Impact of Starch Storage Condition on Glycemic Index and Resistant Starch of Cooked Potato

(Solanum tuberosum) Tubers

Milan Kumar Lal*1†, Awadhesh Kumar2†, Pinky Raigond1, Som Dutt1, Sushil Sudhakar

Changan1, Kumar Nishant Chourasia1, Rahul Kumar Tiwari1, Dharmendra Kumar1, Srigopal

Sharma3, Swarup Kumar Chakrabarti1 and Brajesh Singh1

1
ICAR-Central Potato Research Institute, Shimla, Himachal Pradesh, India;
2
ICAR-National Rice Research Institute, Cuttack, Odisha, India
3
Govind Ballabh Pant University of Agriculture and Technology, Pantnagar, Uttarakhand,

India.

†
These authors contributed equally to this work.

*Corresponding Author: Milan Kumar Lal

Affiliation: Division of Crop Physiology, Biochemistry & PHT

ICAR-Central Potato Research Institute, Shimla, 171001, Himachal Pradesh, India

Email: milan2925@gmail.com

Contact No: +91-9718815448

Abstract

Potato is a modified stem which is rich in the starch. Very often potato is categorized under

high Glycemic Index (GI) food. Consuming high GI foods should be done with moderation to

prevent insulinemic spikes, which can be a preventive measure against diabetes and related

disorders. We had developed a modified, inexpensive and precise high throughput in-vitro

This is the author manuscript accepted for publication and has undergone full peer review but has not
been through the copyediting, typesetting, pagination and proofreading process, which may lead to
differences between this version and the Version of Record. Please cite this article as doi:
10.1002/star.201900281.

This article is protected by copyright. All rights reserved.

method for estimation of GI in potato tubers. Among the six varieties, Kufri Chipsona-3

exhibited the highest GI (83.08) whereas Kufri Jyoti had the lowest value (72.87); the

resistant starch (RS) content being 1.08% (low) and 2.18% (high) respectively. Our study

showed a significant negative correlation (R=-0.88) between GI and RS whereas a negative

non-significant correlation (R=-0.79) was found between GI and amylose content (AC).

Further, starch storage of cooked potato tubers at 4ºC for various periods (up to 48h) resulted

in a significant reduction in GI and increase in RS content. This newly developed protocol for

estimation of GI in potato is a simple, rapid and precise method. This will not only help the

food industry but also breeder to select the low GI genotype for their breeding programme.

Keywords: glycemic index; resistant starch; cooling; amylose; starch hydrolysis index

1. Introduction

Potato ranks third among the highly consumed food crops with more than one billion

consumers in the world. It is an extensively consumed underground vegetable crop for which

the year 2008 AD was named as “The International Year of the Potato”.[1] Apart from

carbohydrates, potato is a rich source of qualitative proteins, phenolic compounds, ascorbic

acid, and minerals like phosphorus, potassium, calcium along with some elemental

micronutrients.[2] The major carbohydrate present in tubers and root crops is starch account

for 16-24% of their weight and remaining to be water, proteins and lipids.[3] The starch of raw

potato is resistant to digestion, but the digestibility has been found to be increased after

cooking.[4] Due to its high digestibility rate, the glycemic load (GL) and glycemic index (GI)

was found to be high which leads to an increase in postprandial glycemic response. Extended

consumption of carbohydrate-rich food with a sedentary lifestyle may increase the risk of

This article is protected by copyright. All rights reserved.

type-2 diabetes mellitus, cardiovascular disease and obesity. The food and processed products

with low GI and GL were recently suggested to reduce the risk of these diseases.[5]

Starch is made up of amylose (10-25%) and amylopectin (80-90%).[2] Based on

digestibility, starch can be divided into three types namely, rapidly digestible starch (RDS,

hydrolyzed within 20 min), slowly digestible starch (SDS, hydrolyzed between 20 min to 2h)

and resistant starch (RS, not hydrolyzed within 2h).[6] Among these, RS is very important due

to its health-promoting properties. RS is starch-based fibre fraction that resists digestion in

the intestine and passes as such into the large intestine.[7] The RS content of cooked potato

varies from 5-10%[6] but can be recovered by cooling that facilitates the retrogradation of

starch which ultimately results in lowering of GI and GL.[8] RS is also termed as dietary fibre

and it has the same physiological effect on the human body as that of dietary fibre which has

been shown to reduce the risk of type-2 diabetes.[9]

Starchy foods like potato, rice, maize, wheat and other food products can be classified

according to their digestibility in the human gut, which is characterized by the rate and

duration of glycemic response.[10] Jenkins et al., (1981)[11] first introduced the term GI as a

parameter to classify foods based on their blood glucose-raising ability. The GI is defined as

the response of blood glucose measured as an area under the curve (AUC) in test food taken

by a person which is expressed as a percentage of the AUC following consumption of

reference food consumed by the same person under standard condition. Literature suggests

that GI of boiled potato ranges from very low (23 for an unspecified cultivar) to very high

(144 for Desiree). There are three levels of GI viz., low (≤ 55), medium (≥ 56-69) and high (≥

