[01] PHA6122 Pharmaceutical Analysis 2 - Introduction to Instrumental Methods of Analysis.pdf

[TRANS] UNIT 1: INTRODUCTION TO PHARMACEUTICAL ANALYSIS ○ Volumetric B. INSTRUMENTAL METHODS...

[TRANS] UNIT 1: INTRODUCTION TO PHARMACEUTICAL ANALYSIS ○ Volumetric B. INSTRUMENTAL METHODS OUTLINE Measurement of physical or chemical properties of the analyte I Instrumental Methods Of Analysis Separation of components: A Analytical Methods ○ Chromatography i Classical Methods ○ Electrophoretic techniques ii Instrumental Methods Quantitative analyses: II Types Of Instrumental Methods Of Analysis ○ Conductivity A Spectrometric ○ Electrode potential i Absorption ○ Light absorption or emission ii Scattering Of Radiation ○ Mass to-charge ratio iii Refraction ○ Fluorescence iv Diffraction v Rotation B Chromatographic CLASSICAL INSTRUMENTAL C Electrochemical D Miscellaneous More suitable for analysis of Ability to perform trace analysis III Analytical Instruments major constituents A Signal Generator B Transductor/Detector Used to certify analytical Most are multi-channel C Signal Processor standards techniques D Read-Out Device IV General Components Of Instrumental Measurement V Calibration Of Instrumental Methods Generally cheaper Shorter analysis time A Comparison With Standards i Direct Comparison More accurate and precise Amenable to automation ii Titration B External Standard Calibration More robust and less More samples may be C Standard Addition Method susceptible to environmental analyzed quickly, less skill D Internal Standard Method fluctuations and training required VI Signals And Noises A Signal B Noise TYPES OF INSTRUMENTAL METHODS OF ANALYSIS i Chemical Noise SPECTROMETRIC ii Instrumental Noise Emission of radiation iii Signal-To-Noise (S/N) Ratio ○ Emission spectroscopy (X-ray, UV, visible, electron, Auger) VII Definition Of Terms ○ Fluorescence emits light immediately Information in DARK BLUE FONT are mentioned from Sir David’s ○ Phosphorescence and luminescence (X-ray, UV and visible) discussion phosphorescence delays the emission of light A. ABSORPTION INSTRUMENTAL METHODS OF ANALYSIS Spectrophotometry and photometry: X-ray, UV, IR ANALYTICAL METHODS Photoacoustic spectroscopy: uses both light and sound 2 classifications: Classical Methods and Instrumental Methods Nuclear magnetic resonance Electron spin resonance spectroscopy A. CLASSICAL METHODS B. SCATTERING OF RADIATION Also called “Wet-chemical Method” Turbidimetry (transmitted light) Old, traditional Nephelometry (scattered light) Measurement depends on the chemical properties of the Raman spectroscopy sample Reagent is made to react completely with the analyte Relationship between the measured signal and analyte C. REFRACTION concentration is determined by chemical stoichiometry Refractometry Separation: Interferometry ○ Precipitation ○ Extraction ○ Distillation D. DIFFRACTION Qualitative analyses: Xray ○ Colors Electron diffraction methods ○ Boiling or Melting points ○ Solubility E. ROTATION ○ Odor Polarimetry ○ Optical Activity (OA) Optical rotatory dispersion ○ Refractive Index (RI) Circular dichroism Quantitative analyses: ○ Gravimetric CHROMATOGRAPHIC BUSTAMANTE, LISCANO | 3A-PH 1  TRANS: UNIT 1 — INTRODUCTION TO PHARMACEUTICAL ANALYSIS High Performance Liquid Chromatography: (mobile phase: GENERAL COMPONENTS OF INSTRUMENTAL liquid) MEASUREMENT Gas Chromatography (mobile phase: gas) GC: gas chroma: gas ELECTROCHEMICAL Electrical potential/chance ○ Potentiometry (POTENTIal = POTENTIometry) Electrical charge ○ Coulometry (ChaRge = CoulometRy) Electrical current ○ Amperometry ○ Polarography (cuRRent = ampeRometRy; polaRogRaphy) also AMPERE means electrical current CALIBRATION OF INSTRUMENTAL METHODS Electrical resistance Calibration determines the relationship between the analytical ○ Conductometry (resistance = CONDUCTOR) response and the analyte concentration. Directly involves AR/AC MISCELLANEOUS analytical response and analytical concentration through Mass-to-charge ratio the signal generator and read out device ○ Mass spectrometry analytical signal and electrical signal are the same signal Thermal characteristics generated. ○ Thermal gravimetry and titrimetry ○ Differential scanning calorimetry COMPARISON WITH STANDARDS ○ Differential thermal analysis A. DIRECT COMPARISON ○ Thermal conductometric methods property of the analyte is compared with standards such that the Radioactivity property being tested matches the standard ○ Activation and isotope dilution methods concentration of the analyte was then equal to the concentration of the standard after dilution ANALYTICAL INSTRUMENTS Example: using standard weights for analytical balance Converts information about the physical or chemical Instruments and standard weights need to be verified. With characteristics of the analyte to information that can be inaccurate readings of the instrument, a correction factor to manipulated and interpreted by man the balance readings has to be applied through + or -. If a 100-gram standard weight is placed on a balance and the indicated weight is 99.98 grams, the error is 0.02 grams. A +0.02 correction should be applied. B. TITRATION the most accurate of all analytical procedures analyte reacts with a standardized reagent (titrant) in a reaction of known stoichiometry amount of the standardized reagent needed to achieve chemical Figure 1. Diagram showing the general components of an instrumental equivalence can then be related to the amount of analyte present measurement ○ Signal generator produces signal that proceeds to EXTERNAL STANDARD CALIBRATION transducer or detector External standard ○ Transducer/detector converts signal to electrical signal used to calibrate instruments and procedures when there are ○ Signal processor translates instrument voice(electric signal) no interference effects from matrix components in the to human language analyte solution prepared separately from the sample A. SIGNAL GENERATOR series of such