Spectrophotometric determination of Anthocyanin Content in Different Fresh Fruits

Abstract:

Anthocyanins are a type of pigment found in fruits that offer health benefit. It belongs to a class of compound called flavonoids that have antioxidant properties. The colour change from pink red to blue violet with change in pH values from 1.0 to 4.5 indicates the presence of anthocyanin. The total anthocyanin content was analyzed by the pH differential method by measuring the absorbance at pH 1.0 and pH 4.5 by UV-Vis spectrophotometer. Anthocyanin content was estimated in five popular fresh fruit samples, viz. apple, cherry, strawberry, blueberry and orange. The results of anthocyanin content were found to be 10 to 410.70 mg/100 g

Keyword:

Anthocyanin, fruits, Spectrophotometry, pH.

Introduction:

Anthocyanins are water-soluble pigments that based on the pH may impart blue, red or purple color. Examples of fruits rich in anthocyanins are blueberry, bilberry, red raspberry, red cabbage, cherry, eggplant peel and dark grapes. Anthocyanins are known to possess antioxidant activity and for the quantification, separation, identification and purification of anthocyanins, the pH differential method has been used to determine the anthocyanin content and the absorbance was measured using UV- spectrophotometer.

Anthocyanins can be used industrially as natural colors and can be used for a wide variety of foods, cosmetics and drugs. As a natural product, anthocyanins are good for health due to their antioxidant properties and may have a role in boosting the immune system. In addition, it has a role in disease prevention and maintaining health. It was found that the percentage of anthocyanins estimated varies in different plant species. This has an impact on the individual intake of anthocyanins which is influenced by climate, cultivation, cultural. Higher anthocyanin intake was associated with an ~8–10% reduction in hypertension risk .

Materials and Method:

Samples, chemicals and reagents:

5 different types of fruits like apple, cherry, strawberry, blueberry, orange were purchased from local market of Kolkata, west Bengal.

Potassium chloride, sodium acetate, ethanol, methanol, petroleum benzene and hydrochloric acid were procured from Merck, India as were of AR Grade.

Instrument /equipment:

Shimadzu UV1900 spectrophotometer, pH meter, sonicator, and analytical balance were used for this analysis.

Procedure:

Preparation of buffers:

pH1.0 buffer (0.025 M potassium chloride) was prepared by using an analytical balance to weigh 1.86g of KCl in a 1000mL beaker. After that, 980mL of distilled water was added and mixed. The pH was measured using the pH meter and adjusted to pH 1.0 using HCl. The solution was transferred to a 1000mL volumetric flask and the volume was made upto 1000 ml using distilled water.

pH 4.5 buffer (0.4M sodium acetate) was prepared by using an analytical balance to weigh 54.43g of CH₃CO₂Na·3H₂O in a 1000mL beaker. After that, 980mL of distilled water was added and mixed. The pH was measured using the pH meter and adjusted to pH4.5 using HCl. The solution was transferred to a 1000mL volumetric flask and the volume was made upto 1000mL using distilled water.

Preparation of Sample:

Approximate 25 to 30 g fresh blended fruits were weighed and extracted with ethanol and 0.1M HCl mixtures (85:15) in (1:2) ratio between puree and solvent and mixed together for 1 hr by using magnetic stirrer. The mixture was filtered through vacuum filtration, collected supernatant solution. The extraction procedure has been done in triplicate.

Preparation of test solutions:

The dilutions were done by using the volumetric pipette to measure and add the test portion in a 50 ml volumetric flask. The dilution factor was measured by diluting the test portion of each fruit by using pH buffer until the absorbance of the spectrophotometer measured at 520 nm was within the linear range (0.2-1.4 AU). Using the calculated dilution factor, the first one with a pH1.0 buffer and the other test solution with pH4.5 buffer.

Spectrophotometric Determination:

The absorbance was measured within 20-50min of preparation for the 2 test solutions pH1.0 and 4.5 using a spectrophotometer at 520 nm. Absorbance was also measured at 700nm for the correction of haze. The diluted test solution that was found to be excessively turbid was filtered before measuring the absorbance using filter papers with<1.2mm pore size to ensure that anthocyanins were not be absorbed. The blank used was distilled water and the test solutions were read against it.