70).[12] Potato is categorized under high GI food as its average GI value is above 70. This call

for an effort to develop low GI varieties and processing techniques to further reduce the GI of

potato. The postprandial blood glucose response is also influenced by the amount of

carbohydrate consumed. Here comes the concept of GL. It is measured as GI multiplied by a

This article is protected by copyright. All rights reserved.

gram of carbohydrate per serving of test food. Each unit of GL represents the equivalent

glycemic effect of one gram of carbohydrate from a reference food such as white bread.[13,14]

GI can be estimated using both in vivo and in vitro methods. The in vivo method has

long been used for the determination of GI in food and food product, but it requires human

subjects. The complication of in vivo method is that the physiological condition of the human

body varies from person to person resulting in the lack of precision and poor reproducibility

of the results.[15] However, in vitro methods do not suffer from such constraints. Moreover,

this method is better as it is quick, cost-effective and large number of samples can be

screened within a short period of time.[16] This will facilitate potato breeders to select the

lines suitable for breeding low GI with high RS potato cultivar, that may be exploited to

provide health benefit in general and diabetics people in particular.[17]

Cooling of boiled potato in a refrigerator for more than 24h lead to an increase in RS

and decrease in GI.[18,19] Recurrent heating and cooling of potato decrease the glycemic

response by slowing digestion rate and absorption of released glucose. Moreover, it also

increases the RS which hinders the absorption of glucose in the small intestine. Refrigeration

of boiled potato for more than 24h increase the formation of RS which cannot be digested in

the small intestine and thus lowering the glycemic impact of potato tuber. Intrinsic factors

like amylose, granule size of starch and molecular order influence starch digestibility along

with the presence of fibres, fats and proteins in carbohydrate food.[20] The external factor such

as the addition of lipid in food and food products affect the retrogradation properties by

forming amylose-lipid complex.[21]Aim of the present study was to develop quick and precise

in vitro method for estimation of GI in potato tuber and also to find out the relationship

among GI, RS and AC. Since potato is consumed in cooked form, another objective was to

find how starch storage condition (time and temperature) affects the GI, GL, RS and other

nutritional profiles of starch in tubers.

This article is protected by copyright. All rights reserved.

2. Material and Method

2.1. Plant material

Six Indian potato varieties, namely Kufri Chipsona-1 (KC1), Kufri Chipsona-3 (KC3)

(used for making chips), Kufri Frysona (KF) (used for making French fries), Kufri Jyoti (KJ),

Kufri Girdhari (KG) and Kufri Pukhraj (KP) (cover maximum area under cultivation in India

and used for table purpose) were procured from Central Potato Research Institute-Regional

Station, Modipuram, India in 2018. The potato tubers were boiled in the open pan until fully

cooked and peels were removed. The boiled tubers were ground to paste and was lyophilized

(Scanvac Cool Safe, Labogene, Denmark). The freeze-dried samples were ground to a fine

powder and passed through a 100 mesh size sieve. All the analysis was carried out in

triplicate.

2.2. Development of an improved protocol for in vitro GI estimation

In vitro method for estimation of starch hydrolysis index (SHI) and GI was improved and

standardized.[22] The digestive enzymes used in our experiment to digest starch were pepsin

(3000U/mL, from porcine gastric mucosa, Sigma-Aldrich, USA), α-amylase (10U/mg solid,

porcine pancreatic, Sigma-Aldrich, USA) and amyloglucosidase (3300U/mL, Himedia

bioscience, India). We followed the protocol of Goñi et al., 1997 with some modification (we

used a dialysis membrane which was used to mimic the human small intestine) and

standardized it for potato. Briefly, potato powder (0.2 g) was boiled with 2 mL of distilled

water for 2 minutes in a boiling tube. This was followed by addition of 5 mL of 0.1M

phosphate buffer (pH 6.9) (MP Biomedicals, Santa Ana, CA) with vigorous shaking. Prior to

the addition of 0.2 mL of pepsin (250 mg/mL) the pH of the solution was brought down to

2.5 with o-phosphoric acid (Merck, Darmstadt, Germany) and placed in an incubator shaker

at 37ºC at 120 rpm for 60 min. Then pH was adjusted to 6.9 with potassium hydroxide (20%)

followed by the addition of 0.3 mL of α-amylase (125mg/mL). The solution was then

This article is protected by copyright. All rights reserved.

transferred to dialysis membrane (dialysis membrane 50 width-24.26 mm, diameter-14.3 mm,

Himedia Biosciences, India, Catalog no. LA387-5MT) of 10 cm length which was placed in

50 mL centrifuge tube containing 40 mL of 0.1M phosphate buffer (pH 6.9). This was

incubated at 37ºC, at 120 rpm for 180 min. At every 30 min interval, 0.5 mL of aliquots were

drawn from the centrifuge tube up to 3 h, and mixed with 1.5 mL of 0.4 M, sodium acetate

buffer (pH 4.75) followed by addition of 40 µL of amyloglucosidase and incubation at 50ºC

for 30 min. The final volume was made to 10 mL with distilled water. Aliquot of 0.3 mL (in

triplicates) were incubated with GOPOD (glucose oxidase-peroxidase) at 50ºC for 30 min.

