 min followed by centrifugation at 10,000 rpm for 30 min. The resulting supernatant was carefully transferred to preweighted Petri dishes and dried in an oven at 103 ± 2 °C. The WSI (%) was calculated as the percentage of the dried supernatant relative to the initial 2.5 g of sample.

Bulk density (g/mL) was measured by transferring 10 g of the sample into a clean, dry 100 mL graduated cylinder. The cylinder was mounted on a ring stand adjusted such that its base was positioned precisely one inch above a solid platform, allowing the sample to settle by gravity. The volume occupied by the sample was recorded without applying external compaction. Bulk density was calculated as the ratio of the sample mass to the volume it occupied in the cylinder.

2.3.2. Hygroscopicity and Moisture Content

The hygroscopic behavior of berberine hydrochloride and BerbiQ was evaluated by uniformly distributing 1 g of each powder sample across Petri dishes to maximize the surface area exposed to ambient humidity. The dishes were placed in a desiccator maintained at 23 °C and 76% relative humidity, the latter controlled using a nitric acid solution. After 90 min of exposure, the samples exhibited a measurable increase in mass due to moisture absorption. Although hygroscopicity is typically characterized by equilibrium moisture content, a comparative analysis was conducted by quantifying the moisture uptake per gram of drying powder following the 90 min exposure period at 76% relative humidity.

Moisture content was determined by the loss on drying (LOD) method. Prior to use, LOD bottles were predried in a hot air oven at 105 °C for 60 min and then allowed to cool in a desiccator at room temperature for 30 min. Subsequently, 1 g of each sample was accurately weighted into the preconditioned LOD bottles. The samples were then dried in the over at 105 °C for 5 h. After drying, the bottles were transferred to the desiccator and cooled for 30 min at room temperature before reweighing. The percentage of moisture content was calculated using eq , based on the weight loss before and after drying.

$$\mathrm{moisture}\mathrm{content}(\%)=\frac{W_{\mathrm{I}}-W_{\mathrm{F}-}}{W_{\mathrm{I}}}\times 100$$ 1

where W _I is the initial weight of the sample and W _F is the final weight of the sample

2.3.3. Degree of Caking (DC)

To evaluate the degree of caking, powder samples were first dried in a hot air oven at 70 °C and then allowed to cool to room temperature. The dried samples were accurately weighted and subsequently transferred to a 420 mm sieve. The sieving was performed for 5 min using a mechanical shaker to ensure uniform agitation. Following sieving, the mass of powder retained on the sieve was recorded. The caking index was then calculated using eq , based on the proportion of material retained relative to the total initial sample weight.

$$\mathrm{DC}(\%)=\frac{b}{a}\times 100$$ 2

where DC represents the degree of caking (%), ‘a’ is the initial amount of powder used in the sieving process, and ‘b’ is the amount of powder remaining on the sieve after sieving.

2.4. Analysis and Characterization of BerbiQ

2.4.1. Analysis of Berberine

The berberine content was determined using Ultra Performance Liquid Chromatography (UPLC), performed on a Shimadzu Prominence P-Series HPLC system equipped with a quaternary gradient pump, autosampler, photodiode array (PDA) detector and controlled by LabSolutions chromatography software (Shimadzu Corp., Japan). The chromatographic separation was achieved using an Eclipse Plus C18 column (5 μm, 250 mm × 4.6 mm). The mobile phase comprised, Solvent A: 0.136 g of potassium dihydrogen phosphate (KH₂PO₄) dissolved in 100 mL of HPLC grade water, with pH adjusted to 2.5 using orthophosphoric acid and Solvent B: HPLC grade acetonitrile. The elusion was carried out using the following gradient program, 0.0–5.0 min elution with 50:50 (v/v) of A/B, and 5.0–10.0 min linear gradient to 60:40 (v/v) of A/B. The flow rate was maintained at 1.0 mL/min, and the column temperature was set to 32 °C and the injection volume was 20 μL. Berberine was detected using a PDA detector at a wavelength of 346 nm, with the retention time of 7.3 min.

2.4.2. Fourier Transform Infrared Spectroscopy (FT-IR) Analysis

The FTIR-ATR spectra, obtained with 64 scans per sample, were recorded by the JASCO ATR-FT/IR-4700 for berberine hydrochloride and BerbiQ in the 400 to 4000 cm^–1 range.

