Spectrochimica Acta Part B 124 (2016) 74–78

Contents lists available at ScienceDirect

Spectrochimica Acta Part B

journal homepage: www.elsevier.com/locate/sab

Analytical note

Ash analysis of flour sample by using laser-induced

breakdown spectroscopy
Gonca Bilge a, Banu Sezer a, Kemal Efe Eseller b, Halil Berberoglu c, Hamit Koksel a, Ismail Hakki Boyaci a,⁎
a
Department of Food Engineering, Hacettepe University, Beytepe 06800, Ankara, Turkey
b
Department of Electrical and Electronics Engineering, Atilim University, 06836 Ankara, Turkey
c
Department of Physics, Polatlı Faculty of Science & Arts, Gazi University, 06900 Polatlı, Ankara, Turkey

a r t i c l e i n f o a b s t r a c t

Article history: Ash content is a measure of total mineral content in flour. It is also an important quality parameter in terms of
Received 15 December 2015 nutritional labeling as well as processing properties of various cereal products. However, laboratory analysis
Received in revised form 24 July 2016 takes a long time (5–6 h) and results in considerable waste of energy. Therefore, the aim of the study was to de-
Accepted 24 August 2016
velop a new method for ash analysis in wheat flour by using laser induced breakdown spectroscopy (LIBS). LIBS is
Available online 24 August 2016
a multi-elemental, quick and simple spectroscopic method. Unlike basic ash analysis method, it has the potential
Keywords:
to analyze a sample in a considerably short time. In the study, wheat flours with different ash contents were an-
LIBS alyzed using LIBS and the spectra were evaluated with partial least squares (PLS) method. The results were cor-
Ash analysis related with the ones taken from standard ash analysis method. Calibration graph showed good linearity with the
Wheat flour ash content between 0.48 and 1.39%, and 0.992 coefficient of determination (R2). Limit of detection for ash anal-
Mineral content ysis was calculated as 0.026%. The results indicated that LIBS is a promising and reliable method with high sen-
sitivity for routine ash analysis in flour samples.
© 2016 Elsevier B.V. All rights reserved.

1. Introduction
The inorganic part of the food that remains after burning at high
temperatures is called ash. Ash content represents the mineral content
of the food and consists of major elements such as Na, K, Ca and Mg,
and trace elements such as Fe, Zn, Cu. Although the average ash content
of various food groups varies from 0.1% to 2.5% on wet weight basis, it
plays an important role in food matrix [1]. Ash content affects the rheo-
logical properties and baking quality and determines nutritional quality
of foods [2]. It is widely used as an index, especially for refinement of
foods such as wheat flour. Ash content is a good indicator to understand
the discrimination of bran and germ from the wheat kernel during flour
milling. This is because bran has approximately twenty times higher ash
content than endosperm. Furthermore, minerals in powder foods are
responsible for increasing hygroscopicity indirectly through water-min-
eral bonds and indirectly mineral-carbohydrate interactions [2].
In order to determine the total mineral content of wheat flour, ash con-
tent is measured. In the basic ash method, well-mixed flour sample is
placed in a muffle furnace at 550 °C for soft wheat flours or 575–590 °C
⁎ Corresponding author.
E-mail addresses: goncabilge@yahoo.com.tr (G. Bilge), sezerrbanu@gmail.com
(B. Sezer), efe.eseller@atilim.edu.tr (K.E. Eseller), halilb@gazi.edu.tr (H. Berberoglu),
koksel@hacettepe.edu.tr (H. Koksel), ihb@hacettepe.edu.tr (I.H. Boyaci).
for hard wheat flours. Sample is incinerated until light gray ash or constant
weight is obtained. After cooling, the sample is weighed and ash content is
calculated on dry basis [4]. This procedure is time consuming (5–6 h) and
causes energy waste. Hence, alternative processes are needed for the ash
analysis.
Ashing is also the first step of elemental analysis with atomic absorp-
tion spectroscopy and inductively coupled mass spectroscopy. Recent
studies have focused on laser induced breakdown spectroscopy (LIBS)
as a new, rapid and in-situ elemental analysis technique [5,6] in which
plasma is created by means of focusing the laser through the lens on
the sample surface. Due to the high temperature, sample is vaporized,
excited and characteristic plasma light is formed. The obtained light is
collected with fiber optics and directed into the spectrometer, and spec-
tral signals are produced. These signals are related with chemical com-
positions of the sample [7]. Usually for optimal results, LIBS does
require at least minimal sample preparation step, especially for pow-
dered samples and is a rapid and in-situ method, which makes it distinct
from atomic absorption spectrometer (AAS) and inductively coupled
plasma-mass spectrometer (ICP-MS). The application field of LIBS has
been expanding gradually, including studies on metallurgy, mining, en-
vironmental analysis and pharmacology [8–10]. However, there are
only a few studies on food applications such as analysis of rice powder,
unpolished-rice flour and starch [11] and Ca in breakfast cereals [12], Na
and NaCl analysis in bakery products [13]. LIBS is also used for ash

