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Original: publisher page

Bin Tian, Xing Tang*, Lynne S. Taylor* · Molecular Pharmaceutics (Just Accepted Manuscript, web 04 Oct 2016) (2016)

Investigating the Correlation between Miscibility and Physical Stability of Amorphous Solid Dispersions Using Fluorescence-based Techniques

Indomethacin's own fluorescence shifts to the blue when the drug sits among other drug molecules; using this signal, the authors located the loading at which indomethacin forms a drug-rich phase in HPMC, HPMCAS and PVP, and found that in dry storage at 40 °C the HPMC and HPMCAS dispersions above about that loading crystallized within days while those below stayed amorphous for more than a year.

Abbreviations

ASD amorphous solid dispersion; DL drug loading (wt% drug in drug + polymer); IDM indomethacin; HPMC hydroxypropyl methylcellulose; HPMCAS hydroxypropyl methylcellulose acetate succinate; PVP polyvinylpyrrolidone; Tg glass transition temperature; DSC differential scanning calorimetry; IR infrared; XRD X-ray diffraction; ssNMR solid-state nuclear magnetic resonance; AAPS amorphous-amorphous phase separation.

Fig. 17: Miscibility range against crystallization onset, HPMC and HPMCAS
Fig. 17 · p.25 · Miscibility range against crystallization onset, HPMC and HPMCASShows. onsets (days) at each loading under bars marking amorphous, miscibility (transition) and crystallization ranges; no points where powders stayed amorphous.Read it as. red circles HPMC, blue squares HPMCAS; the transition band is 50–60% DL for HPMC and 20–30% for HPMCAS.Also supports K4

Story. Why they did it → how → what they found → how it fits together → what is left. Each part opens with the short version; "More detail" under it gives the full account. Grey ids open the evidence behind a sentence.

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Question

Can the autofluorescence of indomethacin detect drug–polymer phase separation in amorphous solid dispersions, and do dispersions that phase separate crystallize faster during storage?

Short answer

Largely yes. A blue-shifted band near 480 nm appeared at 60% DL in HPMC, 30% in HPMCAS and 80% in PVP, and imaging, IR and DSC put the limit within about one 10% step of that. HPMC and HPMCAS powders crystallized within 2–15 days from 60% and 40% DL and stayed amorphous for 13–14 months below; no PVP powder crystallized within 3 months. ·····

Conclusion

For films of indomethacin, the authors read a new emission band near 480 nm, like that of pure amorphous IDM, as the sign of an IDM-rich phase, and place the miscibility limit at 50–60% DL in HPMC, 20–30% in HPMCAS and 70–80% in PVP; fluorescence imaging, IR and DSC "broadly" agree, although DSC finds PVP miscible at all ratios. ····· In dry storage at 40 °C, HPMC powders from 60% DL and HPMCAS powders from 40% DL crystallized within 2–15 days, while lower loadings stayed amorphous for 13–14 months; the authors call this "a good correlation", conclude that drug-rich domains "led to" enhanced susceptibility to crystallization, and recommend checking homogeneity during formulation screening. ··· The scope is one drug, three polymers and dry storage; within it, the 30% HPMCAS powder, the first loading with a drug-rich band, stayed amorphous for 13 months, and PVP powders at loadings with a drug-rich band did not crystallize within 3 months. ··· Two links were not measured: where the crystals nucleate and how much polymer the drug-rich phase holds; and the molecular origin of the 480 nm band is left open (it "could be due to" indomethacin dimers). ··

01Why this study

Known 1The miscibility limit

Above the drug's amorphous solubility in the polymer, a drug-rich phase forms, and crystallization from it is expected.

Qian 2010, ref 7; Purohit 2015, ref 12 ·

Known 2DSC can miss phase separation

Domains below 30 nm, similar Tgs or remixing on heating hide a second Tg.

Meng 2015, ref 16; Baird 2012, ref 19

Known 3Fluorescence reports the surroundings

A probe registers polymer, drug or mixed surroundings; autofluorescent felodipine shifts to the blue in drug-rich droplets.

Purohit 2015, ref 12; Raina 2015, ref 37 ·

Known 4Miscible ASDs are thought more stable

, but this had hardly been tested.

