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X-ray diffraction for food quality evaluation

Chapter · January 2019

DOI: 10.1016/B978-0-12-814217-2.00022-6

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X-ray diffraction for food
quality evaluation 22
Soumya Ranjan Purohit*, Lakshmi E. Jayachandran*, Anu S. Raj*, Debasis Nayak†,
P. Srinivasa Rao*
*Agricultural and Food Engineering Department, Indian Institute of Technology Kharagpur,
Kharagpur, India, †Department of Metallurgical and Materials Engineering, Indian Institute
of Technology Kharagpur, Kharagpur, India

22.1 Introduction
X-rays can be used for quality checking through imaging, diffraction, or scattering
modes. All the techniques are nondestructive, robust, and are used for specific pur-
poses. The X-ray imaging mode system is simple, quick, and is frequently being used
in quality and safety evaluation, within both food science and production. Quality
checks, such as screening of foreign bodies and micro particles, and measuring density
of food, etc., can be done through the X-ray imaging system. Moreover, the diffraction
pattern is also dependent on the composition of samples. On the other hand, X-ray
scattering is used for determination of the structure of complex proteins, and bio-
molecules having a noncrystalline nature. This chapter details X-ray based tools
and their applications for quality evaluation of major foods like cereal, millets, pulses,
dairy, confectionary, and other processed food products. A pictorial image depicting
an overview of the current chapter has been presented as well (Fig. 22.1).

22.1.1 The basic principle and procedures

22.1.1.1 X-ray generation
All the methods mentioned use X-rays, produced by bombarding a target material,
usually tungsten, molybdenum, or copper, with high-energy electrons. The high-
energy electrons are produced by heating a tungsten filament (cathode) with an elec-
tric current. The electrons pass through a tube with a high voltage potential
(30–150 kV) toward an anode. The anode material is the target material, within which
electrons become excited, and which returns to ground state by emitting photons
called X-rays. These generated X-rays are passed through a filter to make them mono-
chromatic (single wavelength) in nature. A schematic representation of the use of
X-rays for different applications has been shown in Fig. 22.2.

Evaluation Technologies for Food Quality. https://doi.org/10.1016/B978-0-12-814217-2.00022-6

© 2019 Elsevier Inc. All rights reserved.
580 Evaluation Technologies for Food Quality

X-ray

X - ray diffraction
Non destructive, robust,
user friendly, quick
,

Fr
ed es

ve uits
se uls

ge an
s
oil l, p

tab d
a

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re
Ce

Trusted
Food quality evaluation tool!
d d
foo sse
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oc

F
an at, pr
Pr

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al otein
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Fig. 22.1 Summary of the chapter.

22.1.1.2 X-ray imaging

In the case of an X-ray imaging system, X-rays pass through the food sample and fall
on a detector. The X-rays collected by the detector are analyzed and micro-
contaminants present inside food are detected (Fig. 22.2). X-ray imaging techniques
have undergone a revolution in increased sensitivity to low Z (atomic number) ele-
ments and organic materials by considering several X-ray aspects, such as phase-
contrast and dark-field imaging [1]. A synergetic study of X-ray transmission, phase
contrast, and dark-field images leads to better detection of foreign bodies in food and a
subsequent increase in its quality.
There are numerous food industries where X-ray imaging is being used for food
quality control, such as: luggage inspection for contraband food products, packaged
foods, poultry inspection, grain inspection, apples, nuts, and many more [2].
Besides several advantages, X-ray imaging also has a few limitations: the shape
and size of the food sample or container gives a nonuniform X-ray detection, which
may mislead as to presence of foreign particles [2, 3]; X-ray imaging is limited to
detection of macro-size particles; and, it cannot detect the presence of nanomaterials
or degraded biomolecules.

22.1.2 X-ray diffraction and small angle X-ray scattering

Both X-ray diffraction (XRD) and small angle X-ray scattering (SAXS) are based on
analysis of the diffracted/scattered intensity of an X-ray beam hitting a sample that var-
ies with change in incident and scattered angle, polarization, and wavelength or energy.
X-ray diffraction for food quality evaluation 581

X-ray imaging Small angle X-ray scattering

X-ray Detector
X-ray Scattered beam
2q
Transmitted beam
Sample Beam stop

Bragg’s law: l = 2d sin q

Sample

Transmitted
X-ray

X-ray diffraction

Diffracted X-ray
Incident X-ray

2d sin q

d q q

d sin q
Bragg’s law: nl = 2d sin q

Fig. 22.2 Schematic representation of X-ray imaging, small angle X-ray scattering (SAXS),
and X-ray diffraction (XRD).

Fig. 22.2 shows how the incident X-rays are passed at a small angle, the scattered pattern
is collected in SAXS, and the diffracted data is collected in XRD for further analysis.
The diffracted/scattered X-rays follow Bragg’s law, shown below.

nλ ¼ 2d sin θ (22.1)

where n is the positive integer, λ is wavelength of incident X-ray, d is lattice separa-

tion, and θ is the scattering angle.
Although XRD and SAXS obey Bragg’s law of diffraction, the latter is preferred
over the former for biological macromolecules, whereas XRD is mostly used for
detection of crystalline polymorphism of the materials.

