chapter-4 rp-hplc method for determination of tapentadol...
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Chapter-4
RP-HPLC method for determination of
tapentadol and its related impurities
4.1 Introduction
Tapentadol, 3-[(1R,
hydrochloride is in a group of drugs called
opioid is sometimes called narcotic. Tapentadol is approved in United States
moderate to severe chronic pain. Tapentadol is a centrally acting analgesic with a dual mode of
action as an agonist of the µ –opioid receptor and a nor epinephrine reuptake inhibitor [1
chemical structure of tapentadol shown in the Figure
Figure
Tapentadol is a white to off
simulated intestinal fluid (SIF), soluble in ethanol, sparingly solub
soluble in 2-propanol.The melting range of tapentadol is 209
Tapentadol is a new synthetic compound.
manufactured as a single (R, R) stereoisomer. Tapentadol shares a 3
amino structural fragment with morphine and its analogues.
(3-methoxyphenyl)-1-propanone by a Mannich reaction
obtain the racemic 3-dimethylamino
[(1R, 2R)-3-(dimethylamino)-1-ethyl-2-methyl propyl]
hydrochloride is in a group of drugs called opioid pain relievers. It is similar to morphine.
narcotic. Tapentadol is approved in United States and
moderate to severe chronic pain. Tapentadol is a centrally acting analgesic with a dual mode of
opioid receptor and a nor epinephrine reuptake inhibitor [1
chemical structure of tapentadol shown in the Figure-4.1
Figure-4.1 Chemical structure of tapentadol
a white to off-white powder. It is freely soluble in water, 0.1N HCL, and
simulated intestinal fluid (SIF), soluble in ethanol, sparingly soluble in methanol and slightly
propanol.The melting range of tapentadol is 209-210 °C.
Tapentadol is a new synthetic compound. It has two chiral centers and
manufactured as a single (R, R) stereoisomer. Tapentadol shares a 3-(3-hydroxyphenyl) propyl
amino structural fragment with morphine and its analogues. Tapentadol was synthesized from 1
propanone by a Mannich reaction using dimethylamine hydrochloride to
dimethylamino-1-(3-methoxyphenyl)-2-methylpropan-1-one(13)[4
methyl propyl] phenol
opioid pain relievers. It is similar to morphine. An
and used to treat
moderate to severe chronic pain. Tapentadol is a centrally acting analgesic with a dual mode of
opioid receptor and a nor epinephrine reuptake inhibitor [1-3].The
is freely soluble in water, 0.1N HCL, and in
le in methanol and slightly
has two chiral centers and can be
hydroxyphenyl) propyl
synthesized from 1-
dimethylamine hydrochloride to
one(13)[4-5]. This
intermediate is then subjected to crystallization-induced diastereomer transformation, a Grignard
reaction, acylation and finally to catalytic hydrogenolysis to give (2R, 3R)-2-methyl-3-(3-
methoxyphenyl)-N, N-dimethylpentanamine. Tapentadol was isolated as the hydrochloride salt.
All polymorphic forms of tapentadol are freely soluble within the physiological pH range.
Tapentadol is designated as Class 1 (high permeability, high solubility) in the biopharmaceutics
classification system. Stability data have demonstrated that tapentadol hydrochloride is a stable
substance. A retest period of 30 months with storage below 25°C has been approved.
Tapentadol has trade names Nucynta and Palexia. In India it is available as TAPAL (by
MSN Labs). It is a centrally acting analgesic with a dual mode of action as an agonist of the µ-
opioid receptor and as a norepinephrine reuptake inhibitor. It is also an agonist of the σ2 receptor,
though the function of this orphan receptor remains controversial. Tapentadol was developed by
Grünenthal in conjunction with Johnson & Johnson Pharmaceutical Research and Development.
It is being marketed as immediate release oral tablets of 50 mg, 75 mg, and 100 mg under the
brand name Nucynta.
Tapentadol is used for the treatment of moderate to severe pain for both acute (following
injury, surgery, etc.) and chronic musculoskeletal pain. It is also specifically indicated for
controlling the pain of diabetic neuropathy when round-the-clock opioid medication is required.
