emerging instrumental analytical techniques used in
TRANSCRIPT
Zagade et al. World Journal of Pharmaceutical Research
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774
EMERGING INSTRUMENTAL ANALYTICAL TECHNIQUES USED
IN PHARMACEUTICAL ANALYSIS: A REVIEW
Pratik R. Zagade*1, Nikhil N. Kumbhar
2, Kirti S. Raut
1 and Rutuja U. Thombare
1
1Department of Pharmaceutical Chemistry, Seth Govind Raghunath Sable College of
Pharmacy, Saswad, Pune.
2Department of Pharmaceutics, Seth Govind Raghunath Sable College of Pharmacy, Saswad,
Pune.
ABSTRACT
These pharmaceuticals would assist their intent best if they may be free
from impurities and are administered in the proper quantity. Hence,
there are having many demanding situations and it may be decreased
by productive use of excipients, which permits formulators to triumph
over these demanding situations. So, various chemical and
instrumental techniques had been evolved to make drugs serve their
purpose at normal intervals which are involved inside the estimation of
drugs. These prescribed drugs may broaden impurities at numerous
steps in their development, transportation and storage, which makes
the pharmaceutical sensitive to be administered hence they need to be
recognized and quantitated. For this analytical instrumentation and
strategies play a crucial role. This review highlights the function of
emerging analytical instrumentation and analytical techniques, which includes titrimetric,
chromatographic, spectroscopic, electrophoretic, and electrochemical and their corresponding
strategies which have been applied inside the analysis of pharmaceuticals for determine the
quality of the drugs.
KEYWORDS: Pharmaceuticals, Analytical Techniques, chromatography, spectroscopy,
analysis.
1. INTRODUCTION
In the present stage, the number of drugs and combination of their drug formulations are
introduced progressively into the market at a high rate. Formulation can be designed
World Journal of Pharmaceutical Research SJIF Impact Factor 8.084
Volume 9, Issue 8, 774-796. Review Article ISSN 2277– 7105
Article Received on
01 June 2020,
Revised on 22 June 2020,
Accepted on 13 July 2020,
DOI: 10.20959/wjpr20208-18173
*Corresponding Author
Pratik R. Zagade
Department of
Pharmaceutical Chemistry,
Seth Govind Raghunath
Sable College of Pharmacy,
Saswad, Pune.
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according to the route of administration.[1]
In these several drugs or formulations may be
either new entities or partial structural modifications of currently ones or novel dosage forms
and multicomponent dosage forms. As here, increasing interest of multicomponent
formulation due to their multiple actions such as greater patient acceptability, quicker relief,
increase potency and fewer side effects. However, in the development and formulation of
dosage forms there are many challenges and it can be minimized by efficacious utilization of
excipients, which sanctions formulators to surmount these challenges. Moreover, Regulatory
guidance provides only constrained details of the requisites for the data sets associated with
the pharmaceutical development[2]
but more detailed information is available for the
toxicological assessment of excipients.[3]
Reactions can occur in the dosage form when the
drug substance is reactive and may be expedited by physical and chemical interaction with
excipients. In some cases, excipients do not interact chemically but stimulate the degradation
of drug substance.
As we know that analytical chemistry is play paramount role in the fields of science and
medicine and it is linked with the disseverment, identification and quantification of the
chemical components. Mostly, it deals with the two aspects of chemical characterization i.e.
qualitative (what it is) and quantitative (how much it is). In addition to qualitative analysis is
diagnosed by way of colour, odour or melting point, beside Quantitative analysis is achieved
by means of quantification of weight or volume. However, different analytical methods are
routinely being utilized for analyzing the drug samples in bulk, pharmaceutical formulation
and organic or biological fluids.
Analytical techniques may be separated into non instrumental and instrumental. Here, in non-
instrumental techniques (wet chemistry methods) are used for separations along with
precipitation, extraction, and distillation. Now, in instrumental methods are constructed for
measuring to some physical properties of substance is evaluated its chemical composition
through the usage of instrument. Newly, the assay strategies within the monographs
encompass chromatography, titrimetry, spectrometry and capillary electrophoresis; also, the
electro analytical strategies can be seen inside the literature.