22.1.2.1 Crystallinity analysis (XRD)

X ray diffraction data can be utilized to understand crystal polymorphism, and the
fraction of crystalline and amorphous content. There are various software applications
available for simulation of diffraction data, such as Crystal Diffract, MAUD, or X’Pert
High Score Plus. Crystallinity is generally determined by considering the area under
582 Evaluation Technologies for Food Quality

crystalline peak and area under total diffractograph. On the other hand, the crystalline
polymorph is determined by using the position of peak at a particular 2θ.

22.1.2.2 Structural analysis (SAXS)

In case of SAXS, the angle of incidence of the X-ray beam is kept small and the
scattered beam is collected for further analysis. SAXS generated data can be simulated
with several software packages, such as the ATSAS2.6.1 software suite [4, 5],
SASTBX [6], or CRYSOL [7] for the precise model of protein-protein complexes
and biomolecules (Fig. 22.3). The scattering intensity I(q) of X-ray is plotted against
scattering angle 2θ or scattering vector q. The values of I(q) and q are calculated by the
following equations.

AðqÞA∗ ðqÞ
I ð qÞ ¼ (22.2)
V
4πsin θ 2π
q¼ ¼ (22.3)
nλ d

where A(q) is the amplitude of scattering vector.

The scattering intensity of X-ray is obtained at the detector end. The SAXS patterns
emerge due to the shape and interaction of the scattering objects. Bio molecules are
disordered in nature, resulting in diffuse scatter around the positions of Bragg reflec-
tions. These diffuse scattering can be modelled by an atomic B factor. In order to sim-
plify the modeling, known bio molecules or proteins present in the result can be
modelled independently with relative weights. However, data with high resolution
is needed for this approach [9]. The high-resolution data of bio-molecules can be
accessed through crystallographic databases, such as protein data bank. After getting
the initial phases, a primary model is built. This model can be refined by changing the
atomic positions and their respective Debye-Waller factors (or B-factors, occurring
due to thermal motion of the atom). The refined model is set to fit the observed dif-
fraction data leading toward a better set of phases [8].

22.2 Recent technology and application of X-ray

diffraction in analysis of foods
22.2.1 Cereals, millets, and pulses
XRD is widely employed in analyzing changes in the structure and crystallinity of
cereal starches following various processing techniques. X ray scattering at a small
angle gives details about the size of the repeating lamellar structure (10–50 nm), while
that at wide angle reveals the structure of the nano particles (1–10 nm), indicative of
the liquid/solid form. Wide-angle X-ray scattering (WAXS) confirms the existence of
different types of starch structures, viz: A, B, C, and V-types (Fig. 22.4). It provides
 100 100
Scattered X-ray
(wavenumber: ks)
Incident X-ray
Scattering vector
(wavenumber: ki) ks 10–1
q = ks – ki 10–1

r 2θ
ki
(A) 10–2
(B)
Sample 0.00 0.15 0.30 0.45 0.60 0.75 0.00 0.15 0.30 0.45 0.60 0.75

q = |q| = 4π sin q/ λ 100 100

10–1 10–1
Form of particle
I(q) ~ q (rod)
–1

~ q–2(plate)
Scattering intensity I(q)

10–2 10–2
Surface structure (C) (D)
Interparticle I(q) ~ q–(6 – ds) 0.00 0.15 0.30 0.45 0.60 0.75 0.00 0.15 0.30 0.45 0.60 0.75
(ds: surface fractal dimension)
100 100
Inter-atomic structure
Size of particle (WAXD)
10–2
I(q) ~ exp (–q2Rg2/3) 10–1

(Rg: radius of gyration)

10–2 10–2

(E) (F)
Scattering angle 2q or scattering vector q 10–3
0.00 0.15 0.30 0.45 0.60 0.75 0.00 0.15 0.30 0.45 0.60 0.75

Fig. 22.3 Scattering vector (q) and typical SAXS scattering intensity data over scattering angle in 2D, and SAXS curves for six example proteins.
Comparison of I(q) calculated by CRYSOL from the all atom structure (blue) and by the two-body model (green). Error bars indicate the simulated
“experimental” error for each bin. The example proteins are: (A) 1ENH 54 residues, (B) 2CRO 71 residues, (C) 2PTH 193 residues, (D) 1PTA 327
residues, (E) 1A12 (chain A) 401 residues, and (F) 1JET 520 residues [8].
584 Evaluation Technologies for Food Quality

A
A

6 8 10 12 14 16 18 20 22 24 26 28 30

Vh

B
Vh
B/Vh
B
B Vh

6 8 10 12 14 16 18 20 22 24 26 28 30
Diffraction angle 2q (degrees)

Fig. 22.4 Diffraction pattern of various starch polymorphs.