Ivanisevic 2010, ref 31

Gap → this paperDoes a drug-rich phase, detected through the drug's own fluorescence, go together with faster crystallization?
More detail

What was already known. The introduction builds on these points:

  • Why ASDs and why polymers. Amorphous drugs dissolve faster and can raise bioavailability but tend to crystallize; dispersing them in an amorphous polymer is the usual remedy, and several products are ASDs. (Hancock 2000, ref 1; Janssens 2009, ref 6)
  • Solubility versus miscibility. Treating the polymer as a solvent, the crystal solubility of the drug in the polymer is the loading below which no crystallization can occur, but it is "virtually always too low" for practical use; the miscibility limit is the amorphous solubility, the highest loading that can stay one homogeneous amorphous phase. Between the two limits there is a driving force for crystallization but not for phase separation, and the drug "may remain" amorphous because nucleation and growth are slow in a glass. (Qian 2010, ref 7; Sun 2010, ref 8; Rubinstein 2003, ref 14) ··
  • Above the miscibility limit. Amorphous–amorphous phase separation forms a drug-rich phase with less polymer, from which crystallization "is anticipated", lowering dissolution and bioavailability. On this basis the authors write that "it is clear" the polymer should be miscible with the drug. (Tian 2014, ref 4; Purohit 2015, ref 12; Zhang 2011, ref 13) ·
  • Measuring miscibility is hard. DSC is the most common test (one Tg = miscible, two = two phases), but domains below 30 nm, similar Tgs or remixing on heating hide separation, and single Tgs for separated systems and two Tgs for miscible ones have both been reported. SEM, TEM and AFM resolve nanoscale domains without identifying them chemically; Raman mapping identifies composition but is slow; XRD, IR and ssNMR are also used. (Vasanthavada 2004, ref 18; Qian 2010, ref 20; Lodge 2006, ref 21; Li 2016, ref 24; Purohit 2015, ref 25; Rumondor 2009, ref 28) ·
  • Fluorescence as a probe. In earlier work an environment-sensitive probe registered a hydrophilic environment in pure polymer, a hydrophobic one in pure amorphous drug and an intermediate one, changing fairly continuously with loading, in miscible mixtures; the emission of autofluorescent felodipine became stronger and shifted to the blue when drug-rich nanodroplets formed in water. (Purohit 2015, ref 12; Raina 2015, ref 37) ·
  • The open link. Miscible ASDs "should" be more stable, but the correlation between miscibility and crystallization tendency had not been studied much; earlier PVP studies tied fast crystallization at high humidity to moisture-induced phase separation or to solvent-dependent miscibility. (Ivanisevic 2010, ref 31; Rumondor 2009, ref 40; Paudel 2012, ref 41) ·

What the paper set out to do. Use the autofluorescence of indomethacin, which should emit differently in a polymer-rich and in a drug-rich environment, together with imaging, IR and DSC, to find the miscibility limit of IDM in HPMC, HPMCAS and PVP; then store dispersions dry at 40 °C and test whether the phase-separated ones crystallize sooner. ·

02How they approached it

One autofluorescent drug in three polymers. Miscibility was read from thin films over 5–95% DL with four methods; stability from bulk powders and films stored over desiccant at 40 °C, so absorbed water could not change miscibility. ·····

IDM–HPMCcellulose ether, Methocel E5Hydroxyl groups that can hydrogen bond to IDM; films 5–95% and powders 10–90% DL; measured. ···
IDM–HPMCAScellulose ester, MF gradeAcetate and succinate groups; IR could not be used because HPMCAS absorbs in the carbonyl region; measured. ·
IDM–PVPK30Strong IR evidence of IDM–PVP interaction; powders stored only 3 months; measured. ··
Controlsreference filmsPure amorphous IDM film, polymer-only films and the bare quartz slide. ·

What was done

Experiment

Drop-cast films, 5–95% DL→emission spectra at 355 nm excitation→emission maximum versus DL.····

Experiment

Same films→fluorescence microscopy→uniform field or domains.···

Experiment

Films on KRS-5→IR carbonyl bands→IDM dimer band at 1709 cm-1.··

↓
Experiment

Rotary-evaporated, cryomilled powders, 10–90% DL→modulated DSC→Tg versus DL.··

↓
Comparison

Four miscibility estimates per polymer side by side (Table 2).

↓
Experiment

Powders at 40 °C over desiccant→XRD up to 3–14 months→crystallization onset (Table 1).·····