Therefore, Drug analysis shows the important role in the development of drugs,
manufacturing along with their therapeutic use. Pharmaceutical industries depend upon
quantitative chemical analysis, to make sure the high-quality, purity and efficacy of the raw
material used and final product obtained meet the desired specifications. Thus, for newly
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delivered pharmaceutical formulations, it becomes vital to increase new analytical techniques
because from time to time the dosage form includes other substances which probably
interfere inside the assay and if not corrected can also impart a systemic error to the assay.
And also, for drug or drug combination might not be authentic in any pharmacopoeias and
thus, no analytical approach for quantification is available. Therefore, different analytical
methods are used to check the excellent requirements of the dosage shape. In addition to
Modern analytical strategies are also playing key function in assessing chemical quality
standards of medicine. Thus, analytical techniques are required for fixing their standards of
medicines and its everyday checking. Thus, the overview highlights a variety of emerging
analytical, chromatographic and instrumental approach developed consisting of
spectrophotometer; HPLC, HPTLC, and GC have wide software in assuring the best and
quantity of pharmaceutical products and those instrumental strategies are simple, precise,
speedy and reproducible as compared to non- instrumental strategies.[4]
2. ANALYTICAL TECHNIQUES
2.1. Titrimetric techniques
In 18th
century, it was year 1835 when French chemist and physicist Joseph Louis Gay-
Lussac invented titrimetric method of analysis, the volumetric technique which results in the
originates the term as titration. Now, there some innovation techniques have been used, i.e.,
spreading of non-aqueous titration method, expanding the applied field of titrimetric methods
to weak acids, bases and potentiometric end points detection enhancing the precision of the
methods. Through the improvement of functional group analysis approaches titrimetric
techniques had been proven to be useful in kinetic measurements which can be in applied to
set up reaction rates. It has a few benefits which include saving time and labor, also greater
precise approach, the reason that is no need of the use of reference standards. For instances,
in the beyond albendozole, captopril and gabapentin in commercial dosage forms become
decided with the aid of titrimetric techniques.[5-7]
The determination of Sparfloxacin the non-
aqueous titration method has been used.[8]
Moreover, to its application in drug determination
titrimetry has been used in the past for the evaluation of degradation products of the
pharmaceuticals.[9]
2.2 Chromatographic techniques
In 1903, Russian-Italian botanist Tswett who invented chromatography technique. During
1970’s, different strategies which includes thin-layer chromatography, open-column
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chromatography and paper chromatography are involved in maximum chemical separations.
Primarily, chromatography is a separation method that separates mixtures in to individual
components by way of the usage of mobile and stationary phase. The constituents of relative
mixture move at different speed for causing them to separate.
The separation of compounds is relying on differential partitioning between the mobile and
stationary phase. When the stationary phase is a solid support of adsorptive nature and mobile
phase is liquid or gaseous phase it is referred to as adsorption Chromatography. Analytical
chromatography is done usually with lesser quantities of material or mixtures and is for
measuring the relative proportions of analytes in a mixture. According to USP
chromatography may be described as a technique by using which solute are separated by
means of a differential migration method in a gadget consisting of two or more phases, one in
every of which move constantly in a given direction.
2.2.1. Thin layer chromatography
As Thin layer chromatography (TLC) is an old technique still it finds a lot of application in
the field of pharmaceutical analysis. In this technique, a solid phase, the adsorbent, is coated
onto a solid support as a thin layer usually on a glass, plastic, or aluminium support.
Several factors determine the efficiency of this kind of chromatographic separation. First the
adsorbent should show extreme selectivity in the direction of the substance being separated in
order to the dissimilarities inside the fee of elution be large. For the separation of any given
mixture depends upon absorptive power these are too strong to adsorb or too weak to adsorb.