Although tapentadol is not indicated for the treatment of non-diabetic neuropathic pain, it is
often used off-label for this purpose. It also has the potential to treat depression like its close
relative, tramadol, but it is not approved for this purpose [6-15].The chronic musculoskeletal
pain were shown in the Figure-4.2
Figure-4.2: Chronic musculoskeletal pain
Tapentadol is available in the United States in both immediate release and extended
release formulations as NUCYNTA® and NUCYNTA ER® respectively, from Janssen
pharmaceuticals. The immediate release formulation is provided in 50 mg (yellow), 75 mg
(orange), and 100 mg (red/orange) tablets, to be used once every 4–6 hours as needed to control
pain, up to a maximum dose of 600 mg per 24 hour period (700 mg on day one). The extended
release formulation is provided in 50 mg (white), 100 mg (very light blue), 150 mg (light blue),
200 mg (blue), and 250 mg (dark blue) dosage. The different dosage forms of tapentadol shown
in the Figure-4.3. The extended release dosage starts at 50 mg twice a day, and is then slowly
titrated to an effective and tolerable dose in the range 100mg to 250mg twice a day, with a
maximum dose of 500 mg (250 mg twice a day) per 24 hour period. The dosage and preparation
used should be frequently re-evaluated by the prescribing physician, and the dose should be
lowered if possible to the lowest effective dose if pain decreases. All preparations of
NUCYNTA® contain only the R, R stereoisomer, which is the weakest isomer in terms of opioid
activity [16-17].
Figure-4.3 Tapentadol tablets
Tapentadol is not official in any pharmacopoeia. A few reports are available in literature
on drug interaction between tapentadol and N-desmethy tapentadol in urine. Since this drug is
being marketed in domestic and international market, the present investigation by the author
describes a rapid, accurate and precise RP-HPLC method for the determination of related
substances in bulk sample and pharmaceutical dosage form. The detector response was linear in
the concentration range of 5.06 – 40.46µg/ml of drug and its related substances. The method was
validated as per ICH guidelines. Accordingly, the aim of present study was to develop a stability
indicating relative substances method by RP-HPLC.
In the available literature, many analytical procedures have been reported for the
quantitative determination of tapentadol in pure form as well as in pharmaceutical dosage
formulation by different analytical techniques like spectrophotometry [18-23] and high
performance liquid chromatography [24-35].
A variety of spectrophotometric methods were developed for the determination of
tapentadol in various pharmaceutical dosage forms. Adithya et.al proposed spectrophotometric
estimation of tapentadol in bulk and its pharmaceutical formulation [18]. Pranathi S. developed
new analytical methods for the estimation of tapentadol in bulk drug and in pharmaceutical
formulations [19]. A first order derivative spectrophotometric method for simultaneous
estimation of paracetamol and tapentadol hydrochloride in tablet dosage form was reported by
Samil d. Desai et.al [20]. Mokhtar et.al proposed spectrophotometric methods for determination
of tapentadol hydrochloride [21].Some other UV spectroscopy methods were reported by
different authors [22-23].
Different high performance liquid chromatography methods are available for determination
of tapentadol in bulk and formulations. Ramanaiah et.al proposed stability indicating RP-LC
method for simultaneous estimation of tapentadol and paracetamol in bulk and its pharmaceutical
formulations [24]. A simple, rapid and cost effective method for the routine analysis of
tapentadol hydrochloride was developed by Gandhi and et.al [25]. Mayur et.al proposed new
HPLC method for tapentadol hydrochloride in formulations [26]. Some more chromatographic
and spectroscopic methods were also reported in literature for tapentadol [27-35].
Though large number assay methods are available in literature for tapentadol, only very
few of them are standard, sensitive and selective. In view of the importance of tapentadol in drug
formulation in the treatment of various chronic diseases, a more simple, sensitive, selective and
robust method is needed for its validation in bulk drug formulations. We are now reporting a
simple sensitive and selective RP-HPLC method for the validation of tapentadol and its related
impurities which is also robust and rugged method. The method is validated as per the ICH
guidelines [36-38].
4.2 Experimental:
4.2.1 Chemicals, Reagents and samples:
The standard, samples of tapentadol and known related substances of tapentadol, such as
methoxy impurity [(2R, 3R)-3-(3-methoxyphenyl)-N,N,2-tri methyl pentanamine] and alcohol
impurity [(2S)-1- dimethylamino)-3-(3-methoxyphenyl)-2-methylpentan-3-ol hydrochloride
were received from Bio-Leo analytical Labs India (P) Ltd. Prasanth nagar, Hyderabad.HPLC
grade methanol, Acetonitrile was purchased from Merck, Mumbai, India. Analytical reagent
grade potassium hydrogen phosphate and potassium hydroxide were purchased from Merck,
Mumbai, India. High purity water was prepared by using Millipore Milli-Q plus water
purification system. The purity of all samples and impurities used in this study was greater than
99%.