The absorbance was measured at 510 nm (Spectrophotometer, Model-4001/A,

ThermoSpectronic, USA). D-glucose (0.2 g) (MP Biomedicals, USA) was taken as a standard

carbohydrate for this experiment. The average value was estimated and used to plot curves

followed by calculating the AUC. The SHI was calculated by dividing the AUC of potato

sample by AUC of glucose, which was expressed as a percentage. The formula suggested by

Goni et al. (1997)[23] was used to calculate the predicted glycemic index (PGI).

PGI = 39.21 + (0.803 x SHI)

Where, PGI: predicted Glycemic Index or Glycemic index, SHI: starch hydrolysis index.

The formula, GL = (GI × Available carbohydrate)/100 was used to calculate the GL for 100g

of potato (http://www.glycemicindex.com/faqsList.php#1).

Where, GL: Glycemic Load, GI: Glycemic index

2.3. Estimation of RS content in potato

RS kit (Megazyme International Ireland Ltd) was used for estimation of RS content in

potato with slight modification. Non-resistant starch was hydrolyzed by incubation of dried

potato powder (0.1g) at 37 ºC with pancreatic α-amylase (10mg/mL) and amyloglucosidase

(300U/mL) in 25 mL of the conical flask for 16h on incubator shaker. After incubation for

16h, the enzymatic reaction was stopped using ethanol and centrifugation was done to

This article is protected by copyright. All rights reserved.

separate non-resistant starch. The residue was purified with 4 mL of 50% ethanol. The

supernatant was discarded and the pellet was dried followed by re-suspension in 2M KOH

and stirred on an ice bath for 25 minutes. Sodium acetate (8 mL, 1.2M, pH-4.3) buffer and

0.1 mL of amyloglucosidase (3300U/mL) were added and incubated at 50ºC for 30 min. An

aliquot (0.1 mL) was taken and was mixed with 3 mL of GOPOD reagent (as provided by RS

kit of Megazyme International, Ireland Ltd) which was then incubated at 50ºC for 20 min.

The RS content was measured at 510 nm and expressed as a percentage (%) of dry matter.

2.4. Estimation of Amylose content (AC)

Amylose content of potato was estimated using the protocol developed by Juliano et al.

1981[24] with modification. Dry potato powder (0.1g) was mixed with 1 mL of absolute

ethanol and 9 mL sodium hydroxide (1 N) and the tube was placed in a boiling water bath (10

min). Then, the volume was made up to 100 mL. An aliquot (5 mL) was mixed with 1 mL

acetic acid (1 N) and 2 mL of iodine solution (1g of iodine and 10 g of KI in 500 mL in

water) in a volumetric flask. This was incubated in dark for 20 min for colour development.

The final volume was made to 100 mL and absorbance was read at 620 nm.

2.5. Estimation of available carbohydrate (ACHO)

ACHO of potato sample were determined using the Megazyme kit (as provided by

ACHO kit of Megazyme International, Ireland Ltd) in freeze-dried potato powder in

triplicate.

2.6. Estimation of total starch content (TSC)

Total starch content was analysed in the potato tuber according to Bjorck 1992.[25] TSC

was analysed in freeze-dried potato powder in triplicate.

2.7. Organoleptic test for texture analysis of potato

The texture analysis by the sensory or organoleptic test was done for all the six varieties

by taking 10 volunteers. The potato tubers were cooked by boiling (15-20 min) and after

This article is protected by copyright. All rights reserved.

cooling were examined. The precautions were taken to wash the mouth before testing the

sample and the final decision about each organoleptic characteristic was taken on consensus.

The texture was decided in four major categories i.e. 1) extremely mealy–floury, 2) medium

to slightly mealy/granular-Mealy, 3) gummy/pasty−waxy and 4) watery or

translucent−soggy. The relationship between the texture and GI were classified as per Henry

et al. 2005 [26]

2.8. Starch storage and cooling experiment

Whole potato tubers were boiled for 30 minutes on an induction stove until the sample

could be pierced through using a fork. Three potato tubers were taken from each variety,

peeled and mashed. The samples were used for in vitro GI determination and RS content. The

effect of retrogradation was studied on SHI, GI and RS by storing the samples in a

refrigerator at 4ºC for 0, 12, 24 and 48 h. The effect of storage was also studied on the

nutritional profile of starch such as AC, ACHO, GL, and TSC by storing the samples in the

refrigerator at 4ºC for 0h (cooked) and 48h (cooled).

2.9. Statistical analysis

Data analysis was also carried using Graph Pad Prism 7 software (GraphPad Software

Inc., California, USA). Analysis of variance (ANOVA), the test of significance and

comparison of means, using the Tukey's test were performed using Graph Pad Prism 7

software. Pearson correlation was used to establish the relationship between HI, GI, RS and

AC. The difference among the treatment in retrogradation experiment was carried out by two-

way ANOVA and Tukey multiple comparisons test was used to compare between the means

of treatments (P ≤ 0.05).

