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Short Communication

Determination of fortified resistant maltodextrin in juicesby High performance liquid chromatography

Alok Sharma, Nikhil Kumar Singh*, Meena Sharma, Rahul Singh, Chandrakant Katiyar

Dabur Research & Development Centre, Dabur India Limited, Sahibabad 201010, Ghaziabad, U.P., India

a r t i c l e i n f o

Article history:

Received 24 February 2013

Accepted 18 March 2013

Available online 26 March 2013

Keywords:

High performance liquid

chromatograph (HPLC)

Resistant maltodextrin & refractive

index detector

Orange citrus punch juice

* Corresponding author.E-mail address: nikhilfoody@gmail.com (

0976-1209/$ e see front matter Copyright ªhttp://dx.doi.org/10.1016/j.ijcas.2013.03.005

a b s t r a c t

Background/objectives: The importance of food fibers has led to the development of a large

and potential market for fiber-rich products and ingredients and in recent years. There is a

trend to find new sources of dietary fiber that can be used in the food industry. Resistant

maltodextrin is a water soluble fiber and short chain polymer of glucose that are resistant

to digestion in the human digestive system.

Method: As per the AOAC method for determination of total dietary fiber the estimation of

low molecular weight resistant maltodextrin involves many tedious steps which make it

lengthy & time consuming.

Results: Being colorless, tasteless and of low calorific value, resistant maltodextrin qualifies

to be a perfect material to used for dietary fiber fortification purposes. Keeping in view the

time of analysis, possibility of analytical errors in multiple processing steps & cost of

analysis, a simple procedure for determination of resistant maltodextrin has been devel-

oped and validate using High performance liquid chromatograph with refractive index

detector (HPLC-RID) in fortified fruit juices in 0range citrus Punch.

Conclusion: For the confirmation of newly developed analytical method several samples of

juices were also analyzed. The method given specifically measures resistant maltodextrin.

Copyright ª 2013, JPR Solutions; Published by Reed Elsevier India Pvt. Ltd. All rights

reserved.

1. Introduction the large intestine. Hydrogenated resistant maltodextrin is

Resistant maltodextrin (RMD) is, by definition, short chain

polymers of glucose that are resistant to digestion in the

human digestive system. It is produced from starch by heat

and enzymatic treatments1 and contains a mixture of oligo-

and poly-glucosides with a random distribution of glycosidic

linkages of 1e2 and 1e3 in addition to 1e4 and 1e6, which

makes itself resistant to digestive enzymes.2The human

digestive system effectively digests only alpha-1,4 linkages,

therefore other linkages render the molecule resistant to

digestion. Most part of RMD is nondigestible and reaches

N.K. Singh).2013, JPR Solutions; Publi

obtained by hydrogenation of RMD to suppress browning.

Both are highly safe3 and low-caloric food ingredients.4

Resistant maltodextrin been shown to increase fecal bulk

by 93% and decrease symptoms of constipation.5 Research

studies have shown that RMD has improvement of glucose

tolerance,6 and attenuation of postprandial blood triglyceride

elevation.7 It was reported that RMD does not inhibit diffusion

of metal ions in vitro.8 RMD is a water soluble fiber, colorless,

tasteless which does not impart any sensorial attribute to

the food product. Since resistant maltodextrin is stable to

temperature variations & high acid conditions, it becomes a

shed by Reed Elsevier India Pvt. Ltd. All rights reserved.

y = 173.15x + 3385.6R² = 0.9999

0

100000

200000

300000

400000

0 1000 2000

Area

Conc.(ppm)

Linearity

Area

Linear (Area)

Fig. 1 e Calibration curve for resistant maltodextrin.

Table 2e Table shown result for the estimation of RMD indifferent samples.