http://dx.doi.org/10.1016/j.sab.2016.08.023
0584-8547/© 2016 Elsevier B.V. All rights reserved.
 G. Bilge et al. / Spectrochimica Acta Part B 124 (2016) 74–78 75

Fig. 1. LIBS experimental setup.

analysis in herbaceous materials [14] and elemental analysis of plant
materials such as Cannabis plant material [15] and Ocimum L. species
[16].
The study is focused on development of a new method for ash anal-
ysis in wheat flour by using LIBS. The aim was to develop a relatively
rapid, practical and economical method compared to the conventional
ones. For this purpose, wheat flours with different ash contents were
analyzed with LIBS, and the spectra were evaluated using chemometric
methods. The results of LIBS were compared with the results of basic ash
method, and mineral matter composition of wheat flour was investigat-
ed with AAS method.
2.3. LIBS instrumentation
LIBS spectra were recorded using a Polaris New Wave 50 mJ
1064 nm Nd:YAG laser (Fremont, CA USA) and Applied Spectra 5 chan-
nel Aurora LIBS spectrometer (Fremont, CA USA). Fig. 1 shows the ex-
perimental setup of the LIBS system. The laser was operated at a
fundamental wavelength of 1064 nm and used for sample ablation. It
was operated in the Q-switched mode at a repetition rate of 4 Hz,
300 ns gate delay and 1.05 ms integration time. The laser energy was
38 mJ/pulse. Pelleted samples were scanned with laser at 7 different re-
gions for each pellet, and 15 shots were performed from each region.

2. Experimental
2.4. Data analysis
2.1. Sample preparation
To evaluate the multi elemental spectrum and reduce the matrix ef-
Wheat flour samples with different ash contents were obtained fect of flour samples, flours with different ash contents were analyzed
from Ankara Metropolitan Municipality Bakery Plant (Ankara Halk using PLS application in the program (Version7.5.2 for Windows 7, Ei-
Ekmek Fab. Ankara, Turkey). Milled flour samples at different fine- genvector ResearchInc., Wenatchee, WA, USA). In this study, 69 differ-
ness were used randomly. The flour samples were dried in a drying ent flours with various ash contents were used for calibration data set;
cabined at 105 °C for 2 h. For LIBS analysis, three pellets were pre- and 16 different flours with various ash contents were used for valida-
pared from each flour sample (400 mg) with a pellet forming press tion data set. Orthogonal signal correction (OSC), standard normal
under 10 tons of force. The diameter and height of pellet is 13 mm
and 2 mm, respectively. Sixty-nine different flour samples with dif-
ferent ash contents were analyzed to obtain data for the calibration
graph. Furthermore, 16 flour samples were analyzed for the valida-
tion of LIBS method.