2.4.3. X-ray Diffraction (XRD) Analysis

The crystalline nature of berberine hydrochloride and BerbiQ was analyzed using powder X-ray diffraction (Bruker D8 Focus) at ambient temperature. The measurements were performed with Cu Kα radiation (λ = 0.1541 nm) over a 2θ range of 4 to 80°, with a scanning speed of 0.08° per minute. Prior to analysis, the samples were vacuum-dried at 60 °C.

2.4.4. Scanning Electron Microscope (SEM) Analysis

The structural morphology of BerbiQ was examined using a scanning electron microscope (Vega3Tescan, Germany). The sample was analyzed at an accelerated voltage of 5 kV, mounted on aluminum stubs with double-sided carbon tape, and sputter-coated with a thin layer of gold using a sputter gold coater.

2.4.5. Transmission Electron Microscopy (TEM) Analysis

The morphology of BerbiQ was evaluated using a transmission electron microscope (TEM, JEM-F200, JEOL, Japan). The sample was subjected to 10 min of sonication, and a few droplets of BerbiQ were placed on a glow-discharged, carbon-coated TEM grid and allowed to dry at ambient temperature.

2.4.6. Differential Scanning Calorimetry (DSC)

The thermal stability behavior of berberine hydrochloride and BerbiQ was assessed using a differential scanning calorimeter (DSC, DSC822e, Mettler Toledo). Approximately 10 mg of the sample was placed in a hermetically sealed aluminum pan and heated at a rate of 2 °C/min from 30 to 400 °C, with nitrogen continuously purged at a flow rate of 40 mL/min. The resulting data were analyzed using TA STARe software.

2.5. Storage Stability

To evaluate the stability of BerbiQ, three distinct temperature settings (4, 25, and 45 °C) were applied over a period of 180 days. Samples were collected at specific intervals, and their berberine concentrations were measured both before and after storage. Each measurement was performed in triplicate. The stability of berberine was calculated using eq .

$$\mathrm{stability}\mathrm{of}\mathrm{berberine}(\%)=\frac{B_{\mathrm{t}}}{B_0}\times 100$$ 3

where B ₀ is the initial concentration of berberine, and B _t represents the concentration of berberine at specific time points during the sampling period.

2.6. Bioavailability Study

2.6.1. Study Design

The study was conducted as a randomized, double-blind, single-dose, two-period, two-way crossover trial involving healthy adult human subjects under fasting conditions. A total of ten healthy participants, aged 18 to 45, who met the inclusion and exclusion criteria, were recruited for the study. The participants were randomly assigned in a 1:1 ratio (N = 5 per group) to receive either berberine or BerbiQ, and subsequently crossed over to the second treatment group after a seven-day washout period. The randomization process was performed using a chart developed by a statistical expert.

2.6.2. Ethical Approval

The study protocol was authorized by a registered ethical committee (Reg. No. ECR/141/Indt/KA/2013/RR-19). The study was conducted in accordance with multiple ethical and regulatory guidelines, including the ICMR Ethical Guidelines for Biomedical Research on Human Participants (2006), ICH (Step 5) Guidance on Good Clinical Practice, Schedule Y (amended version, 2014) of the Central Drugs Standard Control Organization (CDSCO), as well as the standards of Good Laboratory Practice (GLP) and Good Clinical Practice (GCP) for Clinical Research in India. Additionally, the study adhered to the Good Clinical Laboratory Practice (GCLP) guidelines, the Declaration of Helsinki (Fortaleza, October 2013), and the 21 CFR Parts 50, 56, and 320 USFDA guidelines for industry, along with other relevant regulatory requirements. The investigator explained the subjects in understandable language before initiation of study regarding the purpose, procedure and potential hazards and rights of the subjects. All the subjects were provided with written and signed formed consent. The protocol was registered with Clinical Trials Registry India (clinicaltrials.gov) (CTRI/2024/07/070781).

2.6.3. Intervention and Dosage

The bioavailable berberine formulation (BerbiQ) and conventional berberine were provided by Molecules Biolabs Private Limited. To assess bioavailability, human volunteers underwent a minimum 10 h fasting period before treatment. Subjects were then orally administered a single dose of 350 mg capsules containing 250 mg of either BerbiQ or conventional berberine, as per the randomization procedure, along with 250 mL of water. A mouth check was conducted to verify compliance with the administration.