Here, lists of some adsorbents within the order of adsorptive power.[11]
Table 1: Chromatographic absorbents.[10]
Sr.no. Most strong adsorbent Least strong adsorbent
1. Alumina (Al2O3) Silica gel (SiO2)
2. Charcoal (C)
3. Florisil (MgO/SiO2) anhydrous
Thin layer chromatography (TLC) is a suitable technique for the analysis of a large variety of
organic and inorganic materials, due to its distinctive benefits such as minimum sample
clean-up, flexibility in sample characteristic, wide preference of mobile phases, high sample
loading capacity and low cost.[11]
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For analyzing unknown materials in bulk drugs TLC is an effective tool.[12]
It provides a
relatively high degree of declaration that all probable components of the drug are separated.
TLC at the high precision used to quantitative analytical purpose using spot elution and then
followed by spectrophotometric measurement. Determination of some steroid’s TLC has
been applied pioglitazone[13,14]
celecoxib.[15]
TLC performs a vital role inside the early stage
of drug development when records about the impurities and degradation products in drug
product and drug substance is inadequate. Various impurities of pharmaceuticals have been
screened and evaluated by using TLC.[16]
The determination of herbal products i.e.,
Snowdrops (Galanthus L.) is determined by TLC and UV spectrophotometry.[17]
2.2.3. High-performance liquid chromatography (HPLC)
HPLC is one of the advanced technique of liquid chromatography, invented first time in the
year 1980, for bulk drug materials during assay condition.[18]
Here, HPLC technique used for
separating the complex mixture of molecules or substances encountered in biological and
chemical systems, in order to admit better the role of individual molecules.
The specificity of the HPLC technique is superior and simultaneously sufficiently precise and
accurate. At the time of literature survey, it was realized that HPLC has been the maximum
broadly used technique over all of the chromatographic techniques. Table 2 indicates the
assessment of high-performance liquid chromatography of different dosage forms with
mobile phase, which might be taken by a number of the reviewed publications.
In liquid chromatography the selection of detection approach is crucial to guarantee that all
the components are detected. One of the broadly used detectors in HPLC is UV detector
which is able to monitoring several wavelengths concurrently; this is possible because of
applying a multiple wavelength scanning program. As in enough quantity, UV detector
assures all the UV-absorbing components are detected.
PDA i.e., a photodiode array is a coated array of discrete photodiodes on an integrated circuit
(IC) chip for spectroscopy. It is placed at the photograph plane of a spectrometer to permit a
range of wavelengths to be sensed concurrently. When a variable wavelength detector
(VWD) is used with changing wavelength, a sample must be injected numerous times, to
make sure that all of the peaks are detected. In the case of PDA, when it used a wavelength
range can be programed and all the compounds that absorb within this range may be
recognized in a single analysis. PDA detector also can examine pick purity by matching
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spectra inside a peak. PDA detector finds its application in the technique improvement of
Iloperidone in pharmaceuticals.[19]
The refractive index detector is the detector of choice whilst one desires to detect analytes
with restrained or no UV absorption which include sugars, alcohols, carbohydrates, fatty
acids, and polymers. This detector is having the shortest sensitivity amongst all detectors but
suitable at excessive analyte concentrations.
In the pharmaceutical formulation, to evaluate the content material of volgibose Lakshmi and
Rajesh used the refractive index detector.[20]
The electrochemical detector answered to the
substances that are both oxidizable or reducible and the electric output results from an
electron flow induced with the aid of the chemical reaction that takes place at the surface of
the electrode. This detector was carried out recently to investigate the content of glutathione
in human prostate most cancers cells and lung adenocarcinoma cells.[21]
One of the maximum sensitive detectors some of the LC detectors is fluorescence detector.
Commonly, the sensitivity of this detector is 10–1000 times higher as compare to the UV
detector for active UV absorbing materials used as an advantage that measurement of unique
fluorescent species in samples. One of the maximum vital applications of fluorescence is the
estimation of pharmaceuticals.[22]
Over certain period of time, most analyst used the reversed-phase technique with the help of
UV absorbance detection, because this implement the surpasses available analysis time,
reliability, repeatability and sensitivity. Various drugs have been assayed in pharmaceutical
formulations[23,24]
and in biological fluids[25,26]
by use of HPLC. Thus, HPLC provides a
primary service in answering many questions posed by the pharmaceutical industry.
