development of barley-milk based fermented probiotic drink

372 Pages·2015·6.82 MB·English

DEVELOPMENT OF BARLEY-MILK BASED FERMENTED
PROBIOTIC DRINK

THESIS SUBMITTED TO THE
ICAR-NATIONAL DAIRY RESEARCH INSTITUTE, KARNAL
(DEEMED UNIVERSITY)

IN PARTIAL FULFILMENT OF THE REQUIREMENT
FOR THE AWARD OF THE DEGREE OF

DOCTOR OF PHILOSOPHY
IN
DAIRY TECHNOLOGY

BY

KUNAL KUMAR AHUJA
M. Tech. (Dairy Technology)

DAIRY TEHNOLOGY DIVISION
ICAR-NATIONAL DAIRY RESEARCH INSTITUTE
KARNAL-132 001 (HARYANA), INDIA

2015

Regn. No. 1030901

DEDICATED TO MY

BELOVED PARENTS

ABSTRACT
Barley being a rich source of soluble and insoluble dietary fiber also provides good
substrate for the growth of probiotic lactic acid bacteria (LAB). However, presence of
certain anti-nutrients such as phytate and tannins limit its suitability for human
consumption. In the present study, L. plantarum 344 was selected as suitable culture for
the development of barley-milk based fermented probiotic drink amongst five randomly
selected strains of probiotic LAB (L. acidophilus 13, L. casei 297 & 299, L. plantarum
344 and L. rhamnosus RSI3) based on its antinutrient reduction potential, compatibility
with co-culture S. thermophilus 20, and sensory acceptability of barley-milk based
fermented probiotic curd. Based on preliminary trials type of milk and flour were
selected for the preparation of probiotic drink. A central composite rotatable design of
response surface methodology consisting of 3 variables namely barley flour, probiotic
culture and co-culture concentration at 5 levels was adopted to optimize the level of
variables. Overall acceptability score, probiotic count and β-glucan content of fermented
probiotic drink were selected as responses. The optimized drink rated 7.8 (in between
like very much and like moderately) for overall acceptability on 9 point hedonic scale
and contained 8.59 log cfu/ml of probiotic strain, and 0.14 g/100 g β-glucan. Barley flour
concentration was the most critical variable amongst the three, which significantly
(ρ<0.01) affected the overall acceptability, probiotic count and β-glucan content of
probiotic drink. When analysed for antioxidant potential, the optimized probiotic drink
exhibited 0.40 and 0.79 mg/ml trolox equivalent ABTS and DPPH radical scavenging
activity, respectively. Antimicrobial activity of the supernatant obtained from optimized
product was high against tested pathogens, namely, E. coli (22 mm) and S. typhii (20
mm), whereas, moderate activity was observed against S. dysenteriae (16 mm) and S.
aureus (12 mm). Angiotensin-converting enzyme (ACE) inhibitory activity of developed
product was less than 15%. The optimized drink remained best acceptable up to a
period of 9 days when packed in PET bottles and stored at 5±1°C. However, product
packed in LDPE pouches, when compared with the PET bottles had significantly
(ρ<0.01) higher non-LAB count and yeast and mould count. Changes in sensory quality
of stored product in two packages exhibited almost similar trend. Responses from 100
respondents representing potential consumers belonging to mixed age, location and
economic group indicated, 89% of the respondents liked the optimized product with
score closer to ‘liked extremely’ side on 9-point hedonic scale. Cost of production per
200 ml product in PET bottle was estimated to be around `12.09.
िवषय : जौ - दधू आधािरत िकि(cid:214)वत प्रोबायोिटक पेय का िवकािसकरण
