Sappan Wood Extract Research Articles

Investigating the potential of waste oyster shell as a sustainable bio-mordant in natural dyeing

The use of natural bioresources in textile dyeing has attracted significant research interest due to their environmentally friendly and low-toxic nature. This study investigated the utilization of waste oyster shell (WOS) as a bio-mordant combined with sappan wood extract to dye silk fabrics using pre-, meta-, and post-mordanting method. The crystal structure of CaCO3 derived from WOS and their impact on color properties, color strength and color fastness were analyzed. The dyed fabrics displayed a range of shades from light pink to deep red and brown. FTIR analysis confirmed chemical interactions among dye, silk, and WOS mordants. Notably, silk treated with WOS at 800 °C as a pre-mordant showed color strength approximately 2.6 times higher than that of un-mordanted samples. Additionally, bio-mordanted samples exhibited improved color fastness compared to untreated ones (rating 2), with pre-mordanting offering the highest resistance to washing (rating 4–5). These findings shed light on the potential of waste oyster shell as an effective, sustainable alternative to conventional metal mordants in textile dyeing. This study not only support the utilization of waste but also enhances environmental and resource efficiency of the dyeing process.

Fabrication of multifunctional hybrid pigment for color cosmetics based on chitosan-modified palygorskite and sappanwood extract

Consumer perception and market demand have driven the replacement of synthetic colorants with naturally derived alternatives in the cosmetic industry. This study describes a facile way to prepare durable inorganic-organic hybrid pigment with advanced biocompatibility, antibacterial and hydrophobic properties tailored for color cosmetics by initial modification of palygorskite with chitosan to anchor sappanwood dye extract and subsequently coating with amino-modified silicone oil (ASO). The hybrid pigments were characterized by transmittance electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy and the Brunauer–Emmett–Teller method. The results indicated that the sappanwood dye was loaded on chitosan-modified palygorskite via hydrogen bonding and electrostatic interaction. Furthermore, the chitosan-palygorskite/sappanwood hybrid pigment exhibited enhanced biocompatibility and color stability on exposure to different heating temperatures and UVA radiation after subsequent hydrophobic modification with amino-modified silicone oil. Moreover, facial foundation cosmetics based on the chitosan-palygorskite/sappanwood@ASO composites exhibited excellent brightening and skin color corrective effect on human volunteers without any adverse response. And no significant difference was observed in 12 out of 14 sensory evaluation indexes in the comparison of this hybrid pigment-based makeup with two commercially available products. This study provides a new route to stabilize natural botanical colorant for cosmetic use by chitosan-modified clay minerals.

Effect of Foam Mat Drying on Antioxidants in Secang Wood Extract (Caesalpinia Sappan L)

There is a notable increase in interest in utilizing herbal plants, with some herbal products being particularly rich in bioactive compounds, including antioxidants. Antioxidants play a pivotal role in the protection of the body against oxidative stress, through the neutralization of harmful free radicals. One plant extract that exhibits notable antioxidant activity is sappan wood extract (Caesalpinia sappan L). The objective of this study is to assess the impact of foam formulation and temperature on the drying process and antioxidant activity of sappan wood extract, which has been dried by the foam-mat drying method. The research was conducted in three stages, namely extraction, drying and analysis of results. Drying of sappan wood extract was carried out at 40, 60 and 80°C, with different compositions of foaming agent. The results demonstrated that antioxidant activity exhibited a tendency towards stability at a drying temperature of 40°C. However, the highest reduction in water content was observed with drying at 80°C. The most effective foam composition was identified as foam composition 2, 0% CMC + 20% egg white (albumin) + 10%. Conversely, the foam composition with the highest antioxidant activity was identified as foam composition 5, 0.5% CMC + 30% gum Arabic. The most appropriate antioxidant degradation equation for this study was the first-order equation. The most accurate drying rate model for this study was the Lewis drying rate model.