Experiment

Films at 40 °C over desiccant→fluorescence microscopy at 5–30 days→crystals.···

↓
Comparison

Miscibility limit against crystallization onset for HPMC and HPMCAS (Fig. 17).··

More detail

What was done

  1. Films. IDM and polymer at eleven ratios from 5:95 to 95:5 were dissolved at 10 mg/mL in methanol:dichloromethane (1:2, v/v); 30 µL was dropped on a quartz slide (fluorescence) or a KRS-5 window (IR), dried at about 20 °C under nitrogen at 20% RH or less, then overnight under vacuum. Drug-only and polymer-only films served as controls.
  2. Bulk powders. Nine ratios from 10:90 to 90:10 in the same solvent were rotary evaporated at 60 °C, dried overnight under vacuum and cryomilled with liquid nitrogen.
  3. Fluorescence spectra. Fresh films were excited at 355 nm and emission recorded from 410 to 670 nm; spectra were normalized to their maximum and the wavelength of maximum emission plotted against DL (n = 3). Selected films were also annealed for 5 h at Tg − 10 °C. ·
  4. Fluorescence imaging. Films were viewed with 330–380 nm excitation and emission above 420 nm, fresh and after 5, 12 and 26 days (30 days for PVP) at 40 °C; fresh films were checked for birefringence. ··
  5. IR. Transmission spectra (4 cm-1, 64 scans) of the films; the carbonyl region of IDM–HPMC is shown, IDM–PVP data are described but not shown (published before), IDM–HPMCAS could not be analysed. ···
  6. DSC. About 4 mg of fresh powder in a pinholed pan, heated from −10 to 190 °C at 5 °C/min with ±1 °C modulation every 60 s; mean of two runs.
  7. Stability. Powders and films were kept in desiccators over desiccant at 40 °C. Powders were scanned by XRD (5–40° 2θ) at intervals that started at 2–10 days for the high loadings and became monthly, for 14 months (HPMC), 13 months (HPMCAS) or 3 months (PVP); the first time Bragg peaks were seen is the crystallization onset of Table 1. ··
Terms used on this page
  • Miscibility limit: the highest drug loading at which drug and polymer still form one homogeneous amorphous phase; above it, amorphous–amorphous phase separation gives a drug-rich and a polymer-rich phase.
  • Emission maximum: the wavelength at which the fluorescence spectrum peaks; a shift to shorter wavelength (blue shift) here means a less polar, drug-rich surrounding.
  • Cyclic dimer band: the IR band at 1709 cm-1 from two IDM carboxylic acid groups hydrogen bonded to each other; it fades when a polymer breaks these dimers.
  • Bragg peaks: sharp XRD reflections of crystals on top of the broad amorphous halo.
  • Crystallization onset: here, the first storage time at which Bragg peaks were seen; it is bounded by the previous sampling time.
Fig. 1: Structures of indomethacin, HPMC, PVP K30 and HPMCAS
Fig. 1 · p.5 · Structures of indomethacin, HPMC, PVP K30 and HPMCASShows. IDM with its carboxylic acid and indole ring; substituents of HPMC and HPMCAS.

03Main findings

1

Indomethacin's emission reports its surroundings

Pure amorphous IDM peaks near 476 nm; IDM dispersed at 5% DL in HPMC, HPMCAS or PVP peaks near 508–520 nm.

Fig. 2: Emission of pure amorphous IDM against 5% DL films
Fig. 2 · p.9 · Emission of pure amorphous IDM against 5% DL filmsShows. pure IDM peaks near 476 nm; IDM in HPMC, HPMCAS and PVP near 508–520 nm; polymers and quartz emit little.Also supports K1
More detail

The pure amorphous IDM film emitted strongly with a maximum near 476 nm (intensity about 400). The 5% DL films emitted at longer wavelength: about 510–515 nm with HPMC (about 255), about 508 nm with HPMCAS (about 250) and a weak, broad band near 515–520 nm with PVP (about 87). The neat polymer films gave only weak emission near 435–440 nm (about 48–65) and the quartz slide about 15, so the IDM emission dominates the film spectra. All values are read from Fig. 2. The authors take this blue shift of the pure drug as showing that IDM emission is sensitive to its local environment and can probe miscibility ("indicates"). Their premise is that IDM dispersed in a polymer sits in a more polar environment than in pure amorphous IDM. Fresh and annealed films gave similar spectra, which the authors read as an unchanged miscibility after annealing (the printed sentence lacks a "not"). ··

2

A drug-rich band appears abruptly at a loading set by the polymer

The emission maximum drops from about 511 to 488 nm between 50 and 60% DL in HPMC, from about 513 to 480 nm between 20 and 30% in HPMCAS, and from about 511 to 476 nm between 70 and 80% in PVP. ·····