2.2. Ash analysis

3–5 g of flour samples were weighted into ashing dishes. Then the
samples were placed in a muffle furnace at 550 °C for soft wheat flours
or 575–590 °C for hard wheat flours. They were incinerated until light
gray ash or constant weight was obtained. After cooling, the samples
were weighted, and ash contents were calculated on dry basis (Eq.
(1)) [4].

dry weight of residue

%Ash ¼  100 ð1Þ
sample weight Fig. 2. LIBS spectra of flour samples with different ash content.
76 G. Bilge et al. / Spectrochimica Acta Part B 124 (2016) 74–78

Table 1 related to all or some of the minerals in the structure of flour. For in-
Possible contributions to the identified spectral peaks from various elements in the 350– stance, spectral line intensity of one element can increase while spectral
800 nm spectral region.
line intensity of another element can decrease with increasing ash con-
Central wavelength (nm) Possible element tent. To overcome this, emission intensities were normalized. For each
388.205 Fe I (388.327) spectral line, signals to fully averaged peak intensity calculations were
393.360 Ca II (393.366), Ca I (393.529) made by averaging the fully integrated peak intensity and normalizing
396.821 Ca II (396.8469) the full spectrum (Fig. 2). The method has been applied as a reliable an-
422.700 Ca I (422.672), Ca II (422.815)
alytical method [18,19] as it provides a linear relation between intensi-
443.548 Ca I (443.569)
445.517 Ca I (445.478) ties of atomic spectral lines and ash content of many types of materials.
471.388 Ca III (471.626) Wheat flour contains P, K, S, Ca, Mg, Fe, Zn, and Cu. All of the related
473.598 Ca III (473.668) elements in Fig. 2 are presented in Table 1. To confirm the LIBS elemen-
516.528 Fe I (516. 748), Mg I (516.732) tal analysis, AAS was used for flour samples, and the results are present-
518.95 Ca I (518.885), Na II (518.715), Mg I (518.360)
527.096 Fe I (527.035), Na II (527.572), S I (527.899)
ed in Table 2. The table indicates that the flour samples with low ash
558.909 Ca III (557.9076), P II (558.327) content have lower elemental contents and this is quite obvious in K,
589.031 Na I (588.995) Mg, Ca, Na and Zn contents (Table 2). In the literature, all mineral com-
610.298 Ca I (610.272), S II (610.226), Zn II (610.249) position of wheat flour was analyzed using LIBS, and the applicability of
612.231 Ca III (612.011), Ca I (612.221), Fe I (612.790)
LIBS was also confirmed with a reference elemental analysis method,
616.26 Ca I (616.217)
643.982 Ca I (643.907), Fe I (643.875) namely X-ray fluorescence spectrometer (XRF) [20]. LIBS can measure
645.04 Na II (645.076) all elements in the flour, which makes it applicable for the measurement
646.268 Ca I (646.256) of ash content representing the total mineral content. For this reason,
656.23 Fe II (656.220) the obtained LIBS spectra were correlated with the results of ash analy-
714.785 Ca I (714.815), Na IV (714.2), Fe I (714.814)
sis by using PLS method. Fig. 3 demonstrates the calibration and valida-
744.769 N I (744.229)
747.328 Zn II (747.877), N I (746.831) tion graphs of wheat flour. The ash calibration model with the lowest
766.568 K I (766.489) root meat square error calibration (RMSEC: 0.032) and the highest coef-
769.893 K I (769.896) ficient of determination R2 (0.999) was obtained with OSC, SNV, vari-
777.224 Ca III (777.710)
ance (std) scaling, class centroid centering ve scaling pre-processing
