Journal of Chromatographic Science 2015;53:1346– 1352
doi:10.1093/chromsci/bmv023 Advance Access publication April 1, 2015
Determination of Curcuminoids in Curcuma longa Linn. by UPLC/Q-TOF–MS: An
Application in Turmeric Cultivation
Kamran Ashraf1, Mohd Mujeeb1*, Altaf Ahmad2, Niyaz Ahmad3 and Mohd Amir1
1
Bioactive Natural Product Laboratory, Department of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Jamia Hamdard, New
Delhi, 110062, India, 2Molecular Ecology Laboratory, Department of Botany, Faculty of Science, Jamia Hamdard, New Delhi, 110062,
India, and 3Department of Pharmaceutics, Faculty of Pharmacy, Jamia Hamdard, New Delhi, 110062, India
*Author to whom correspondence should be addressed. Email: drmmujeeb12@gmail.com
Received 25 February 2014; revised 6 December 2014
Cucuma longa Linn. (Fam—Zingiberaceae) is a valued medicinal plant
contains curcuminoids (curcumin, demethoxycurcumin and bisdeme-
thoxycurcumin) as major bioactive constituents. Previously reported
analytical methods for analysis of curcuminoids were found to suffer
from low resolution, lower sensitivity and longer analytical times. In
this study, a rapid, sensitive, selective high-throughput ultra high per-
formance liquid chromatography-tandem mass spectrometry (UPLC/
Q-TOF–MS) method was developed and validated for the quantifica-
tion of curcuminoids with an aim to reduce analysis time and enhance
efficiency. UPLC/Q-TOF – MS analysis showed large variation
(1.408 – 5.027% w/w) of curcuminoids among different samples
with respect to their occurrence of metabolite and their concentra-
tion. The results showed that Erode (south province) contains highest
quantity of curcuminoids and concluded to be the superior varieties.
The results obtained here could be valuable for devising strategies for
cultivating this medicinal plant.
Introduction
Turmeric (Curcuma longa Linn.) belongs to the family
Zingiberaceae comprises more than 80 species of rhizomatous
perennial herbs and has extensive occurrence in the tropics
of Asia Africa and Australia (1). India is the largest producer,
consumer and exporter of turmeric in the world and contains
highest diversity of 40 species (2). The major and character-
istic active components of the turmeric are three different
curcuminoids pigments, curcumin (C) 1,7-bis-(4-hydroxy-3-
methoxyphenyl)-1,6-heptadiene-3,5-dione], demethoxycurcumin
(DMC) [1-(4-hydroxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)-
1,6-heptadiene-3,5-dione] and bisdemethoxycurcumin (BDMC)
[1,7-bis(4-hydroxy phenyl)-1,6-heptadiene-3, 5-dione] (3 – 5).
The chemical structures of these curcuminoids derivatives are
shown in Figure 1. Globally curcuminoids are gaining importance
as a budding source of new drug(s) to combat a variety of ailments
as it contains molecules endorsed with antifungal properties (6),
anti-inflammatory, hepatoprotective, antitumor, antiviral (7) and
anticancer activities (8).
In recent days different methods for the quantitative estima-
tion of the curcuminoids have been reported and it was found
that the contents of curcuminoids in turmeric were varying a
lot while employing high performance liquid chromatography
(HPLC) (6) and high performance thin layer chromatography
(HPTLC) (2). These techniques need re-evaluation using a very
robust and sensitive latest analytical technique like UPLC/
Q-TOF –MS for the analysis of basic drugs. A chromatographic as-
sessment between HPLC and HPTLC methods has also been
# The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com
Article
Downloaded from https://academic.oup.com/chromsci/article/53/8/1346/312947 by guest on 31 May 2023
reported for quality control (QC) of curcuminoids, but very
few studies have been reported to determine curcuminoids in
different curcuma species or in the same species of samples col-
lected from different cultivation regions and this information is
substantially important for the QC of related herbal medicines
(9). Chromatographic methods such as HPTLC (2, 10), HPLC
(6) and GC –MS (11) have also been reported for the quantifica-
tion of curcuminoids, but these methods were found to suffer
from low resolution, lower sensitivity/selectivity and longer ana-
lytical times. Karasz et al. (12) also reported the HPLC method
for the separation of curcuminoids by using a fluorescence
detector. In general all these methods are not stable enough to
get reproducible results, led to lack of baseline separation even-
tually resulted in impure peaks. It is therefore necessary to
develop a rapid and reliable method for the quantification of
three curcuminoids in C. longa.
In the area of fast chromatographic separation, recently ultra
high performance liquid chromatography coupled with tandem
mass spectroscopy has been established to be one of the prom-
ising developments and found to be an extremely sensitive and
specific technique for the analysis of basic drugs (13).
The UPLC system is specifically constructed in such a way that
it can operate with the fast injection cycles, low injection vol-
umes, negligible carryover and temperature control (4 – 408C),
extra stability over a broad range of pH (1 – 12) which together
contributes to rapid and sensitive analysis. Plumb et al. (14) re-
ported that simultaneous performance of orthogonal quadrupole
time-of-flight mass spectrometry (Q-TOF – MS) in combination
with UPLC helpful in structure elucidation and identification of
fragmentation patterns of the compounds. Consequently, as a re-
sult, ultra performance liquid chromatography coupled with tan-
dem mass spectroscopy has been proven to be a powerful
hyphenated technique for analytical investigations (15).
The high medicinal value of C. longa and its rising demand en-
couraged authors to investigate the chromatographic variability
profile among the different accessions. This may lead to an im-
portant documentation for determination of variability among
the accessions. So this study has undertaken to estimate metabol-
ic variability by UPLC/Q-TOF –MS among different accessions of
turmeric collected from all major ecological zones of India.
Experimental
Chemicals and reagents
All samples of C. longa were collected from different geograph-
ical regions of India (Table I) and were identified by taxonomist
Prof. M.P. Sharma, Department of Botany, Faculty of Science,
Jamia Hamdard, New Delhi. LC – MS grade acetonitrile ( purity
99.98%), methanol ( purity 99.99%), ammonium formate ( purity
99.95%) and formic acid ( purity 99.98) were purchased from
Fluka analytical, Sigma-Aldrich Corporation (St. Louis, MO USA).
All analytical standard of curcumin, demethoxycurcumin and bis-
demethoxycurcumin ( purity .98%) were purchased from Ms.
Chroma Dex, Anna CA. Water used in the entire analysis was of
LC–MS grade. Other chemicals used were of analytical grade ob-
tained from commercial sources.
Instruments
Ultra high performance liquid chromatography was performed
formate (70 : 30; v/v) in a gradient mode, which was degassed
previously, was employed for UPLC analysis. The pH of mobile
phase was default. The flow rate of the mobile phase was main-
tained at 0.25 mL/min and 10 mL of sample solution was injected
in each run. The total chromatographic run time was 5.0 min.
The column and auto-sampler were maintained at 40 and 48C,
respectively.
TOF–MS conditions
Mass spectrometry was performed on a Waters Synapt Q-TOF
Premier (Micromass MS Technologies, Manchester, UK) mass
spectrometer. The capillary voltage, sampling cone voltage, ex-