2.6.4. Sample Collection

Blood samples were collected through an indwelling intravenous cannula inserted into a vein in the forearm. A 5 mL blood sample was taken 1 h before dosing for the predose phase, and subsequent samples were collected at 0.5, 1, 2, 6, 12, 24, and 48 h postdosing. A total of eight 5 mL blood samples were collected into prelabeled K2EDTA vacutainers. Heparin-lock was employed to prevent blood clotting in the indwelling cannula. Prior to each sample collection, 0.5 mL blood was discarded to eliminate any potential interference from saline and heparin. The total volume of blood collected, including the 12 mL required for screening and the 0.5 mL discarded before each sample, did not exceed 86 mL per subject over the course of the study. No additional blood samples were taken for repeat laboratory analyses.

2.6.5. Sample Separation and Preparation

All blood samples were initially stored in a thermo-insulated box with wet ice following collection and subsequently transferred to the sample processing room. There, they were centrifuged at 4000 ± 50 rpm for 10 min at temperatures ranging from 2 to 8 °C to separate the plasma. The isolated plasma was then evenly distributed into appropriately labeled polypropylene tubes.

2.6.6. Sample Preparation and Quantification of Berberine in Plasma

The separated plasma samples were mixed with 1 mL of acetonitrile containing 3% acetic acid, vortexed for 5 min, and then centrifuged for 10 min. The supernatant was evaporated under nitrogen, and subsequently, 1 mL of methanol was added, followed by vortexing. The samples were then filtered immediately through a 0.2 μm filter and analyzed using UPLC.

2.7. Statistical Analysis

All statistical analyses were performed using SPSS version 16.0. Data are presented as mean ± SD, and analysis of variance (ANOVA) was employed for data evaluation. Pairwise differences were assessed using a paired t test, with statistical significance set at P < 0.05.

3. Results and Discussion

3.1. Physical Parameters

The berberine used for comparison in this study was purified berberine hydrochloride, obtained from B. aristata root extract and standardized to ∼98% purity. This unformulated berberine hydrochloride served as a baseline reference to evaluate the impact of formulation through OMICS technology.

3.1.1. Water Solubility Index and Bulk Density

The water solubility index (WSI) of purified berberine hydrochloride and the formulated BerbiQ was assessed to evaluate the enhancement in aqueous dispersibility. As shown in Table , berberine hydrochloride exhibited a WSI of 7.11  ±  1.46%, while the BerbiQ formulation showed a significantly improved WSI of 12.96  ±  2.58%, indicating a nearly 2-fold increase in solubility. This enhancement may be attributed to the advanced OMICS technology-based complexation, where berberine hydrochloride was synergistically integrated with silymarin and coconut milk powder. The lipids and proteins in coconut milk, along with the nanomilled silymarin extract, likely facilitated hydrogen bonding, hydrophobic interactions, and encapsulation, thus promoting greater molecular dispersion and solubility in aqueous systems. While berberine hydrochloride already possesses better solubility than the free base form, the observed increase in WSI of BerbiQ highlights the additional contribution of lipid–protein encapsulation and nanoparticle stabilization in the final product.

1. Physical Properties of Berberine Hydrochloride and BerbiQ.

parameters	berberine hydrochloride	BerbiQ
water solubility index (%)	7.11 ± 1.46	12.96 ± 2.58
bulk density (g/mL)	0.47 ± 0.08	0.39 ± 0.06
moisture content (%)	4.21 ± 0.82	1.28 ± 0.27
hygroscopicity (g/g)	0.23 ± 0.05	0.58 ± 0.14
degree of caking (%)	1.45 ± 0.34	7.74 ± 1.42

Bulk density is a critical property of powder and granular products, influencing storage efficiency and ease of transportation. The lower bulk density observed in BerbiQ can be attributed to the presence of coconut milk proteins and lipids, which are comparatively bigger than berberine hydrochloride. BerbiQ demonstrated a lower bulk density (0.39 g/mL) compared to berberine hydrochloride (0.47 g/mL) while exhibiting a higher water solubility index (Table ). The observed increase in soluble content was associated with a decrease in bulk density, highlighting an inverse relationship between bulk density and solubility. ,

3.1.2. Moisture Content and Hygroscopicity

The moisture content of purified berberine hydrochloride and the formulated BerbiQ was found to be 1.28 and 4.21%, respectively (Table ). The increased moisture content in BerbiQ is attributed to the presence of hydrophilic components such as coconut milk proteins and lipids, which are known to interact with water molecules and retain ambient moisture. Similarly, hygroscopicity, a critical indicator of powder stability and storage sensitivity, was markedly higher in BerbiQ (0.58 g/g) compared to berberine hydrochloride (0.23 g/g). This result is consistent with the greater moisture-holding capacity of amphiphilic biomacromolecules present in coconut milk powder used during the OMICS-based complexation process.