quantitative and qualitative information on the micro- and macro structures of the
native starch compound. Starch, being a semicrystalline polymer with low and imper-
fect crystallinity, undergoes two phase transitions involving: (i) glass transition affect-
ing the amorphous phase; and (ii) melting of crystallites involved in the amylopectin
branches. A wide variety of studies have utilized this technique for analyzing
processing induced changes in various vegetal sources of starches.
XRD has been employed in analyzing the effect of parboiling on the sound (SK) and
fungal infested rice kernels (FIK) in nonissuable rice from a storage warehouse [10].
Predominant peaks were observed in the diffractogram of parboiled sound rice at dif-
fraction angles of 20, 15.3, and 18.4 degrees, while these peaks were subdued/under
developed in parboiled FIK (Fig. 22.5). Thus, the study reported a significantly lower
percentage of crystallinity in FIK (8.69%) than in SK (16.81%); an implication of the
higher impact of parboiling on FIK affecting its physico-chemical and microbiological
qualities adversely. Further, the higher water absorption capacity of pregelatinized rice
flour in comparison to raw rice flour (Fig. 22.6) has been reported by the same group of
authors, and the lower crystallinity or the amorphous state of starch (pregelatinized)
X-ray diffraction for food quality evaluation 585

3000

SK
2500

2000
Intensity

1500
FIK

1000

500

5 10 15 20 25 30 35 40
2q

Fig. 22.5 Variation in X-ray scattering between sound kernel and fungal infested kernel.

Fig. 22.6 X-ray scattering of raw and pregelatinized rice.

exhibited rapid moisture absorption as compared to the native ungelatinized sample

(raw rice flour). Moreover, the lower percentage of crystallinity in the pregelatinized
sample was deduced from X-ray diffractogram by taking the ratio of area under prom-
inent peak and the area under the whole curve, multiplied by 100 [11].
586 Evaluation Technologies for Food Quality

The effect of extrusion cooking on flour crystallinity has been studied on corn flour
systems [12]. This research reported that the diffraction pattern of the extruded flour
changed to the V-type pattern as against the A-type pattern, characteristic of the cereal
starches indicating a loss in starch crystallinity. This was elucidated by the appearance
of diffraction peaks at reflection angles of 7 and 13 degrees, featuring the existence of
V-type patterns in the extruded flour. The V-type starches are formed as a result of the
complexing between the amylose and other compounds such as iodine, alcohols, or
lipids that occurs during hydrothermal treatments or high temperature processing
[13]. Previous studies have also reported the disappearance of A-type starch structures
and the formation of V-type hydrates upon extrusion of wheat-almond flour blends
above 90°C [14]. This could be due to the destruction of the starch structure under
the impact of the intense shear field within an extruder. The effect of high pressure
processing on buckwheat starch revealed a loss in crystallinity as a result of pressure
induced gelatinization [15]. A novel quantification method based on XRD has been
developed, focusing on the diffractogram at a reflection angle of 5.5 degrees to eval-
uate samples with a high moisture content, such as cooked rice [16]. The method has
been successfully employed for studying cooked rice retrogradation. The above-
mentioned method eliminates the tedious sample preparation procedures for XRD
involving dehydration and powdering of the samples, as well as post processing anal-
ysis, such as mathematical peak separation procedures for determination of crystallin-
ity. Difference in crystallinity of rice pasta produced by conventional extrusion and
extrusion cooking was also elucidated using the XRD technique [17].
Structural variations in processed finger millet following hydrothermal, decortica-
tion, expansion, and popping treatments have been conducted [18] and significant
changes were reported in the crystal morphology of finger millet following all the
above mentioned processing conditions, indicated by the disappearance of all the three
major peaks observed in native millet.
Diffraction patterns have been obtained for both starch and proteins isolated from
cowpea and groundnut flour. The flour characteristics, such as the crystallinity index
and degree of crystallization, have been deduced and compared for both flour samples
[19]. Soybean flours under XRD exhibited C-type diffraction pattern, with more
B-type polymorphs than A-type and the percentage crystallinity varying from 27%
to 36%. XRD, in conjunction with a deconvoluted method, has been successfully used
to estimate the relative crystallinity of A-type and B-type polymorphs in chickpea
starch [20].

22.2.2 Dairy products

Bovine milk with 3.5%–5% lipid represents the complexity of natural fats, with more
than 400 fatty acids and 200 triglycerols (TG) [21]. TG constitute the majority, 97%–
98%, of lipid content in milk. TG exhibits polymorphism because of the difference in
lateral packing of the hydrocarbon chains of fatty acids and different possibility in
longitudinal stacking of lamellar molecules. This variation in level of organization
is identified with short and long spacings of X-Ray diffraction (XRD) at wide
(WXRD) and small (SXRD) angles, respectively [22]. The longitudinal stacking is
X-ray diffraction for food quality evaluation 587