This article is protected by copyright. All rights reserved.

 3. Result and Discussion

3.1. Improved in vitro GI method for potato and its validation

We improved in vitro protocol for estimation of GI in the potato which is the modified

version of the method developed earlier for different crops.[22,27] Here we used dialysis

membrane to mimic the small intestinal wall and avoid the need to inactivate the enzyme as it

is restricted within the membrane. The maltose released as a result of starch digestion was

further hydrolysed with amyloglucosidase to one glucose molecule. The predicted GI value

of six varieties viz., KC1, KC3, KF, KG, KJ and KP using Goñi’s protocol was 87.53, 88.03,

82.21, 81.56, 76.86, and 81.78 respectively. The formula used for the estimation of predicted

GI was PGI = 39.21 + (0.803 x HI)]. However, when we used the dialysis membrane in our

modified protocol the predicted GI values changed. The change in the GI value was due to

the enzyme (α-amylase) and pH condition provided inside the dialysis membrane which

mimics the diffusion process of the small intestine. The predicted GI values of six varieties

after using dialysis membrane was 82.30, 83.08, 76.49, 77.17, 72.87, and 75.64 in KC1, KC3,

KF, KG, KJ, and KP respectively. There was a significant change in the GI values when both

the method was performed. Our protocol showed a strong significant positive correlation with

the Goñi’s protocol (Linear equation, Y = 0.9326x + 0.525, R2 = 0.9641) (Fig. S1). This

modified protocol for determination of GI is cheap, simple, requires easily available

chemicals and equipment (like spectrophotometer) and unlike other in vivo methods, it is

independent of human or animal subject requirement in the experiment, which was also

advocated by other authors. [20,27–29]

3.2. Relationship among GI, RS and AC in potato

When the protocol of Goñi’s et al., 1997[22] was used, the GI in six potato varieties

varied from 88.03 (KC3) to 76.86 (KJ). Though the trend was the same, the absolute values

of GI was different from Goñi’s protocol and modified protocol (Fig. S1). The estimated GI

This article is protected by copyright. All rights reserved.

value of modified protocol varied significantly from 83.08 to 72.87 in cooked tubers of KC3

and KJ respectively (Table 1). The SHI was calculated as the percentage of total glucose

released from the samples as compared to that released from standard glucose (0–180

min).[22] The SHI (area under the curve with respect to that of the reference food sample) was

positive and significantly correlated with GI of potato (Table 2, Table S1). This was also

concordant with Kumar et al., 2018 [27] where they worked in rice.

The RS content was varied significantly among the six varieties. The maximum RS

was observed in KJ (2.18%) with the lowest GI (72.87±0.09) whereas KC3 (1.08%) had the

minimum RS and the highest GI (83.08±0.15) (Table 1). These results were supported by

statistical analysis where RS and GI were highly significant and negatively correlated (R =-

0.88) (p<0.05, Table 2, Table S1).Previous work in different crops reported similar

results.[27,30] From these results, it was evidence that the varieties with low RS content

showed a higher rate of starch hydrolysis which led to higher GI. Whereas the variety

containing higher RS was digested slowly in in vitro conditions. A similar phenomenon

occurs under in vivo condition, where potato with high RS and low GI helped in maintaining

the blood glucose level with decreased insulin response.[31]

We observed that AC was maximum in KJ (22.32%) and minimum in KC3 (18.43%)

(Table 1). AC was found to be negatively correlated with GI (R = -0.79), but was non-

significant (Table 2; Table S1). These results are concordant with the previous data where

Fitzgerald et al. 2011[32] also reported a negative correlation between the predicted GI and

AC. However Kumar et al. 2018[27] reported that GI was poorly correlated with AC (R = -

0.185) in rice, this may be due to the presence of RS type 5 in rice as starch granules are

physically trapped within the food matrix. However, in potato, there is the presence of RS 3.

In rice, amylose forms complex with lipid and make it thermally stable which is the

characteristic of RS type 5.[33]

This article is protected by copyright. All rights reserved.

 3.3. Relationship between RS and AC in potato

The AC alone does not contribute to lowering GI in potato. There are other factors which

are also responsible for lowering GI value along with amylose interaction. Amylose form

complex with lipid component present in the potato which leads to decrease in the glycemic

responses.[34]
[35]
In raw potato, the RS content can be as high as 70% but it decreases while cooking

and can come down to 5 to 10%.[6] Miller et al., 1992 and Leeman et al., 2008 provided

conclusive evidence of the positive correlation between AC and RS enhancement. Our result

shows a positive correlation between RS and AC (R = 0.68) but was not significant (Table 2;

Table S1). The results were concordant with Kumar et al. 2018.[27] This indicates that the

more the amylose content the more is the RS and the lower is GI and vice versa.