Fig. 3: IDM–HPMC films: normalized spectra and emission maximum versus loading
Fig. 3 · p.9 · IDM–HPMC films: normalized spectra and emission maximum versus loadingShows. the maximum stays near 511–515 nm up to 50% DL and drops to about 488 nm at 60% (wide error bar), then about 476–483 nm.Also supports K1
More detail
  • HPMC. From 5 to 50% DL the maximum stayed near 511–515 nm (text: "~515 nm"), with a slight fall at higher loading. At 60% a new peak near 480 nm appeared, like that of pure IDM, while a shoulder near 515 nm remained; the maximum fell to about 488 nm, with a wide spread between the three 60% films (about 478–499 nm). From 70 to 95% it stayed at about 476–483 nm. · The authors read the new peak as IDM in an IDM-rich, less polar phase next to IDM still in the polymer, so that IDM–HPMC is miscible up to 50% and the limit "appears to be" between 50 and 60% DL. ··
  • HPMCAS. The maximum was about 510–513 nm at 5–20% DL and about 480 nm at 30%, staying near 477–483 nm up to 95%. The authors place the limit between 20 and 30% ("appears"). ·
  • PVP. The maximum drifted from about 518 nm at 5% to about 507–511 nm at 60–70% DL, then dropped to about 476 nm at 80% and stayed there at 90 and 95%. The ~480 nm band "is not observed until" 80% DL; Table 2 gives a spectroscopic limit of 70–80% for PVP. ··
  • Reading. At higher loadings the spectra "appear to have contributions" from IDM in two environments, the polymer matrix and an IDM-rich phase; a gradual drift is what a miscible system would give, an abrupt change what phase separation would give. · The molecular origin of the band is not settled: it "could be due to" more IDM dimers or other IDM–IDM contacts forming with the drug-rich phase. Because the spectra report the local environment, the authors consider the approach "potentially sensitive" to domains too small to image.
Fig. 4: IDM–HPMCAS films: normalized spectra and emission maximum versus loading
Fig. 4 · p.10 · IDM–HPMCAS films: normalized spectra and emission maximum versus loadingShows. the drop from about 513 to 480 nm between 20 and 30% DL.Also supports K2
Fig. 5: IDM–PVP films: normalized spectra and emission maximum versus loading
Fig. 5 · p.10 · IDM–PVP films: normalized spectra and emission maximum versus loadingShows. a gradual drift from about 518 to 507–511 nm up to 70% DL, then a drop to about 476 nm at 80%.
3

Imaging, IR and DSC put the limit within about one step of the spectra

HPMCAS films look heterogeneous from 30% DL, HPMC films from 60% and PVP films from 70%; in HPMC the IDM dimer band returns at 60%; DSC Tg stops falling at about 50% (HPMC) and 30% (HPMCAS) but falls linearly for PVP. Table 2 ranks the limits HPMCAS < HPMC < PVP. ·····

Fig. 7: Fresh IDM–HPMCAS films imaged by fluorescence, 5–95% DL
Fig. 7 · p.12 · Fresh IDM–HPMCAS films imaged by fluorescence, 5–95% DLShows. uniform at 5–10%, dark branched marks at 20%, a dark network at 30%, large dark domains at 60–70%, interconnected patterns at 80–95%.Also supports K3
Table 2: Miscibility of the three systems by four methods
Table 2 · p.23 · Miscibility of the three systems by four methodsShows. DSC, IR, imaging and spectroscopic limits side by side.Also supports K2
More detail
  • Imaging (fresh films). IDM–HPMC films were uniform from 5 to 50% DL and showed green and dark domains from 60%, read as IDM-rich (green, fluorescent) and HPMC-rich (dark) phases; "the miscibility of IDM in HPMC appears to be exceeded" around 60%. IDM–HPMCAS films were uniform at 5 and 10%, "reasonably homogeneous" at 20%, partly heterogeneous at 30% and clearly separated above, giving a limit of about 20%. IDM–PVP films were clearly heterogeneous from 70% DL. No film showed birefringence. ······· The images themselves show some features below these limits: dark branched marks in the 20% HPMCAS film, sparse dark spots in the 30% and 40% HPMC films and a network of thin dark lines in the 60% PVP film. Imaging cannot resolve domains smaller than about 300–500 nm.
  • IR. Amorphous IDM shows carbonyl bands at 1733, 1709 and 1683 cm-1, assigned to free acid, cyclic-dimer acid and amide carbonyls (ref 32). In the 5% HPMC film the 1709 cm-1 band vanished and the 1733 cm-1 band grew: HPMC breaks the IDM dimers ("indicate"). The spectra changed little up to 50% DL; from 60% the 1709 cm-1 band returned and grew with loading, "consistent with" an IDM-rich phase, giving an IR limit around 50%. In Fig. 9 the 40% and 50% traces already sit between the 5–30% traces and the 60% trace near 1705–1709 cm-1. ···· For PVP, films with 5–80% DL lacked the 1733 and 1709 cm-1 bands and showed new bands at 1724 and 1640 cm-1, a strong IDM–PVP interaction; above 80% the spectra resembled pure IDM. The authors estimate the IR limit as "at least 80%" and attribute it to the strong interaction ("can be attributed"). ··
  • DSC. Each powder showed a single Tg. For HPMC, Tg fell steeply from about 395 K at 10% DL to about 328 K at 40%, then stayed at about 320–324 K from 50 to 80%, close to pure IDM (about 320 K); the authors suggest a second Tg may be missing because separated domains remix on heating, and call the DSC limit of about 50% "not definitive". For HPMCAS, Tg fell from about 369 K at 10% to about 324 K at 30% and stayed at about 317–322 K above. For PVP, Tg fell almost linearly from about 412 K at 10% to about 328 K at 90%. All values are read from Fig. 10. ····
  • Table 2. The authors summarize: DSC 40–50% (HPMC), 20–30% (HPMCAS), all ratios (PVP); IR about 50%, not done, about 80%; imaging about 50%, about 20%, about 70%; spectroscopy 50–60%, 20–30%, 70–80%. They see "good alignment" and conclude that fluorescence spectroscopy and imaging "appear to be useful techniques" for autofluorescent drugs. · The table mixes last-miscible and first-separated loadings between columns.
Fig. 6: Fresh IDM–HPMC films imaged by fluorescence, 5–95% DL
Fig. 6 · p.11 · Fresh IDM–HPMC films imaged by fluorescence, 5–95% DLShows. uniform green up to 50% with sparse dark spots at 30–40%; speckle at 60%; dark domains from 70%.
Fig. 8: Fresh IDM–PVP films imaged by fluorescence, 5–95% DL
Fig. 8 · p.13 · Fresh IDM–PVP films imaged by fluorescence, 5–95% DLShows. uniform up to 50%, thin dark lines at 60%, dark specks at 70%, mottled texture at 80–95%.
Fig. 9: IR carbonyl region of IDM–HPMC films, 5–100% DL
Fig. 9 · p.14 · IR carbonyl region of IDM–HPMC films, 5–100% DLShows. the 1709 cm-1 dimer band absent at low loading and returning from 60%; 40% and 50% traces intermediate.
Fig. 10: Tg of the bulk powders versus loading
Fig. 10 · p.15 · Tg of the bulk powders versus loadingShows. HPMC and HPMCAS Tg level off near pure IDM (about 320 K) from about 50% and 30% DL; PVP falls almost linearly.
4

