*5.2.2.4 Fastness*

*B. orellana* reportedly has moderate to poor light fastness, but moderate to excellent fastness to washing, rubbing, and perspiration.

#### *5.2.2.5 Functional properties of annatto related to textiles application*

Extract of annatto has remarkable antimicrobial and antioxidant properties and a study revealed that the annatto dye had a bactericidal effect and could reduce *E. coli* activity [46]. Ethanolic extract form seeds showed broad spectrum antibacterial activity against *Bacillus subtilis*, *Staphylococcus aureus*, *Streptococcus pyogenes*, *Salmonella typhi*, *Pseudomonas aeruginos*a, *Escherichia coli,* and *Candida albicans* [47]. When compared to the traditional method, annatto dye extracted by the ultrasound aided technique has higher antibacterial and antioxidant activities [48]. Grampositive bacteria show more sensitivity to annatto dye extracted by conventional and ultrasound-assisted extraction methods than gram-negative bacteria and. *B. subtilis* showed the lowest sensitivity toward annatto dye, while *Escherichia coli* gave the highest sensitivity. Annatto dye extracted by UAE showed a bactericidal effect against *Salmonella enteritidis* [48]. Annatto extracted from annatto seeds (*Bixa Orellana*) using ultrasound technique was used to color biodegradable films based on poly (3-hydroxybutyrate). Photo-degradation under UVA exposure of colored films showed improvement as suggested by the SEM micrographs, and the film also showed good thermal stability as confirmed by the thermo gravimetric analysis [49].

### *5.2.2.6 Case study-5*

Cotton, wool, and silk were dyed with an aqueous extract of the *B. orellana* seed powder using the one-step simultaneous sequence and the two-step pre-mordanting sequence by the ultrasound technique **[**50**]**. Enzymes were used along with tannic acid to treat the fabric; cellulose and amylase for cotton and protease for silk and wool. The exhaustion of color on the fiber ranged from 55 to 63% for the ultrasound technique, while it was lower ranging between 42 and 46% for the conventional exhaust dyeing procedure for all three fibers. Because bixin and norbixin are polar and acidic, they have a stronger affinity for wool and silk (protein fibers) which have more polar groups than cotton (cellulosic fiber). Enzyme treatment increased

#### **Figure 1.**

*Effect of enzyme treatment on the color related properties (L\*, a\* and b\*) of cotton, wool, and silk were dyed with an aqueous extract of the* Bixa orellana *seeds.*

the surface color strength (K/S) for all the fibers and lowered the L\* values (indicating darker shades). It also slightly increased a\* and b\* values of all the dyed fibers indicating redder and yellower i.e. probably an orangish tone (**Figure 1**).

#### *5.2.2.7 Case study-6*

Eco-friendly bamboo fiber was dyed with *B. orellana* using potash alum as mordant. The dyeing procedures variables of time, temperature, and pH were optimisation for the pre-mordanting process. An increase in time from 15 min to 60 min increased the color yield and a sharp liner increase in the surface color depth (K/S) was also observed with an increase in temperature from ambient temperature


*Colorimetric Measurement and Functional Analysis of Selective Natural Colorants… DOI: http://dx.doi.org/10.5772/intechopen.102473*

#### **Table 8.**

*K/S (color strength) and other color related parameters of bamboo fabric pre-mordanted with potash alum and dyed with aqueous extract of annatto seeds (*Bixa orellana*) under variable conditions of dyeing.*

to 80°C. The K/S value of the mordanted bamboo fabric was significantly higher when dyeing was carried out under alkaline conditions. The optimum conditions of dyeing of the potash-alum pre-mordanted bamboo fabric with aqueous extract of seeds of annatto (*B. orellana*) was thus reported as dyeing time 60 min, dyeing temperature 80°C, and pH–12. Varied shades of orange (sherbet-orange to gingerorange and apricot colors) were obtained which were dark as indicated by negative L\* values. The shades were redder as indicated by positive a*\** values and yellower as reflected by positive b\* values (**Table 8**). The yellower tint (b\*) was more pronounced compared to the redness tint (a*\**) in most cases [51].