3.4. Effect of maturity and texture on GI

The sensory or organoleptic test analysis was done taking 10 volunteers for all the six

cultivars. It was found that KC1 and KC3 were classified as floury texture and have a high

GI. However, KF, KG and KJ all have inherent waxy property, out of which KF and KJ have

intermediately high GI whereas KG have a high GI (Table 3). Henry et al. 2005[26] also found

a strong correlation between the amylose content, waxy loci and GI which indicated that

amylose is the major constituent that affects GI. The six varieties also showed variation in GI

and texture. High GI variety had floury texture, whereas the variety which has intermediately

high GI had waxy texture (Table 3). Similar results were reported, where it was observed that

waxy cultivars exhibiting high moisture and low starch had medium GI in potato, whereas the

floury potato exhibiting low moisture and high starch had high a GI.[26] All six varieties are

grown in India is of medium duration (90 to 100 days), and GI of varieties comes under high

and intermediately high GI.

This article is protected by copyright. All rights reserved.

 3.5. Effect of retrogradation on GI and RS

Processing condition such as cooling can influence the glycemic impact of cooked

potatoes through retrogradation of starch polymers.[37,38] Low field nuclear magnetic

resonance (LF-NMR) studies reported that retrogradation is more extensive in boiled potatoes

after 24-48h of cooling than after 1-2h.[39] Our result showed that cooling/retrogradation had

significant GI lowering effect when cooked potatoes were stored for 0h, 12h, 24h and 48h

(Fig. S2). The maximum GI lowering effect was seen after 48 h of cooling. Our results

showed that GI ranged from 72.87 to 83.08, 66.67 to 78.28, 63.19 to 75.89, 61.63 to 69 for 0,

12, 24 and 48h of retrogradation respectively (Fig. 1(b); Table S2(a)). These results were

concordant with previous studies.[4,40] A similar trend was observed for the SHI of six

cultivars of potato (Fig. 1(a)). The GI decreased significantly in all cultivars (p<0.05) and the

decrease was maximum (22%) and minimum (13.44%) in KC3 and KG respectively (Table

S2(a)). Studies suggests that GI of potato was decreased significantly by 25% when cooled

and again reheated for consumption.[41] The retrogradation may change the starch structure

under low temperature, which ultimately leads to a lowering of GI and GL.[4,8]

The RS content was found to increase in the boiled potatoes when cooled at 4ºC,

however, the maximum effect was observed after 48h. The RS content of potato varieties

ranged from 1.08% to 2.18%, 1.80% to 3.66%, 2.32% to 4.13%, 2.55% to 4.52% for 0, 12, 24

and 48h of retrogradation respectively (Fig. 1(c); Table S2(b)). We found that there was a

significant increase in RS, up to 152.59% in KG after 48h of storage (p<0.05) (Fig. 1(a);

Table 2(b)). These results are concordant with those of Monro et al., 2009[38], where they

reported an increase in RS from 3-7% to 5-11% after storage of cooked potatoes at 4°C for

44h. During cooling, the gelatinized starch molecules begin to realign and RS formation is

enhanced. RS may be recovered by boiling potatoes and cooling or retrogradation (Fig. 3)

[8]
which significantly reduce GI . GI of potato was decreased significantly by 25% when

This article is protected by copyright. All rights reserved.

cooled and again reheated for consumption.[41] Storage of starch under low temperature

increases the resistant starch from 1.18% to 4.63%, when kept for 24 h in the refrigerator.[40]

This practice can be easily followed at home on a daily basis to get more dietary fibre.

3.6. Effect of storage on starch characteristics

The GI and GL was also affected by starch properties like RS, AC, ACHO and

TSC.[38,40,42,43] We report that the AC varied from 18.43% to 22.32 % and 19.96% to 23.99%

in cooked and cooled potato tubers respectively (Fig. 2(c)). It was observed that AC increased

significantly after cooling in all the storage treatments as compared to non-retrograded

samples (Table S4(c); p<0.05). When tubers were cooked there was the disruption in the

granules of which form a gel. However, cooling increases the firmness in gel and disrupted

granule re-associate.[31] The ACHO varied significantly from 10.52 (KP) to 14.59 g (KG) and

9.28 (KP) to 12.24 g (KG) per 100g of dry weight of cooked and retrograded samples

respectively (Fig 2(d); Table S4(d)). As discussed earlier GL is dependent on GI and

ACHO.[40,44] Therefore the food with lower ACHO contributes to low GL, despite of its high

GI value. The TSC in cooked and retrograded sample varied from 57.74% (KF) to 71.03%

(KJ) and 54.93% (KF) to 67.96 % (KC1) respectively (Fig 2(f)). It was found that there was

significant decrease in TSC in retrograded treatment in all the six cultivars (Table S4(f);

p<0.05), except KC3, where TSC was non-significant between treatment. The minimum

decrease in TSC observed when potato was retrograded was 2.2% (KC3) and the maximum

was 6.4% (KF). The decrease in TSC during retrogradation was may be due to re-association

and reformation of amylose and amylopectin into an ordered structure.[4] It is to be noted that

KC3 is a chipping potato variety whereas KF is extensively used for making French fries.