Downloaded from https://academic.oup.com/chromsci/article/53/8/1346/312947 by guest on 31 May 2023

with a Waters ACQUITY UPLC system (Serial No. F09 UPB
920M; Model Code UPB; Waters Corporation, MA, USA) 150
equipped with a binary solvent manager, an auto-sampler, col-
umn manager and a tunable MS detector (Synapt; Waters, Serial
No. JAA 272; Micromass Limited, Manchester, UK). Membrane fil-
ters of 0.22 mm (Millipore) were used for filtration of mobile
phases.
Chromatographic conditions
Chromatographic separation was performed on a Waters
ACQUITY UPLC BEH C8 (100.0  2.1 mm; 1.7 mm) column.
The mobile phase consisted of acetonitrile –10 mM ammonium
traction cone voltage, source temperature, desolvation tempera-
ture, cone gas flow, desolvation gas flow, trap gas flow and source
gas flow were set to 3 kV, 35 V, 4 V, 1208C, 3508C, 50 L/h, 500 L/h,
1.50 mL/min and 1.50 mL/min, respectively, for all three com-
pounds. The Q-TOF Premier was operated in negative ionization
mode with resolution over 8500 mass with 1.0 min scan time.
Quantitation was performed using MS – MS transitions, m/z
367.0894/217.1410 for curcumin (Figure 2A and B), 337.3505/
175.1325 for demethoxycurcumin (Figure 2C and D) and
307.0344/145.1024 for bisdemethoxycurcumin (Figure 2E and F),
with a scan time of 1.0 min and 0.02 s inter-scans per transition.
The optimum values for compound-dependent parameters such
as trap collision energy and transfer collision energy were set to
17 and 4 V for curcumin, 16 and 4 V for demethoxycurcumin
and 18 and 4 V for bisdemethoxycurcumin to obtain the fragmen-
tation information.
The accurate mass and composition of the precursor ions and
fragment ions were calculated using the Mass Lynx V 4.3 soft-
ware installed in the instrument. Argon was employed as the col-
lision gas at a pressure of 5.3  1025 torr.