methods and 7 latent variables. Root Mean Square Error of Cross Valida-
tion (RMSECV) was obtained as 0.129, and root mean square error pre-
diction (RMSEP) value of the validation model was obtained as 0.063.
variate (SNV), variance (std) scaling, class centroid centering and scal- Flour ash does contain a considerable amount of oxygen because
ing pre-processing methods were used for ash prediction in flour. combustion forms oxides of the minerals. Significant amount of carbon
is also present in the form of fixed carbon. LIBS analysis can be per-
2.5. Atomic absorption analysis formed directly on flour sample without ashing, thus LIBS also measures
C, H and O elements which are at minimum levels in whole spectrum. In
Inorganic matter composition of flour was analyzed using AAS as a the study, PLS method was used for evaluation of the spectra and PLS
reference method. Sample preparation step was performed with acid analyzes the changing major parameters and forms a model by exclud-
digestion according to EPA Method 3051A [17]. 0.3 g of flour samples ing the remaining almost constant parameters such as H (656.282 nm),
were placed in a fluorocarbon polymer vessels, and 10 ml of concentrat- C (247.856, 392.069 and 833.515 nm) and O (715.670, 777.539 nm)
ed HNO3 was added. The samples were extracted through heating with lines [21]. Therefore, it is not necessary to exclude the C, H and O lines
CEM Corp. MARS laboratory microwave unit (Matthews, NC, USA). After from the full spectrum for ash analysis. The results are very promising
cooling, the vessel contents were filtered with Whatman No. 1 filter as can be deduced from the coefficient of determination for calibration
paper and diluted in 100 ml of deionized water. AAS were recorded and validation graphs.
with the Thermo Scientific ICE 3000 Series Atomic Absorption Spec- To evaluate the measurement sensitivity and precision, relative
trometer (Cambridge, UK). standard error (RSD), relative error of prediction (REP), and limit of de-
tection (LOD) values of PLS method were calculated using the following
3. Results and discussion equations [22].
RSD, REP and LOD values are calculated as 4%, 6.5% and 0.03%, which
In this study, wheat flour pellets were analyzed using LIBS. LIBS spec- indicates that LIBS is a reliable and sensitive method for ash analysis in
tra were recorded through laser-scanning of seven regions (15 shots/re- flour samples and has good prediction ability. Conventional ash analysis
gion for each pellet). For each flour sample, 3 pellets were analyzed with method is based on gravimetric analysis in which the mass of analyte is
LIBS, and 105 spectra were obtained from each pellet. Then, 315 spectra weighed, and it provides information only about total mineral content,
from 3 pellets were used to obtain the average spectrum for that sam- while LIBS offers data on both total mineral content and mineral compo-
ple. Full spectrum of wheat flour is presented in Fig. 2. sition. Besides, ashing method is troublesome and inconvenient for in-
LIBS spectra were normalized to eliminate the matrix effect of sam- dustrial applications as it requires a long analysis time. In the
ples. There is no exact/direct relation between ash content of flours and literature, ash analysis by using LIBS was performed for online determi-
intensity of atomic emission lines since high or low ash content can be nation of coal quality on moving belt conveyers in coal industry [23].

