Figure 755 Intraplate errors in PAMPA measurement in 2% DOPC model in .NET framework

Generate DataMatrix in .NET framework Figure 755 Intraplate errors in PAMPA measurement in 2% DOPC model
Figure 755 Intraplate errors in PAMPA measurement in 2% DOPC model
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UV Spectral Data
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The use of direct UV spectrophotometry to measure sample concentrations in pharmaceutical research is uncommon, presumably because of the prevalence and attractiveness of HPLC and LC/MS methods Consequently, most researchers are unfamiliar with how useful direct UV can be The UV method is much faster than the other methods, and this is very important in high-throughput screening If samples are highly impure or decompose readily, the UV method is inappropriate to use LC/MS has been demonstrated to be a suitable detection system
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TABLE 721 Approximate Intraplate Errors in PAMPA Measurementa Pe 10 6 cm=s <001 01 05 1 5 10 20 30 50
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%CV >100% 60% 25% 15% 10% 10% 15% 20% 25%
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Based on $ 6000 measurements of > 200 different compounds using the 2% DOPC/dodecane (model 10) PAMPA system
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PERMEABILITY
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Pe (10-6 cm/s units)
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Pe (10-6 cm/s units)
VERAPAMIL
Pe (10-6 cm/s units)
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Interplate errors in 2% DOPC model (pH 74) over 12 months
under those conditions [556] When used carefully, LC/MS produces excellent results However, when LC/MS data-taking is driven very rapidly (eg, 20 min/ plate), disappointing results have been noted in collaborative studies [data not shown] Figures 757a c show the acceptor, donor, and reference spectra of 48 mM propranolol at the end of 15 h PAMPA assay using 20% wt/vol soy lecithin in dodecane The sum of the donor (3 mM) and the acceptor <1 mM well concentrations indicates that 45 mM is lost to the membrane In the absence of sink-creating surfactant, only a trace of propranolol reached the acceptor wells at the end of 15 h, with 94% of the compound trapped in the membrane, compared to 19% in the 2% wt/vol DOPC case (Table 75) The effective permeability in 20% soy dropped to 1:8 10 6 cm=s, compared to the DOPC value of 10:2 10 6 cm=s With surfactant-created sink condition in the acceptor compartment, the amount of propranolol reaching the acceptor wells is dramatically increased (Fig 757d),
PAMPA: 50 MODEL LIPID SYSTEMS
OPTICAL DENSITY (45 mm pathlength)
(a) OPTICAL DENSITY
00010 00008 00006 00004 00002 00000 -00002 -00004 250 275 300 325 350 375 400 425 450
(d) WITHOUT ACCEPTOR SINK (15 hr)
0012 0010 0008 0006 0004 0002 0000
250 275 300 325 350
WITH ACCEPTOR SINK (3 hr)
WAVELENGTH (nm) (b) (e) OPTICAL DENSITY
004 003 002 001 000
275 300 325 350 250
WAVELENGTH (nm)
OPTICAL DENSITY
0005 0004 0003 0002 0001 0000
WAVELENGTH (nm)
WAVELENGTH (nm)
(c) OPTICAL DENSITY
010 008 006 004 002 000
250 275 300 325 350
(f) OPTICAL DENSITY
010 008 006 004 002 000
250 275 300 325 350
WAVELENGTH (nm)
WAVELENGTH (nm)
Figure 757 UV spectra of propranolol: (a,d) acceptor wells; (b,e) donor wells; (c,f) reference wells (pH 74, 47 mm)
with the concomitant decrease in membrane retention from 94% to 41% Furthermore, the effective permeability rises to 25:1 10 6 cm=s, more than a 10-fold increase, presumably due to the desorption effect of the surfactant Only 3 h permeation time was used in the case (Figs 757d f) With such a sink at work, one can lower the permeation time to less than 2 h and still obtain very useful UV spectra This is good for high-throughput requirements Figure 757a shows that reproducible absorbances can be measured with optical density (OD) values as low as 00008, based on a spectrophotometric pathlength
PERMEABILITY
of 045 cm The baseline noise (OD in the range 350 500 nm in Fig 757a) is estimated to be about 00002 OD units
78 781
THE OPTIMIZED PAMPA MODEL FOR THE GUT Components of the Ideal GIT Model
The examination of over 50 PAMPA lipid models has led to an optimized model for gastrointestinal tract (GIT) absorption Table 722 shows six properties of the GIT, which distinguish it from the blood brain barrier (BBB) environment 1 The in vitro measurements of permeability by the cultured-cell or PAMPA model underestimate true membrane permeability, because of the UWL, which ranges in thickness from 1500 to 2500 mm The corresponding in vivo value is 30 100 mm in the GIT and nil in the BBB (Table 722) The consequence of this is that highly permeable molecules are (aqueous) diffusion limited in the in vitro assays, whereas the membrane-limited permeation is operative in the in vivo case Correcting the in vitro data for the UWL effect is important for both GIT and BBB absorption modeling 2 The in vivo environment of the GIT is characterized by a pH gradient; the pH value is constant at 74 in the receiving compartment (blood), and varying in the donor compartment (lumen) from $5 to $8 from the start to the end of the small intestine In contrast, the BBB has a constant iso-pH 74 Modeling the two environments requires proper pH adjustment in the in vitro model, as indicated in Table 722 3 The receiver compartment in the GIT has a strong sink condition, effected by serum proteins In contrast, the BBB does not have a strong sink condition In the GIT, lipophilic molecules are swept away from the site of absorption; in