Figure 1. Chemical structure of curcuminoids.
Preparation of samples
Eight different fresh rhizomes of turmeric collected from differ-
ent geographical locations were dried, powdered and passed
through sieve # 60 so as to obtain uniform powder. For the prep-
aration of analysis samples, 2.0 g of powdered rhizomes were re-
fluxed with methanol for 2 h at a temperature of 708C. The
aliquots obtained of different samples were filtered through
membrane filter and evaporated to dryness on a water bath.
The residues obtained were reconstituted in LC–MS grade meth-
anol and further transferred to a 10-mL of volumetric flask to fi-
nally make up the volume. Appropriate dilutions were made
before LC–MS analysis.

Table I
List of Different Accessions of C. longa Used in This Study

Code no. Cultivation regions (Provinces) Latitude, longitude Source Time of collections
T1 Trivandrum (Kerala) 88290 1500 N, 768570 900 E Local farmer 21st Oct. 2011
T2 Lucknow (UP) 268500 49.200 N, 808560 49.200 E Herbal Garden, Integral University 25th Oct. 2011
T3 Nashik (Maharashtra) 20800 000 N, 738460 4800 E Local farmer 13th Nov. 2011
T4 Delhi (Delhi) 288360 3600 N, 778130 4800 E Herbal garden, Hamdard University 19th Dec. 2011
T5 Erode (Tamil nadu) 118210 000 N, 778440 000 E Local farmer 1st Nov. 2011
T6 Guwahati (Assam) 268110 000 N, 918440 000 E Local farmer 23th Nov 2011
T7 Surat (Gujrat) 268110 000 N, 918440 000 E Local farmer 6th Jan 2012
T8 Patna (Bihar) 258360 39.600 N, 85880 38.400 E Local farmer 12th Jan 2012

Determination of Curcuminoids in Curcuma longa Linn. 1347

Calibration standards and QC sample preparation
The stock solution of 1,000 mg/mL concentration for curcumin,
demethoxycurcumin and bisdemethoxycurcumin were pre-
pared by dissolving 1,000 mg of the compound, made up to a
final volume of 1 mL in LC – MS grade methanol and sonicated
at 308C for 30 min. Working stock solutions were obtained by
step-wise dilution of the stock solution in methanol and were fil-
tered through 0.22 mm membrane filter before UPLC analysis.
Calibration curve standards consisting of a set of 10 non-zero
concentrations were prepared by dissolving curcumin, deme-
thoxycurcumin and bisdemethoxycurcumin in methanol to
yield a concentration range of 1 –1,000 ng/mL (1, 5, 10, 20, 50,

Downloaded from https://academic.oup.com/chromsci/article/53/8/1346/312947 by guest on 31 May 2023

Figure 2. (A) The mass spectrum (MS) of curcumin showing precursor ion spectra ( protonated precursor [M–H] – ions at m/z 367.0894) along with fragmentation transitions; (B)
the mass spectrum MSMS of curcumin showing product ion spectra (major fragmentized product ion mass spectra at m/z 217.1410); (C) the MS of demethoxycurcumin showing
precursor ion spectra ( protonated precursor [M –H] – ions at m/z 337.3505) along with fragmentation transitions; (D) the mass spectrum MSMS of demethoxycurcumin showing
product ion spectra (major fragmentized product ion mass spectra at m/z 175.1325); (E) the mass spectrum (MS) of bisdemethoxycurcumin showing precursor ion spectra
( protonated precursor [M– H] – ions at m/z 307.0344) along with fragmentation transitions and (F) the mass spectrum MSMS of bisdemethoxycurcumin showing product ion
spectra (major fragmentized product ion mass spectra at m/z 145.1024).