Table 2
Mineral compositions of flour samples with different ash content.

Samples ash% K (μg g−1) Mg (μg g−1) Ca (μg g−1) Na (μg g−1) Zn (μg g−1) Total major mineral (μg g−1)

Sample 1 0.5523 1820.08 ± 11.77 368.02 ± 1.51 231.30 ± 20.19 20.45 ± 6.62 9.04 ± 0.97 2448.89
Sample 2 0.6500 2071.63 ± 21.62 429.46 ± 26.98 255.69 ± 10.47 22.88 ± 6.50 9.98 ± 1.59 2789.64
Sample 3 0.7067 2356.12 ± 16.73 423.08 ± 4.24 282.34 ± 9.88 25.42 ± 5.90 10.47 ± 2.64 3097.43
Sample 4 1.3900 3142.33 ± 49.93 1182.74 ± 0.58 338.58 ± 2.02 50.72 ± 7.84 24.59 ± 0.39 4738.96
 G. Bilge et al. / Spectrochimica Acta Part B 124 (2016) 74–78
Fig. 3. PLS calibration and validation graphs of flour samples with different ash contents.
However, the present study is a novel study on food applications which
has made use of LIBS for ash analysis in flour samples. The results of the
study are very promising for food industry because conventional ash
analysis method is both expensive and time consuming. The developed
method is reliable, rapid, economical and in-situ, which makes it very
appealing for food industry. Ashing is not only for analysis of total min-
eral content but also for first step sample preparation before specific el-
ement analysis. However, dry ashing procedure is not applicable for
volatile trace elements analysis such as As, Hg and sometimes Fe ele-
ment due to high temperature [24]. LIBS provides data on both ash con-
tent and mineral composition including the trace elements. Ash analysis
has a crucial role for understanding the milling performance of flour
(grade of flour) and the type of wheat, which makes it indispensable
for the manufacturers. Besides, ash content is a quality control parame-
ter for many other food products such as meat, fruit juices, tea, and
cheese. In these foods, ash content indicates the mineral content and
saltiness of the food and determines the physicochemical properties of
it. Consequently, ash analysis by LIBS can be a premise method for
other food products if the homogeneity problem of sample and standard
sample preparation difficulties are overcome [25,26].
4. Conclusion
Ash analysis is a routine but a significant flour quality analysis which
takes a long time and causes energy waste. Therefore, the aim of the
study was to develop a new method for ash analysis. LIBS is a multi-el-
emental, quick and simple spectroscopic method. Unlike basic ash anal-
ysis method, LIBS has the potential to analyze a flour sample only in a
few seconds. Performance of this new technique was evaluated in com-
parison with basic ash method. Wheat flour samples with different ash
contents were analyzed using LIBS, and the spectra were evaluated with
partial least squares method. The results were correlated with the re-
sults taken from standard ash analysis method. Calibration graph
showed good linearity with the ash content between 0.48 and 1.39%
with 0.992 coefficient of determination. Limit of detection for ash anal-
ysis was calculated as 0.026%. The results indicated that LIBS is a prom-
ising and reliable method for routine ash analysis in flour samples
analyzed rapidly. Compared to the conventional ash analysis method,
LIBS has a lot of advantages in terms of duration, cost and sample prep-
aration. Furthermore, as it requires only a very short analysis time,
much higher number of flour samples can be analyzed in rapidly by
using LIBS. This method seems to be a useful tool for quality control lab-
oratories of flour mills and other food processing companies.
77
Acknowledgement
This research is partly supported by Republic of Turkey, Ministry
of Science, Industry and Technology, research grant Project No.
0875.STZ.2015.
References
[1] M.R. Marshall, Ash Analysis, in: S.S. Nielsen (Ed.), Food Analysis, Springer, New
York, NY, USA 2010, p. 105.
[2] P. Schuck, R. Jeantet, A. Dolivet, Analytical Methods for Food and Dairy Powders,
Wiley-Blackwell, Oxford, UK, 2012.
[4] A.A.f.C. Chemistry, AACC International Method 08-01.01 Ash—Basic Method,
1999.
[5] D.M. Díaz Pace, C.A. D'Angelo, D. Bertuccelli, G. Bertuccelli, Analysis of heavy metals
in liquids using Laser Induced Breakdown Spectroscopy by liquid-to-solid matrix