Research of the Dyeing Properties of Lychee Leaves and Sappan Wood Extract on Silk-Linen Fabric

Synthetic dyes are the primary colouring materials for textile dyeing; however, the residual industrial wastewater from the dyeing process contains complex concentrations of chemical substances, which result in toxicity and environmental pollution with severe negative impacts on ecology and health. This study adopts an eco-friendly perspective, utilizing a mixture of lychee leaves and sappan wood extracts for dyeing silk-linen fabrics. It explores the impacts of dyeing time, mordants, and dyeing methods on the colour quality of the samples. A computer colourimeter was used to compare the colour effects of different dye solution ratios on silk-linen fabrics, and the colour fastness to water, perspiration, and dry rubbing of the dyed fabrics was tested. The results show that the optimized dyeing process for silk-linen fabrics using a mixed solution of lychee leaves and sappan wood includes the following findings: Dyeing time has no significant impact on the CLE L*, a*, and b* values. The highest L* value and colour brightness are achieved under conditions without mordants. By mixing dye solutions in different proportions and treating them with mordants, the colour gamut of silk-linen fabrics has been expanded. The dyed colours are similar to the main popular colours of 2024, making them the trend colours of this year; the samples' fastness performance tests indicate that the colour fastness to water, perspiration, and dry rubbing essentially meets the requirements for textile fabrics. The use of lychee leaves and sappan wood for fabric dyeing is not only environmentally friendly but also possesses significant health-beneficial properties, offering potential commercial value.

THE EFFECTIVENESS OF SAPPAN WOOD EXTRACT (Caesalpinia sappan L.) AS AN ALTERNATIVE MATERIAL FOR DENTAL PLAQUE IDENTIFICATION

The main factor of periodontal disease is dental plaque, which is attached to the surface of the teeth and consists of microorganisms originating from saliva and food debris. Dental plaque has a transparent colour, so a disclosing agent is needed to identify it. Sappan wood (Caesalpinia sappan L.) is a plant that contains brazilin and flavonoids that can be used for natural colouring. This study aims to determine the effectiveness of sappan wood extract (Caesalpinia sappan L.) as an alternative material for dental plaque identification. The experiment was conducted by measuring the plaque index of 20 people who did not clean their teeth for 12 hours for two periods. The first period of examination used a tablet disclosing agent, and the second period used 75% sappan wood extract. The research data were analyzed for normality with the Shapiro-Wilk dan Paired T-test. The result showed a significant difference in the mean plaque index score in the first period (1.3470) and the second period (1.0375) (p<0.05). It was concluded that staining dental plaque using a disclosing agent is more effective than 75% sappan wood extract (Caesalpinia sappan L.)

AKTIVITAS ANTIBAKTERI EKSTRAK KAYU SECANG ( Caesalpinia sappan L.) TERHADAP BAKTERI Edwardsiella tarda DAN Edwardsiella ictaluri PATOGEN BUDIDAYA PERIKANAN

The disease caused by pathogenic bacteria is one of the main problems in aquaculture in the Jambi Province. One of the diseases that occurs in cultivated catfish is due to infection by bacteria Edwardsiella tarda and Edwardsiella ictaluri, which cause Edwardsiellosis with a mortality rate above 50%. The antibacterial properties of the sappanwood extract (Caesalpinia sappan, L.) can be used to overcome problems in aquaculture, while reducing the use of antibiotics that can have negative impacts. This research aims to explore the antibacterial potential of sappanwood extract (Caesalpinia sappan, L.), a local plant believed to possess strong antimicrobial activity. The study began with isolating bacteria from catfish that showing symptoms of the disease, identifying the test bacteria, followed by extracting sappanwood using 96% ethanol solvent. Subsequently, the antibacterial activity was tested using the disc diffusion method with extract concentrations of 1%, 5%, and 10%. The research results indicate that sappanwood extract at a concentration of 5% has a very strong ability to inhibit the growth of E. tarda and E. ictaluri bacteria. The inhibition zones produced show significant antibacterial activity equivalent to the antibiotic ciprofloxacin used as a positive control.

Kajian Antibakteri Ekstrak Etanol Kayu Secang (Caesalpinia sappan L.) dan Pemanfaatannya Pada Udang Vannamei (Liptopenaeus vannamei) Segar

Vannamei shrimp (Liptopenaeus vannamei) is a prominent fishery commodity due to its high consumption demand. However, susceptibility to bacterial damage and quality deterioration poses challenges. This study investigates the antibacterial potential of sappan wood extract in preserving shrimp shelf life. The research explores optimal extraction parameters, including solvent type and maceration duration, to assess yield percentage and antibacterial efficacy against Escherichia coli, Salmonella sp., and Vibrio parahaemolyticus. Utilizing a Complete Randomized Block Design (CRBD) in two phases, the study identifies 90% ethanol solvent and 6 hours maceration (P4M6) as the most effective treatment, yielding 7.32%. The extract addition significantly reduces shrimp microbial count, highlighting its potential in food preservation.