छात्र का नाम : कुणाल कुमार आहूजा प्रमखु सलाहकार : डॉ० आशीष कुमार िसहं
सारांश
जौ, घुलनशील एवं अघुलनशील आहार रेशाओं के समद्धृ  (cid:304)ोत होने के साथ‐साथ प्रोबायोिटक जीवाणुओं के िवकास 
के िलए अ(cid:205)छा  अधः (cid:232)तर प्रदान करता है I लेिकन, कुछ पोषक िवरोधी त(cid:215)व, जैसे िक फाइटेट एवं टैिनन की 
उपि(cid:232)थित जौ के मानवीय उपभोग को सीिमत करती है I वतमर् ान अ(cid:218)ययन म(cid:581), एल० (cid:220)लांटेरम एनसीडीसी344 को 
पांच लैिक्टक अ(cid:224)ल जीवाणुओं (एल० एिसडोिफलस एनसीडीसी13; एल० केजाइ एनसीडीसी297; एल०
केजाइ एनसीडीसी 299; एल० (cid:220)लांटेरम एनसीडीसी344 एवं एल० रहेमनोसस आरएसआई3 म(cid:581) से पोषक 
िवरोधी त(cid:215)व(cid:585) के पराभव;  सह-जामन (एस० थम(cid:574)िफलस 20)  के साथ संगतता; एवं जौ ‐ दधू  सयुंक्त आधािरत 
िकि(cid:214)वत प्रोिबओिटक के संवेदी मू(cid:227)यांकन के आधार पर, जौ दधू  आधािरत प्रोबायोिटक पेय के िवकािसकरण के 
िलए चयिनत िकया गया I प्रारंिभक परीक्षण(cid:585) के आधार पर प्रोबायोिटक पेय के अनुकूलन के िलए जौ के
आटे, एव ं दधू के प्रकार का चयन िकया गया I तीन चर जैसे िक, जौ का आटा; प्रोबायोिटक संरोपण; एवं
सह-जामन के संरोपण का अनुकूलन पांच (cid:232)तर वाले प्रितिक्रया-सह-कायप्रर् णाली के क(cid:581)द्रीय-समग्र-घूणनर् -
योग्य-पिरक(cid:227)पना (cid:622)वारा िकया गया I अनुकूलन के िलए, (cid:232)वीकायतर् ांक (कुल िमलाकर); प्रोबायोिटक की
िगनती; एवं प्रोबायोिटक पेय म(cid:581) बीटा-ग्लूकन का (cid:232)तर को प्रितिकयार्(cid:215)मक चर के (cid:510)प म(cid:581) चुना गया I 
अनुकूिलत पेय को 9-िब(cid:219)दओु ं वाले हेडोिनक पैमाने पर 7.8 मू(cid:227)यांिकत िकया गया, िजसमे प्रोबायोिटक
की िगनती 8.59 प्रित िमली०, एवं बीटा-ग्लूकन 0.14 ग्रा० प्रित 100 ग्रा० िनिहत थे I तीनो चर(cid:585) म(cid:581) जौ
के आटे की सांद्रता सबसे मह(cid:215)वपूण र् थी िजसने साथकर् (ρ<0.01) (cid:510)प से (cid:232)वीकायतर् ांक (कुल िमलाकर);
प्रोिबओिटक की िगनती, एवं बीटा-ग्लूकन के (cid:232)तर को प्रभािवत िकया I ऑक्सीकरण-रोधी गितिविध की
क्षमता का िव(cid:230)लेषण करने पर, अनुकूिलत प्रोबायोिटक पेय ने 0.40 एवं 0.79 िमग्रा० प्रित ग्रा० ट्रोलॉक्स
के बराबर क्रमशः डी० पी० पी० एच० एवं ए० बी० टी० एस० के मुक्तकण(cid:585) को समािजतर् िकया I
अनुकूिलत प्रोबायोिटक पेय के सतह-तरल की रोगाणरु ोधी क्षमता का िव(cid:230)लेषण करने पर  इ० कोलाई (22
िममी०) एवं से(cid:227)मोनेला टाइफी (20 िममी०) के िव(cid:510)द्ध अिधक गितिविध एव ं िशजेला िडसे(cid:219)ट्री (16 िममी०)
एव ं (cid:232)टेफ़० ओरीअस (12 िममी०) के िव(cid:509)द्ध म(cid:218)(cid:225)यम गितिविध पायी गयी I िवकिसत उ(cid:215)पाद की ACE
िनरोधी क्षमता 15 प्रितशत से कम थी I अनुकूिलत पेय पी० ई० टी० बोतल(cid:585) म(cid:581) 9 िदन(cid:585) तक सबसे अिधक 
(cid:232)वीकाय र् बना रहा, जबिक एल० डी० पी० ई० थैली म(cid:581) पैक िकये गए पेय म(cid:581) गैर‐ लैिक्टक अ(cid:224)ल जीवाणुओं, एवं 
खमीर की संख्या अिधक (ρ<0.01) पायी गयी I  भ(cid:214)डारण के दौरान दोन(cid:585) प्रकार के पैकेज म(cid:581) संवेदी गुणव(cid:215)ता म(cid:581) 
पिरवतनर्   लगभग  सामान  प्रिविृत  म(cid:581)  पाये  गए I  िविभ(cid:219)न  आयु,  (cid:232)थान,  आिथकर्   समूह  वाले  100  संभािवत 
उपभोक्ताओ ंकी प्रितिक्रयाओं का िव(cid:230)लेषण करने पर 89 प्रितशत उपभोक्ताओ ंको अनुकूिलत पेय 9‐िब(cid:219)दओु ं वाले 
हेडोिनक पैमाने पर “अ(cid:215)यंत पसंद आया” के करीब लगा I पी० ई० टी० बोतल(cid:585) म(cid:581) पैक िकये गए 200 िमली०
उ(cid:215)पाद की लागत लगभग 12.09 ` आंकी गयी I
CONTENTS
Chapter Title Page
No.
1.0 INTRODUCTION … 1-4
2.0 REVIEW OF LITERATURE … 5-46
2.1 Barley … 5
2.1.1 Classification of Barley … 5
2.1.2 Nutritional and other health benefits of barley … 6