HPMC and HPMCAS powders above the limit crystallize within days

At 40 °C over desiccant, HPMC powders crystallized after 15 days at 60% DL and 6 days at 70–90%; HPMCAS powders after 10, 6, 8, 4, 2 and 4 days at 40–90%. HPMC up to 50% and HPMCAS up to 30% DL stayed X-ray amorphous for 14 and 13 months.

Table 1: Crystallization onset of HPMC and HPMCAS powders at 40 °C
Table 1 · p.18 · Crystallization onset of HPMC and HPMCAS powders at 40 °CShows. "a" (amorphous) up to 50% HPMC and 30% HPMCAS; 2–15 days above.Also supports K4
More detail

The onset values come from repeated XRD scans: an onset is the first scan with Bragg peaks, so it is bounded by the previous scan (for example 10 and 15 days for 60% HPMC). ··· The authors conclude that IDM–HPMC powders are more stable than IDM–HPMCAS powders ("demonstrate"). Fig. 17 lays the onsets under bands marking each polymer's amorphous, miscibility and crystallization ranges: for HPMC the transition band spans 50–60% DL, for HPMCAS 20–30%. The authors state that dispersions found miscible have "excellent stability" and those with a detected drug-rich phase crystallize at short times, "a good correlation" (abstract). For HPMC, 50% DL is both the last loading without a drug-rich band and the last amorphous powder, and 60% the first with both a drug-rich band and crystals. For HPMCAS the first loading with a drug-rich phase by spectra, images and DSC (30%) stayed amorphous for 13 months, and crystallization began at 40%. ····· The 40% HPMCAS crystals give peaks near 11.6, 16.7, 21.8 and 26.6° 2θ, while the 80–90% crystals show a fuller pattern from about 8.5° upward; the authors do not identify the crystal forms. One unexplained sharp reflection near 31.7° 2θ appears in the 10% and 30% HPMC traces from about 1–2 months. ·

Fig. 12: XRD of IDM–HPMCAS powders during storage at 40 °C
Fig. 12 · p.17 · XRD of IDM–HPMCAS powders during storage at 40 °CShows. halos only at 10–30% DL for 13 months; Bragg peaks from 2–10 days at 40–90%.
5

PVP dispersions resisted crystallization

No PVP powder showed Bragg peaks within 3 months at any loading up to 90%; PVP films crystallized only at 80% DL and above. ·