4.2.2 Instrumentation:
For initial method development studies Waters prominence HPLC system was employed.
This was equipped with a quaternary UFLC LC-20AD pump, DGU-20A5 degasser, SPD-M20A
diode array detector, SIL-20AC auto sampler,CTO-20AC column oven and CBM-20A
communications bus module. Agilent 1200 series high pressure liquid chromatographic
instrument provided with Auto sampler and VWD UV detector with thermostatted column
compartment connected with EZ Chrom software was employed for the validation of the drug
and its related impurities. The analysis was carried out on Zodiac C18 250 x 4.6mm, column with
5µm particle size.
4.2.3 Standard and sample solutions:
4.2.3.1. Preparation of standard solution:
Accurately weighed and transferred 29.0 mg of tapentadol working standard into a 25 ml
clean and dry volumetric flask. 5.0ml of diluent were added and sonicate to dissolve the drug.
The solution was finally made up to the Volume with acetonitrile: methanol (1:1v/v) used as
diluent. 2.0ml of this solution was further diluted to 100ml with the diluent.
4.2.3.2 Sample preparation:
100.0 mg of tapentadol sample was sonicated with 5ml of diluent for 15minutes in a 50ml
dry volumetric flask and made up to the Volume with diluent. The resultant solution was filtered
through a 0.45 micron filter and the first 5 ml were discarded.
4.3 Evaluation of system suitability:
The system suitability was evaluated by injecting a known Volume of sample containing a
known amount of tapentadol into chromatograph and calculated the number of theoretical plates
and asymmetry of the chromatogram. When the number of theoretical plates is >3000 and tailing
factor of tapentadol peak is not more than 2.0, column is found suitable for analysis.
4.4 Results and discussion:
4.4.1 Method development and optimization:
The main objective of this work is to develop a simple and rapid method for the
determination tapentadol and its related impurities in tablet formulations by using reverse phase
high performance liquid chromatography. The method development was initiated with solubility
study of tapentadol. Based on these studies, acetonitrile: methanol (1:1v/v) mixture was chosen
as diluent for the preparation of samples solutions. From the molecular formula of tapentadol it
was notice that it is polar in nature due to the presence of one –OH group. Hence, a non polar C18
column containing octadecyl chemically bonded to porous silica stationary phase was selected
for developing reverse phase high performance liquid chromatogram. The tapentadol solution
has pH 9.0. Therefore, a buffer solution of pH about ±2.0 with respect the observed pH 7.0 is
most suitable for validation study. Hence, buffer solution of pH 7.0 was chosen for
chromatographic studies. From the molecular structure, it was observed that, there is
chromophore group (Phenyl group) was present in tapentadol. Hence there is possibility for its
UV–Visible detection. The UV experiment was performed for maximum absorbance of
tapentadol and finally concluded that tapentadol shows maximum absorbance at 220nm.
To arrive at the optimal chromatographic conditions for the determination of tapentadol
and its related substances, various trails were performed with different chromatographic
conditions and the results obtained were analyzed
Trail-1
The first trail method was performed by using gradient mode, buffer solution (pH 7.0) as
mobile phase-A and acetonitrie-methanol mixture (3:7v/v) as mobile phase-B. The injection
Volume was 10µL.
Column : Zodiac C18 250X4.6 mm with 5 µm particle size
Pump mode : Gradient
Flow rate : 1.0 ml/min
Detection wavelength : UV , 220 nm
Injection Volume : 10µL
Column temperature : 450C
Run time : 55 min
Buffer : Dissolved 6.8 mg of potassium dihydrogen ortho phosphate into 1000 ml of
Milli-Q water and the pH is adjusted pH 7± 0.005 with 10% KOH.
Mobile phase A : Buffer (pH-7.0)
Mobile phase B : Acetonitrile and methanol (3:7 v/v)
In this trail there is no elution of tapentadol isomer impurities peaks up to run time of 55 minutes
and asymmetry of tapentadol peak was more than 2.0
Trail-2
To overcome the limitations of the above trail method, the experiment was repeated by
changing the mobile phase-B composition using buffer solution-acetonitrile-methanol mixture in
1:2:7(v/v) ratio and reducing the injection Volume from 10µL to 5µL.The remaining conditions
same as above.