measured by XRD at small angles 0 degree <θ < 5 degrees, and is related to the num-
ber of chains stacked one upon another in the crystalline cell. For TG, this thickness, or
long spacing (LS), generally takes value 2 (2L) or 3 (3L), with distances varying in the
range 40–50 and 55–70 Å, respectively. This provides information about the repeat
distance of methyl end groups in TG [23]. Short spacings (SS), observable with wide
angle XRD (8.5 degrees < θ < 13 degrees), represent the cross sectional packing of
aliphatic chains. SS characterizes the polymorphic forms in crystalline subcells [24].
Crystallization behavior of milk fat is influenced by composition of the medium,
processing condition, such as cooling time and crystallization temperature, mixing, or
shearing rate, etc. XRD had been exclusively used by the authors as complementary to
differential scanning calorimetry (DSC) for studying the crystalline structures and
thermal behavior of cream and anhydrous milk fat (AMF). SXRD shows that unstable
α form is initially formed upon quenching the cream at 8°C, which subsequently
disappears with time and is composed of two different lamellar arrangements: 2L
(47 Å) and 3L (70.4 Å). On studying the dry fractionation of milk fat, Lopez et al.
[25] reported that three longitudinal organizations, corresponding to three different
types of lamellar structures, are successively formed on crystallization of milk fat.
The initial crystalline structures are of double chain length (2L) with thickness
47.3 Å and 41.6 Å, respectively, followed by five sharp XRD peaks in triple chain
length (3L) of thickness 72.1 Å. The three lamellar structures formed during cooling
of milk fat correspond to unstable hexagonal packing of α type.
Temperature dependent XRD studies on AMF by Lambert et al. [26] explained the
coexistence of two different groups of TG in crystalline form. On comparison with TG
in cream, Lopez et al. [22] concluded that TG in AMF was disordered during crystal-
lization due to interface curvature in emulsion droplets.
Owing to the health benefits, unsaturated fatty acids (UFA) are preferred over sat-
urated fatty acids. Bugeat et al. [27] elucidated consequences of increased UFA com-
position in the crystallization properties and melting behavior of TG in milk, with
XRD. The results showed that on cooling, higher crystallization temperature TG solid-
ify in α2L (45–49 Å) structures and low crystallization temperature TGs in α3L
(75.5 Å) structures in UFA enriched TG.
XRD has been employed for the characterization of milk fat in processed cheese
products [28]. With AMF as a control, the researchers showed that the dispersion of
milk fat as cream and embedding fat globules into the casein matrix resulted in a
higher proportion of β polymorph. The study suggested that inclusion of milk fat glob-
ules in the protein matrix or the presence of other ingredients increases the ratio of β
polymorph to β0 form. Rapid cooling of cheese products resulted in the formation of β0
polymorph.

22.2.3 Chocolate
Chocolate is a complex mixture of sugars and cocoa solids (dispersed hydrophilic
phase) in fat rich cocoa butter (continuous phase). Macroscopic properties and sensory
qualities of chocolate are dependent on the crystalline structure of fat. X-ray diffrac-
tion has been employed in chocolate processing to study the polymorphs of fats and
588 Evaluation Technologies for Food Quality

Table 22.1 Polymorphic forms of cocoa butter [29].

Form Melting point (°C) XRD peak (Å)

I (γ) 14 4.18
II (α) 20 4.2
III (β0 ) 22 4.2
IV (β0 ) 24 4.13 and 4.32
V (β) 30 4.58
VI (β) 32 4.59

crystallinity of sugars. The specific polymeric forms of cocoa butter identified by

XRD are listed in Table 22.1.
The relationship between the crystallization behavior and structure of cocoa butter
was elucidated with the help of XRD [30]. According to the study, below 15°C meta-
stable forms of γ and α dominated, whereas between 15°C and 20°C the mixture
nucleated initially to α form and further transformed to stable phases. The researchers
also proposed the lifetime of different polymorphs with respect to temperature. The
existence of β0 phase ahead of formation of β polymorph and the inability of cocoa
butter crystals to be nucleated directly from melt to β form were delineated from this
research.
XRD aided in studying the crystallization of cocoa butter equivalents with mango
kernel fat and palm oil mid-fraction [31]. On formulation of reduced fat chocolate with
limonene [32], XRD readings proposed that the presence of limonene enhanced the
formation of lower polymorphic forms of fat on cooling and accelerated to stable
forms. The study demonstrated the formation of α and β0 IV forms in the initial stage
to βV upon storage and further transformation to βVI.
XRD served as a validation technique for modeling the phase change of cocoa but-
ter during rapid cooling (>100°C/min) of chocolate [29]. Crystal formation in velvet
chocolate and polymorphic transformation during the velvet formation, identified
through XRD, revealed the presence of βV form with 3L spacing, similar to normal
chocolate [33].
Temperature variation during transport and storage lead to chocolate bloom. XRD
assisted studies related to blooming of cocoa butter include researches by [34, 35], etc.
The major limitation on employing XRD for studies on crystal polymorphism in
chocolate is the presence of sugar. The sugars present in chocolate have specific
XRD patterns, which overlap with the signals of cocoa butter. The signals from sugar
have strong peaks at 4.52 Å (similar to β form) and 4.71 Å, interfering with the signals
from fat. Techniques developed to overcome this hurdle include glassifying the sugar
[36] or solid-liquid extraction for removing sugar. Guthrie et al. [37] proposed an
XRD technique that focuses on peaks specific to form V and VI of cocoa butter to
differentiate between them. Diffraction peaks corresponding to lattice spacing of
3.98 Å and 3.70 Å were examined to study phase of dark chocolate without sugar
removal.
X-ray diffraction for food quality evaluation 589