3.1.3. Degree of Caking

The degree of caking was found to be higher for BerbiQ (7.44%) compared to berberine hydrochloride (3.45%) (Table ). The increased caking tendency observed in BerbiQ is primarily attributed to the inclusion of coconut milk proteins and lipids, which are known to be hydrophilic and capable of absorbing ambient moisture. , These components facilitate the formation of liquid bridges between particles upon moisture adsorption, leading to particle adhesion and aggregation. Additionally, the incorporation of high molecular weight synergistic molecules such as silymarin and the processing steps involved in nanomilling and encapsulation contribute to changes in particle surface energy and flowability. Since the degree of caking is closely related to both moisture content and hygroscopicity, the elevated caking behavior of BerbiQ is consistent with its higher moisture content (4.2%) and hygroscopicity (0.58 g/g) compared to berberine hydrochloride.

3.2. Characterization of BerbiQ

3.2.1. FT-IR Studies

The FT-IR spectra of berberine hydrochloride and BerbiQ (Figure ) were analyzed, and the major absorption bands and their corresponding functional group assignments are summarized in Table . This comparative analysis confirms that the characteristic functional groups of berberine hydrochloride are preserved in BerbiQ, with minor shifts indicating hydrogen bonding and successful complexation mediated by OMICS technology. In the FT-IR spectrum of berberine hydrochloride (Figure (a)), a peak observed at 710 cm^–1 corresponds to the disubstituted aromatic ring, while the peaks at 896 and 975 cm^–1 are attributed to the aromatic cyclic ether and cyclic ether groups of the berberine hydrochloride molecule, respectively. A peak at 924 cm^–1 corresponds to the deformation of the aromatic carbon–hydrogen group. The presence of an aromatic oxide group (Arc-O–CH₃) in berberine hydrochloride is confirmed by two strong symmetric peaks at 1032 and 1062 cm^–1, along with two strong asymmetric peaks at 1226 and 1269 cm^–1. Five prominent and sharp peaks observed at 1362, 1387, 1502, 1566, and 1598 cm^–1 are attributed to the nitrogen-containing six-membered heteroaromatic structure of berberine hydrochloride. Additionally, a peak at 1458 cm^–1 corresponds to the bending vibration of the asymmetric −CH₃ group. The absorption bands observed at 1619 and 1632 cm^–1 correspond to the stretching vibrations of the quaternary iminium ion (–CN–) and heterocyclic amine (C–N) groups, respectively, from the aromatic ring of berberine. The peak at 2845 cm^–1 corresponds to the aliphatic C–H stretching of the methoxy group, while two additional peaks at 2911 and 2946 cm^–1 are attributed to the C–H stretching vibrations of alkanes. The large and broad peak observed between 3100 and 3600 cm^–1 corresponds to the hydroxyl (O–H) functional groups in alcohol and phenol derivatives. However, in BerbiQ (Figure (b)), this band became broader (3090 to 3680 cm^–1), suggesting that the band is not solely attributed to phenolic or hydroxyl derivatives, but also to hydrogen bonding interactions between berberine hydrochloride and the coconut proteins and lipids. Furthermore, the FTIR spectra of the prepared BerbiQ exhibited characteristic peaks closely resembling those of berberine hydrochloride, thereby confirming the effective complexation of berberine hydrochloride within the BerbiQ formulation via hydrogen bonding, facilitated by OMICS technology.

2.

FT-IR spectra of (a) berberine hydrochloride (b) BerbiQ.