external standards containing the analyte in known Energy source: light (electromagnetic radiation) concentrations is prepared Chemical system - sample/matter ○ Interacting with the stimulus from the energy source ○ What you want to test/the sample Signal: output/reaction Results in the production of an analytical signal reflecting the presence and usually the concentration of the analyte B. TRANSDUCTOR/DETECTOR Transforms the analytical signal produced by the signal generator into an electrical signal Converter C. SIGNAL PROCESSOR Modies and “cleans up” the electrical signal to make it more convenient to interpret calibration curve is prepared by plotting the data or by fitting them to a suitable mathematical equation, such as the D. READ-OUT DEVICE slope-intercept form used in the method of linear least squares converts the electrical signal to a form usable to the the relationship between concentration and signal analyst. BUSTAMANTE, LISCANO | 3A-PH 2  TRANS: UNIT 1 — INTRODUCTION TO PHARMACEUTICAL ANALYSIS response signal is then obtained for the sample and used to ** Internal standard method: the reference material is added directly predict the unknown analyte concentration from the calibration to the sample and the comparison is made within the same spectrum curve or best- fit equation or plane. response signal is obtained (absorbance, peak height, peak External standard method: the reference material is in a separate area) as a function of known analyte concentration solution and the comparison is made between two separate spectra. ** has a non-substantial matrix effect SIGNALS AND NOISES STANDARD ADDITION METHOD SIGNAL useful for analyzing complex samples in which the likelihood of analytical measurement that carries information about the matrix effects is substantial analyte that is of interest to the scientist Spiking - adding one or more increments of a standard solution It involves: to sample aliquots containing identical volumes Absorbance peak area peak location peak height retention time NOISE [INTERFERENCE] analytical measurement made up of extraneous information that is unwanted it degrades the accuracy and precision of an analysis and also places a lower limit on the amount of analyte that can be detected A. CHEMICAL NOISE INTERNAL STANDARD METHOD arise from a host of uncontrollable variables that affect the Internal standard chemistry of the system being analyzed substance that is added in a constant amount to all This includes: samples, blanks, and calibration standards in an analysis undetected variations in temperature or pressure calibration involves plotting the ratio of the analyte signal to the fluctuations in relative humidity internal-standard signal as a function of the analyte concentration vibrations that lead to stratification of powdered solids of the standards changes in light intensity laboratory fumes B. INSTRUMENTAL NOISE this ratio for the sample is then used to obtain their analyte associated with each component of an instrument concentration from a calibration curve Discrepancy possibilities: device is calibrated, but one result became deviated = may imply improper preparation. device is calibrated, results from trials were consistent, but theoretical yield was different from the actual yield = sample was not as expected since theoretical yields are only based on ideal conditions. C. SIGNAL-TO-NOISE (S/N) RATIO equation that indicates the magnitude of an experimental effect above the effect of experimental error due to chance fluctuations A good S/N ratio is above 1 a ratio less than 1 may indicate that there are more interference than signal produced. BUSTAMANTE, LISCANO | 3A-PH 3  TRANS: UNIT 1 — INTRODUCTION TO PHARMACEUTICAL ANALYSIS DEFINITION OF TERMS A. INTERNATIONAL COUNCIL FOR HARMONISATION (ICH) an international non-profit organization that aims to develop guidelines via a process of scientific consensus with regulatory and industry experts working together B. OUT-OF-CONTROL PROCESS A process in which variations among the observed sampling results cannot be attributed to a constant system of chance causes C. OUT OF SPECIFICATIONS (OOS) RESULT a result that falls outside established acceptance criteria which have been established in official compendia and/or by company documentation CASE EXAMPLE: A manufacturing process consistently produces products with varying weights, even after adjustments are made. Out of Specific result D.OUT OF TREND (OOT) RESULT Specification that fails to a time-dependent result which falls outside a prediction interval or s Results meet the fails a statistical process control criterion criteria. CASE EXAMPLE: A product’s potency falls below the minimum concentration specified. E. STANDARD Out of Trend A result that a result that falls outside established acceptance criteria which Results deviates from have been established in official compendia and/or by company an expected documentation trend over F. SPECIFICATION time. a list of tests, references to analytical procedures, and appropriate acceptance criteria that are numerical limits, ranges, or other criteria for the test described establishes the set of criteria to which a material should conform to be considered acceptable for its intended use G. SYSTEM SUITABILITY TEST used to verify that the test system will perform in accordance with CASE EXAMPLE: the criteria set forth in the procedure Temperature readings from a the tests are based on the concept that the equipment, reactor consistently increase electronics, analytical operations, and samples analyzed over time; this may indicate a constitute an integral system that can be evaluated as such potential equipment malfunction or process deviation. [ADDITIONAL] DIFFERENES OF OOC, OOS, OOT REFERENCES Point of Error Example Out of Indicates a Notes from Batch 2026 – 3APH ⋆౨ৎ˚⟡˖ ࣪. Control systemic Process problem in the process University of Santo Tomas PowerPoint Presentation. BUSTAMANTE, LISCANO | 3A-PH 4