The GL reported being decreased significantly in all the six varieties (Table S4(e);

p<0.05). It ranged from 15.91 (KP) to 22.52 (KG) and 11.92 (KP) to 16.53 (KG) in cooked

and retrograded samples, respectively (Fig. 2(a)). Pinhero et al., 2016[42] also reported the

This article is protected by copyright. All rights reserved.

range of GL both in cooked and retrograded potato from 2 to 20 in fourteen cultivars of

potato. However, the maximum reduction of GL was found in KF (31.1%) and minimum in

KP (25.1%) as compared to cooked tubers. According to Brennen 2005[45] GL values, less

than 10 is considered as low, between 11 to 19 as medium and higher than 20 is considered as

high glycemic food. Higher is the GL, more is the insulinogenic effect of food.[46] Due to the

decrease in GI as well as ACHO in retrograded potato, GL also decreased significantly in all

six varieties as compared to cooked tubers (Table S4(e); p<0.05).

4. Conclusion

The aforementioned in vitro method for GI, which is standardized for potato will be

helpful to screen a large number of varieties/germplasm. This will also help breeders to

evaluate the GI of their variety/hybrids in a very short course of time. The finding of a

negative correlation between GI and RS and with AC will help breeder and molecular

biologist to develop variety high in AC. Moreover, the lowering of GI and increasing in RS

due to storage would be helpful for the general population to reduce the intake of high GI

foods and gain the benefits like avoiding type 2 diabetes, obesity and cancer if they follow a

healthy lifestyle.

Acknowledgement

This work has been supported by ICAR-Central Potato Research Institute, Shimla and

ICAR-National Rice Research Institute, Cuttack. The authors also acknowledge contributions

of Puja, Archana Panda, Chandrashekhar Sahu and Lopamudra Nayak of ICAR-NRRI for

their constant support in conducting the experiment.

Conflict of interest

The authors confirm that this manuscript has no conflict of interest.

This article is protected by copyright. All rights reserved.

Reference

[1] “FAOSTAT,” can be found under http://www.fao.org/faostat/en/#home, n.d.

[2] P. Raigond, B. Singh, S. Dutt, Dalamu, A. Joshi, Indian J. Hortic. 2017, 74, 103.

[3] R. Hoover, Carbohydr. Polym. 2001, 45, 253.

[4] J. H. Dupuis, Q. Liu, Am. J. Potato Res. 2019, 96, 127.

[5] D. J. A. Jenkins, C. W. Kendall, L. S. Augustin, S. Franceschi, M. Hamidi, A.

Marchie, A. L. Jenkins, M. Axelsen, in Am. J. Clin. Nutr., Narnia, 2002, pp. 266S-

273S.

[6] H. N. Englyst, S. M. Kingman, J. H. Cummings, in Eur. J. Clin. Nutr., 1992, pp. S33-

50.

[7] I. Tetlow, Agronomy 2018, 8, 81.

[8] J. H. Dupuis, Z. H. Lu, R. Y. Yada, Q. Liu, Int. J. Food Sci. Technol. 2016, 51, 2233.

[9] M. Champ, L. Martin, L. Noah, M. Grams, FOOD Sci. Technol. YORK-MARCEL

DEKKER- 1999, 169.

[10] J. Singh, A. Dartois, L. Kaur, Trends Food Sci. Technol. 2010, 21, 168.

[11] D. J. A. Jenkins, T. M. S. Wolever, R. H. Taylor, H. Barker, H. Fielden, J. M.

Baldwin, A. C. Bowling, H. C. Newman, A. L. Jenkins, D. V Goff, Am. J. Clin. Nutr.

1981, 34, 362.

[12] T. M. S. Wolever, Eur. J. Clin. Nutr. 2013, 67, 1229.

[13] J. C. Brand-Miller, Nutr. Rev. 2003, 61, S49.

[14] J. W. Woolnough, J. A. Monro, C. S. Brennan, A. R. Bird, Int. J. Food Sci. Technol.

2008, 43, 2245.

[15] S. Hirsch, G. Barrera, L. Leiva, M. Pía De La Maza, D. Bunout, Nutr Hosp 2013, 28,

541.

[16] K. L. Ek, S. Wang, L. Copeland, J. C. Brand-Miller, Br. J. Nutr. 2014, 111, 699.

This article is protected by copyright. All rights reserved.

[17] V. Buono, A. Paradiso, F. Serio, M. Gonnella, L. De Gara, P. Santamaria, J. food

Compos. Anal. 2009, 22, 556.

[18] A. . . . kerberg, H. G. M. M. Liljeberg, . . Granfeldt, A. . rews, I. M. .

j rck, I. M. . j rck, J. Nutr. 1998, 128, 651.

[19] M. Leeman, E. Östman, I. Björck, Eur. J. Clin. Nutr. 2008, 62, 87.

[20] A. C. Dona, G. Pages, R. G. Gilbert, P. W. Kuchel, Carbohydr. Polym. 2010, 80, 599.

[21] R. M. Mariscal‐ Moreno, J. de D. Figueroa‐ Cárdenas, D. Santiago‐ Ramos, P.

Rayas‐ Duarte, Int. J. Food Sci. Technol. 2019, 54, 1651.