In-vitro Anti-inflammatory Activity for Combination of Ethanol Extract from Sappan Wood (Caesalpinia sappan L.) and Red Ginger Rhizome (Zingiber officinale Roxb.)

Sappan wood and red ginger rhizome have been utilized as herbal medicine due to their anti-inflammatory, anti-oxidant, anti-microbial, analgesic, and immunomodulatory activities. Previous study revealed that sappan wood and red ginger rhizome showed good anti-inflammatory activity. However, the studies are focused on single extract. There are no research about combining the sappan wood extract and red ginger rhizome extract for antiinflammatory agent. Therefore, this study was conducted to determine the anti-inflammatory activity of the combination of sappan wood and red ginger rhizome ethanol extracts for antiinflammatory properties using red blood cell (RBC) membrane stabilization test. The tested samples consisted of sappan wood ethanol extract, red ginger rhizome ethanol extract, combination formulas (F1, F2, and F3), and positive control (diclofenac sodium). F1 comprises a 1:1 mixture of ethanol extract of sappan wood and red ginger rhizome, while F2 and F3 are composed of 2:1 and 1:2, respectively. The IC50 values for sappan wood ethanol extract, red ginger rhizome ethanol extract, and formulas F1, F2, and F3 were 47.63, 181.26, 101.93, 94.98, and 104.98 ppm, respectively. Statistical analysis using the one-way Anova followed by the Games-Howell post-hoc test revealed a statistically significant difference (p < 0.05) in the IC50 of each sample. For single extracts, sappan wood is better as an anti-inflammatory. Meanwhile, in the formulations, Formula F2 is superior as an anti-inflammatory agent

Pharmaceutical activity of sappan wood extract (Caesalpinia sappan L.) for treating Escherichia coli infection in piglets.

Escherichia coli infection is one of the major diarrheal diseases resulting in the loss of pigs at a young age. This research investigated the antimicrobial activity of Caesalpinia sappan wood extract against E. coli infection as an antibiotic replacement. E. coli was cultured from diarrheal piglets and then used to find the minimal inhibition concentration (MIC). Caesalpinia sappan wood extract (500 mg/kg) was used for the treatment of diarrheal piglets compared to antibiotics (enrofloxacin 5 mg/kg) by oral administration. Another three groups of diarrheal piglets were used supplemented feed with 1% and 2% extract compared with commercial feed. Subsequently, E. coli enumeration, fecal shape, fecal color, and growth rate were recorded from day 1 to 7. Based on the results, C. sappan wood extract could inhibit E. coli growth at a MIC of 16-34 mg/ml. The number of colonies did not significantly differ between C. sappan wood extract and enrofloxacin treatment groups. A supplemented feed with 1% and 2% C. sappan wood extract could improve the fecal shape and fecal score compared to the control group, albeit only in suckling pigs. There were significant differences from the control group on days 4, 5, 6, and 7 (p < 0.05). However, the average daily gain did not significantly differ among the three groups. The results indicate that C. sappan wood extract could improve diarrheal signs in suckling pigs and can be used as a replacement for antibiotics for organic pig production.

The efficacy of topically applied Sappan wood (Caesalpinia sappan L.) ethanol extract during incision wound healing in albino rats.