2. FT-IR Spectral Assignments of Berberine Hydrochloride and BerbiQ.

wavenumber (cm–1)	functional group/vibration	assignment description	observed in
2946, 2911, 2845	C–H stretching (alkanes and methoxy)	aliphatic C–H and OCH3 groups	berberine hydrochloride
2943, 2914, 2844	C–H stretching (alkanes and methoxy)	similar bands retained in complex	BerbiQ
3100–3600	O–H stretching (broad)	phenols/alcohols	berberine hydrochloride
3090–3680	O–H stretching (broadened)	hydrogen bonding in complex	BerbiQ
1632, 1619	CN and C–N stretching	quaternary iminium and heterocyclic amine groups	berberine hydrochloride
1633, 1622	CN and C–N stretching	slightly shifted due to complexation	BerbiQ
1598, 1566, 1502	aromatic CC/heteroaromatic ring stretching	nitrogen-containing six-membered ring	berberine hydrochloride
1595, 1567, 1503	aromatic CC/heteroaromatic ring stretching	retained in BerbiQ	BerbiQ
1458	–CH3 asymmetric bending	methyl group bending vibration	both
1387, 1362	C–N/CH3 bending	heteroaromatic contributions	both
1269, 1226	asymmetric C–O–C stretching	aromatic oxide (ArC–OCH3)	berberine hydrochloride
1270, 1228	asymmetric C–O–C stretching	retained with minor shift in BerbiQ	BerbiQ
1103, 1140	C–O–C stretching (ethers)	cyclic/aromatic ether groups	berberine hydrochloride
1104, 1123	C–O–C stretching	present in BerbiQ with similar patterns	BerbiQ
1032, 1062	symmetric C–O–C (ether)	aromatic oxide group (ArC–OCH3)	berberine hydrochloride
1032, 974	symmetric C–O–C (ether)	retained with minor variation	BerbiQ
924	aromatic C–H deformation	substituted aromatic ring	berberine hydrochloride
923	aromatic C–H deformation	slightly shifted	BerbiQ
896, 975	cyclic ether/aromatic cyclic ether	signature of berberine structure	berberine hydrochloride
895	cyclic ether	present in BerbiQ	BerbiQ
710, 712	disubstituted aromatic ring	ring out-of-plane bending	both

3.2.2. XRD Studies

The XRD patterns collected between 5 and 30° 2θ (Figure  ) reveal distinct crystalline signatures for berberine hydrochloride and the BerbiQ complex. Berberine hydrochloride (Figure a) displays sharp diffraction peaks at 9.2, 11.6, 13.0, 13.8, 16.4, 21.0, 25.5, and 26.3° 2θ values. These peaks correspond to the Miller planes (100), (110), (111), (200), (210), (220), (311), and (222), respectively matching previously reported crystallographic features of berberine hydrochloride. BerbiQ formulation (Figure b) retains reflections at 9.1° (100), 13.0° (111), 16.4° (210), 21.0° (220), and 25.5° (311). ,,− However, in the BerbiQ formulation (Figure b), the relative peak heights corresponding to characteristic berberine planes were significantly reduced or broadened, indicating partial loss of long-range order and reduced crystallinity. In particular, the peaks at 11.6, 13.8, and 26.3°, prominent in pure berberine hydrochloride, are absent or overwhelmed by amorphous background in BerbiQ, suggesting effective complexation and partial amorphization. The observed reduction in peak sharpness and intensity in BerbiQ is indicative of decreased crystallinity, a sign that the berberine hydrochloride molecules have been successfully integrated into the formulation matrix. This structural alteration is commonly associated with enhanced solubility and dissolution rates, which likely supports the improved bioavailability in OMICS-enabled delivery strategies.

3.

XRD pattern of (a) berberine hydrochloride, (b) BerbiQ.

3.2.3. SEM Analysis

The surface morphology and shape of BerbiQ were examined through SEM analysis. SEM images (Figure ) clearly revealed that the BerbiQ formulation exhibited spherical shapes, well-dispersed particles with no aggregation, and a smooth surface, indicating successful complexation of berberine hydrochloride through OMICS technology. The particle size of BerbiQ was determined using ImageJ software, and the results showed a nearly uniform distribution with a narrow size range of 130 to 350 nm, yielding a mean particle size of 199.19 ± 64.24 nm.

4.

Scanning electron micrographs of BerbiQ particles at different magnifications: (a) 1 μm, (b) 500 nm, and (c) 200 nm, revealing spherical morphology and uniform nanoscale distribution. (d) Particle size distribution histogram confirms a narrow size range with an average particle size around 200 nm, indicating efficient OMICS technology.

3.2.4. TEM Analysis

The morphological characteristics of BerbiQ were further analyzed using TEM, as depicted in Figure . The BerbiQ formulation displayed nanosized particles with a smooth, spherical shape, free from agglomeration, indicating the stability of the formulation. The mean particle size of BerbiQ was 184 ± 31 nm, which is consistent with the results obtained from SEM analysis (Figure ).

5.

Transmission electron microscopic photographs of BerbiQ with different magnifications.

