Mobilní analyzátory a jejich využití ve farmaceutickém průmyslu Tomáš Černohorský Univerzita Pardubice Fakulta chemicko-technologická Ústav environmentálního a chemického inženýrství Centralizovaný rozvojový projekt MŠMT č. C29: Integrovaný systém vzdělávání v oblasti výskytu a eliminace reziduí léčiv v životním prostředí
Mobilní analyzátory aktuálně využívané a perspektivní technologie Ramanova spektrometrie, SERS Ramanova spektrometrie FT IR spektrometrie NIR spektrometrie IMS spektrometrie Hmotnostní spektrometrie Spojení separačních technik a hmotnostní spektrometrie Mikrofluidické chipové technologie
Mobilní analyzátory obory využití ve farmaceutickém průmyslu Kontrola vstupních surovin (RMID Raw Material ID) Mezioperační kontrola Výstupní kontrola Kontrola padělků léčiv a substandardních produktů Screeninkové metody pro kontrolu znečištění v odpadních vodách
Ramanova spektrometrie, FT IR a NIR spektrometrie Raman characterizes materials by the frequencies of their molecular vibrations Mid-infrared (MIR aka FTIR) Near-infrared (NIR)
IR Absorption versus Raman Absorption/Reflectance all wavelengths in some wavelengths absorbed/reflected Raman Scattering single wavelength (laser) multiple wavelengths emitted
amount of light scattered Molecular Vibrations Raman fingerprint of the molecule Shifted wavelength of light scattered
What is Raman?
Introduction to FTIR Spectroscopy FTIR is based on measurement of the wavelengths that are absorbed as a result of the interaction with the sample. Absorbed wavelengths = Unique Fingerprint
Molecular Excitation Molecules can be excited when they are exposed to energy the more energy, the more excited they become. With a gentle nudge they become vibrationally excited (think wobbling bowl of jello). vibrational enery levels Energy (wavenumbers) 20,000 first electronic excited state 4,000 vibrational enery levels 0 electronic ground state
Molecular Excitation vibrational enery levels Energy (wavenumbers) 20,000 first electronic excited state 12,738 4,000 vibrational enery levels 0 IR absorption (e.g., FTIR) electronic ground state
Molecular Excitation vibrational enery levels first electronic excited state Rayleigh 12,738 Raman virtual excited state 785nm excitation Energy (wavenumbers) 20,000 4,000 vibrational enery levels 0 IR absorption electronic ground state
ABSORPTION v. RAMAN KIC K FTIR La se rp rov i de st he Raman ground state
Raman Spectrum Rayleigh scatter 4000 3500 Raman scatter 3000 2500 2000 1500 1000 500 0-500 0 500 1000 1500 2000 Raman shift (cm-1) {change in energy} 2500 3000
FTIR, Raman Spectra
Dextrose, Monohydrate FTIR NIR excellent selectivity requires contact / prep less selectivity non-contact / no prep * RAMAN typical range excellent selectivity non-contact / no prep 500 1000 1500 2000 2500 3000 3500 5000 wavenumbers (cm-1) 6000 7000 8000 9000 10000
-1 [ x 1] 0 Citric Acid C itr ic A c id ( G r o u n d ) N IR.s p c 7 6 Near-Infrared Good A r b itr a r y 5 4 3 2 1 0-1 -2 [ x 1] 0 1.1 4.5 C it r ic 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 Wavelength (micrometers) 2.0 2.1 2.2 2.3 2.4 A c id. s p c 4.0 (FTIR) 3.5 A b s o rb a n c e Mid-Infrared 3.0 2.5 2.0 1.5 Better 1.0 0.5 4 [ x ]1 0 4000 C it r ic 3500 3000 2500 2000 Wavenumber (cm-1) 1500 1000 A c id. s p c 6.0 5.5 5.0 Best 4.5 A r b itr a r y Raman 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 2800 2600 2400 2200 2000 1800 1600 1400 Wavenumber (cm-1) 1200 1000 800 600 400 200
Fluorescence Fluorescence Is an optical phenomenon where a molecule is electronically excited by absorbing photons The molecule then relaxes by emitting photons Drives the operation of TV s, fluorescent lights, etc. Can also be problematic in Raman spectroscopy laser
Fluorescence Looks like this the broad background is due to fluorescence. It is an additional source of noise, and if present, will slow down the data acquisition (in extreme cases, even prohibiting data acquisition) counts spectrum of animal-derived monoglycerides example moderate fluorescence, but suitable for material ID spectrum of soy protein example of extreme fluorescence 500 1000 1500 2000 Raman shift ( cm-1) 2500 3000
Mobilní analyzátory Identifikace vstupních surovin
From Spectrum to Identification Scan spectrum Match Library spectrum Identification!