Medical manifestations in the form of incisions, burns, and trauma will trigger a natural wound-healing process that involves complex interactions between cells. Brazilin and other secondary metabolites found in Sappan wood have numerous bioactive qualities, including anti-inflammatory, anti-cancer, and antioxidant properties. This study aimed to investigate the efficacy of Sappan wood (Caesalpinia Sappan L.) ethanol extract topically on the incision wound healing of albino rats. Twenty male rats were randomly assigned into five groups with four replications, i.e., (C-) was treated ointment-based, (C+) was treated with 10% povidone-iodine, (T1, T2, and T3 groups) were treated with Sappan wood extract concentration for 6.5%, 15%, and 30%, respectively. The treatment was topically administered to wounded areas twice a day for 15 days. Wound healing was evaluated histologically as the following parameters collagen deposition, polymorphonuclear neutrophils (PMN), angiogenesis, and fibrosis degree using H&E staining. IL-2 level was evaluated using the enzyme-linked immunosorbent assay (ELISA) method. Wound length reduction was calculated on days 8 and 15. As a result, the 6.5% (T1), 15% (T2), and 30% (T3) Sappan wood extract groups were improved significantly (p < 0.05) compared to ointment-based (C-) and povidone-iodine (C+) groups on the collagen deposition, PMN, angiogenesis, fibrosis degree, and IL-2 level. In particular, the 6.5% (T1) Sappan wood extract group was highlighted significantly (p < 0.05) compared to other groups, evidenced by the improvisation of wound healing parameters and reduction of wound length on days 8 and 15. In conclusion, a 6.5% Sappan wood extract revealed its applicability to improve incision wound healing in albino rats.

Anti-inflammatory activity of sappan wood extract cream (Caesalpinia sappan) in porphyromonas gingivalis-induced gingivitis rats models

Background: Gingivitis is a dysbiotic condition characterized by persistent inflammation caused by a disease-associated multispecies bacterial population that has established itself in the subgingival region. Aim: The objectives of this study is to analyze the anti-inflammatory activity of sappan wood extract cream (SWC) in Porphyromonas gingivalis induced-periodontitis rats model. Methods: In this study, rats were infected to the gingiva with P. gingivalis as an inducer for gingivitis rats model for 14 days. The SWC were treated topically into the infected tissue. The inflammation and angiogenesis of gingival tissue was analyzed using histological examination. An immunohistochemistry (IHC) assay was used to analyze the Nuclear Factor kappa-B (NF-kB) protein expression. Masson Trichrome (MT) assay was used to analyze the histopathology of collagen Scores. The Tumor Necrosis Factor-α (TNF-α), Interleukin-1 β (IL-1β), IL-6, and P38 genes expression were measured using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Results: The induction of P. gingivalis -induced the gingivitis in rats model indicated by reddish gingiva and bacterial plaque. MT tests showed that SWC increased collagen density, HE test showed increased angiogenesis and in contrast, lowered inflammation score. IHC test showed that SWC decreased the expression of NF-kB protein. The TNF-α, IL-1β, IL-6, and P38 gene expression were also decreased due to the SWC treatment. Conclusion: Sappan wood extract cream proves that it has anti-inflammatory activity which has the potential to prevent or treat gingivitis

Microcrystalline Cellulose Isolation and Impregnation with Sappan Wood Extracts as Antioxidant Dietary Fiber for Bread Preparation.

Microcrystalline cellulose (MCC) has gained considerable attention as a functional ingredient in bread making. This work demonstrates the isolation of MCC from sugar cane bagasse (SCB) for preparing bread. The effect of MCC on bread attributes and antioxidant activity by impregnation with sappan wood extract (SAP) was evaluated. The highest crystallinity index and suitable size of MCC were achieved at 85 °C under 90 min hydrolysis condition. Increasing MCC/SAP levels in bread showed a significant increase in bread color with decreases in the specific volume and baking loss. There was a positive correlation between bread texture and the MCC/SAP level. The addition of MCC/SAP interfered with the bread hardness. Low MCC/SAP levels have no effect on springiness and cohesiveness; however, 4% MCC/SAP has significantly decreased these attributes, with the highest antioxidant activity and phenolic content. Therefore, MCC can be functionalized with SAP as an antioxidant fiber additive for health benefits in bakery products.