1348 Ashraf et al.


Figure. 2 Continued
100, 200, 400, 800 and 1,000 ng/mL). However, QC samples
were prepared at three levels; 800 ng/mL (HQC, high quality
control), 400 ng/mL (MQC, middle quality control) and
5.0 ng/mL (LQC, low quality control). All the solutions were
stored at 2 –88C until use.
Validation of UPLC method
The developed method was validated according to USFDA guide-
lines (16). Calibration curve standards consisting of a set of 10
non-zero concentrations were prepared by dissolving curcumin,
demethoxycurcumin and bisdemethoxycurcumin in methanol
to yield a concentration range of 1 – 1,000 ng/mL (1, 5, 10, 20,
50, 100, 200, 400, 800 and 1,000 ng/mL). Peak area versus con-
centration of analytes were utilized for the construction of cali-
bration curves using weighted (1/x 2) linear least squares
regression of the curcuminoids. The lower limit of quantification
(LLOQ) is the lowest concentration of the calibration curve,
which could be calculated with satisfactory accuracy and preci-
sion. The limit of detection (LOD) and limit of quantification
(LOQ) for curcumin, demethoxycurcumin and bisdemethoxy-
curcumin were determined at a signal-to-noise ratio of 3 : 1 and
10 : 1, respectively, by injecting a series of dilute solutions with
known concentrations. The recovery of curcumin, demethoxy-
curcumin and bisdemethoxycurcumin were performed at LQC,
MQC and HQC levels. Evaluation was done by comparing the
mean area response of six replicates of extracted samples (spiked
before extraction) to that of unextracted samples (spiked after
extraction) at each QC level. Determination of intra-day accuracy
and precision and replicate analysis of turmeric samples were
performed on the same day. The run consisted of a calibration
curve and six replicates of LLOQ, LQC, MQC and HQC samples.
Downloaded from https://academic.oup.com/chromsci/article/53/8/1346/312947 by guest on 31 May 2023
The inter-day accuracy and precision were also evaluated by anal-
ysis of six precision and accuracy batches on three consecutive
validation days. The intra-assay and inter-assay accuracy (% recov-
ery of the method) was determined as follows: % recovery ¼
(mean determined concentration/nominal concentration) 
100. The intra-assay and inter-assay precision (% relative standard
deviation or RSD) of the method was calculated from the mean
measured concentrations as follows: % RSD ¼ (SD of mean mea-
sured concentration/mean measured concentration)  100.
Results
UPLC/Q-TOF–MS method analysis
Initially, the curcumin, demethoxycurcumin and bisdemethoxy-
curcumin were analyzed on a BEH C18 column using acetoni-
trile – ammonium formate (50 : 50) as a mobile phase at a flow
rate of 0.3 mL/min and a column temperature of 408C. Under
these conditions, the shapes of the peaks were not sharp.
Subsequent trials were made using different amounts of acetoni-
trile, pH and temperature. The best separation was achieved on
the same column at 408C. Mobile phase consisting of acetonitrile
and ammonium formate buffer (10 mM, pH 5) in the ratio of 70 :
30 (v/v) at a flow rate of 0.25 mL/min was found to be suitable
during optimization. Ammonium formate was found to have
the ability to promote ionization of analytes, improve the peak
symmetry and was easily miscible with organic solvent; hence,
it was selected as a suitable buffer for the mobile phase.
Chromatogram showed the retention time of curcumin, deme-
thoxycurcumin and bisdemethoxycurcumin were 2.18, 2.35
and 1.84 min, respectively (Figure 3). The MS full scan spectra
for all analytes showed protonated precursor [M – H]2 ions at
the negative mode for m/z 367.0894/217.1410 for curcumin,
Determination of Curcuminoids in Curcuma longa Linn. 1349
 Downloaded from https://academic.oup.com/chromsci/article/53/8/1346/312947 by guest on 31 May 2023
Figure 3. A Typical chromatogram of standard curcumin, demethoxycurcumin and bisdemethoxycurcumin showing RT at 2.18, 2.35 and 1.84 min.