3.2.5. DSC Studies

The DSC thermogram of berberine hydrochloride revealed several endothermic peaks (Figure (a)). The endothermic peak observed around 90 °C corresponds to the dehydration of the berberine hydrochloride molecule. The endothermic peaks at 127 and 139 °C are associated with the decomposition and separation of the CO group, while the sharp endothermic peak at 191 °C corresponds to the melting point of berberine, further confirming its crystalline nature. The presence of a semistable polymorph in the berberine structure is suggested by the exothermic peak at 204 °C followed by an endothermic peak at 283 °C. In the case of BerbiQ, the endothermic peaks at 128 and 186 °C, corresponding to berberine (Figure (b)), are broader with reduced intensity and a slight shift, indicating the successful complexation of berberine via OMICS technology.

6.

DSC thermogram of (a) berberine hydrochloride and (b) BerbiQ.

3.3. Storage Stability

The storage stability of bioactive-oriented formulations is a crucial factor in assessing their suitability for use in nutraceuticals, functional foods, and dietary supplements. No changes in color or physical appearance of the stored BerbiQ were observed during the 180-day study conducted at 4, 25, and 45 °C. The berberine content under different stability conditions is summarized in Table . The results indicated no statistically significant variations in berberine content after 180 days of storage at these temperatures, confirming the stability of the BerbiQ formulation.

3. Berberine Content in the BerbiQ under Different Stability Conditions.

	berberine content (%)
stability condition (°C)	initial	30 days	60 days	90 days	120 days	150 days	180 days
4 ± 2	99.85 ± 0.37	99.72 ± 0.47	99.49 ± 0.36	99.28 ± 0.51	98.85 ± 0.56	98.64 ± 0.45	98.41 ± 0.77
25 ± 2	99.85 ± 0.42	99.38 ± 0.39	98.95 ± 0.41	98.52 ± 0.62	98.16 ± 0.71	97.85 ± 0.56	97.36 ± 0.68
45 ± 2	99.85 ± 0.45	99.02 ± 0.59	98.22 ± 0.56	97.84 ± 0.44	97.38 ± 0.78	97.09 ± 0.61	96.82 ± 0.78

3.4. Bioavailability Study

This study was designed as a randomized, double-blind, single-dose (350 mg capsule containing 250 mg of berberine), two-period, two-way crossover trial to evaluate and compare the oral bioavailability of berberine in BerbiQ and conventional berberine formulations. A total of 10 eligible healthy subjects (6 male and 4 female) under fasting conditions participated in the study. The demographics and baseline characteristics of the study participants are presented in Table . No adverse effects were reported by any of the subjects throughout the study.

4. Subject Characteristics.

parameters	mean ± SD	min	max
number of subjects	10 (M = 6, F = 4)	-	-
age	30.60 ± 4.01	24	37
height (cm)	166.59 ± 8.88	156.40	176.70
weight (kg)	58.64 ± 2.59	54.60	62.10
systolic blood pressure (mmHg)	123.20 ± 3.68	118	126
diastolic blood pressure (mmHg)	73.80 ± 3.19	70	78
heart rate (beats per minutes)	74.40 ± 2.07	70	78
respiratory rate (breaths per minutes)	18 ± 0	18	18
body temperature (°F)	95.87 ± 0.28	95.40	96.40

Table summarizes the total average pharmacokinetic parameters (mean ± SD) for both the BerbiQ and conventional berberine groups, derived from plasma concentrations of berberine, along with corresponding P-values. The mean concentration of BerbiQ and conventional berberine as a function of time is depicted in Figure . The maximum plasma concentration (C _max) and the area under the curve (AUC_0–48) over 48 h postadministration for BerbiQ were 1.21 ± 0.28 ng/mL and 29.89 ± 4.10 ng·h/mL, respectively. In contrast, the C _max and AUC_0–48 values for conventional berberine were 0.29 ± 0.06 ng/mL and 7.01 ± 0.94 ng·h/mL, respectively (Table ). BerbiQ demonstrated a significantly (P < 0.001) higher bioavailability than conventional berberine, with a 4.26-fold increase based on AUC_0–48. The absorption of berberine in BerbiQ was 4.10-fold higher than that of conventional berberine, with statistical significance observed at the P < 0.01 level, as indicated by the C _max values. The results clearly demonstrate that BerbiQ exhibits enhanced bioavailability, attributed to the effective complexation and sustained release of berberine facilitated by OMICS technology. AUC_0‑∞ represents the area under the plasma concentration versus time curve from time zero to infinity, calculated as AUC_0‑∞ = AUC_0‑t + C_t /K _el, where C_t is the last measurable concentration and K _el is the terminal elimination rate constant (Table ). AUC_0‑∞ values were 41.95 ± 9.41 ng.h/mL for BerbiQ and 8.75 ± 1.73 ng·h/mL for conventional berberine, clearly demonstrating that BerbiQ exhibits superior bioavailability compared to conventional berberine. This enhancement is attributed to the sustained release and effective complexation of berberine via OMICS technology.