However, the risks of HPLC consist of solvents, free of columns and a lack of long-time
period reproducibility because of the nature of column packing. Liquid chromatography
linked with mass spectrometry (LC–MS) is taken to consideration as one of the maximum
crucial techniques of the last decade of 20th century. Within pharmaceutical industry the
analytical field in many stages of quality control and assurance is depends on the HPLC
method.[27,28]
Currently, HPLC-MS has been used for determination of purity of drugs.[29-33]
In
addition to its utility in analyzing the drugs HPLC alone and with hyphenated technique have
been used to analyze the impurities of the pharmaceuticals[34,35]
and degradation
products.[36,37]
Development of HPLC methods for simultaneous analysis of antiretrovirals in
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pharmaceutical formulations.[38]
Using Rp-Hplc method In vitro metabolic stability study of
new cyclen based antimalarial drugs.[39]
Table 2: Different dosage forms with mobile phase used during high performance liquid
chromatography.[40-47]
Sr. No. Name of drug Mobile phase
1.
Guaiphenesin phenylephedrine HCl,
Paracetamol, chlorpheniramine maleate and
bromhexine HCl
Methanol and acetonitrile (3:2)
2. Clarithromycin powder Acetonitrile 0.035 mol l-1 pott. Di-
hydrogen ortho-phosphate
3. Phenylepherine, paracetamol, caffeine and
chlorpheniramine
Acetonitrile, MeoH, 10mm
phosphate buffer 16:22:62v/v
4. Ibuprofen and diphenhydramine Hcl -
5. Codeine phosphate, ephedrine HCl and
chlorpheniramine maleate
Methanol, glacial acetic acid,
triethylamine (980:15:6 v/v)
6. Pseudoephedrine HCl, fexofenadine HCl
cetirizine .2 HCl
Tea solution; MeoH; acetonitrile
(50:20:30, v/v/v).
7.
Phenylpropanolamine HCl, Caffeine,
paracetamol, glycerylguaiacolate;
chlorpheniramine maleate
Consisting of a mixture of acnion
pair solution (15:85, v/v)
8. Telmisartan, amlodipine besylate and
hydrochloro-thiazide
Orthophosphoric acid: ACN
(60:40) CHCl3: MeoH formic acid
85:15:5
2.2.3 High performance thin layer chromatography (HPTLC)
Further advancement of the technique, high performance thin layer chromatography
(HPTLC) arrive as an important technique in analysis of drug. HPTLC is a rapid separation
technique and flexible sufficient to investigate a various range of samples. One of the
advantages of this technique is it is simple to handle and requires a short analysis time to
analyze the complex mixtures or the crude sample cleanup. Also, this can be evaluates the
entire chromatogram with a variety of parameters without time limits. Furthermore, there is
simultaneous but independent development of more than one samples and standards on each
plate, leading to an accelerated reliability of results. This technique has been used to
quantitate drugs such as alfuzosin,[48]
ethinyl estradiol and cyproterone,[49]
Aceclofenac,
paracetamol and tramadol.[50]
Literature survey emphasizes that the importance of this
technique along with their challenges and opportunities of the HPTLC.[51,52]
Densitometric
analysis is also done by HPTLC method for Amlodipine, Hydrochlorothiazide, Lisinopril and
Valsartan.[53]
HPTLC Fingerprinting for Quality Control of an Herbal Drug - The Case of
Angelica gigas Root can be done.[54]
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Table 3 shows different steps applicable to this technique during method development,
validation and quantitative analysis i.e., estimation of some category of drugs are as follows-
Table 3: Estimation of some category of drugs by using HPTLC technique.[55-66]
Sr.no. Category of drugs Drugs
1. Analgesics/anti-inflammatory
Paracetamol/piroxicam
Ibuprofen
Dexibuprofen
2. Cardiovascular agent
NevibololHCl
Ranolazine
Telmisartan/ramipril
3. Antibiotics/anti infective Azithromycin
Cefetamet
4. Antiretroviral Emtricitabine/tenofovir
Lamivudine/stavudine/ Nevirapine
5. Antipsychotic/anticonvulsant/Anxiolytic Duloxetine HCl
Risperidone
2.2.4. Gas chromatography (GC)
Moving in advance with any other chromatographic technique, gas chromatography is an
effective separation approach for detection of volatile organic compounds. Combining
separation and online detection permits accurate quantitative evaluation of complex mixtures,
together with traces of compounds down to parts per trillions in some particular cases. Gas
liquid chromatography carries a huge role in the analysis of pharmaceutical product.[67]
The
production of high-molecular mass products inclusive of polypeptides, or thermally volatile
antibiotics bounds the scope of this approach.
