1. INTRODUCTION:

A food product's flavor is one of the most crucial factors in determining consumer acceptance, and vanillin is the most well-liked and often utilized flavoring agent on the planet. It has two kinds of chemical structures. They are Methyl vanillin laid out in Fig.1(a) and Ethyl vanillin laid out in Fig.1(b). Methyl vanillin can be founded in nature, Ethyl vanillin is an artificial fragrance. Application of vanillin. The percentage of vanillin used in different fields i.e; Approximately 50% of vanillin used is as a food additive, more than 20% as a medical intermediate, and remaining 30% for other uses. Available as Crystalline powder form sources Orchid vanilla planifolia. Due to its high price, only a small percentage of organically generated vanillin is employed in foods and beverages. According to a survey, plant-derived vanillin accounted for just 0.2% of the market's needs. The majority of the vanillin produced was made through biotechnological or synthetic methods. Vanillin concentration in custard powder should not exceed 70mg kg^-1, as per FDA standards. The excess ingestion may lead to liver, kidney and spleen damage or impairment in their function. As a result of its potential risks to food safety, vanillin should be found. Natural and artificial food flavouring have the same chemistry, however artificial flavouring compounds are thought to have negative health effects including the potential for carcinogenicity, which can cause the development of cancer, genotoxicity, which can cause chromosome damage, and neurotoxicity, which can harm nerve tissues. The intention of the current study was to undertake a conductometric evaluation of the vanillin content added to several brands of vanilla-flavored custard powder. There are various methods for determination of vanillin content they are Capillary Eelctrophoresis^1,10, Gas Chromatography², Hyphenated techniques (i.e, Gas chromatography-Mass spectroscopy, Liquid chromatography - Mass spectroscopy)^3,4, Spectroscopic method, High pressure liquid chromatography⁵^,⁷^,1¹^,1², HPTLC⁶, colorimetry^13,15-22,24, Electrochemical methods (Conductometry and Voltametry)¹⁴^,23.

2. MATERIALS AND METHODS:

2.1Instruments used: pH meter, Conductivity Meter of make-Mettler Toledo (model: S 210) were made use for assessing the test.

2.2 Reagent and Chemicals: NaOH, HCl, Methanol, Na₂CO_3,n-Hexane, CuSO₄, DDW Vanilla flavored custard powder.

2.3 Optimization of titrant: Vanillin is a weak acid due to its phenolic activity. Hence, we can go with acid - base conductometric titration. As we know that NaOH is most abundant and easily available so, NaOH is selected as titrant. Then we tried with different concentrations of NaOH (0.1M, 0.05 M, 0.001 M conductometric titrations are performed in order to estimate vanillin content in custard powder, observation and graph are shown in the (Table I).

2.4 Standardization of 0.001M NaOH: For this procedure it was utilized 0.001 M HCl, standard and 0.001 M NaOH. To standardize the NaOH Solution, 10mL of standard HCl were pipetted into a 100mL beaker containing water, the conductivity cell was immersed. After stirring for two minutes after each addition of the reagent solution, the conductance was measured (Table II). The endpoint was established using a graph of conductance versus the volume of additional titrant displayed in Fig. 1. From that the actual molarity of NaOH (i.e., 0.00105) was calculated.

2.5 Extraction of vanillin from custard powder: Inorder to estimate vanillin content in custard powder first we need to extract vanillin from vanilla flavoured custard powder.Methanol was used to extract the vanillin from the vanilla-flavored custard powder. To 10g of the sample (custard powder), 40mL of methanol was added, and the mixture was then ultrasonically processed for 15 minutes. 4 mL of saturated copper sulphate solution was added afterward to precipitate the milk protein in the sample. Whatman filter paper No. 42 was used to filter the outcome. After being extracted once with 40mL hexane and being acidified with 20mL 6 M HCl, the filtrate of the custard powder samples was treated with 20mL 15% (w/v) aqueous sodium carbonate. From the above extracted sample pipette out 50mL in the beaker containing 100mL of double distilled water.