22.2.4 Fruit juice powders

Fruit powders, rich in sugar, often encounter drawbacks in terms of their functional
properties, such as stickiness, solubility, and hygroscopicity. This is often accompa-
nied by difficulties in the packaging and utilization of the fruit juice powders. X-ray
diffraction is one of the most reliable techniques in analyzing the powder quality of
dehydrated fruit juices. A hollow diffractogram, with diffused and large peaks, is char-
acteristic of amorphous content, indicating a disorderly arrangement of molecules;
whereas, sharp and defined peaks are a feature of the highly ordered molecular
arrangement of a crystalline state.
Diffractogram of spray dried blueberry juice (BJ) with added maltodextrin (MX)
exhibited broad peaks 2θ ¼ 15 degrees and no defined peaks at different levels of
water activity [38, 39]. When inulin was added to the BJ, an abrupt transition from
amorphous to crystalline state was observed at water activity above 0.4. This study
confirmed the ability of MX in conserving the amorphous state of BJ, indicative of
a better product performance. XRD has revealed the amorphous state of encapsulated
passion fruit juice for over 25 days of storage life as loose and dry powder, ensuring
better vitamin C stability [40].The structural stability of dried figs stored under dif-
ferent relative humidity conditions has also been successfully evaluated using wide
angle XRD [41]. The effectiveness of ultrasound assisted osmotic dehydration of
strawberries in retaining better powder characteristics, similar to conventional air dry-
ing, with minimal sugar retention has also been studied using XRD techniques [42].
This technique has been used to detect the presence of food ingredients, such as in
freeze dried acerola samples, wherein the semi defined peaks were indicative of
the presence of sugars and gum Arabic [43].
A comparison of the nature of crystallinity has been made to analyze the structural
properties in freeze dried papaya, guava, and sapota powders [44]. Defined peaks were
observed at 18, 19, and 15 degrees at 2θ angle, which is an indication of well-retained
semicrystalline structures after the juice dehydration. Degree of gelatinization in
durian flour was estimated from the ratio of diffraction peak area to total diffraction
area [45, 46] (Table 22.2).

22.2.5 Cooking oils, organogels, and margarines

Oils, usually liquids at room temperature, are mainly composed of triacylglycerols
(TAGs), and are widely used in cooking, as well as in many ready to eat products,
salad dressings, spreads etc. Many of these food products undergo deterioration in sen-
sorial properties due to the crystallization of the oil components, especially TAGs.
This phenomenon is more prominent during chilled or frozen storage.
XRD, in combination with DSC, provides valuable information on the crystalliza-
tion enthalpy and polymorphic forms of oil fats under different storage conditions.
The induction period of oils during storage could be estimated by the technique of
XRD, by recording the time until which a visible change in the diffractogram was
observed. The induction period of oil crystallization was determined to be 8.5 h for
oil stored isothermally at 17°C, while it was observed to be 10 h in the case of
590 Evaluation Technologies for Food Quality

Table 22.2 Analysis of fruit juice powders by X-ray powder diffraction (recent studies).

Processing
Product techniques Operational conditions Reference

Strawberry Ultrasound assisted 40 kV, 30 mA, 2θ ¼ 0–60 [42]

powder osmotic dehydration degrees, step size—0.02
degrees
Blueberry Spray drying 45 kV, 40 mA, 2θ ¼ 10–100 [38]
juice powder degrees, step size—0.016
degrees
Acerola Freeze drying 2θ ¼ 5–120 degrees, step [43]
powder size—0.02 degrees
Mango Foam mat drying 45 kV, 30 mA, 2θ ¼ 5–50 [46]
powder degrees, step size—0.02
degrees
Durian flour Microwave drying and 40 kV, 40 mA, 2θ ¼ 4–30 [45]
hot air drying degrees, step size—0.02
degrees
Persimmon Spray drying NA [47]
powder

oil stored at 25°C. Also, the presence of peaks in WAXS and its absence in SAXS
indicated that the TAG existed in liquid form only, especially at 17°C [48].
Polymorphism in sunflower oil and extra virgin oil has been studied by researchers
using XRD [49, 50]. The XRD studies in zero trans fat shortening, based on palm
stearin-rice bran oil, revealed that it predominantly comprised of β0 form (most stable)
and less of the undesirable β form. This could be an effective alternative to partially
hydrogenated fats used in food processing [51, 52]. Similar studies have been con-
ducted for the development of zero trans margarine from soybean oil-soy trisaturate
blends [52] and pine nut oil-palm stearin blends with predominant β0 crystals [53].
Thus, XRD finds application in revealing the molecular buildup of crystalline parti-
cles aiding in the production of zero trans margarines with desirable properties. XRD
has also been used to study the oil binding capacity and entanglement of the gel struc-
ture in organogel prepared with combined cinnamic acid-rice bran oil mixtures at
varying concentrations of cinnamic acid [54].

22.3 Miscellaneous applications

Curcumin-β-cyclodextrin inclusion complex prepared by co-extrusion is a natural dye
used as food color. The series of thin and intense lines indicating the crystallinity in
curcumin, as well as in β-cyclodextrin, remained unchanged in a simple mixture;
while in a coprecipitated mixture, some of the spectral lines of curcumin disappeared
while new lines appeared, suggesting the formation of a new solid crystalline phase
corresponding to an inclusion complex of the same nature [55].
X-ray diffraction for food quality evaluation 591

Antimicrobial coating, prepared using chitosan and polyvinyl alcohol (PVA), is

very effective in extending the shelf life of minimally processed fresh food products.
XRD is a very powerful tool for studying the miscibility and morphology of antimi-
crobial film [56]. While the diffraction peak of chitosan is observed between the
reflection angles of 10 and 20 degrees, the corresponding peaks of chitosan-PVA mix-
ture was observed between 11 and 48 degrees, clearly indicating that the crystallinity
of chitosan-PVA is much higher than that in chitosan. Similar studies have been con-
ducted on starch based edible packaging films produced from corn starch or amylo-
maize [57]. XRD is also one of the most widely adopted techniques for the
characterization of nanocomposites, which promote the usage of biopolymers in food
packaging. It is highly effective in characterizing the degree of intercalation, exfoli-
ation, and dispersion of biopolymers [58].