Column : Zodiac C18 250X4.6 mm with 5 µm particle size
Pump mode : Gradient
Flow rate : 1.0 ml/min
Detection wavelength : UV , 220 nm
Injection Volume : 5µL
Column temperature : 450C
Run time : 55 min
Buffer : Dissolved 6.8 mg of potassium dihydrogen ortho phosphate into 1000 ml of
Milli-Q water and the pH is adjusted pH 7± 0.005 with 10% KOH.
Mobile phase A : Buffer (pH-7.0)
Mobile phase B : Buffer, acetonitrile and methanol (1:2:7 v/v)
In this trail also there was no elution of tapentadol isomer impurity peaks up to run time 55
minutes but other impurities like alcohol and methoxy impurities were well separated from
tapentadol and method given excellent peak shape with tailing of tapentadol peak 1.16.
Trail-3
To achieve the separation of tapentadol impurities, the mobile phase-B composition was
changed by using buffer solution-acetonitrile-methanol mixture in 3:2:5(v/v) ratio and the
experiment was repeated under the optima conditions.
Column : Zodiac C18 250X4.6 mm with 5 µm particle size
Pump mode : Gradient
Flow rate : 1.0 ml/min
Detection wavelength : UV , 220 nm
Injection Volume : 5µL
Column temperature : 450C
Run time : 55 min
Buffer : Dissolved 6.8 mg of Potassium dihydrogen ortho phosphate into 1000 ml of
Milli-Q water mixes to dissolve adjust pH 7± 0.005 with 10% KOH.
Mobile phase A : Buffer (pH-7.0)
Mobile phase B : Buffer, Acetonitrile and methanol (3:2:5 v/v)
In this trail tapentadol peak was eluted in 14 minutes and the isomer impurity of tapentadol was
well separated from the main tapentadol peak.
Finally, satisfactory separation with better peak shape was achieved within a reasonable
retention time with gradient mode with flow rate 1.0mL/min at temperature 450C.
4.5 Method validation:
In order to determine the related substances of tapentadol, the method was validated as
per the ICH guidelines individually in terms of system suitability, specificity, precision,
accuracy, linearity, robustness, limit of detection and limit of quantification (LOD and LOQ) and
solution stability.
4.5.1 System suitability test:
System suitability was studied by injecting diluent as blank, placebo, standard solution
and tapentadol solution into the HPLC system and good resolution was obtained between
impurities and tapentadol. The system suitability results were given in the Table-4.1.The system
suitability chromatogram of tapentadol in bulk and formulation form shown in the Figure-4.4
Table-4.1. Results of system suitability
System suitability parameters Result Acceptance criteria as per USP
Theoretical plates 18819 NLT 3000
Tailing factor 1.16 NMT 2.0
4.5.2 Specificity of method with related substances:
Specificity is the ability of the method to accurately measure the analyte response in the
presence of all potential sample components. The response of the analyte in test mixtures
containing the analyte and all potential components is compared with the response of a solution
containing only analyte.For specificity determination, solution containing diluent and all related
substances of tapentadol such as methoxy impurity [(2R, 3R)-3-(3-methoxyphenyl)-N, N, 2-tri
methyl pentanamine] and alcohol impurity [(2S)-1-dimethylamino)-3-(3-methoxyphenyl)-2-
methylpentan-3-ol hydrochloride were prepared by mixing in suitable proportions. Then diluent,
standard preparation, sample preparation, sample spiked with impurities were injected with the
chromatograph and the peak homogeneity was verified for tapentadol and its related substances
using EZ Chrom software. The summary of specificity experiment results are shown in the
Table-4.2.The specificity chromatogram is given in the Figure-4.5.
Table-4.2: Results of specificity experiment
S.No.
Name of the
impurity/analyte
Peak purity
RT(Individual)
RT (Spiked sample)
1 Tapentadol 1.00000 13.99 13.99
2 Methoxy impurity 1.00000 39.75 39.76
3 Alcohol impurity 1.00000 30.89 30.85
4 Isomer-1 impurity 0.99290 16.05 15.96
5 Isomer-2 impurity 1.00000 16.98 16.94
4.5.3 Precision:
4.5.3.1 System precision (Repeatability):
Six replicate aliquots of sample solution of tapentadol spiked with impurities were
injected into RP-HPLC system and the chromatograms were recorded for checking the
performance of the system under the chromatographic conditions on the day tested and
calculated the relative standard deviation of the percentage of impurity. The RSD values
obtained were 0.49% and 0.25% for methoxy impurity and alcohol impurity respectively. This
showed the good precision of the system. The system precision results are shown in Table-4.3.