3. RESULTS AND DISCUSSION:

Sodium hydroxide is a strong base that reacts with strong acid (Hydrochloric acid), to give a neutral salt, the reaction depicted in Fig.2.The titration of NaOH was made to reach a standardization of this reagent ; the end point of conductance of NaOH with volume of 0.001 M HCl standard was analyzed and plotted , the respected graph in Fig.3 was meant to be ideal.The sodium ions, which have a significantly lower conductance than the hydrogen ions that were initially present in the solution, replace the hydrogen ions.That is, a salt formed has smaller conductance then the strong acid from which it was made.After the neutralization, the conductance increases,since the OH- ions are no longer used up in the reaction.From the graph shown in Fig.4, 10.5 mL was identified as end point. Vanillin and NaOH's reaction mechanism appears in Fig.5 for the availability of H⁺ions. It means these much amount of NaOH is required to neutralize the reaction. The final volume from the extract was titrated as the standardization in increments of 0.5 mL up to the end analysis (Table III).Graphs plotted were figured in Fig.6, Fig.7 and Fig. 8, corresponds to Custard powder-1, 2, 3. The end points were determined and the concentration of unknown sample was calculated.

3.1 Calculating molarity of NaOH: The molarity of NaOH was calculated according to the equation. M1V1 = M2V2 where,

V1 = Volume of NaOH ( i.e, 10.5mL)

M1 = Molarity of NaOH

M2 = Molarity of HCl( i.e, 0.001M)

V2 = Volume of HCl (i.e 10mL)

so,

M NaoH = M HCl * V HCl / V NaOH

= 0.001 * 10/10.5

Concentration of NaOH = 0.00105 M

The same approach was used to analyze the concentration of vanillin using 0.00105 M NaOH as a titrant.In this case,however, the component of the conductivity is different the observed when strong base reacted with weak acid;the OH- ions deprotonate vanillin, giving as a result a molecule with a negative charge.This Charge effects the conductance of the solution , since this property is dependent on charges present in the system,causing an increase in the conductance with the addition of NaOH in the reaction.Thus, with more OH-.more vanillin will be deprotonated,and a higher conductance will be reported.The following observation shows the behaviour of vanillin sample when titrate with NaOH,previously standardized.

3.2 Calculation:

PRODUCT-1

1000mL of 1M NaOH = 152 grams of vanillin

3.5mL of 0.00105 M NaOH = “ A “ grams of vanillin

i.e., A= 0.000532 gm for 10 grams of custard powder

10gms of custard powder contain 0.000558 gms

100gms of custard powder contain -----------?

= 100*0.000558/ 10

100 grams of custard powder = 0.00558 gms

PRODUCT-2

1000mL of 1M NaOH = 152grams of vanillin

4 mL of 0.00105 M NaOH = “ A “ grams of vanillin

i.e., A= 0.000638 gm for 10grams of custard powder

10gms of custard powder contain 0.000638gms

100 gms of custard powder contain -----------?

= 100*0.000638/ 10

100 grams of custard powder = 0.00638 gms

PRODUCT-3

1000mL of 1M NaOH = 152grams of vanillin

3.5mL of 0.00105 M NaOH = “A“ grams of vanillin

i.e., A= 0.000532 gm for 10 grams of custard powder

10 gms of custard powder contain 0.000558 gms

100 gms of custard powder contain -----------?

= 100*0.000558/ 10

100 grams of custard powder = 0.00558 gms

The various locally available vanilla flavoured custard powder-1, 2, 3 samples were tested for vanillin determination by conductometry. The results (Table IV) were found that the amount of vanillin is within the limits. The measurement of vanillin in custard powder has also been effectively accomplished using the suggested technique.

4. ACKNOWLEDGEMENT:

The authors are thankful to The Management, HMVS, Hyderabad for furnishing the resources required for the completion of this experimental task.

5. CONFLICTS OF INTEREST:

The authors declare conflicts of interest as none.

6. ABBREVIATIONS LIST:

gms – grams

mL – milli liter

NaOH -Sodium hydroxide

HCl – Hydrochloric acid

M – Molarity

Methanol - MeOH

Sodium carbonate - Na₂CO₃

n-Hexane: C₆H₁₄

Copper sulfate: CuSO₄

Double distilled water: DDW

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Received on 29.08.2023 Modified on 17.11.2023

Research J. Science and Tech. 2024; 16(1):6-10.

DOI: 10.52711/2349-2988.2024.00002