Calculations:

The anthocyanin pigment concentration (as cyanidin-3-glucoside equivalents) was calculated using the following equation:

Anthocyanin concentration : A x MW x DF x10³V

s x l x 26900

Where,

A= (A520nm-A700nm)pH1.0-(A520nm-A700nm)pH4.5

MW=449.2 gmoL G¹for cyanidin-3-glucoside

DF =Dilution factor

L = Path length (cm)

s = Sample weight in g

e =26,900molar extinction coefficient, in L/mol cm for cyanidin-3-glucoside

103 = Factor for conversion from g to mg

Results and Discussions:

The estimation of anthocyanin content was carried out in fresh fruits such as apple, cherry, strawberry, blueberry and oranges. The results were incorporated in Table 1.

Table 1: anthocyanin content of fresh fruits

Types of Fruits	Anthocyanin content (mg/100g)
Apple	23.5
Cherry	135.4
Strawberry	40.8
Blueberry	410.70
Orange	10

Conclusion:

Thus, the method is successfully adopted for estimation of anthocyanin contents of different fresh fruits samples it was been observed that blueberry has highest amount anthocyanin. The method was found to be rapid, accurate and cost effective, hence, may be adopted for routine analysis.

Contributed by : Payel De .

Testimonial

Mitra S.K. have added value to our business process by effective inspection, strict quality control and timely response. It is a pleasant experience to work with “Team Mitra S.K.”. We hope the same service level will continue in the future too....

Sirsendu Mukherjee

Chief Operation, FAMD, Tata Steel Limited..

Mitra SK Inspection Gulf LLC has been appointed by us and the services provided by them has been up to the mark and as per our expectations....

Ajay Mathur

Sr. General Manager,, Emirates Trading Agency L.L.C..

Mitra S. K. Private Limited is skilled in analysis and laboratory working. They understand operating requirements. They bring a winning combination of experience and commitment to excellence. During our rigorous selection process, Mitra S. K. showed us their flexibility and significant capabilities in testing and supervision field. They have deep understanding of the testing, strong customer care capabilities and their global delivery footprints....

Amar Singh Gohil

Mines Manager, Hindustan Zinc Limited, a Vedanta Company.

The Inspection services provided by Mitra S. K. Inspection Gulf has been prompt and accurate and has been in the line with our expectations....

Naser Bustami

General Manager, Stevin Rock L.L.C..

We would like to express our high appreciation of the business association our company had with Mitra S. K. as a testing and inspection house for their quality of service, good documentation and reporting during the job ....

R. Jeyakumar

Executive Vice President (Works),, DCW Limited.

We would like to express our high appreciation for PT Mitra S.K. Analisa Testama as a professional and reliable Testing and Inspection house for their quality services rendered which currently differentiates them from their competitors. they we selected by us due to high quality of services, accurate results, fast response, full support to give documentation and reporting of detail condition during the job....

Tulus Situmeang

AVP – Coal Quality & Sales Compliance Head,, PT Indo Tambangraya Megah Tbk (BANPU).

Their job was excellent and showed Mitra S.K. differential from others....

Geraldo Pimenta

Laboratory Head, MMX.

Mount Gibson engaged Mitra S.K. Private Limited (Mitra S.K.) in mid-2013 to observe and inspect the procedures for unloading of iron ore from vessels in Chinese ports, and the sampling of the ore. Mitra S.K. personnel have since that time overseen numerous shipment movements and cargo discharges, and provided reports thereon. We have found the service offered by Mitra S.K. to be very reliable and professional, with reports being timely and informative.....

Konstantin Gertsen

Shipping Administrator, Mount Gibson Iron Limited.

We’ve been pleased with the improvement we have seen in CIQ results, and Mitra S.K’s reports have certainly helped us to resolve any disputes with our customers in a more timely manner….....

Brian Mcdonald

Senior Director – Asia Pacific Iron Ore Sales,, Cliffs Natural Resources Pty Ltd..

Write to Us

LATEST blog

Mineral Beneficiation and Upgradation

Posted on December 03 2024 By Mitra S.K ADMIN

What if your old PCBs (Printed Circuit Boards) hold untapped Treasure?

Posted on October 17 2024 By Mitra S.K ADMIN

Per- and polyfluoroalkyl substances (PFAS), commonly referred to as "forever chemicals," have emerged as a significant environmental and health concern

Posted on October 04 2024 By Mitra S.K ADMIN

If Your Starch Helping or Holding You Back?

Are your mineral transportation operations hindered by freezing in cold climates? Wondering how MSK tackled this challenge?