Table-4.3: Precision study of tapentadol
4.5.3.2 Intermediate Precision (Ruggedness):
The intermediate precision also called as ruggedness, is the inter-day variation. It is
defined as the degree of reproducibility obtained by following the same procedure as mentioned
for method precision experiment. The ruggedness of the test method was demonstrated by
carrying out precision study in six preparations of sample, a single batch sample by different
analysts, different columns, on different days and using different instruments and calculated the
percentage of impurities. The percentage of relative standard deviation for impurities from six
spiked sample preparations were found to be 0.16% for methoxy impurity and 0.22% for alcohol
impurity showing high ruggedness of the proposed method. The validated intermediate precision
results are given in the Tables-4.4.
Table-4.4: Statistical data of intermediate precision
S.No. Methoxy impurity(%w/w) Alcohol impurity(%w/w)
1 1.00 1.00
2 1.00 1.00
3 0.99 1.00
4 1.00 1.00
5 0.99 1.00
6 1.00 1.00
Average 1.00 1.00
SD: 0.00 0.00
% RSD 0.49 0.25
4.5.4 Accuracy:
The accuracy of the proposed method was tested by rearing sample solutions with known
quantities of methoxy impurity, alcohol impurity at the level of LOQ, 50%, 100%, 150% and
200% of target concentration (i.e., 1.0 % of test concentration.). The chromatograms were
recorded and the recovery percentages were evaluated from the peak areas. The results are
shown in Tables-4.5-4.6.
Table-4.5: Accuracy results of methoxy impurity
Spike level
Amount Added
Amount Recovered
% Recovery
% of mean
S.No. Methoxy impurity(%w/w) Alcohol impurity(%w/w)
1 1.00 1.00
2 1.00 1.00
3 1.00 1.00
4 1.00 1.00
5 1.00 1.00
6 1.00 1.00
Average 1.00 1.00
SD: 0.00 0.00
% RSD 0.16 0.22
(ppm) (ppm) recovery
LOQ level
0.441 0.433 98.25
98.84 0.441 0.439 99.56
0.441 0.435 98.71
50%
10.03 10.40 103.76
104.32 10.03 10.51 104.88
10.03 10.42 103.96
100%
20.05 20.66 103.04
102.88 20.05 20.60 102.72
20.05 20.52 102.35
150%
30.08 31.12 1003.47
103.05 30.08 30.86 102.62
30.08 31.18 103.67
200%
40.10 39.98 99.71
99.42 40.10 39.76 99.14
40.10 39.70 99.01
Table-4.6: Accuracy results of alcohol impurity
Spike level
Amount Added
Amount Recovered
% Recovery
% of mean
(ppm) (ppm) recovery
LOQ level
0.40 0.402 99.78
99.38 0.40 0.402 99.64
0.40 0.989 98.71
50%
10.08 10.35 102.77
102.68 10.08 10.39 103.08
10.08 10.30 102.19
100%
20.15 20.25 100.49
100.17 20.15 20.263 100.53
20.15 20.04 99.47
150%
30.23 31.26 103.42
103.05 30.23 31.36 103.74
30.23 30.82 101.97
200%
40.30 41.02 101.78
101.01 40.30 40.40 100.25
40.30 41.12 102.03
For methoxy impurity, the recovery percentage was calculated and obtained in the range
98.84 to 104.32%. For alcohol impurity, these values were obtained in the range 99.38-103.05%.
All the recovery values indicate good accuracy of the proposed method.
4.5.5 Linearity:
The concentration ranges in which tapentadol, its methoxy impurity and its alcohol
impurity can be determined with good accuracy were evaluated by preparing calibration plots
between the concentration of the analyte and the peak areas. Six different aliquots of standard
solutions (LOQ, 25, 50,100,150 and 200% of test concentration) were injected into the RP-
HPLC chromatograph and the chromatograms were recorded. Plot of peak areas of the resultant
chromatograms were plotted against the analyte. The experimental data obtained are shown in
Tables-4.7, 4.8, 4.9 and 4.10. The resultant linear plots obtained for these data are given in
Figure-4.6
Table-4.7: Results of linearity experiment of tapentadol
S.No.