Table II Table III

Calibration Curves, LOD and LOQ of the Investigated Components (n ¼ 6) Intra-day and Inter-day Variation and Accuracy of Curcuminoids by the Proposed Method (n ¼ 6) by
UPLC/Q-TOF –MS
Component Linearity Regression equation, LOD LOQ
(ng/mL) y ¼ (mx þ c) (ng/mL) (ng/mL) Compound Mean (ng/mL) SD % RSDa % Accuracyb
Curcumin 1 –1,000 1864x – 1492 0.318 1.000 Intra-day
r ¼ 0.999 Curcumin
Demethoxycurcumin 1 –1,000 1676x – 183.2 0.337 1.068 LLQQC 0.988 0.035 3.584 98.8
r ¼ 0.998 LQC 4.945 0.115 2.325 98.9
Bisdemethoxycurcumin 1 –1,000 1016x – 592.8 0.382 1.279 MQC 399.424 7.228 1.809 99.85
r ¼ 0.999 HQC 798.236 10.68 1.337 99.77
Demethoxycurcumin
y is the peak area, x is the concentration, m is the slope and c is the intercept of the regression line, LLQQC 0.985 0.038 3.907 98.53
respectively. LQC 4.956 0.11 2.219 99.12
MQC 399.338 6.831 1.71 99.83
HQC 802.897 10.543 1.313 100.36
Bisdemethoxycurcumin
337.3505/175.1325 for demethoxycurcumin and 307.0344/ LLQQC 1.02 0.04 4.77 102
LQC 4.983 0.145 2.909 99.66
145.1024 for bisdemethoxycurcumin. MQC 398.81 6.727 1.68 99.7
HQC 798.931 9.892 1.238 99.86
Inter-day
Curcumin
Validation parameter LLQQC 1.01 0.042 4.158 101
The linear calibration plot was obtained over the concentration LQC 5.021 0.144 2.867 100.42
MQC 399.552 4.34 1.264 99.88
range of 1 –1,000 ng/mL for curcumin, demethoxycurcumin, bis- HQC 801.75 6.603 0.959 100.21
demethoxycurcumin. Good linear correlations (r . 0.998) were Demethoxycurcumin
obtained between peak areas and concentration using the least LLQQC 0.991 0.03 3.118 99.15
LQC 5.06 0.11 2.173 101.2
squares linear regression model. The linearity data are summa- MQC 399.318 4.637 1.161 99.82
rized in Table II. HQC 803.029 4.784 0.595 100.37
Bisdemethoxycurcumin
The LOQ for curcumin, demethoxycurcumin and bisdemethox- LLQQC 0.985 0.038 3.857 98.5
ycurcumin were 1.000, 1.068 and 1.279 ng/mL, respectively LQC 5.029 0.151 3.002 100.58
(Table II). Six precision and accuracy batches were run to check MQC 398.165 5.527 1.388 99.54
HQC 798.614 9.892 0.624 99.82
intra-day, inter-day precision and accuracy. The intra-day precision
and accuracy were noted and ranged 1.337–3.584% and 98.80– a
Expressed as % RSD ¼ (SD/mean)  100.
b
99.85% for curcumin, 1.313 – 3.907% and 98.53 – 100.36% for Calculated as (mean determined concentration/nominal concentration)  100.

