1. INTRODUCTION TO DERIVATIVE: SPECTROSCOPY:

Spectrophotometric methods are still frequently used and are the most common. Due to the equipment's general availability, the procedures' simplicity, the methodology's speed, and its accuracy, spectrophotometric techniques continue to be popular.¹Derivative spectrophotometry is playing a very important role in the multi-component investigation of mixtures using UV-VIS molecular absorption spectrophotometry under computer-controlled apparatus.²

Derivative spectroscopy has been widely used as a method for quantitative analysis, characterization, and quality control in the fields of pharmacology, biomedicine, and agriculture. It consists of the change from regular spectra to their first, second, or higher subordination of spectra. It is typical to find D0 spectra, also known as essential zero order.³

Figure 1: Zero, first, second and higher order derivatives.⁴

The objective with which derivative methods used in analytical chemistry are^4,5:

· Spectral differentiation

· Spectral resolution enhancement

· Quantitative analysis

1.1 Measurement Techniques of the Derivative Spectroscopy:

The derivative spectra value is measured using the zero-crossing method, graphic measurement, and numerical measurement, respectively.⁴

2. DERIVATIVE SPECTRA:

In quantitative analysis, derivative spectra increase the difference between spectra to separate overlapping bands. The Savitzky-Golay digital algorithm technique is most typically used to obtain derivative spectra.

2.1 The Way of obtaining the derivative orders:

A spectrum that is normal or zero order can be transformed using derivative spectroscopy into its first, second, or higher derivative spectrum. The transformation of derivative spectra can be clearly understood in terms of an ideal absorption band, which is represented by a Gaussian band. When absorbance is plotted versus wavelength, a graph that should have gone to zero on the ordinate instead displays a peak with maxima and minima as well as points of inflection.⁴

Figure 2: Derivative Spectra.^[4]

2.1.1 First-order derivative spectrum:

It is obtained by derivatizing the zero-request range once.³

2.1.2 Second-order derivative spectrum:

It can be obtained by twice derivatizing the zero-request range.³

2.1.3 Third-order derivative spectrum:

The second range of requests and the third request subsidiary range compare the unique bend's scatter capacity.³

2.1.4 Fourth-order derivative spectrum:

In comparison to the first band, it has a more refined focal top and is rearranged range of the second request. The fourth subsidiary restricted groups are not inflexible.³

3. MULTI-COMPONENT ANALYSIS:

Derivative spectrophotometry has found many uses in the investigation of multicomponent samples. This approach is based on the use of derivative spectra that result from zero-order derivatization of UV-VIS absorption spectra. The Beer law in derivative form assumes the following form:

Dn = dnA/ dλn = dnε/ dλn cl

where D is the n-order derivative at wavelength λ, l is the thickness of the absorption layer, and ε is the molar absorption coefficient. The derivative spectrum of a mixture is the total of the derivative spectra of each component as long as the additivity law is maintained:

Dn mix = Dn 1 + Dn 2 +···+ Dn n

where the value of the n-order derivative of the mixture at analytical wavelength, Dn 1, Dn 2, ..., Dn x are the values of the n-order derivative at an analytical wavelength of 1st, 2nd, ..., xth component of the mixture. The "zero-crossing technique" is the name given to this method of quantitative determination. It enables the determination of a few analytes in a sample simultaneously. When compared to the conventional approach, the extra characteristic of derivative spectrophotometry is the dependence of the derivatization result on the form of zero-order spectra. While broad even powerful zero-order signals are flattened and ultimately zeroed by derivatization, analyte signals that are in the basic spectrum are amplified. This characteristic enhances the determination's selectivity and enables the background's impact to be eliminated.⁶

4. APPLICATIONS:

Quality control is a crucial pharmaceutical industry activity since medications must be marketed as safe and therapeutically effective formulations and free from Impurities.^7,8Modern innovative medications are produced quickly, necessitating the development of more advanced analytical methods for evaluating them. Pharmaceutical analysis and formulation are both concerned with the stability testing^9,10,11of active ingredients and pharmaceutical products as part of the control process.