Level
Amount of
tapentadol(ppm)
Average peak areas of
tapentadol
1 LOQ 0.20 34013
2 25.0% 5.06 677567
3 50.0% 10.12 1431271
4 100.0% 20.23 2771905
5 150.0% 30.35 4165940
6 200.0% 40.46 5575700
Table-4.8: Results of linearity experiment of methoxy impurity
S.No.
Level
Amount of methoxy
impurity (ppm)
Average peak areas of
methoxy impurity
1 LOQ 0.44 57863
2 25.0% 5.01 537163
3 50.0% 10.03 1200327
4 100.0% 20.05 2522604
5 150.0% 30.08 3778187
6 200.0% 40.10 4969776
Table-4.9: Results of linearity experiment of alcohol impurity
S.No.
Level
Amount of alcohol
impurity (ppm)
Average peak areas of
alcohol impurity
1 LOQ 0.40 42499
2 25.0% 5.6 547837
3 50.0% 10.11 1254668
4 100.0% 20.22 2632847
5 150.0% 30.33 3880785
6 200.0% 40.44 5196770
The data was subjected to statistatical analysis and the results of these analyses are
presented in Table-4.10. The values of different statistatical analysis like correlation coeffient,
Y- intercept, residual sum square and relative standard deviation conforms high precision and
accuracy of the proposed method.
Table-4.10: Summarized statistical results of tapentadol and its impurities
Tapentadol Methoxy impurity Alcohol impurity
Correlation coefficient 1.0000 0.9998 0.9998
Slope 137485 125667 129897
% of Y-Intercept 4.9599 4.9816 4.9337
Residual sum square 0.9999 0.9995 0.9996
Residual standard deviation 23251 47583 46133
4.5.6 Robustness:
The robustness of an analytical method is a measure of its capacity to remain unaffected
by small changes but deliberate variations in method parameters and provides an indication of its
reliability during normal usage. The study was carried out with respect to change in flow rate,
column temperature and pH of buffer. The chromatographic conditions were maintained same as
per test method in each case. From the obtained results, it was observed that there was no much
variation in retention time, theoretical plates and asymmetry for tapentadol peak, obtained at
different deliberately varied conditions from the test method. Hence method is robust for all the
varied conditions. The complete robustness results are shown in Table-4.11
Table-4.11: Robustness results of tapentadol
Robust Condition Theoretical plates Asymmetry
As per test method 17421 1.07
Flow changed to 0.9ml/min 18321 1.05
Flow changed to 1.1ml/min 17227 1.04
Column Temperature changed to 40°C 16827 1.04
Column Temperature changed to 50°C 19184 1.04
Buffer pH changed to 6.8 16942 1.08
Buffer pH changed to 7.2 18016 1.04
4.5.7 Solution Stability:
The stability of sample solution was tested by recording the chromatograms of freshly
prepared sample solutions of tapentadol at different time intervals i.e. after 24 hours and 48
hours by keeping the sample temperature at 25°C. The % difference in the % of impurities from
those at different time intervals was found as zero. From this observation, it was concluded that
sample solution was stable for 48 hours at ambient temperature (25°C). The solution stability
results are shown in Table-4.12
Table-4.12: Solution stability results
Impurity name Initial After 24 hours After 48 hours
% of Desfluoro impurity 0.00 0.00 0.00
% of difference - NIL NIL
% of Related compound-A 0.13 0.12 0.14
% of difference - 0.01 0.01
% Benzylated impurity 0.00 0.00 0.00
% of difference - NIL NIL
% of total impurities 0.13 0.12 0.14
% of difference - 0.01 0.01
4.5.8 Limit of detection and limit of quantification:
The detection limit and limit of quantification of tapentadol and related substance were
determined by diluting known concentrations. The simplest method to calculate limit of
detection and limit of quantification is signal to noise ratio method. The quantification limits and
detection limits of tapentadol and its impurities are given in the Tables- 4.13-4.14.
Table-4.13: Limit of quantification results of tapentadol and its impurities
S.No.
Name of the component
S/N ratio
% level of component with respect
to sample concentration
1 Tapentadol 10.10 0.01
2 Methoxy impurity 10.38 0.02
3 Alcohol impurity 10.20 0.02
Table-4.14: Limit of detection results of tapentadol and its impurities
S.No.
Name of the component
S/N ratio
% level of component with respect
to sample concentration
1 Tapentadol 2.91 0.003
2 Methoxy impurity 3.32 0.007
3 Alcohol impurity 3.42 0.0062
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