Spectrum on skin Jonathan Hadgraft & Majella Lane
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1 Spectrum on skin Jonathan Hadgraft & Majella Lane The School of Pharmacy, University of London
2 radiofrequency microwave far infra red uv x rays γ rays log ν / Hz λ 3km 3m 30cm 3mm 0.03mm 300nm 3nm 3pm
3 radiofrequency microwave NMR ESR far infra red IR neutrons uv x rays γ rays log ν / Hz λ 3km 3m 30cm 3mm 0.03mm 300nm 3nm 3pm
4 radiofrequency microwave far infra red uv x rays γ rays log ν / Hz λ 3km 3m 30cm 3mm 0.03mm 300nm 3nm 3pm
5
6 Skin structure Major barrier is in stratum corneum (15 μm thick) transcellular intercellular follicular eccrine Role of lipids Corneodesmosomes
7 Composition & structure of intercellular channels Michaels et al Albery & Hadgraft Elias Boddé Potts & Francoeur Talreja et al. ceramides (50%) cholesteryl sulphate (5%) cholesterol (25%) free fatty acids (15%) Boddé H. et al Int. J. Pharm. 53 (1989) 13-24
8 Cryo-sectioning and cryo-tem Freezing 20 ms -180C nm thick vitreous section No cryo protectants Lars Norlen
9 Hydrogen bonding with ceramides HO HO NH O HO O HO O HO NH HO HO HO NH O HO O O N HO O HO NH HO O N esr neutron scattering & monolayers Hadgraft, J. et al. Int. J. Pharm. (1997) 141, Gay, C.L. et al. (1989) Int J. Pharm. 49, 39-45
10 Biophysical techniques Infra red Diffusion studies Imaging Diagnosis NMR OTTER TEWL MS
11 Fourier Transform Infra Red
12 Diffusion studies saturated solution membrane Zn Se crystal peak area control Azone from ir source to detector peak area time (mins) 400 Transcutol: water (50:50) control time (mins) 1200 Harrison, J. et al. Pharm. Res. (1996) 13,
13 ATR FTIR benzoic acid octanol: silicone membrane 12 C=O peak area pre-treatment no pre-treatment Time (hours) Dias, Hadgraft, Raghavan & Tetteh. J. Pharm. Sci. 93 (2004)
14 Something for nothing: chemometrics InSight
15 Octanol C=O peak area pre-treatment no pre-treatment Water vapour Silicone membrane Time (hours) Software enables separation of methyl and ethyl paraben Benzoic acid Dias, Hadgraft, Raghavan & Tetteh. J. Pharm. Sci. 93 (2004) Russeau
16 9.5 CN Absorbance CNP in water CNP in 5050 Pgwater CNP in wet PGL Time (min) McAuley, Lane
17 Silicone membrane: pools? 2 CN peak absorbance (AU) Propylene glycol PEG 400 IPM IPIS ipm ipm 21 h Time (mins) Profiles analysed using Scientist and solutions to Fick s Laws, modelling pg Kazarian, McAuley, Lad, Lane
18 Infra red imaging: chemometrics 50 x 50 µm Kazarian
19 Sampling the skin: tape stripping Determination of drug concentration in the stratum corneum (SC) by sequential removal of thin layers of SC at the same site with adhesive tape. Industrial University collaboration Guy et al.
20 Distribution profile of active across the stratum corneum (SC) C x Evaluation of the rate and extent of active penetration into the stratum corneum -the rate-determining barrier to skin permeation. How much? AUC x/h How fast? How far? Measure drug concentration profile as a function of position in the SC Required: (i) amount on each strip, (ii) penetration depth into SC Guy et al.