5. Average Pharmacokinetic Variables from Plasma Berberine of BerbiQ and Conventional Berberine.

PK parameters	BerbiQ	conventional berberine	P value
C max (ng/mL)	1.21 ± 0.28	0.29 ± 0.06	<0.01
AUC0‑t  = AUC0–48h (ng·h/mL)	29.89 ± 4.10	7.01 ± 0.94	<0.001
AUC0‑∞ (ng·h/mL)	41.95 ± 9.41	8.75 ± 1.73	<0.001
T max (h)	3.20 ± 1.93	5.20 ± 1.67	<0.05
t 1/2 (h)	25.49 ± 8.02	16.25 ± 5.52	<0.05
K el	0.03 ± 0.01	0.05 ± 0.01	<0.05

7.

Mean plasma concentrations (ng/mL) of BerbiQ compared with Conventional Berberine. All the values stated are mean ± SD.

The time to reach the maximum plasma concentration (T _max) for BerbiQ was 3.20 ± 1.93 h, which was significantly shorter than the T _max of conventional berberine (5.20 ± 1.67 h) with a P-value of <0.05 (Table ). Although the T _max for BerbiQ was achieved earlier than conventional berberine following oral administration, BerbiQ demonstrated a 4.26-fold higher AUC_0‑t and a 4.10-fold higher C _max compared to conventional berberine. Additionally, the sustained release profile of berberine in BerbiQ suggests more efficient absorption, which can be attributed to the effective complexation of berberine in BerbiQ using OMICS technology. The elimination rate constant (K _el) was 0.03 ± 0.01 for BerbiQ and 0.05 ± 0.01 for conventional berberine (Table ). The lower K _el value observed for BerbiQ indicates a slower elimination rate, suggesting an extended absorption period of berberine. This effect can be attributed to the sustained release of berberine from the BerbiQ formulation, resulting from its effective complexation by OMICS technology. The terminal half-life (t _1/2) was 25.49 ± 8.02 h for BerbiQ and 16.25 ± 5.52 h for conventional berberine (Table ). The longer t _1/2 observed for BerbiQ indicates a prolonged availability of berberine, which can be attributed to the effective complexation of berberine in the BerbiQ formulation through OMICS technology.

Table provides a summary of published pharmacokinetic data from studies on different formulations. In a study by Alolga et al. the effects of gut microbiota metabolism on the pharmacokinetics of berberine were investigated in healthy male individuals of African and Chinese volunteers. The study included 20 nonsmoking male volunteers, comprising 10 Africans and 10 Chinese individuals. Each volunteer received a single dose of 600 mg of berberine hydrochloride, equivalent to a net dose of 543 mg. The C _max values were significantly higher in the African group (0.16 ng/mL) compared to the Chinese group (0.06 ng/mL). Similarly, the AUC values were significantly higher in the African group (0.96 ng·h/mL) compared to the Chinese group (0.47 ng·h/mL), with no notable difference in T _max, which remained at 4 h for both groups. Another study by Blöcher et al. examined the impact of OCT1 and CYP2D6 polymorphisms on berberine pharmacokinetics in 44 human participants, who were administered a single oral dose of 1000 mg of berberine. The C _max and AUC values for all participants were 0.43 ng/mL and 9.27 ng·h/mL, respectively, with a T _max of 3 h.