[22] I. Goñi, A. Garc\’\ia-Alonso, F. Saura-Calixto, Nutr. Res. 1997, 17, 427.

[23] I. Goñi, L. Bravo, J. A. Larrauri, F. Saura Calixto, F. S. Calixto, Food Chem. 1997, 59,

269.

[24] B. O. Juliano, C. M. Perez, A. B. Blakeney, T. Castillo, N. Kongseree, B. Laignelet, E.

T. Lapis, V. V. S. Murty, C. M. Paule, B. D. Webb, Starch-Stärke 1981, 33, 157.

[25] I. M. E. Björck, M. A. Siljeström, J. Sci. Food Agric. 1992, 58, 541.

[26] C. J. K. Henry, H. J. Lightowler, C. M. Strik, M. Storey, Br. J. Nutr. 2005, 94, 917.

[27] A. Kumar, U. Sahoo, B. Baisakha, O. A. Okpani, U. Ngangkham, C. Parameswaran,

N. Basak, G. Kumar, S. G. Sharma, J. Cereal Sci. 2018, 79, 348.

[28] D. J. A. Jenkins, D. G. Popovich, C. W. C. Kendall, A. V. Rao, T. M. S. Wolever, N.

Tariq, L. U. Thompson, S. C. Cunnane, Scand. J. Gastroenterol. 1997, 32, 10.

[29] H. Araya, P. Contreras, M. Alvina, G. Vera, N. Pak, Eur. J. Clin. Nutr. 2002, 56, 735.

[30] J. H. Cummings, H. N. Englyst, Am. J. Clin. Nutr. 1995, 61, 938S.

[31] K. L. Ek, J. Brand-Miller, L. Copeland, Food Chem. 2012, 133, 1230.

[32] M. A. Fitzgerald, S. Rahman, A. P. Resurreccion, J. Concepcion, V. D. Daygon, S. S.

Dipti, K. A. Kabir, B. Klingner, M. K. Morell, A. R. Bird, Rice 2011, 4, 66.

[33] B. Sawicka, P. Das Gupta, Acta Sci. Pol. Ser. Agric. 2018, 17, 153.

This article is protected by copyright. All rights reserved.

[34] S. S. Jayanty, K. Diganta, B. Raven, Am. J. Potato Res. 2019, 96, 183.

[35] A. García-Alonso, I. Goñi, Nahrung - Food 2000, 44, 19.

[36] J. B. Miller, E. Pang, L. Bramall, Am. J. Clin. Nutr. 1992, 56, 1034.

[37] K. Lin Ek, S. Wang, J. Brand-Miller, L. Copeland, Food Funct. 2014, 5, 2509.

[38] J. Monro, S. Mishra, E. Blandford, J. Anderson, R. Genet, J. Food Compos. Anal.

2009, 22, 539.

[39] E. Micklander, A. K. Thybo, F. van den Berg, LWT-Food Sci. Technol. 2008, 41,

1710.

[40] B. Nayak, J. De J. Berrios, J. Tang, Food Res. Int. 2014, 56, 35.

[41] R. Tahvonen, R. M. Hietanen, J. Sihvonen, E. Salminen, J. Food Compos. Anal. 2006,

19, 372.

[42] R. G. Pinhero, R. N. Waduge, Q. Liu, J. A. Sullivan, R. Tsao, B. Bizimungu, R. Y.

Yada, Food Chem. 2016, 203, 356.

[43] S. Mishra, J. Monro, D. Hedderley, Starch/Staerke 2008, 60, 500.

[44] A. Zurbau, A. L. Jenkins, E. Jovanovski, F. Au-Yeung, E. A. Bateman, C. Brissette, T.

M. S. Wolever, A. Hanna, V. Vuksan, Eur. J. Clin. Nutr. 2019, 73, 79.

[45] C. S. Brennan, Mol. Nutr. Food Res. 2005, 49, 560.

[46] F. S. Atkinson, K. Foster-Powell, J. C. Brand-Miller, Diabetes Care 2008, 31, 2281.

This article is protected by copyright. All rights reserved.

Fig. 1: (a) Variation in starch hydrolysis index (SHI) of six potato varieties after 0, 12, 24 and

48 h of storage at 4°C. (b) Variation in Glycemic Index (GI) of six potato varieties after 0, 12,

24 and 48 h of storage at 4°C. (c) Variation in Resistant Starch (RS) of six potato varieties

after 0, 12, 24 and 48 h of storage at 4°C.

*Data represents mean±SEM for triplicate.

This article is protected by copyright. All rights reserved.

KC1: Kufri Chipsona 1; KC3: Kufri Chipsona 3; KF: Kufri Frysona; KG: Kufri Girdhari; KJ:

Kufri Jyoti; KP: Kufri Pukhraj.

Fig. 2: (a) Effect of storage on the GI of six potato varieties. (b) Effect of storage on RS of

six potato varieties. (c) Effect of storage on AC of six potato varieties. (d) Effect of storage

on available CHO of six potato varieties. (e) Effect of storage on GL of six potato varieties.

(f) Effect of storage on the TSC of six potato varieties.