22.4 Conclusion and outlook

The X-ray scattering and diffraction method is a user friendly and advanced nonde-
structive technology, which has great potential to be employed for quality analysis of
food. For a long time, this technique has held a firm position in the field of material
science; however, in recent years this technique has been widely used in food analysis
as well. As discussed in this chapter, a wide range of food and food products can be
tested for their characteristics and discriminated from other reference or model foods.
Though in many research applications, this technology has a special place in quality
analysis of samples, there is still a need for development of a dedicated method for
quality analysis in short time frames. There is a need to couple this technology and
advanced statistical methods to develop rapid and reliable quality analysis methods.

References
[1] R. Fitzgerald, Phase-sensitive X-ray imaging, Phys. Today 53 (2000) 23–26.
[2] R.P. Haff, N. Toyofuku, X-ray detection of defects and contaminants in the food industry,
Sens. & Instrumen. Food Qual. 2 (2008) 262–273.
[3] M.S. Nielsen, T. Lauridsen, L.B. Christensen, R. Feidenhans, X-ray dark-field imaging for
detection of foreign bodies in food, Food Control 30 (2013) 531–535.
[4] M.V. Petoukhov, P.V. Konarev, A.G. Kikhney, D.I. Svergun, ATSAS 2.1—towards auto-
mated and web-supported small-angle scattering data analysis, J. Appl. Crystallogr.
40 (2007) s223–s228.
[5] C.E. Schindler, S.J. de Vries, A. Sasse, M. Zacharias, SAXS data alone can generate high-
quality models of protein-protein complexes, Structure 24 (2016) 1387–1397.
[6] H. Liu, A. Hexemer, P.H. Zwart, The Small Angle Scattering ToolBox (SASTBX): an
open-source software for biomolecular small-angle scattering, J. Appl. Crystallogr.
45 (2012) 587–593.
[7] D. Svergun, C. Barberato, M.H. Koch, CRYSOL—a program to evaluate X-ray solution
scattering of biological macromolecules from atomic coordinates, J. Appl. Crystallogr.
28 (1995) 768–773.
[8] K. Stovgaard, C. Andreetta, J. Ferkinghoff-Borg, T. Hamelryck, Calculation of accurate
small angle X-ray scattering curves from coarse-grained protein models, BMC Bioinform.
11 (2010) 429.
592 Evaluation Technologies for Food Quality

[9] L. Feigin, D.I. Svergun, G.W. Taylor, General principles of small-angle diffraction,
in: Structure Analysis by Small-Angle X-Ray and Neutron Scattering, Springer, 1987,
pp. 25–55.
[10] S.R. Purohit, P.S. Rao, Water absorption and gelatinization kinetics of non-issuable rice
and its characterization, J. Food Meas. Charact. 11 (2017) 2110–2118.
[11] S.R. Purohit, P.S. Rao, Modelling and analysis of moisture sorption isotherm of raw and
pregelatinized rice flour and its crystalline status prediction, Food Anal. Methods
10 (2016) 1914–1921.
[12] L.C. Oliveira, J.H.T. Barros, C.M. Rosell, C.J. Steel, Physical and thermal properties and
X-ray diffraction of corn flour systems as affected by whole grain wheat flour and extru-
sion conditions, Starch-St€arke 69 (2017) 1600299.
[13] P.L. Bail, H. Bizot, M. Ollivon, G. Keller, C. Bourgaux, A. B, Monitoring the crystalli-
zation of amylose–lipid complexes during maize starch melting by synchrotron X-ray dif-
fraction, Biopolymers 50 (1999) 99–110.
[14] M.A. Brennan, E. Derbyshire, B.K. Tiwari, C.S. Brennan, Ready-to-eat snack products:
the role of extrusion technology in developing consumer acceptable and nutritious snacks,
Int. J. Food Sci. Technol. 48 (2013) 893–902.
[15] Z. Zhou, X. Ren, F. Wang, J. Li, X. Si, R. Cao, et al., High pressure processing manipulated
buckwheat antioxidant activity, anti-adipogenic properties and starch digestibility,
J. Cereal Sci. 66 (2015) 31–36.
[16] A. Nagataki, H. Tomita, Y. Himeda, T. Takemori, M. Fukuoka, A quantification method
of retrogradation for cooked rice based on a single isolated peak in X-ray diffraction,
J. Cereal Sci. 79 (2018) 80–85.
[17] A. Marti, K. Seetharaman, M.A. Pagani, Rice-based pasta: a comparison between conven-
tional pasta-making and extrusion-cooking, J. Cereal Sci. 52 (2010) 404–409.
[18] U. Dharmaraj, P. Parameswara, R. Somashekar, N.G. Malleshi, Effect of processing on the
microstructure of finger millet by X-ray diffraction and scanning electron microscopy,
J. Food Sci. Technol. 51 (Mar 2014) 494–502.
[19] G.K. Kaptso, N.Y. Njintang, M.G.M. Nguemtchouin, A.F. Amungwa, J. Scher,
J. Hounhouigan, et al., Characterization of morphology and structural and thermal prop-
erties of legume flours: cowpea (Vigna unguiculata L. Walp) and bambara groundnut
(Vigna subterranea L. Verdc.) Varieties, Int. J. Food Eng. 12 (2016) 139–152.
[20] Y. Sun, H. Ye, B. Hu, W. Wang, S. Lei, X. Wang, et al., Changes in crystal structure of
chickpea starch samples during processing treatments: an X-ray diffraction and starch
moisture analysis study, Carbohydr. Polym. 121 (May 05 2015) 169–174.
[21] J. Gresti, M. Bugaut, C. Maniongui, J. Bezard, Composition of molecular species of
triacylglycerols in bovine milk fat, J. Dairy Sci. 76 (1993) 1850–1869.
[22] C. Lopez, P. Lesieur, C. Bourgaux, G. Keller, M. Ollivon, Thermal and structural behavior
of milk fat: 2. Crystalline forms obtained by slow cooling of cream, J. Colloid Interface
Sci. 240 (2001) 150–161.
[23] A.G. Marangoni, L.H. Wesdorp, Structure and Properties of Fat Crystal Networks, CRC
Press, 2012.
[24] C. Lopez, P. Lesieur, G. Keller, M. Ollivon, Thermal and structural behavior of milk fat: 1.
Unstable species of cream, J. Colloid Interface Sci. 229 (2000) 62–71.
[25] A.D. Lopez, C.D. Mathers, M. Ezzati, D.T. Jamison, C.J. Murray, Global and regional
burden of disease and risk factors, 2001: systematic analysis of population health data,
Lancet 367 (2006) 1747–1757.
[26] A. Lambert, F. Bougrioua, O. Abbas, M. Courty, M. El Marssi, V. Faivre, et al., Temperature
dependent Raman and X-ray diffraction studies of anhydrous milk fat, Food Chem. (2017).
X-ray diffraction for food quality evaluation 593