Derivative spectrophotometry as a powerful method for efficiently eliminating sample matrix and as a potential application for speedy simultaneous multicomponent determination. For single-component pharmaceutical dosage forms and multicomponent pharmaceutical dosage forms, respectively, the analytical characteristics of derivative spectrophotometry methods established in the preceding five years are briefly described in Tables 1 and 2.⁵

4.1 Inorganic Analysis:

For the simultaneous measurement of trace elements with identical chemical characteristics present in mixtures at various concentration levels, Derivative spectrometry is particularly commonly used in water and inorganic chemistry due to its enhanced selectivity and sensitivity compared to standard spectrophotometry. The first and second order derivatives are typically utilised for this purpose, however higher order derivatives may offer more trustworthy results described in Tables 3.^39-52

Table 1. Multiple Compound Determination by Derivative Spectrometry.^12-26

Compound	Matrix	Wavelength		Solvent	Linearity (µg/ml)	Ref
Compound	Matrix	Λ1	Λ2	Solvent	Linearity (µg/ml)	Ref
Atenolol Amlodipine	Tablet	251 nm	264 nm	Water	5-50 5-45	12
Chlorpheniramine Maleate Diphenhydramine Hydrochloride	Physical Mixture	265 nm	251 nm	Methanol	4.5-9.5	13
Ciprofloxacin Fluocinolone	Otic Solution	278 nm	238 nm	Methanol	3-15	14
Diclofenac Sodium Nicotinamide	Tablet	276 nm	214 nm	Methanol	5-80 10-140	15
Gallic Acid Ascorbic Acid	Solution	213 nm	265 nm	Distilled Water	1-30 3-35	16
Glibenclamide Metformin	Tablet	232 nm	225 nm	Methanol	1-80 1-10	17
Finasteride Tadalafil	Capsule	238 nm	292 nm	Methanol	10-14 3-40	18
Hydrochlorothiazide Triamterene	Tablet	280.5 nm	379 nm	Methanol	0.64-28.8 0.8-19.2	19
Sulphadoxine trimethoprim	Veterinary	289.2 nm	230.5 nm	Ethanol	15-200 40-300	20
Dextromethorphan HBr Glyceryl Guaiacolate	Syrup	236.6 nm	285 nm	Methanol Water(50:50)	40-120 20-50	21
Propranolol Hydrochloride Rosuvastatin Calcium	Synthetic mixture	239.43 nm	249.03 nm	Distilled water	1-24	22
Sacubitril Valsartan	Tablet	247 nm	233 nm	Methanol	4.9-24.5 5.1-25.5	23
Teneligliptin Metformin	Tablet	219 nm	223 nm	Methanol	1-100	24
Empagliflozin Metformin Hydrochloride	Tablet	224 nm	232 nm	Methanol	5-30	25
Etoricoxib Paracetamol	Tablet	248 nm	258.4 nm	Methanol	1-8 5.42-43.3	26
Isoniazid Ciprofloxacin	Tablet	262 nm	274 nm	Distilled water	2-24 2-22	27

Table 2. Individual Determination of Compound by Derivative Spectrometry^.28-38

Compound	Matrix	Wavelength (Λ)	Solvent	Linearity (µg/ml)	Ref
Ivabradine Hydrochloride	Tablet	286 nm	Water	10-30	28
Atorvastatin	Tablet	237 nm	Methanol	5-20	29
Bumetanide	Tablet	262 nm	Distilled Water	10-70	30
Clarithromycin	Tablet	268 nm	Methanol	5-60	31
Ciprofloxacin	Ophthalmic Solution	278.9 nm	Methanol	3.0-28.0	32
Metformin Hydrochloride	Tablet	225 nm	Distilled Water	1-14	33
Apremilast	Tablet	230 nm	Methanol	2-12	34
Capecitabine	Tablet	280 nm	Distilled Water	40-80	35
Rizatriptan Benzoate	Tablet	225 nm	Distilled Water	0.1-360	36
Emtricitabine	Tablet	269 nm	Distilled Water	1-20	37
Deferiprone	Capsules	278 nm	Distilled Water	2-10	38

Table 3: Analysis of Ions, Clinical, Forensic, Food, Biological Compounds by Derivative Spectrometry