More detail

All IDM–PVP powders stayed X-ray amorphous over 3 months, so the authors suggest PVP dispersions "may have the best physical stability", in particular at high loading; PVP films showed no crystals from 5 to 70% DL within 30 days. ··· Films at 80–95% DL crystallized: crystals at 30 days in the 80% film, at 12 days in the 90% film and faint clusters already at 5 days in the 95% film (text: "after longer time periods (30 days)"). · The authors suggest fluorescence microscopy "may be more sensitive" than XRD to small amounts of crystal, especially at the film surface. The fluorescence spectra and images point to a drug-rich phase at 70–80% DL, whereas earlier ssNMR found IDM–PVP miscible at about 5 nm for 10–50 wt% polymer; the authors offer two hedged explanations: a different preparation method ("One possible explanation") or a spectrum that tracks IDM dimers rather than a separate phase ("might be"), which however "does not explain" the heterogeneous images. ···· The PVP powders at loadings with a drug-rich band therefore did not crystallize within the 3 months tested. ··

Fig. 13: XRD of IDM–PVP powders during storage at 40 °C
Fig. 13 · p.18 · XRD of IDM–PVP powders during storage at 40 °CShows. halos only at every loading up to 3 months.Also supports K5
Fig. 16: IDM–PVP films during storage at 40 °C
Fig. 16 · p.21 · IDM–PVP films during storage at 40 °CShows. no crystals at 5–70% DL by 30 days; crystals at 80% (30 days), 90% (12 days) and 95% (from 5 days).

04Further observations

  • Films crystallized from 50% DL in both cellulose polymers within 26 days, and HPMCAS films earlier and more heavily. ··
  • Film and powder thresholds differ by one step: the 50% HPMC film crystallized, the powder did not; the 40% HPMCAS film did not, the powder did. ····
  • Every powder showed one Tg, even where other methods saw two phases. ·
  • The HPMCAS crystals at 40% DL and at 80–90% DL give different XRD patterns.
More detail
  • Film crystallization. In HPMC films no crystals appeared from 5 to 40% DL within 26 days; at 50–90% clusters of bright crystals were seen at 12 and 26 days (none at 5 days), and the 95% film already showed crystalline features at 5 days. In HPMCAS films no crystal clusters appeared up to 40% DL; at 60% and 80% many crystals were present at 5 days, at 70%, 90% and 95% from 12 days, and at 50% scattered crystals at 26 days. The authors call the HPMCAS films earlier and more crystallized, "consistent with" XRD. ·· Many later images were taken at lower magnification (100 µm scale bar against 20 µm).
  • Films versus powders. The 50% HPMC film crystallized by 12 days although the 50% powder stayed amorphous for 14 months and the spectra placed 50% as miscible; the 40% HPMCAS film showed no crystals at 26 days although the 40% powder crystallized by 10 days. Films and powders were also made by different routes (drop casting at about 20 °C against rotary evaporation at 60 °C and cryomilling). ·······
  • One Tg. No powder showed two Tgs; for HPMC at 50–90% DL the authors suggest remixing of separated domains on heating ("might result from"). ·
  • Crystal patterns. The 40% HPMCAS powder gave a few peaks (about 11.6, 16.7, 21.8 and 26.6° 2θ), the 80% and 90% powders many (about 8.5, 11.9, 14.5, 18.5, 22.0, 22.7, 26.5 and 28.3°), read from Fig. 12.
  • No birefringence and no annealing effect. Fresh films showed no birefringence, and annealing for 5 h at Tg − 10 °C left the spectra unchanged. ·
Fig. 14: IDM–HPMC films during storage at 40 °C
Fig. 14 · p.19 · IDM–HPMC films during storage at 40 °CShows. no crystals at 5–40% DL by 26 days; crystal clusters from 12 days at 50–90%.
Fig. 15: IDM–HPMCAS films during storage at 40 °C
Fig. 15 · p.20 · IDM–HPMCAS films during storage at 40 °CShows. no crystal clusters at 5–40% DL; many crystals from 5 days at 60% and 80%.

05How the authors connect the findings

1In a miscible film, IDM is surrounded by polymer, a more polar environment, and emits near 510–515 nm. ·
↓
2Above the miscibility limit the film separates into a drug-rich and a polymer-rich phase; IDM in the drug-rich phase emits like pure amorphous IDM, near 480 nm. ·
↓
Below the limitOne homogeneous phase↓the polymer inhibits crystallization kinetically↓amorphous for 13–14 months.··
Above the limitA drug-rich phase with less polymer↓weaker crystallization inhibition↓crystals within days.··
What sets the switch point. The polymer: strong IDM–PVP hydrogen bonding goes with the highest limit ("can be attributed to"), and HPMCAS gives the lowest. ·
Conclusion drawn. Miscibility "is indeed an important" ASD property (abstract: "a good correlation", "suggesting").
More detail
  1. IDM disperses in the polymer of a miscible film; its emission then reports a polar, polymer-rich environment, with a maximum near 510–515 nm that changes only gradually with loading. In HPMC the IR shows that the polymer breaks IDM's carboxylic acid dimers. ···
  2. When the loading exceeds the miscibility limit, amorphous–amorphous phase separation creates an IDM-rich phase. Part of the IDM then emits like pure amorphous IDM (about 480 nm), domains appear in the images, and in HPMC the IDM dimer band returns. ···