This article is protected by copyright. All rights reserved.

*a b values in a column with the same superscript do not differ significantly (p < 0.05).

GI: Glycemic Load; RS: Resistant Starch; AC: Amylose Content; CHO: Total Carbohydrate

Content; GL: Glycemic Load; TSC: Total Starch Content; KC1: Kufri Chipsona 1; KC3:

Kufri Chipsona 3; KF: Kufri Frysona; KG: Kufri Girdhari; KJ: Kufri Jyoti; KP: Kufri

Pukhraj.

Fig. 3: Overview of the effect of starch storage (cooling) in potato from 0 to 48 h on

Glycemic Index and Resistant Starch

Denotes decrease in the value of GI after cooling as compared to control

Denotes increase in value of RS after cooling as compared to control

This article is protected by copyright. All rights reserved.

Table 1: SHI, GI (both Goni’s Protocol and Modified Protocol), RS, TSC, AC, ACHO, GL of the six
potato variety

Vari SHI GI SHI GI RS TSC AC (%) ACHO GL

ety (Goni’s (Goni’s (Modifi (Modifi Conte (%) (mg/10
Protocol) Protoco ed ed nt (%) 0g dry
l) Protoco weight)
Protoco l)
l)
KC1 87.53± 54.63± 82.30± 1.20±0 71.04± 19.27± 11.52± 19.14±
60.17±0.68 0.54 0.18 0.06 .07 0.31 0.19 0.13 0.24

KC3 88.03± 53.66± 83.08± 1.08±0 65.34± 18.43± 12.60± 20.74±

60.79±0.73 0.58 0.08 0.15 .02 0.58 0.08 0.13 0.21

KF 82.21± 46.43± 76.49± 1.95±0 57.74± 20.10± 11.69± 17.89±

53.55±0.49 0.39 0.21 0.17 .03 0.32 0.12 0.03 0.03

KG 81.56± 47.27± 77.17± 1.35±0 69.78± 21.47± 14.59± 22.52±

52.74±0.57 0.46 0.54 0.43 .02 0.35 0.18 0.18 0.18

KJ 76.86± 41.92± 72.87± 2.18±0 71.04± 22.32± 13.78± 20.09±

46.89±0.38 0.30 0.11 0.09 .01 0.36 0.23 0.14 0.22

KP 81.78± 45.37± 75.64± 1.60±0 60.22± 19.43± 10.52± 15.92±

53.01±0.32 0.25 0.76 0.61 .01 0.52 0.14 0.19 0.38

Mea 54.52 83 48.21 77.92 1.56 65.86 20.17 12.45 19.38

n
SD 5.21 4.18 4.95 3.97 3.96 5.27 1.33 1.52 2.10

P <0.001 <0.001
valu * *
e

Modified Protocol compared with Goni’s Protocol, *t-test for two paired samples are significant at the
level of P value < 0.001. The value in bold are statistically significant at the level of P value < 0.001.
SHI: Starch Hydrolysis Index, GI: Glycemic Index; RS: Resistant Starch; TSC: Total Starch Content;
AC; Amylose Content; ACHO: Total Carbohydrate Content; GL: Glycemic Load; KC1: Kufri
Chipsona 1; KC3: Kufri Chipsona 3; KF: Kufri Frysona; KG: Kufri Girdhari; KJ: Kufri Jyoti; KP:
Kufri Pukhraj.

This article is protected by copyright. All rights reserved.

Table 2: Pearson correlation matrix between GI, SHI, RS and AC

Variable SHI GI RS AC

SHI 1

GI 1 1

RS -0.88** -0.88** 1

AC -0.79 -0.79 0.68 1

Mean of six varieties taken for correlation studies

**Significant at P value <0.05

SHI: Starch Hydrolysis Index, GI: Glycemic Index, RS: Resistant Starch, AC; Amylose Content

Table 3: GI classification and characterization of potato variety of ICAR-CPRI

Potato Variety GI classification Crop Type Texture

KC1 High Medium duration Floury

KC3 High Medium duration Floury

KF Intermediately High Medium duration Waxy

KG High Medium duration Waxy

KJ Intermediately High Medium duration Waxy

KP Intermediately High Medium duration Waxy

ICAR-CPRI: Indian Council of Agricultural Research- Central Potato Research Institute, KC1: Kufri

Chipsona 1, KC3: Kufri Chipsona 3, KF: Kufri Frysona, KG: Kufri Girdhari, KJ: Kufri Jyoti, KP:

Kufri Pukhraj.

This article is protected by copyright. All rights reserved.

Graphical Abstarct

An improved method of in-vitro estimation of the Glycemic index (GI) was developed for
potato. The low-temperature storage up to 48 h increased amylose and resistant starch, but
starch hydrolysis index, GI, glycemic load, total starch and available carbohydrate was
decreased. This improved method will help potato breeders and food industries to select low
GI varieties and food. Cooking and low-temperature storage of potato tuber for up to 48 h
may help to maintain blood sugar level of health-conscious and diabetic person.

This article is protected by copyright. All rights reserved.