Ions	Matrix	Wavelength (Λ)	Solvent	Linearity (µg/ml)	Reff
Pb (II)	Environmental Water	510 nm	Dithizone	30	39
Cd (II)	Environmental Water	542 nm	Dithizone	30	39
Zn (II)	Environmental Water	520 nm	Dithizone	30	39
Iron (III)	Cement	580 nm	Ethanol Water 50:50	1.6-5.58	40
Aluminium (III)	Cement	540 nm	Ethanol Water 50:50	0.269-2.15	40
Co(II)	Traces	598 nm	1.0% Sodium Chloride	0.15-2.0	40
Fe(II)	Traces	567 nm	1.0% Sodium Chloride	0.05-0.75	40
DOC	Soil Water	300 nm	Distilled Water	0-100	41
Clinical and Forensic Analysis
Cefuroxime	Urine	292.5 nm	0.1 N Sodium Hydroxide	2 – 10	42
Cefadroxil	Urine	267 nm	0.1 N Sodium Hydroxide	2 – 10	43
Lamivudine	Serum	265 nm	Methanol	1 – 50	44
Zidovudine	Serum	271.6 nm	Methanol	2 – 100	45
Naproxen	Plasma	328.2 nm	Methanol	5.0 – 100	46
Piroxicam	Plasma	343.5 nm	0.1 N Hydrochloric Acid	0.5 – 12	47
Rofecoxib	Plasma	316 nm	Methanol	5.8 – 26.2	48
Urea	Urine	338 nm	Distilled Water	0.1-1.5	49
Food Analysis
Luwak Coffee	Mixture	221 nm	Water	0-500	50
Non-Luwak Coffee	Mixture	233 nm	Water	0-500	50
Caffeic Acid	Mixture	297 nm	Water	0-500	50
Chlorogenic Acid	Mixture	360 nm	Water	0-500	50
Vanillic Acid	Mixture	259 nm	Water	0-500	50
Analysis of Biological Compounds
Tryptophan	Mixture	236.1 nm	Distilled Water	0.1–20.0	51
Tyrosine	Mixture	222.5 nm	Distilled Water	1.0– 50.0	51
Phenylalanine	Mixture	218.9 nm	Distilled Water	1.0– 45	51
Oxyhaemoglobin	Pigment Mixtures	415 nm	Distilled Water	25-2000	52
Methaemoglobin	Pigment Mixtures	405 nm	Distilled Water	25-1000	52

Table 4: Analysis Colorants and Pesticides in Beverages by Derivative Spectrometry⁵³

Compound	Matrix	Wavelength (Λ)	Solvent	Linearity(µg/ml)	Reff
Yellow	Binary Mixtures	400 nm	Distilled Water	0.025-0.125	53
Red	Binary Mixtures	530 nm	Distilled Water	0.025-0.125	53
Scarlet	Binary Mixtures	500 nm	Distilled Water	0.025-0.125	53
Navy	Binary Mixtures	620 nm	Distilled Water	0.025-0.125	53
Blue	Binary Mixtures	600 nm	Distilled Water	0.025-0.125	53
Analysis of Pesticides in Beverages
Luvon	Coffee	388 nm	n- hexane	1-3	54
Luvon	Tea	388 nm	n- hexane	1-3	54
Luvon	Fruity	388 nm	n- hexane	1-3	54
Luvon	Mountain Dew	577 nm	n- hexane	1-3	54
Luvon	Alcohol	259 nm	n- hexane	1-3	54

6. CONCLUSION:

The simultaneous identification of two or more compounds in the same sample without prior chemical separation is one of the classic analytical challenges. The examination of mixtures by UV-VIS molecular absorption spectrophotometry involves a significant amount of derivative approaches. This aid quantitative analysis in resolving band overlaps. While multivariate calibration is the best method for more complicated mixes, derivative strategies have been highly effective in resolving binary and ternary mixtures. For the quantitative examination of multi-component mixtures, the application of the spectrophotometric derivative approach provides an effective instrument. This method has been widely used in the last few decades in a variety of fields, including gas detection, organic analysis, inorganic analysis, biological analysis, environmental analysis, food analysis, cosmetics analysis, and dye and colour analysis. It has also been used to solve the problem of interference caused by substances other than analytes that are frequently present in pharmaceutical formulations or for the combination of two or more drug substances. DS is a method for resolving overlapped spectra, to put it briefly. It is determined by zero-order absorption spectrum bandwidth.

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Received on 25.11.2022 Modified on 23.01.2023

Asian J. Pharm. Ana. 2023; 13(2):108-114.

DOI: 10.52711/2231-5675.2023.00019

Investigated reaction	Characteristics of the method
Degradation of indomethacin in an alkaline solution	The monitoring of the degradation product using its four derivative spectra at 360 nm
Acidic hydrolysis of lorazepam	The kinetics of hydrolysis was observed by monitoring the main degradation product. It was assayed using the first derivative values at 231.6 nm
Photochemical degradation of nisoldipine	The first and the second derivative spectrophotometric methods at 285 and 291 nm were proposed for the investigation of the photodegradation reaction
Acidic hydrolysis of nordazepam	The amplitude between 244–251 nm of the fourth-order derivative spectra of nordazepam was used
Decomposition of omeprazole in aqueous solution	The values at 313 nm of the first derivative spectra of omeprazole were used
The photodegradation of thioridazine	The second derivative spectra at 280 nm were used for examination of the kinetics of degradation of thioridazine