1350 Ashraf et al.

Table IV
Contents (mg/g) of Curcuminoids in C. longa (n ¼ 6)
Samples Curcumin Demethoxycurcumin Bisdemethoxycurcumin
(mg/g) (mg/g) (mg/g)
Trivandrum 15.96 + 0.191 11.21 + 0.145 6.87 + 0.032
Lucknow 7.98 + 0.044 6.07 + 0.044 6.30 + 0.179
Nashik 10.01 + 0.112 7.50 + 0.164 5.58 + 0.087
Delhi 9.01 + 0.131 7.91 + 0.155 5.57 + 0.140
Erode 19.25 + 0.202 16.20 + 0.153 14.82 + 0.261
Guwahati 13.98 + 0.012 9.81 + 0.131 6.35 + 0.123
Surat 5.84 + 0.025 4.04 + 0.062 4.2 + 0.019
Patna 17.01 + 0.180 12.40 + 0.166 9.70 + 0.011
demethoxycurcumin and 1.238 – 4.77% and 99.66 – 102.00% for
bisdemethoxycurcumin, respectively. The inter-day precision
and accuracy were ranged 0.959 – 4.158% and 99.88 – 101.00%
for curcumin, 0.595–3.118% and 99.15–101.2% for demethoxy-
curcumin and 0.624–3.857% and 98.5–100.58% for bisdemethox-
ycurcumin, respectively. The %RSD and accuracy are summarized
in Table III. The recovery for curcumin, demethoxycurcumin and
bisdemethoxycurcumin was calculated by comparing the peak
areas of samples, which were pre-spiked with analytes at low
(25%), medium (50%) and high concentration (75%) levels with
peak areas of analytes representing 100% extraction of samples
at low, medium and high concentration levels.
The mean extraction recovery of curcumin, demethoxycurcu-
min and bisdemethoxycurcumin were found to be 98.61, 97.97
and 98.43%, respectively. There were no significant interferences
noted at the retention times of analytes.
For ruggedness study, one complete precision and accuracy
batch was processed and analyzed by different analysts using dif-
ferent columns and different set of solutions. The mean accuracy
and precision (%RSD) (n ¼ 6) were ranged 99.20– 100.21% and
0.959 – 3.06 for curcumin, 99.33 – 100.66% and 0.78 – 2.81 for
demethoxycurcumin and 98.6 – 99.6% and 0.95 – 3.18 for bisde-
methoxycurcumin, respectively, for all QC levels.
The findings of UPLC/Q-TOF–MS analysis showed that the con-
tents of three curcuminoids in C. longa were quite variable, con-
tents of three curcuminoids in most C. longa samples were
curcumin . demethoxycurcumin . bisdemethoxycurcumin ex-
cept the sample collected from Lucknow and Surat (Table IV).
Curcumin content in all extracts was found in the range of
5.84 + 0.025–19.25 + 0.202 (mg/g) while the content of deme-
thoxycurcumin and bisdemethoxycurcumin were found in the
ranges of 4.04 + 0.062–16.20 + 0.153 (mg/g) and 4.2 + 0.019–
14.82 + 0.261 (mg/g), respectively.
Discussion
The proposed research was carried out for quantification of cur-
cuminoids within eight accessions of C. longa using UPLC/
Q-TOF – MS analysis. Three curcuminoids used as a standard in
this study showed separate peaks at 2.18, 2.35 and 1.84 min of
curcumin, demethoxycurcumin, and bisdemethoxycurcumin.
The quantification of the analytes was achieved by using selective
reaction monitoring (SRM) scan mode which makes the pro-
posed method most satisfactory. Previously reported analytical
methods like HPLC –UV for the quantification and determination
of curcuminoids have several disadvantages, such as poor
separation, poor resolution and complex solvent mixtures.
These methods are also not selective and rapid, and complicated
gradient elution is required. In the proposed method, the linear-
Total
curcuminoids ity was in the range between 1 and 1,000 ng/mL which makes
(mg/g) the method most suitable for the trace quantification of analytes.
34.04 + 0.212 This technique offers improved quality data in terms of increased
20.35 + 0.135 detection limits and chromatographic resolution with greater
23.09 + 0.147 sensitivity. The advantages of our developed method over
22.49 + 0.161
50.27 + 0.245 previous techniques are the short analysis time (5 min), high sen-
30.14 + 0.129 sitivity (LLOQ: 1.000, 1.068 and 1.279) ng/mL for curcumin,
14.08 + 0.098
39.11 + 0.116
demethoxycurcumin and bisdemethoxycurcumin, respectively,
and simple extraction procedure. This developed method can
also be used for the quantification of individual curcuminoids
for routine analysis. In this experiment, highest and lowest
Downloaded from https://academic.oup.com/chromsci/article/53/8/1346/312947 by guest on 31 May 2023
percentage of curcuminoids present in C. longa were found to
be present in Erode (south province) and Surat (west province),
respectively. Our results are supported by Ashraf et al. (2) who