2.1.2.1 Barley and cardio vascular disease … 8
2.1.2.2 Barley and diabetes melitus … 9
2.1.2.3 Barley and obesity … 10
2.1.3 Issues related to human consumption of barley … 11
2.1.3.1 Anti-nutrients in Barley … 11
2.1.3.1.1 Phytate … 11
2.1.3.1.2 Total Phenols … 12
2.1.3.1.3 Digestive enzyme inhibitors … 13
2.1.3.2 Other issues limiting human consumption … 13
of barley
2.1.4 Processing treatments to enhance the nutritional … 14
quality of barley
2.1.4.1 Pearling, milling and refining … 14
2.1.4.2 Soaking … 15
2.1.4.3 Fermentation … 16
2.1.4.4 Germination … 18
2.1.5 Effects of processing on barley β-glucan … 20
2.2 Milk nutrients for well being … 20
2.3 Fermented milks … 21
2.4 Cereal-milk based fermented foods … 22
2.4.1 Rabadi … 22
2.4.2 Tarhana … 23
2.4.3 Kishk … 23
2.4.4 Curd rice … 24
2.4.5 Cereal based low fat yoghurt … 25
2.4.6 Medida … 25
2.4.7 Akpan … 25
2.4.8 Bircher Muesli … 25
2.4.9 Crowdies … 25
2.4.10 Other composite fermented milk … 26
2.5 Barley based fermented foods … 26
2.5.1 Injera … 26
2.5.2 Aara … 27
2.5.3 Barley tempeh … 27
2.6 Shelf-life of fermented milks … 28
2.7 Shelf-life of cereal-milk based fermented milk … 28
2.8 Probiotics, prebiotics and synbiotics … 29
2.8.1 Type of probiotics … 31
2.8.2 Health benefits of probiotics … 31
2.8.2.1 Antimicrobial activity and control of … 32
gastrointestinal infection
2.8.2.2 Effectiveness against diarrhea … 32
2.8.2.3 Improvement in lactose metabolism … 33
2.8.2.4 Antimutagenic properties … 34
2.8.2.5 Anticarcinogenic properties … 34
2.8.2.6 Reduction in serum cholesterol … 34
2.8.2.7 Helicobacter pylori infection reduction … 35
2.8.2.8 Improvement in inflammatory bowel … 35
disease
2.8.2.9 Immune system stimulation … 35
2.8.3 Use of lactobacilli as probiotics … 36
2.8.3.1 Lactobacillus acidophilus … 36
2.8.3.2 Lactobacillus casei … 37
2.8.3.3 Lactobacillus plantarum … 38
2.8.3.4 Lactobacillus rhamnosus … 39
2.8.4 Therapeutic dose requirements … 40
2.9 Cereal-milk based probiotic products … 41
2.10 Commercial potential of milk-cereal based fermented foods … 43
2.11 Effective Packaging … 44
2.12 Sensory evaluation … 45
2.13 Hedonic scale … 45
2.14 Central composite rotary experiment … 46
3.0 MATERIALS AND METHODS … 47-102
3.1 MATERIALS / INGREDIENTS … 47
3.1.1 Milk … 47
3.1.2 Skim milk powder … 47
3.1.3 Barley … 47
3.1.4 Salt and spices … 47
3.1.5 Chemicals … 48
3.1.6 Bacterial cultures … 48
3.1.7 Packaging material … 48
3.2 METHODS … 49
3.2.1 Analysis of Milk … 49
3.2.2 Preparation of barley flour … 49
3.2.3 Analysis of barley flour … 51
3.2.3.1 Moisture content … 51
3.2.3.2 Ash content … 51
3.2.3.3 Protein content … 51
3.2.3.4 Crude fat … 52
3.2.3.5 Crude fibre … 53
3.2.3.6 Total starch content … 53
3.2.3.6.1 Preparation of reagents for … 54
estimation of total starch
content
3.2.3.6.2 Estimation of total starch … 54
3.2.3.7 Total Carbohydrate … 56
3.2.3.7.1 Preparation of reagents for … 56
total carbohydrate estimation
3.2.3.7.2 Estimation of total … 56
carbohydrates
3.2.3.8 Reducing sugar … 56
3.2.3.9 Glucose content … 57
3.2.3.10 β-glucan … 58
3.2.3.10.1 Preparation of reagents and … 59
working solution
3.2.3.10.2 Standard assay procedure … 59
for β-glucan
3.2.3.11 Total dietary fiber … 60
3.2.3.11.1 Standard assay procedure … 61
for total dietary fibre
3.2.3.12 Phytic acid … 62
3.2.3.12.1 Standard assay procedure … 63
for phytate
3.2.3.12.2 Phytic acid content … 64
estimation