21 Lipid extraction lipid disorder Peak height (area) 0.45 C-H stretching asymmetric C-H stretching symmetric Peak shift 0.40 Absorbance O-H bending C=O Amide I Amide II Wavenumbers (cm -1 ) Dias, Guy, Hadgraft. Lane
22 Lipid extraction (peak area) 1.2 d-decanol normalised C-H peak area initial control d-octanol d-hexanol min Stratum corneum depth (μm) 55 min TS Both D-octanol and D-hexanol extract the lipids after 55 min of exposure D-decanol does not extract the SC lipids Dias, Guy, Hadgraft. Lane
23 Lipid disorder (peak shift) 10 C-H peak frequency shift (cm-1) initial control d-hexanol d-decanol d-octanol min Stratum corneum depth (μm) 55 min TS All vehicles produce a blue shift after 30 min of exposure D-decanol disordering effect is maintained for 55 min and in ~2 μm of the SC. Dias, Guy, Hadgraft. Lane
24 Solvent uptake (peak area) and lipid disorder (peak shift) C-H2 asymmetric stretching frequency shift (cm -1 ) solvent uptake volunteer 1 volunteer 2 volunteer 3 volunteer 4 volunteer 5 volunteer 6 volunteer 7 volunteer 8 Relationship between the solvent uptake (normalised C-D peak area) and the frequency shift for each volunteer, after exposure to D- decanol for 30 min, 55 min and tape stripping. Dias, Guy, Hadgraft. Lane
25 Diagnostics? Psoriasis UVA treatment Absorbance Absorbance Wavenumbers (cm-1) Wavenumbers (cm-1) Pre-treatment Peak areas C-H asymmetric = 0.07 C-H symmetric = 0.03 Post-treatment Peak areas C-H asymmetric = 0.18 C-H symmetric = 0.07 Watkinson & Burgess 2002.
26 Clinical dosing: 2 mg/cm 2 dynamic 20 μm Skin lipids 20% volume: limited solubility capacity In vitro experiments difficult to mimic dose
27 What does solvent deposited solid chemical look like on skin surface? Environmental Scanning Electron Micrographs (ESEM) 200 μm * * No deposited chemical 10 μ g/cm 2 4-cyanophenol in acetone Bunge
28 Particle size and contact with intercellular channels no contact contact no contact Bunge
29 Permeation of propylene glycol and drug: effect of the amount of gel applied Flux (ng/cm2/hr) Flux PG (µg/cm2/hr) Time (hours) 0 Drug: 10mg/cm2 Drug: 40mg/cm2 PG: 10mg/cm2 PG: 40mg/cm2 Trottet et al. Int. J. Pharm. 274 (2004)
30
31 Mass spectroscopy Royal Society fellow Dr Simona Francese Malcolm Clench SHU
32 GARField - a magnet for planar samples Fourier Transform gives 1D projection B o : Magnetic field (0.7 T) G y : Field Gradient (17 T/m) B 1 : Radio Frequency field Magnet Pole G y B 1 B 0 Skin Marker tape Glass slide RF Sensor Coil P. M. Glover, P. S. Aptaker, J. R. Bowler, E. Ciampi, P. J. McDonald J. Magn. Reson. (1996) 139, 90.
33 Skin dehydration in vitro Amplitude min 51 min 136 min 238 min 340 min 493 min Skin Profile direction Position (μm) Glass Slide sensor M Dias, J Hadgraft, P M Glover and P J McDonald, J. Phys. D: Appl. Phys. (2003)
34 Comparison with imaging studies Time course of kinetics similar Chan & Kazarian
35 In vivo lower arm & hand
36 OTTER Pulsed laser excitation Infra red emission signal Air Sample Opto Thermal Transient Emitted Radiation Imhof, London South Bank University
37 Permeation into nails in vivo Glycerol Concentration Nail hydration profile Control 25 Concentration [a.u.] min 8min 10min Concentration [a.u.] Left Thumb Nail Right Thumb Nail Depth [um] Depth [um] Hirata, Imhof, Xiao, Zheng, McAuley, Lane
38 Hydration profiles in vivo laser confocal Raman Cork, Matts et al.
39 Skin structure Major barrier is in stratum corneum (15 μm thick) transcellular intercellular follicular eccrine Role of lipids Corneodesmosomes
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