conversion, Spectrochim. Acta B At. Spectrosc. 61 (2006) 929–933.
[6] R.A. Multari, D.A. Cremers, J.A.M. Dupre, J.E. Gustafson, Detection of biological con-
taminants on foods and food surfaces using laser-induced breakdown spectroscopy
(LIBS), J. Agric. Food Chem. 61 (2013) 8687–8694.
[7] F. Anabitarte, A. Cobo, J.M. Lopez-Higuera, Laser-induced breakdown spectroscopy:
fundamentals, applications, and challenges, ISRN Spectroscopy (2012) 1–12.
[8] D.A. Rusak, B.C. Castle, B.W. Smith, J.D. Winefordner, Fundamentals and applications
of laser-induced breakdown spectroscopy, Crit. Rev. Anal. Chem. 27 (1997)
257–290.
[9] L. St-Onge, E. Kwong, M. Sabsabi, E.B. Vadas, Quantitative analysis of pharmaceutical
products by laser-induced breakdown spectroscopy, Spectrochim. Acta B At.
Spectrosc. 57 (2002) 1131–1140.
[10] E. Tognoni, V. Palleschi, M. Corsi, G. Cristoforetti, Quantitative micro-analysis by
laser-induced breakdown spectroscopy: a review of the experimental approaches,
Spectrochim. Acta B At. Spectrosc. 57 (2002) 1115–1130.
[11] A. Khumaeni, Z.S. Lie, H. Niki, K.H. Kurniawan, E. Tjoeng, Y.I. Lee, K. Kurihara, Y.
Deguchi, K. Kagawa, Direct analysis of powder samples using transversely excited
atmospheric CO2 laser-induced gas plasma at 1 atm, Anal. Bioanal. Chem. 400
(2011) 3279–3287.
[12] E.C. Ferreira, E.A. Menezes, W.O. Matos, D.M.B.P. Milori, A.R.A. Nogueira, L. Martin-
Neto, Determination of Ca in breakfast cereals by laser induced breakdown spec-
troscopy, Food Control 21 (2010) 1327–1330.
[13] G. Bilge, İ.H. Boyacı, K.E. Eseller, U. Tamer, S. Çakır, Analysis of bakery products by
laser-induced breakdown spectroscopy, Food Chem. 181 (2015) 186–190.
[14] T.L. Westover, R.M. Emerson, Rapid analysis of inorganic species in herbaceous ma-
terials using Laser-Induced Breakdown Spectroscopy, Ind. Biotechnol. 11 (2015)
322–330.
[15] M.M. El-Deftar, J. Robertson, S. Foster, C. Lennard, Evaluation of elemental profiling
methods, including laser-induced breakdown spectroscopy (LIBS), for the differen-
tiation of Cannabis plant material grown in different nutrient solutions, Forensic Sci.
Int. 251 (2015) 95–106.
[16] D.K. Tripathi, A. Pathak, D. Chauhan, N. Dubey, A. Rai, R. Prasad, An efficient ap-
proach of Laser Induced Breakdown Spectroscopy (LIBS) and ICAP-AES to detect
the elemental profile of Ocimum L. species, Biocatalysis and Agricultural Biotechnol-
ogy 4 (2015) 471–479.
[17] EPA, Method 3051 microwave assisted acid digestion of sediments, sludges, soils
and oils, EPA, 1994 (Washington, District of Columbia).
78 G. Bilge et al. / Spectrochimica Acta Part B 124 (2016) 74–78
[18] B.C. Windom, P.K. Diwakar, D.W. Hahn, Dual-pulse Laser Induced Breakdown Spec-
troscopy for analysis of gaseous and aerosol systems: plasma-analyte interactions,
Spectrochim. Acta B At. Spectrosc. 61 (2006) 788–796.
[19] J.E. Carranza, D.W. Hahn, Sampling statistics and considerations for single-shot anal-
ysis using laser-induced breakdown spectroscopy, Spectrochim. Acta B At.
Spectrosc. 57 (2002) 779–790.
[20] L.C. Peruchi, L.C. Nunes, G.G.A. de Carvalho, M.B.B. Guerra, E. de Almeida, I.A.
Rufini, D. Santos Jr., F.J. Krug, Determination of inorganic nutrients in wheat
flour by laser-induced breakdown spectroscopy and energy dispersive X-ray
fluorescence spectrometry, Spectrochim. Acta B At. Spectrosc. 100 (2014)
129–136.
[21] N.I.o.S.a. Technology, NIST Atomic Spectra Database Lines Form, 2016.
[22] M.A. Gondal, Z.S. Seddigi, M.M. Nasr, B. Gondal, Spectroscopic detection of health
hazardous contaminants in lipstick using Laser Induced Breakdown Spectroscopy,
J. Hazard. Mater. 175 (2010) 726–732.
[23] M. Gaft, E. Dvir, H. Modiano, U. Schone, Laser Induced Breakdown Spectroscopy ma-
chine for online ash analyses in coal, Spectrochim. Acta B At. Spectrosc. 63 (2008)
1177–1182.
[24] S.S. Nielsen, Food Analysis, Springer, 2010.
[25] G. Bilge, B. Sezer, K.E. Eseller, H. Berberoğlu, A. Topçu, İH. Boyacı, Determination of
whey adulteration in milk powder by using laser induced breakdown spectroscopy,
Food Chem 212 (2016) 183–188.
[26] G. Bilge, H.M. Velioglu, B. Sezer, K.E. Eseller, I.H. Boyaci, Identification of meat species
by using laser-induced breakdown spectroscopy, Meat Sci. 119 (2016) 118–122.