Specificity dextrose monohydrate dextrose anhydrous fructose sucrose lactose monohydrate glucosamine HCl trehalose 500 1000 Raman shift ( cm-1) 21 1500 2000
Identity Testing: Is this material what it claims to be?
Traditional RMID Workflow 23
The handheld Raman Solution for RMID Workflow Typically < 6 months 24
Progression of Raman Instruments 25
Thin-Pack Spectrometer, Handheld Example Hermetically Sealed Enclosure for CCD CCD Chip and TE Cooler Raman Probe Imaging mirrors Grating Grating-Stabilized Laser Collimating and Focusing Optics Lasing substrate 26 Focusing Optics
microphazir Rx 27
MATERIAL A MATERIAL C MATERIAL B
System must be a slave to the objective Factors influencing the measurement and measurement uncertainty INSTRUMENT measurement MATERIAL ENVIRONMENT uncertainty NECESSITATES SUFFICIENT RESOLVING POWER 2000-dimensional
Example of Instrumental Factors
MATERIAL A MATERIAL C MATERIAL B
Material Identification (which one is it?) unknown pentaethylene glycol tetraethylene glycol triethylene glycol 400 600 800 1000 1200 1400 Raman shift ( cm-1) 1600
Material Identification (which one is it?) pentaethylene glycol tetraethylene glycol triethylene glycol 400 600 800 1000 1200 1400 Raman shift ( cm-1) 1600
Material Identification (which one is it?) pentaethylene glycol tetraethylene glycol triethylene glycol 400 600 800 1000 1200 1400 Raman shift ( cm-1) 1600
Material Identification (which one is it?) pentaethylene glycol tetraethylene glycol triethylene glycol 400 600 800 1000 1200 1400 Raman shift ( cm-1) 1600
Reality tends to look like this MATERIAL B MATERIAL A ENORMOUS class separation between materials MATERIAL C
TruScan System Logic reference (library) measurement determine if aspects of measurement statistically contradict reference model NO PASS (no Raman evidence to believe otherwise) 37 YES Suspect (Raman evidence of discrepancy)
Decisive Results p-value = 0.57 ephedrine HCl (measured) ephedrine HCl (reference) 500 1000 1500 2000 2500 p-value = 3.1x10-12 ephedrine HCl (measured) pseudoephedrine HCl (reference) 500 38 1000 1500 2000 2500
Authentication of phenol Reactive mass-classifier (Pr(D H)) 39 HQI phenol 0.45 0.9981 benzaldehyde 0.000000000000000000000000000015 0.9902 benzoic acid 0.00000000000000000000000000000000011 0.9827 benzyl alcohol 0.0000000000000000000000000000000000012 0.9714 toluene 0.0000000000000000000000000000000000000054 0.9890
Mobilní analyzátory mezioperační kontrola Raman FT IR NIR IMS viz. dále
Mobilní analyzátory detekce padělků léčiv a substandardních produktů Primární technika Ramanova spektrometrie Doplňková technika NIR Doplňková technika digitální mikroskopie a analýza obrazu
Is this what it is SUPPOSED to be? 42
Counterfeit Drug Prevalence Estimates Russia 10% EU 1% US 1% Lebanon 35% Mexico 10% Source: 2006 WHO Report China 8% Columbia 5% India 20% Nigeria 16% Kenya 30% Indonesia 25%
What is a counterfeit drug? No consistent, universally agreed-upon definition exists, making regulation and enforcement difficult. WHO s definition: A counterfeit drug is a drug that has been deliberately and fraudulently mislabeled with respect to identity and/or source. Counterfeiting can apply to both branded and generic products and may include products with the correct ingredients or with the wrong ingredients, without active ingredients, with insufficient active ingredients or with fake packaging.
Counterfeit vs. Substandard drugs Low quality product (degraded, insufficient ingredients, incorrect ratio of ingredients, wrong ingredients, lack of registration or documentation) SUBSTANDARD DRUGS COUNTERFEI T DRUGS Willfully mislabeled as to identity or source
A Complex Problem Substandard medicines: Products whose composition and ingredients do not meet the correct scientific specifications, and are potentially dangerous Counterfeit (falsified) medicines: Products deliberately and fraudulently mislabeled with respect to identity and/or source.
Which is which? Real and Fake Tamiflu..Can you tell the difference?
Authentic Viagra tablet, 100mg dose Titanium oxide(anatase) Viagra 100mg Sildenafil Lactose 500 1000 1500 Raman shift ( cm-1) 48 2000 2500
Counterfeit Viagra breakdown Unknown Calcium sulfate hemihydrate Titanium oxide, anatase Sildenafil 500 1000 1500 2000 Raman shift ( cm-1) 49 2500