Currently, assay of drugs carried out with help of gas chromatography such as isotretinion,[68]
cranberry products[69]
and employed in the evaluation of residual solvents in betamethasone
valerate.[70]
For analysis of impurities of pharmaceuticals gas chromatography plays an
important role. Recently, to estimate the process related impurities of the pharmaceuticals GC
has been applied,[71-73]
residual solvents listed as impurity by the International Conference of
Harmonization (ICH) are screened by the GC using a variety of detectors.[74-75]
2.3. SPECTROSCOPIC TECHNIQUES
2.3.1. Spectrophotometry
Another valuable group of strategies which find a critical region in pharmacopoeias are
spectrophotometric techniques based totally on natural UV absorption and chemical
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reactions.[76]
Spectrophotometry is nothing but the quantitative measurement of the reflection
or transmission properties of a material with the help of wavelength.
The advantages of these techniques are low time and laborious. The precision of these
techniques is also excellent. The use of UV–Vis spectrophotometry particularly applied in the
analysis of pharmaceutical dosage form has increased quickly over the last few years.[77-78]
The colorimetric methods are usually depends on the following aspects:
Oxidation-reduction process.
Complex-formation reaction.
A catalytic effect.
It is essential to mention that colorimetric techniques are often used for the assay of bulk
materials. For example, the blue tetrazolium assay is used for the evaluation of corticosteroid
drug formulations.[79]
The colorimetric method is likewise exploited for the dedication of
cardiac glycosides and is provided in European Pharmacopoeia. Several approaches the usage
of spectrophotometry for determination of active pharmaceutical ingredients in bulk drug and
formulations were stated and information of these methods are recorded in Table 4.
Derivative spectroscopy uses first or uppermost derivatives of absorbance with respect to
wavelength for qualitative research and estimation. The concept of derivatizing spectral
records became first offered inside the 1950s, while it was shown to have many advantages.
After all, the approach received little consideration primarily due to the complexity of
generating derivative spectra the with the assist of early UV–Visible spectrophotometers. The
preface of microcomputers inside the late 1970’s made it typically convincing to apply
mathematical strategies to generate derivative spectra quickly, effortlessly and reproducibly.
About the application of UV spectrophotometry, the derivative technique has observed its
applications. Additionally, it can be observed in fluorescence spectrometry,
atomic
absorption, fluorimetry and infrared.[80]
The use of derivative spectrometry is not restricted to
important cases, however may be of advantage every time quantitative study of normal
spectra is uncertain. Disadvantage is additionally associated with derivative methods; the
differential degrades the signal-to-noise ratio, in order that some form of smoothing is
required conjunction with differentiation.
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Table 4: Quantitative analysis of drugs in pharmaceutical formulations by UV–visible
spectrophotometric procedures.[81-96]
Sr.no. Name of drug Dosage form
1. Amiloride Tablet
2. Aceclofenac and Paracetamol Tablet
3. Dapsone Tablet
4. Aceclofenac and Tizanidine Tablet
5. Atenolol, Ibuprofen and Famotidine Tablet
6. Pantoprazole and Domperidone Tablet
7. Norfloxacin And Ornidazole Tablet
8. Acetaminophen and Ascorbic acid Tablet
9. Famotidine and Domperidone Tablet
10. Acetaminophen and Tramadol Tablet
11. Amoxicillin and Cloxacillin Capsule
12. Amitriptyline and chlordiazepoxide Tablet
13. Telmisartanand Ramipril Tablet
14. Benzoic acid Salicylic acid Tablet
15. Caffeine and Paracetamol Tablet
16. Metronidazole and Amoxicillin Mixture
2.3.2. Near infrared spectroscopy (NIRS)
Near infrared spectroscopy (NIRS) is a rapid and non-destructive method that gives multi
thing analysis of just about any matrix. In current years, NIR spectroscopy has gained a huge
appreciation in the pharmaceutical enterprise for raw material testing, product quality control
and technique monitoring. The increasing pharmaceutical interest in NIR spectroscopy is
probably a direct impact on consequence of its major advantages over different analytical
techniques, namely, an easy sample preparation without any pretreatments, the probability of
keeping apart the sample size position by using use of fiber optic probes, and the expectancy
of chemical and physical sample parameters from one single spectrum. The leading
pharmacopoeias have generally approved NIR techniques. The European Pharmacopoeia in
chapter 2.2.40[97]
and United States pharmacopoeias[98]
forward the suitability of NIR
instrumentation for application in pharmaceutical testing.