S. no. Name of the product (sample) RMD (% w/w)

1. Apple pineapple juice 3.03%

2 Banana strawberry juice 1.19%

3. Mango milk 3.02%

4. Green apple juice 1.53%

5. Multi fruit juice 1.35%

i n t e rn a t i o n a l j o u rn a l o f c h em i c a l a n d an a l y t i c a l s c i e n c e 4 ( 2 0 1 3 ) 3 9e4 340

preferable option for fortification as dietary fiber to maintain

the digestive health. Most of the resistant maltodextrin in the

food products today however is purposefully manufactured

from starch by treatment with heat and acid or heat and en-

zymes. Analytically, there are not much references available

on determination of fortified dietary fibers in fruit juices

except for AOAC which has a methodology for Total Dietary

Fiber in Foods Containing Resistant Maltodextrin (AOAC

2001.03) which is applicable on fruit juice. As per the AOAC

method when samples are digested using the human stimu-

lating enzymes of the dietary fiber tests, only insoluble dietary

fibers & high molecular weight soluble dietary fiber get

precipitated and collected on sintered glass crucible, whereas

the low molecular weight soluble resistant maltodextrin

passes through it. The method AOAC 2000.03 suggests a

number of steps including size exclusion chromatography for

estimation of resistantmaltodextrin. Thus a simple procedure

has been developed using High performance liquid chro-

matograph with refractive Index detector which can be used

in routine for determination is resistant maltodextrin in for-

tified fruit juices. For the verification of this analytical method

several samples of different juices i.e., apple pineapple juice,

banana strawberry juice, Mango milk, green apple juice and

multi fruit juice were also analyzed.

2. Materials and methods

2.1. Standards and chemicals

All the chemicals used in the experiments were of known

purity and analytical or HPLC grade.

Table 1 e Recovery of the resistant maltodextrin at different le

Recoverylevel

Resist

Amount present(%w/w)

Amounta added(%w/w)

Tot

80% (A1) 1.40 1.082

100% (B1) 1.40 1.350

120% (C1) 1.40 1.810

Mean

RSD

a Average determined on three replicate determinations.

2.2. Materials

Resistant maltodextrin (Fibersol 2, referred as FS2) is product

of Matsutani Chemical Industry Co., Ltd. (Hyogo, Japan).

Dietary fiber content of FS2 served for this study was 94%,

respectively.8

2.3. Instrumentation

The HPLC system (Agilent 1200 series, USA) consisted of a

refractive index detector (RID/G1362A), Quaternary pump

(G1311A),onlinedegasser (G332A),anauto-injector (ALSG1329A),

thermostat(G1316A) and an HPLC column (Hyper REZ XP carbo-

hydrate column 300 mm � 4.6 mm, 8 mm particle size, Themo-

scientific,USA).DataanalysiswascarriedoutusingChemstation

software (Agilent, USA).

2.4. Chromatographic conditions

The chromatographic elution was carried out in isocratic

mode using a mobile phase consisting of water (HPLC Grade).

The analysis was performed while maintaining the column

temperature at 80 �C, RID temperature at 40 �C at a flow rate of

1.0 mLmin-1. The run-time of the analysis was approximately

15 min.

2.5. Samples

Sample of ‘orange citrus punch (Active Fiber) of Batch No NB

1001Awere provided by the Dabur Research and Development

Centre. Sample of ‘orange citrus punch (Active Fiber) of Batch

No NB 1001A were provided by the Dabur Research and

Development Centre for the study. Apple pineapple juice,

banana strawberry juice, Mango milk, green apple juice and

vels. The experimental findings are given below.

ant maltodextrin

al amount presenta

(%w/w)

Amount recovereda

(%w/w)Recovery(% w/w)

2.482 2.487 100.2

2.750 2.757 100.2

3.210 3.210 100.0

100.1

0.25

Fig.2e

HPLC

pro

filesofresistantm

altodextrin

fortifica

tion

juices.

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i n t e rn a t i o n a l j o u rn a l o f c h em i c a l a n d an a l y t i c a l s c i e n c e 4 ( 2 0 1 3 ) 3 9e4 342

multi fruit juice were also taken for the verification of the

study.