[27] S. Bugeat, J. Perez, V. Briard-Bion, P. Pradel, A. Ferlay, C. Bourgaux, et al., Unsaturated

fatty acid enriched vs. control milk triacylglycerols: solid and liquid TAG phases exam-
ined by synchrotron radiation X-ray diffraction coupled with DSC, Food Res. Int.
67 (2015) 91–101.
[28] P.R. Ramel, A.G. Marangoni, Characterization of the polymorphism of milk fat within
processed cheese products, Food Struct. 12 (2017) 15–25.
[29] B.J. Le Reverend, P.J. Fryer, S. Coles, S. Bakalis, A method to qualify and quantify the
crystalline state of cocoa butter in industrial chocolate, J. Am. Oil Chem. Soc. 87 (2010)
239–246.
[30] A.G. Marangoni, S.E. McGauley, Relationship between crystallization behavior and struc-
ture in cocoa butter, Cryst. Growth Des. 3 (2003) 95–108.
[31] S. Sonwai, P. Kaphueakngam, A. Flood, Blending of mango kernel fat and palm oil mid-
fraction to obtain cocoa butter equivalent, J. Food Sci. Technol. 51 (2014) 2357–2369.
[32] J. Ray, W. MacNaughtan, P.S. Chong, J. Vieira, B. Wolf, The effect of limonene on the
crystallization of cocoa butter, J. Am. Oil Chem. Soc. 89 (2012) 437–445.
[33] L. Bayes-Garcı́a, T. Calvet, M.A.n. Cuevas-Diarte, E. Rovira, S. Ueno, K. Sato, New tex-
tures of chocolate are formed by polymorphic crystallization and template effects: velvet
chocolate, Cryst. Growth Des. 15 (2015) 4045–4054.
[34] S. Hodge, D. Rousseau, Fat bloom formation and characterization in milk chocolate
observed by atomic force microscopy, J. Am. Oil Chem. Soc. 79 (2002) 1115–1121.
[35] Y. Kinta, T. Hatta, Morphology of chocolate fat bloom, in: Cocoa Butter and Related
Compounds, Elsevier, 2012, pp. 195–212.
[36] C. Loisel, G. Keller, G. Lecq, C. Bourgaux, M. Ollivon, Phase transitions and polymor-
phism of cocoa butter, J. Am. Oil Chem. Soc. 75 (1998) 425–439.
[37] J. Guthrie, D. Reid, K.B. Walsh, Assessment of internal quality attributes of mandarin
fruit. 2. NIR calibration model robustness, Aust. J. Agric. Res. 56 (2005) 417–426.
[38] S.B. Araujo-Diaz, C. Leyva-Porras, P. Aguirre-Banuelos, C. Alvarez-Salas, Z. Saavedra-
Leos, Evaluation of the physical properties and conservation of the antioxidants content,
employing inulin and maltodextrin in the spray drying of blueberry juice, Carbohydr.
Polym. 167 (Jul 01 2017) 317–325.
[39] D.G. Stevenson, R.K. Doorenbos, J.-l. Jane, G.E. Inglett, Structures and functional prop-
erties of starch from seeds of three soybean (Glycine max (L.) Merr.) varieties*, Starch-
St€arke 58 (2006) 509–519.
[40] D. Borrmann, A.P.T.R. Pierucci, S.G.F. Leite, M.H.M.d.R. Leão, Microencapsulation of
passion fruit (Passiflora) juice with n-octenylsuccinate-derivatised starch using spray-
drying, Food Bioprod. Process. 91 (2013) 23–27.
[41] F. Badii, A. Farahnaky, H. Behmadi, Effect of storage relative humidity on physical sta-
bility of dried fig, J. Food Process. Preserv. 38 (2014) 477–483.
[42] E. Amami, W. Khezami, S. Mezrigui, L.S. Badwaik, A.K. Bejar, C.T. Perez, et al., Effect
of ultrasound-assisted osmotic dehydration pretreatment on the convective drying of
strawberry, Ultrason. Sonochem. 36 (May 2017) 286–300.
[43] F.P. De Moraes, A.C. Gonçalves, T.B. Verı́ssimo Miguel, K.C. Borges, R.T.P. Correia,
Freeze dried acerola (Malpighia emarginata) pulp and pomace: physicochemical attri-
butes, phytochemical content and stability during storage, J. Food Ind. 1 (2017) 17–38.
[44] K.A. Athmaselvi, C. Kumar, M. Balasubramanian, I. Roy, Thermal, structural, and phys-
ical properties of freeze dried tropical fruit powder, J. Food Process. 2014 (2014) 1–10.
[45] S. Bai-Ngew, N. Therdthai, P. Dhamvithee, W. Zhou, Effect of microwave vacuum drying
and hot air drying on the physicochemical properties of durian flour, Int. J. Food Sci.
Technol. 50 (2015) 305–312.
594 Evaluation Technologies for Food Quality