NIR spectroscopy in mixture with multivariate data analysis opens many exciting approaches
in pharmaceutical analysis, each qualitatively and quantitatively. A wide variety of
publications describing quantitative NIR measurements of active component in intact pills
have been recorded.[99-101]
In addition to the research articles many evaluation articles have
been published bringing up the utility of the NIRS in pharmaceutical analysis.[102]
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2.3.3. Nuclear magnetic resonance spectroscopy (NMR)
Since the primary report appeared in 1996 describing the utilization of magnetic resonance
chemical analysis to screen for the drug molecules, the sphere of magnetic resonance
primarily based screening has proceeded promptly.[103]
Recently magnetic resonance finds its
application in measurement in order to work out the impurity of the drug[104]
, characterization
of the composition of the drug products and in quantitation of medication in pharmaceutical
formula- tions and biological fluids[105]
, several reviews on the appliance of magnetic
resonance in pharmaceuticals are revealed.[106-107]
2.3.4. Fluorimetry and phosphorimetry
The pharmaceutical industries unendingly search for the sensitive analytical techniques
exploitation the small samples. Fluorescence spectroscopy is one amongst the techniques that
serve the purpose of high sensitivity while not the loss of specificity or precision. A gradual
increase within the variety of articles on the application of fluorimetry[108-109]
and
phosphorimetry[110]
in measurement of varied medication in dose forms and biological fluids
has been detected within the recent past.
2.5. Electrochemical methods
The application of electrochemical techniques in the analysis of drugs and pharmaceuticals
has increased greatly over thelast few years. An amberlite XAD-2 and titanium dioxide
nanoparticles modified glassy carbon paste was developed for the determination of
imipramine, trimipramine and desipramine. The electrochemical behavior of these drugs was
investigated using cyclic voltammetry, chronocoulometry, electrochemical impedence
spectroscopy and adsorptive stripping differential pulse voltammetry.[111]
The
electrochemically initiated formation of capsaicin-benzocaine adduct causes a linear decrease
in the voltammetric signal corresponding to capsaicin which correlates to the added
concentration of benzocaine.[112]
The electrochemical behavior of clioquinol, a molecule with
a large spectrum of clinical applications, was studied by cyclic, differential pulse and square-
wave voltammetry over a wide pH range using a glassy carbon electrode.[113]
Here the
application of various electrochemical modes in the analysis of drugs and pharmaceuticals is
presented in Table no.5
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Table 5: Determination of drug by electrochemical Techniques.[114-120]
Technique Drugs determined Remark
Voltametry b-blocker drugs Nafion-coated glassy carbon electrode
Secnidazole Cathodic adsorptive stripping voltammetry
Polarography Nifedipine
Ciclopirox olamine
Amperometry Diclofenac
Verapamil
Potentiometry N-acetyl-L-cysteine
3. CONCLUSION
The main aim of the pharmaceutical drugs is to serve the human to make them free from
potential illness or prevention of the disease. For the medication to serve its meant purpose
they should be free from impurity or different interference that might dangerous to humans.
This review is aimed toward focusing the role of various analytical instruments within the
assay of pharmaceuticals and giving an intensive literature survey of the instrumentation
concerned in pharmaceutical analysis.
4. ACKNOWLEDGEMENTS
The author thank to teachers for providing the guidance during review work.
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