6. Pharmacokinetic Properties of Various Formulations of Berberine Reported in the Literature.

			pharmacokinetic parameters
formulations/bioactives	dose	no. of subjects	C max (ng/mL)	C max per mg berberine	AUC (ng·h/mL)	AUC per mg berberine	T max (h)	T 1/2 (h)	refs
berberine	543 mg in 600 mg	10	0.16	0.00029	0.96	0.0018	4	3.95
berberine	543 mg in 600 mg	10	0.06	0.00011	0.47	0.0009	4	4.00
berberine extract	1000 mg (2 × 500 mg)	44	0.43	0.00043	9.27	0.0093	3	-
berberine ursodeoxycholate (BUDCA)	250 mg	12	0.40	0.0016	2.9	0.0116	3.5	9.0
BUDCA	500 mg	12	0.40	0.0008	3.3	0.0066	4.0	10.6
BUDCA	1000 mg	14	0.90	0.0009	7.2	0.0072	4.0	7.8
berberine chloride	300 mg	12	0.33	0.0011	3.83	0.0128	3.96	30.60
berberine chloride +40 g simvastatin	300 mg	12	0.38	0.0013	3.83	0.0128	3.13	28.76
berberine chloride +200 g fenofibrate	300 mg	12	0.44	0.0015	4.48	0.0149	3.63	27.04
BerbiQ-Bioavailable berberine formulation	250 mg	10	1.21	0.0048	29.89	0.1196	3.20	25.49	current study

^aAsian.

^bChinese.

A double-blind, randomized, placebo-controlled, dose-ranging study was conducted to compare three different doses (250, 500, and 1000 mg) of berberine ursodeoxycholate (BUDCA) with a placebo in a cohort of 50 subjects. The AUC values for the 250, 500, and 1000 mg doses of BUDCA were 2.9, 3.3, and 7.2 ng·h/mL, respectively. Similarly, the C _max values were 0.40 ng/mL for both the 250 and 500 mg doses, while the 1000 mg dose showed an increased C _max value of 0.90 ng/mL. Both AUC and C _max values exhibited a dose-dependent increase, with higher doses corresponding to higher values for both pharmacokinetic parameters. Li et al. conducted an open label, randomized, parallel study to evaluate the pharmacokinetic interactions and tolerability of berberine chloride when coadministered with simvastatin and fenofibrate in 60 healthy Chinese subjects. The study involved different treatment groups, particularly 300 mg of berberine chloride alone, 300 mg of berberine chloride with 40 mg of simvastatin, and 300 mg of berberine chloride with 200 mg of fenofibrate. The C _max values for the combination of berberine with simvastatin and fenofibrate were slightly elevated, recorded at 0.38 ng/mL and 0.44 ng/mL, respectively, compared to berberine chloride alone (0.33 ng/mL). Additionally, the AUC values were higher for the combined formulations than for berberine chloride alone, suggesting a potential synergistic effect.

Making pharmacokinetic comparisons between different berberine formulations is challenging due to variations in the amount of berberine, significant differences in total product mass, and substantial discrepancies in the absorption of berberine. The data were compared based on C _max and AUC per mg of berberine administered across various pharmacokinetic studies (Table ). These results were also compared with those obtained from the current study, which investigated the bioavailable berberine formulation, BerbiQ. The data indicate that the C _max and AUC per mg of berberine in all other formulations were lower, suggesting that BerbiQ has superior bioavailability compared to the other formulations. Furthermore, BerbiQ demonstrated a 4.26-fold increase in bioavailability over conventional berberine, attributed to the effective complexation and sustained release facilitated by OMICS technology.

4. Conclusions

A novel bioavailable berberine formulation, BerbiQ, was developed using OMICS technology by complexing berberine hydrochloride with synergistic molecules, particularly silymarin, to enhance its efficacy. The formulation process incorporated coconut milk containing proteins and lipids via an advanced bionanotechnology approach. SEM and TEM analyses revealed that BerbiQ exhibited spherical, well-dispersed particles with a smooth surface and no aggregation, confirming the successful complexation of berberine hydrochloride through OMICS technology. Further characterization using IR, XRD, and DSC analyses, along with stability studies, substantiated the effective complexation and stability of berberine hydrochloride within the BerbiQ formulation. The study also demonstrated that the novel BerbiQ formulation significantly enhances the oral bioavailability of berberine compared to the conventional berberine formulation. This enhancement is evident from the 4.26-fold increase in AUC and the 4.10-fold increase in C _max observed for BerbiQ. Additionally, BerbiQ exhibited a shorter T _max, a lower elimination rate constant (K _el), and an extended terminal half-life (t _1/2), collectively indicating sustained release and prolonged systemic availability of berberine. These pharmacokinetic improvements are attributed to the advanced complexation and sustained-release capabilities of OMICS technology employed in the BerbiQ formulation. The results suggest that BerbiQ offers a more efficient delivery system for berberine, potentially reducing the required dose and minimizing associated side effects, thus making it a promising candidate for various therapeutic applications.

The authors declare no competing financial interest.