Potential of Sappan Wood, Purple Cabbage and Beetroot Extract in Sperm Staining

Examination of sperm morphology is an essential criterion for evaluating male fertility. This examination can be done by staining sperm cells with various techniques to facilitate the visualization of sperm cells. Several other methods that can be used to evaluate sperm morphology are Eosin-nigrosin, AgNO3 staining, Papanicolaou, Diff Quick and Giemsa. However, using these synthetic dyes can harm the environment and water sources because they prefer to decompose. The natural dye derived from environmentally friendly plants expected to minimize hazardous waste. This study aimed to identify and compare the potency of several natural dyes derived from purple cabbage, sappan wood and beetroot, which have not been studied in sperm studies. Sperm obtained from 30 men in the campus area and underwent 3-5 days of abstinence from intercourse in preparation for sampling. The sperm stained with natural dye, and their quality compared with WHO standard Papanicolaou. Extracts of sappan wood, purple cabbage and beetroot are made with various mordant alum, ethanol and acetic acid compositions. The study's results showed that the presence of mordant increased the staining quality of sperm with sappan wood and purple cabbage extracts. The use of acid improves the quality of sperm staining with beet extract. There was no different between dyeing with beetroot extract in various solvent compositions and dyeing with Papanicolaou to color all sperm components (p&gt; 0.05). The results showed that beetroot extract has a high potential to evaluate sperm morphology.

Assessment of Teratogenic Effects of Sappan Wood (Caesalpinia Sappan L.) Extract In Rats (Rattus novergicus)

Treatment using plant materials has been widely researched and observed to assist in curing a disease. Medicinal products derived from herbal plants are proven useful as drugs must follow guidelines not to cause acute or chronic toxicity. The purpose of this study was to investigate the teratogenic potential of an ethanolic sappan wood extract on the growth of unborn rats. Six groups of pregnant rats are created: Aquadest was administered to the Control group, whereas the Treatment groups received 100, 200, 300, 400, or 500 mg.kg-1 BW of an ethanolic extract of Sappan wood. On the twenty-first day of gestation, pregnant rats were CO2-euthanized and delivered vaginally. Rats' body weight during pregnancy was observed, along with fetal growth measurements, viscoelastic examination, and skeletal development. Wistar rats' reproductive systems, fetal body weight, body length, and tail length were all unaffected by ethanol extract of Sappan wood at doses from 100 mg/kg to 500 mg/kg. In fetuse rat from the control group and the five dosage groups, no skeletal development had taken place and there were no obvious external abnormalities. Even at the maximum dose of 500 mg, sappan wood extract demonstrated no teratogenic effects on the development of fetal rats.kg-1 BW.

Designing Bio-Based Color Sensor from Myofibrillar-Protein-Based Edible Film Incorporated with Sappan Wood (Caesalpinia sappan L.) Extract for Smart Food Packaging

The use of intelligent sensor-based packaging in food products allows the quick evaluation of food quality. This study aimed to design a color sensor from surimi utilizing sappan wood extract (SWE) for intelligent food packaging. The myofibrillar-protein-based edible film incorporated the SWE at different concentrations (0.15%, 0.25%, and 0.35%). The physical properties of the sappan wood–surimi edible film (SSEF) were analyzed, and the color changes at various pH levels and soaking times under different conditions were assessed. According to the results, different concentrations of the SWE significantly affected the thickness, transparency, tensile strength, and elongation at break of the film. However, the values were still within the acceptable range. The pH values increased the film’s darkness, redness, and blueness. Based on the evaluation of the SSEF under different soaking conditions, the color changes of the film in acidic conditions were more stable than in neutral and alkaline conditions. This study showed that SSEF can be used as intelligent food packaging. It can act as a color sensor due to its sensitivity to the changes in the pH condition of the product.

Physicochemical Characteristics of Edible Film Sodium Caseinate with Sappan Wood Extract Addition

Physicochemical Characteristics of Edible Film Sodium Caseinate with Sappan Wood Extract Addition

Antibacterial Activity Test of Secang Wood (Caesalpinia sappan L.) Ethanol Extract Against Streptococcus mutans

Dental caries is caused by the Streptococcus mutans bacteria, which causes a food deposit to harden and adhere to the tooth surface. Cavities will develop if food residue is not removed. Herbal plants, specifically sappan wood, can be used as an antibiotic. The purpose of this study was to look at the secondary metabolite content of sappan wood extract, antibacterial activity, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of sappan wood extract against Streptococcus mutans bacteria. This research method includes the maceration of sappan wood extract with 96% ethanol solvent, phytochemical screening of sappan wood extract, inhibition zone test with paper disc method, and MIC and MBC tests. Qualitative testing revealed that the sappan wood extract contained alkaloids, flavonoids, tannins, and saponins. Flavonoids, tannins, and saponins are all constituents of plants. Secang wood extract has antibacterial activity against Streptococcus mutans, as evidenced by the formation of inhibition zones at concentrations of 20%, 40%, 60%, 80%, and 100%, namely 8.75±0.354; 11±1.41;, 12.75±0.354; 16.25±0.354; and 17.5±0.000 mm. Sappan wood ethanol extract had a MIC of 12.5% against Streptococcus mutans and an MBC of 25%.