Loading below the limit. The dispersion is one phase; the polymer inhibits crystallization kinetically although the glass is metastable → the HPMC (up to 50%) and HPMCAS (up to 30%) powders stay X-ray amorphous for 14 and 13 months. ··

Loading above the limit. The drug-rich phase holds less polymer, so crystallization inhibition is "less effective" and crystallization is "anticipated to occur more readily" → the HPMC (from 60%) and HPMCAS (from 40%) powders crystallize within 2–15 days. ·· The authors write that drug-rich domains "led to" this enhanced susceptibility.

What sets the switch point. Drug–polymer interaction: PVP forms strong hydrogen bonds with IDM and has the largest miscibility range; HPMC, whose hydroxyl groups also break IDM dimers, comes next; HPMCAS has the lowest limit. The authors link the high PVP limit to the strong interaction ("can be attributed to"); they do not explain the HPMCAS limit. ····

Conclusion drawn. ASDs found miscible had "excellent stability to crystallization", and the "observed correlation" between miscibility and robustness means homogeneity should be evaluated during formulation screening. ·

06What it means for formulation

Stated by the authors
  • Check the homogeneity of candidate dispersions during formulation screening.
  • Formulate homogeneous dispersions to obtain ASDs that resist crystallization in storage.
  • For an autofluorescent drug, the drug's own emission can screen miscibility without added probes. ·
AI reading, not stated in the paper
  • For indomethacin, PVP K30 tolerates far higher loadings than HPMCAS before a drug-rich phase forms.
  • The detected miscibility limit worked as a conservative ceiling for dry storage: below it nothing crystallized in over a year.
More detail
Stated by the authors
  • The correlation "highlights the need to evaluate the homogeneity" of formulations during screening.
  • The results are "particularly important" for HPMCAS and, "to some extent", HPMC, where the IDM-rich phase appears at commercially relevant loadings (20–30% and 50–60% DL).
  • The study "reinforces the importance" of formulating homogeneous dispersions to obtain ASDs resistant to crystallization in storage.
  • Fluorescence spectroscopy and imaging "appear to be useful techniques" for the phase behaviour of autofluorescent drugs (abstract: "show promise").
  • The polymer should be miscible with the drug ("it is clear").
AI reading, not stated in the paper
  • Polymer choice shifted the safe IDM loading threefold (about 20–30% in HPMCAS, 50% in HPMC, 70% or more in PVP); a polymer that hydrogen bonds to the drug's acid group gave the widest window here.
  • Because the first phase-separated HPMCAS loading (30%) still stayed amorphous for 13 months, a loading just above a detected limit is not automatically unstable in dry storage; the limit is a safe side, not a cliff edge.
  • The film screen is fast (drop casting and one spectrum per loading) but was made by a different route from the powders; a screen should be run on material made like the product.
  • Humidity was excluded on purpose; water lowers miscibility in many systems, so dry results are an upper bound for humid storage.

07Open questions

Raised by the authors
  • What causes the 480 nm band: a separate drug-rich phase or IDM dimers forming with loading? ·
  • Why do PVP films look phase separated at high loading while ssNMR found IDM–PVP miscible? ·
Not covered by this paper (AI reading)
  • Where the crystals nucleate, and how much polymer the drug-rich phase holds, were not measured. ·
  • PVP powders were followed for only 3 months. ·
More detail
Raised by the authors
  • The molecular origin of the spectral change "has not been elucidated"; IDM dimers or other IDM–IDM interactions are candidates, and "Clearly more investigations are warranted". ·
  • The IDM–PVP discrepancy at high loading may come from preparation method or from dimer-sensitive spectra; neither was tested. ··
  • Whether fluorescence microscopy detects surface crystals in films that XRD of powders misses.
Not covered by this paper (AI reading)
  • No direct evidence that crystals start in the drug-rich domains; the link between phase separation and crystallization is a correlation across loadings, not a test at fixed loading. ·
  • The composition of the drug-rich and polymer-rich phases was not measured.
  • PVP powders were followed for 3 months against 13–14 months for HPMC and HPMCAS, so the PVP compositions with a drug-rich band were not tested on the same time scale. ·
  • Miscibility was measured on films and stability mainly on powders made differently; the thresholds of the two sample types differ by one step. ·
  • Humid storage, other drugs and other polymer grades were not studied.