Posted on April 30 2024 By Mitra S.K ADMIN

Understanding the Impact of Indirect Greenhouse Gases on Global Warming

Investigation of Physico-chemical properties of Refined Palm oil during frying with special reference to in vitro lipid peroxidation as TBARS

Posted on November 28 2022 By Mitra S.K ADMIN

Spectrophotometric determination of Anthocyanin Content in Different Fresh Fruits

Testimonial

Mitra S.K. have added value to our business process by effective inspection, strict quality control and timely response. It is a pleasant experience to work with “Team Mitra S.K.”. We hope the same service level will continue in the future too....

Sirsendu Mukherjee

Mitra SK Inspection Gulf LLC has been appointed by us and the services provided by them has been up to the mark and as per our expectations....

Ajay Mathur

Amar Singh Gohil

The Inspection services provided by Mitra S. K. Inspection Gulf has been prompt and accurate and has been in the line with our expectations....

Naser Bustami

We would like to express our high appreciation of the business association our company had with Mitra S. K. as a testing and inspection house for their quality of service, good documentation and reporting during the job ....

R. Jeyakumar

Tulus Situmeang

Their job was excellent and showed Mitra S.K. differential from others....

Geraldo Pimenta

Konstantin Gertsen

We’ve been pleased with the improvement we have seen in CIQ results, and Mitra S.K’s reports have certainly helped us to resolve any disputes with our customers in a more timely manner….....

Brian Mcdonald

Write to Us

LATEST blog

Mineral Beneficiation and Upgradation

What if your old PCBs (Printed Circuit Boards) hold untapped Treasure?

Per- and polyfluoroalkyl substances (PFAS), commonly referred to as "forever chemicals," have emerged as a significant environmental and health concern

If Your Starch Helping or Holding You Back?

Understanding PM100 in Mining Activities

Unlocking the Potential of Spent Grain: A Sustainable Approach

Spot Emission Measurement

Opacity Meters

Effluent Treatment Plant

Baseline study in E I A process

Performance guarantee and performance evaluation

Terrestrial Flora and Fauna

Aquatic Flora and Fauna

Mineralogy of Manganese Ore: Beneficiation and Beyond: MSK's Solution to African Mine

Are your mineral transportation operations hindered by freezing in cold climates? Wondering how MSK tackled this challenge?

Understanding the Impact of Indirect Greenhouse Gases on Global Warming

Phase transformation of silica in Coal and Biomass burning processes and its implications on human health

MODIFIED GRAVIMETRIC ESTIMATION OF COBALT IN BLACK MASS

Method development for the estimation of Coumarin content in different Cinnamomum species

Rapid determination of Tyramine and Histamine in Tuna Fish using HPLC-FLD

MICROPLASTIC IN MILK. AN EMERGING THREAT

Estimation of Proline in Honey by High Performance Liquid Chromatography

DNA based approach for the detection of GMO

Development of an HPLC-FLD method for rapid determination of Tyramine and Histamine in Tuna Fish

Determination of Acrylic acid emissions from industrial process by HPLC

COMPARATIVE STUDY OF NANO-FERTILIZER & CONVENTIONAL FERTILIZER WITH RESPECT TO SUSTAINED NUTRIENT RELEASE

Assessing leaching potential of Electronic Waste

Analysis of Red Mud

Determination of gold using ICP-OES after activated carbon enrichment

Determination of platinum group metals using nickel sulfide (NiS) fire assay

QUANTIFICATION OF POTASSIUM IN NPK FERTILIZER BY GRAVIMETRY

Rapid Phase Composition Study of Portland cement-Clinker by Powder X-ray Diffraction Technique and its Impact

Recycling of lead acid batteries

Spectrophotometric determination of phosphorous in ferro alloys, manganese ore and iron ore

A Novel Technique to Identify Mineral Oil Contaminants in Vegetable Oil

Alpha amylase enzyme activity of various grains with special reference to sprouting

A degradation study of beta carotene in infant food and raw vegetables by ozone treatment

Environmental impacts of firecrackers during festivals, events - a lab study

Spectrophotometric determination of Anthocyanin Content in Different Fresh Fruits

EXTRACTION AND ISOLATION OF PIPERINE FROM BLACK PEPPER

Investigation of Physico-chemical properties of Refined Palm oil during frying with special reference to in vitro lipid peroxidation as TBARS

Nitrogen Profile in Fertilizer by IR Absorption Spectroscopy

Protein Digestibility Corrected Amino Acid Score (PDCAAS)- A method to evaluate quality of protein

Determination of Organic carbon and Inorganic carbon of Biomass

Analysis of Anti-Nutritional Factors of Wine

Determination of Ozone Generation Efficiency of UV Lamp by Chemical Method

Our clients