#### *5.2.3 Application of turmeric in food coloration*

Annatto (E-160B) is a natural yellow-orange dye obtained from *B. orellana*, with less toxicity. Compared to synthetic food colorants, it typically demonstrates superior biodegradability and compatibility with the environment [52]. Annatto colorants are used extensively in the food industry, particularly in the processing of dairy and meat products.

#### *5.2.3.1 Color produced*

Annatto gives a yellow to orange-red shade on food.

#### *5.2.3.2 Extraction & Application conditions*

Annatto is water-soluble and can be mixed with sugar powder or potassium carbonate. The pigment is not heated stable. Moreover, there is a considerable loss of pigment due to deep-fat frying at high temperatures (> 200°C). It is stable at a pH 5.0–10.

#### *5.2.3.3 Functional properties related to food application*

Extract of annatto seed possess antimicrobial properties and decrease the growth and activity of *B. subtilis, Staphylococcus aureus, Streptococcus Clostridium* *thermophilus, perfringens, Lactobacillus casei, Lactococcuslactis, Candida albicans, Candida famata, Rodotorula* species, *Aspergillus* species, and *Neurosporacrassa*. The seed extract showed strong inhibition of triglycerides oxidation in rapeseed oil [53] and norbixin was able to inhibit oxidative deterioration of olive oil [18]. Annatto seeds' antioxidant capabilities aid in the treatment of cardiovascular disorders and have been found to decrease the oxidation of low-density lipoprotein in vitro (LDL). Extracts can protect DNA from oxidative damage thereby controlling serious consequences in some age-related cancers.

#### *5.2.3.4 Case study-7*

Color from annatto seeds is safe for human consumption compared to the synthetic colorants commonly used in sweetmeats. *Jilebi* and *jangri* are popular Indian sweetmeats colored in various shades of yellow and red. Bixin extracted from annatto seeds by a dual solvent method (washing the seeds with a non-polar solvent like hexane and then recovery of the dye in acetone) was used to color *jilebi* and *jangri* and its effectiveness was compared to the commercially available counterparts that were colored using synthetic food colorants [54]. The dye was converted to norbixin using potassium hydroxide (alkaline pH) and diluted with potassiun carbonate (K2CO3) to get its water-soluble form. Two water-soluble formulations of the colorant (bixin), one having potassium carbonate (with norbixin content, 10.6%) and the other with sugar (with norbixin content, 11.24%) were prepared. The first formulation with potassium carbonate was used to prepare the batter for the sweetmeat, while the other sugar formulation was used for further sweetening the sweetmeat after frying. The bixin extracts were applied in varying concentrations in both the formulations for the preparation of *jileb*i and *jangri*, and the effect in terms of red and yellow color units using Lovibond tintomer was evaluated. The effect of frying temperatures (142°C and 172°C) on the color value of both *jilebi* and *jangri* were also evaluated. With the increase in the norbixin concentrations, the R values indicated an increasing trend; while the Y values were higher and constant. Commercial *jalebi*s' R values matched the R and Y values of an annatto solution comprising containing 2.5 and 5 mg/kg of norbixin for sugar-based formulations; while potassium carbonate-based formulations, 5 mg/kg of norbixin gave comparable results (R values) though Y values were higher (**Table 9**). For *jangris*, 40 mg/kg of norbixin gave comparable R and Y values for


#### **Table 9.**

*Tinctometer color values of commercial* jalebis *and* jangri*s, and of the solutions containing optimized concentrations of norbixin with potassium carbonate and sugar.*

#### *Colorimetric Measurement and Functional Analysis of Selective Natural Colorants… DOI: http://dx.doi.org/10.5772/intechopen.102473*

both sugar and potassium carbonate-based formulations. Soluble annatto dye sugar powder formulation gave much better results with lower concentrations of dye as compared to soluble annatto dye potassium carbonate formulation. Water-soluble annatto dye sugar solutions at concentrations 5 mg/kg were found to be optimum for *jalebi* and 30 mg/kg for *jangri* as the colored developed was similar that those found in the commercial counterparts. Excellent color matching was observed and no difference in redness and yellowness in the color of the product were reported due to the effect of high temperature used in frying.