reported that highest curcumin content was found in the sample
of Erode by HPTLC analysis.
Conclusions
In this experiment, we reported the variability in concentration
of curcuminoids in samples collected from different geographi-
cal regions. Variability in concentration of curcuminoids could
be due to changes in different environmental conditions of
Indian subcontinent. The results showed that maximum and min-
imum amount of curcuminoids were found to be present in the
sample of Erode (south province) and Surat (west province), re-
spectively (Table IV). The developed method was completely val-
idated and showed satisfactory data for all the parameters tested.
This result would provide an important input in determining ef-
ficient management strategies for cultivation of this medicinal
plant. Erode could be considered as the desirable cultivation re-
gion for the production of turmeric at large scale.
Acknowledgments
The authors are grateful to the Honb’le Vice Chancellor, Dr G.N.
Qazi, Jamia Hamdard, New Delhi, for providing the experimental
facilities for this research study. K.A. is also thankful to University
Grant Commission (UGC), Government of India, New Delhi, for
providing research fellowship (SRF).
References
1. Purseglove, J.W., Brown, E.G., Green, C.L., Robbins, S.R. Spices, Vol. I.
Longman, London, New York, (1981).
2. Ashraf, K., Mujeeb, M., Ahmad, A., Amir, M., Mallick, M.N., Sharma, D.;
Validated HPTLC analysis method for quantification of variability in
content of curcumin in Curcuma longa L (turmeric) collected
from different geographical region of India; Asian Pacific Journal
of Tropical Biomedicine, (2012); 2: 584–588.
3. Bong, P.H.; Spectral and photophysical behaviors of curcumin and
curcuminoids; Korean Chemical Society, (2000); 21: 81 –86.
4. Jayaprakasha, G.K., Rao, L.J.M., Sakariah, K.K.; Improved HPLC method
for the determination of curcumin, demethoxycurcumin, and bisde-
methoxycurcumin; Journal of Agriculture Food Chemistry, (2002);
50: 3668–3672.
5. Verghese, J.; Isolation of curcumin from Curcuma longa L. Rhizome;
Flavour and Fragrance Journal, (1993); 8(6): 315– 319.
Determination of Curcuminoids in Curcuma longa Linn. 1351
 6. Afaq, F., Adhami, V.M., Ahmad, N.; Botanical antioxidants for chemo-
prevention of photo carcinogenesis; Frontiers in Bioscience, (2002);
7: 784–792.
7. Ammon, H.P., Wahl, M.; Pharmacology of Curcuma longa; Planta
Medica, (1991); 57: 1– 7.
8. Polasa, K., Raghuram, T.C., Krishna, T.P.; Turmeric (Curcuma longa L.)
induced reduction in urinary mutagens; Food Chemical Toxicology,
(1991); 29: 699–706.
9. Rui, L., Cheng, X., Min, Y., Hui-Fang, L., Xing, Z., De-An, G.; Qualitative
and quantitative analysis of curcuminoids in herbal medicines derived
from Curcuma species; Food Chemistry, (2011); 126: 1890–1895.
10. Paramasivam, M., Poi, R., Banerjee, H., Bandyopadhyay, A.; High-
performance thin layer chromatographic method for quantitative
determination of curcuminoids in Curcuma longa germplasm;
Food Chemistry, (2009); 113: 640– 644.
11. Asghari, G., Mostajeran, A., Shebli, M.; Curcuminoid and essential oil
components of turmeric at different stages of growth cultivated in
Iran; Research in Pharmaceutical Sciences, (2009); 4(1): 55 –61.
1352 Ashraf et al.
12. Karasz, A.B., DeCocca, F., Bokus, L.; Detection of turmeric in foods
by rapid fluorimetric method and by improved spot test;
Journal Association Official Analytical Chemistry, (1973); 56:
626– 628.
13. Nguyen, D.T., Guillarme, D., Heinisch, S., Barrioulet, M.P., Rocca, S.,
Rudaz, S., et al.; High throughput liquid chromatography with
sub-2 micron particles at high pressure and high temperature;
Journal of Chromatography A, (2007); 1167: 76– 84.
14. Plumb, R., Castro-Perez, J., Gragner, J., Beattie, I., Joncour, K., Wright,
A.; Ultra performance liquid chromatography coupled to quardru-
pole – orthogonal time of flight mass spectrometery; Rapid
Communication Mass Spectroscopy, (2004); 19: 2331– 2337.
15. Novakov, L., Matysov, L., Solich, P.; Advantages of applications of UPLC
in pharmaceutical analysis; Talanta, (2006); 68: 908– 918.
16. Food and Drug Administration (2001). Guidance for Industry
Bioanalytical Method Validation. http://www.fda.gov/downloads/
Downloaded from https://academic.oup.com/chromsci/article/53/8/1346/312947 by guest on 31 May 2023
Drugs/GuidanceComplianceRegulatory Infor mation/Guidances/
UCM070107.