2.6. Procedure

About 2 g of sample was weighed into a 25 ml volumetric flask

which was dissolved and diluted to 25 ml with mobile phase.

Samples were filtered using 0.45 micron nylon filter, before

injecting into the chromatograph.

2.7. Preparation of reference solutions

About 500 mg of standard was dissolved and made up to

volume 50 ml with mobile phase. This was treated as stock

solution and the subsequent dilution was prepared in mobile

phase by taking 5 mL aliquot from stock solution and diluting

up to 50 mL with mobile phase.

2.8. Statistics

Calculations were done on % (w/w) basis.

Area of sampleArea of standard

� concentration of standard ðmg=mlÞweight of sample

� volumemade1000

� 100

3. Method validation

3.1. Precision (system and method)

Repeatability of the sample application and measurement of

peak area were carried out by analyzing the sample in six

replicates. Precision was expressed in terms of percent rela-

tive standard deviation (%RSD) observed for the peak area and

the resistant maltodextrin content in six determinations. The

%RSD for six replicate injections of the standard of resistant

maltodextrin andmeasurement of peak areas was found to be

0.17% and that between the six determinations was observed

to be 0.70%. Such low RSD indicate an excellent precision for

the method.

3.2. Specificity

The specificity of the method was carried out to evaluate the

interference of matrix on resistant maltodextrin peak. The

peak for resistant maltodextrin in the sample was confirmed

by comparing the retention times of the sample peakwith that

of the standard.

3.3. Calibration curve and linearity

The calibration curve was generated from six concentration

levels, i.e., 450, 720, 900, 1080, 1350,, and 1800 mg/ml and the

corresponding peak areas. It demonstrated an excellent line-

arity ina rangeof 451e1805 mg/ml of resistantmaltodextrin. The

linear equation for the calibration curvewas y¼ 173.15x�3385.6

with a correlation coefficient of 0.9999. Fig. 1 displays the cali-

bration curve for resistant maltodextrin.

3.4. Accuracy

The pre-analyzed samples were spiked with extra 80%, 100%,

and 120% of the actual content of resistant maltodextrin

found in the juices by addition of the resistant maltodextrin.

The mixtures thus obtained were reanalyzed by the proposed

method. The experiment was conducted in triplicate for all

the three levels. This was done to check for the recovery of the

resistant maltodextrin at different levels. The experimental

findings are given below (Table 1).

4. Results and discussion

The method given above specifically measures resistant

maltodextrin in several juices (Table 2). The accuracy of the

method was determined as 100% which indicates the excel-

lence of method. Considering HPLC as a powerful tool in the

analysis of complex matrices,9 a new simple and efficient

method is developed on HPLC (Fig. 2). The method is also

found suitable for routine estimation of resistant maltodex-

trin. Most of the laboratories use AOAC 2001.03 method for

determination of resistant maltodextrin10 in juices and other

food commodities which is very lengthy and time consuming

with respect to the various steps of analysis involved

including one where evaporation of water at 50 �C is required

which renders the method lengthy & tedious for routine

purposes as compared to the newly developedmethod. It also

suggests size exclusion chromatography which is not very

popular among small manufacturers for cost and utilization

reason.

For estimating total dietary fiber, the analysis can be per-

formed in parts where the insoluble and high molecular

weight soluble dietary fibers are estimated as per AOAC 985.29

method and the low molecular weight resistant maltodextrin

can be determined by the proposed method. The final result

for total dietary fiber (TDF) would be the sum of the values

obtained in these two analyses.

In this study, method for determination of Resistant malto-

dextrin in orange citrus punch has been validated and other

juice also studied for the confirmation of the study. Thus it can

be stated that the proposedmethod is a simplified with proven

accuracy and precision which can be used for routine deter-

mination of resistant maltodextrin content in the sample of

fruit and vegetable juices fortified with resistant maltodextrin.

Conflicts of interest

All authors have none to declare.

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