Parametric Study and Characterization of Sappan Wood (Caesalpinia Sappan Linn) Natural Red Colorant Extract with Ultrasonic Assisted Extraction Method

In this study, the optimization and characterization of the colorant extraction process from sappan wood were carried out using the ultrasound-assisted extraction method. Sappan is proposed as a source of natural material wealth, producing a natural red colorant that is being needed by the community in the food sector. Based on this research, the optimum operating conditions for the extraction of sappan wood were obtained using a frequency of 40 kHz, a temperature of 60 °C, a ratio of 0.0050 g mL-1, an extraction time of 20 min, and the use of 60% ethanol solvent. Sappan wood extract created a yellow to reddish orange color at acidic pH (2-6), red at neutral pH (7) and shifted towards purplish red with increasing pH. The extract was qualitatively positive containing quinone, flavonoid, quinone, and tannin compounds, as well as several phenolic compounds detected in the Gas Chromatography Mass Spectroscopy qualitative test.

Antioxidant Activity, Physicochemical Characterisation and Antibacterial Properties of Caspian Sea Yoghurt Enriched with Ginger and Sappanwood Extracts

Caspian Sea yoghurt has lower acidity and is thicker than common yoghurt. Caspian Sea yoghurt contains probiotic bacteria that improve the balance of intestinal microflora. This research aimed to determine the antioxidant activity of Caspian sea yoghurt enriched with two different herbs, ginger and sappanwood, at concentrations of 5, 10 and 15% w/v. The pH, total phenol, antioxidant, total LAB, and antibacterial activity of the enriched yoghurt were measured to determine the best of the two products. The results showed that the enrichment of the yoghurt with sappanwood and ginger extracts increased lactic acid bacteria (LAB) growth compared with the control (yoghurt without plant extracts). Further analysis showed that the product that contains 15% sappanwood extract was judged to be the best formulation with a pH of 5.01 and a total LAB of 3.78 x 107 CFU/mL. It also exhibited antibacterial activity against B. cereus and E. Coli with zones of inhibition of 8.00 mm and 8.39 mm, respectively. The total phenol content was 5.48 mg GAE/g of the sample and 50.08% antioxidant activity. In conclusion, Caspian sea yoghurt enriched with herbs showed increased antioxidant and antibacterial activities and may have better potential in health maintenance.

Karakteristik kimia serta organoleptik es krim dengan penambahan puree semangka (Citrullus lanatus Schard) dan ekstrak kayu secang (Caesalpinia sappan Linn)

Watermelon (Citrullus lanatus Schard) contains 93.4% low calories and water, 0.5% protein, 5.3% carbohydrates, 0.2% fat 0.1% fiber, 0.5% ash and vitamins (A, B and C). The usage of watermelon is not optimal in food innovation products due to commonly consume as fresh fruit. Ice cream is one of the food that has an increase in consumers every year but innovated as functional product should be analyzed. Secang wood (Caesalpinia sappan Linn) has potency as an addition to the antioxidant content in ice cream. The addition of watermelon puree and sappan wood extract into the formulation can increase the nutritional value of the resulting ice cream. The purpose of this study was to determine the effect of adding watermelon puree and sappan wood extract on ice cream characteristics. The study was conducted with 2 factors resulted 9 combination treatments using a factorial Randomized Complete Block Design (RCBD). Each treatment was repeated 3 times. Data were analyzed using analysis of variance (ANOVA) and Duncan's. The results of this study indicated that the addition of sappan wood extract to ice cream had an effect on the antioxidant, sugar content, color intensity of brightness L and intensity of yellow color b+. Meanwhile, the parameters of fat content, protein content, melting power, color intensity a+ had no significant effect. The addition of watermelon puree concentration had no significant effect on the protein content, a+ color intensity and color organoleptic. There was an interaction between watermelon puree and secang wood extract the antioxidant, sugar content, fat content and color intensity of L and b+. While there was not interact between these two ingredients on the parameters of protein content, melting power and color intensity b+.