[46] A.M. Chaux-Gutierrez, E.J. Perez-Monterroza, V.R.N. Telis, M.A. Mauro, The physical
and morphological characteristics of mango powder (Mangifera indica L. cv Tommy
Atkins) produced by foam mat drying, Food Biophys. 12 (2016) 69–77.
[47] J. Du, Z.-Z. Ge, Z. Xu, B. Zou, Y. Zhang, C.-M. Li, Comparison of the efficiency of five
different drying carriers on the spray drying of persimmon pulp powders, Dry. Technol.
32 (2014) 1157–1166.
[48] Y. Miyagawa, K. Shintani, K. Katsuki, K. Nakagawa, S. Adachi, Thermal and structural
changes of rapeseed oil during isothermal storage at low temperature, Food Struct.
11 (2017) 8–15.
[49] S. Calligaris, G. Arrighetti, L. Barba, M.C. Nicoli, Phase transition of sunflower oil as
affected by the oxidation level, J. Am. Oil Chem. Soc. 85 (2008) 591–598.
[50] E. Chiavaro, Crystal polymorph structure determined for extra virgin olive oil, Eur. J.
Lipid Sci. Technol. 115 (2013) 267–269.
[51] P.N. Mayamol, T. Samuel, C. Balachandran, A. Sundaresan, C. Arumughan, Zero-trans
shortening using palm stearin and rice bran oil, J. Am. Oil Chem. Soc. 81 (2004) 407–413.
[52] G.R. List, E.A. Emken, W.F. Kwolek, T.D. Simpson, H.J. Dutton, “Zerotrans” margarines:
preparation, structure, and properties of interesterified soybean oil-soy trisaturate blends,
J. Am. Oil Chem. Soc. 54 (1977) 408–413.
[53] P. Adhikari, X.-M. Zhu, A. Gautam, J.-A. Shin, J.-N. Hu, J.-H. Lee, et al., Scaled-up pro-
duction of zero-trans margarine fat using pine nut oil and palm stearin, Food Chem.
119 (2010) 1332–1338.
[54] X. Li, A.S.M. Saleh, P. Wang, Q. Wang, S. Yang, M. Zhu, et al., Characterization of organ-
ogel prepared from rice bran oil with cinnamic acid, Food Biophys. 12 (2017) 356–364.
[55] C.S. Mangolim, C. Moriwaki, A.C. Nogueira, F. Sato, M.L. Baesso, A.M. Neto, et al., Cur-
cumin-beta-cyclodextrin inclusion complex: stability, solubility, characterisation by
FT-IR, FT-Raman, X-ray diffraction and photoacoustic spectroscopy, and food applica-
tion, Food Chem. 153 (Jun 15 2014) 361–370.
[56] S. Tripathi, G.K. Mehrotra, P.K. Dutta, Physicochemical and bioactivity of cross-linked
chitosan-PVA film for food packaging applications, Int. J. Biol. Macromol. 45 (Nov
01 2009) 372–376.
[57] M.N.M. Marı́a A Garcı́a, N.E. Zaritzky, L. Plata, Microstructural characterization of plas-
ticized starch-based films, Starch-St€arke 52 (2000) 118–124.
[58] A. Arora, G.W. Padua, Review: nanocomposites in food packaging, J. Food Sci.
75 (Jan–Feb 2010) R43–R49.

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