08How solid

AI assessment, not a statement made in the paper
Moderate

Four methods agree on the miscibility ranking and the 13–14-month XRD record is clear, but the correlation rests on two polymers, pairs film miscibility with powder stability, and has exceptions at 30% HPMCAS and for PVP. ····

More detail
What holds
  • The emission maximum drops abruptly, by about 23–35 nm, at one loading per polymer, with small replicate scatter away from the boundary. ··
  • Imaging, IR (HPMC) and DSC point to the same ranking, HPMCAS < HPMC < PVP, within about one 10% step. ·····
  • For HPMC the last homogeneous and last amorphous loading coincide (50%), and the 13–14-month XRD record separates stable from fast-crystallizing powders sharply. ·
What weakens it
  • For HPMCAS the first phase-separated loading (30%) stayed amorphous; for PVP no phase-separated powder crystallized within 3 months. ·
  • Miscibility comes mainly from drop-cast films, stability from rotary-evaporated powders; film and powder thresholds differ. ·
  • The origin of the 480 nm band is unresolved; the authors themselves offer a dimer explanation. ·
  • Table 2 mixes conventions, image judgements at the thresholds are visual, and most numbers are plot readings. ··
  • Onsets are bounded by sampling intervals, and PVP powders were followed only 3 months. ·

09Figure guide

Every figure and table of the paper, what it is there for, and what to check on the original. Key figures are shown with the story.

FigureWhat it does in the paperSupportsCheck (AI note)
Fig. 1 (p.5)key figureDefines the systems: indomethacin and the three polymers with their substituents.···0.2Text calls the ring a "tryptophan ring"; the drawing shows an indole.
Fig. 2 (p.9)key figureShows that pure amorphous IDM emits further to the blue than IDM dispersed at 5% DL, the basis of the probe.F1, K1Caption omits the PVP traces shown in the legend.
Fig. 3 (p.9)key figureLocates the HPMC limit: emission maximum drops between 50 and 60% DL.F2, K1The 60% point has a wide error bar.
Fig. 4 (p.10)key figureLocates the HPMCAS limit between 20 and 30% DL.F2, K2
Fig. 5 (p.10)key figureLocates the PVP limit between 70 and 80% DL.F2
Fig. 6 (p.11)key figureImaging of fresh HPMC films: domains from 60% DL.F3Sparse dark spots at 30–40% DL.
Fig. 7 (p.12)key figureImaging of fresh HPMCAS films: heterogeneity from 30% DL.F3, K3Dark branched marks already at 20% DL.
Fig. 8 (p.13)key figureImaging of fresh PVP films: heterogeneity from 70% DL.F3Thin dark line network at 60% DL.
Fig. 9 (p.14)key figureIR of HPMC films: dimer band absent at low loading, back from 60% DL.F340% and 50% traces already intermediate.
Fig. 10 (p.15)key figureTg versus loading: plateaus for HPMC and HPMCAS, linear for PVP.F3No HPMC point visible at 90% DL.
Fig. 11 (p.16)XRD of HPMC powders: amorphous to 50% DL for 14 months, peaks from 6–15 days above.F4One unexplained sharp reflection near 31.7° in 10% and 30% traces.
Fig. 12 (p.17)key figureXRD of HPMCAS powders: amorphous to 30% DL for 13 months, peaks within 2–10 days above.F4Onsets bounded by sampling times.
Fig. 13 (p.18)key figureXRD of PVP powders: amorphous at all loadings for 3 months.F5, K5Only 3 months of storage.
Table 1 (p.18)key figureThe stability result: crystallization onset per loading for HPMC and HPMCAS.F4, K4Abstract thresholds worded "less than" and "higher than" 50% and 30%.
Fig. 14 (p.19)key figureHPMC films in storage: crystals from 50% DL within 12 days.0.450% film crystallized, 50% powder did not.
Fig. 15 (p.20)key figureHPMCAS films in storage: crystals from 50% DL, earlier than HPMC.0.440% film stayed crystal-free while the 40% powder crystallized.
Fig. 16 (p.21)key figurePVP films in storage: crystals only at 80–95% DL.F5Crystals at 12 days (90%) and 5 days (95%), not only at 30 days.
Table 2 (p.23)key figureSummary of the four miscibility estimates per polymer.F3, K2Mixed last-miscible and first-separated conventions.
Fig. 17 (p.25)key figureThe correlation: miscibility bands drawn over the crystallization onsets.·header, K430% HPMCAS lies in the miscibility band but stayed amorphous.
Table of Contents Graphic (p.30)Repeats Fig. 17.