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7 protocols using 874 oven sample processor

1

Hydration and Swelling of Dry Human Amniotic Membrane

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Dry HAM was cut into circles of 5 mm diameter. All samples were hydrated in normal saline until they reached an equilibrium state. The remaining water was blotted with absorption paper. The swelling ratio of the HAM was determined by measuring the diameters of the circles after 5 days.
The water absorption capacity of the HAM was determined by comparing the water content before and after the hydration. The water content in the HAM was measured by the well-established Karl-Fischer titration technique54 (link). The dry HAM was evaluated by a coulometric method (Mettler Toledo, Metrohm 852 Titrando Karl Fisher Titrator with Metrohm 874 Oven Sample Processor, Germany), and the fully hydrated HAM was evaluated by a volumetric method (Mettler Toledo, Metrohm 852 Titrando Karl Fisher Titrator Metrohm 860 KF Thermoprep, Germany).
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2

Hydration Kinetics of MXF-HA Hydrogels

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Hydrogel absorbs water and maintain its shape in its hydrated state. Round cuts (diameter of 5 mm) of dry MXF-HA were hydrated in normal saline. The diameter and weight of hydrated MXF-HA were measured sequentially at 1, 5, 15 and 30 min. When reaching the equilibrium state, the diameter and weight of MXF-HA no longer changed and the time reaching this state was recorded.
The water content of MXF-HA was measured by Karl-Fischer titration40 (link). The coulometric method (Mettler Toledo, Metrohm 852 Titrando Karl Fisher Titrator with Metrohm 874 Oven Sample Processor, Germany) was used for the water content measurement of dry MXF-HA and the volumetric method (Mettler Toledo, Metrohm 852 Titrando Karl Fisher Titrator Metrohm 860 KF Thermoprep, Germany) was used to determine the water content of fully hydrated MXF-HA.
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3

Water Content Analysis of Crystalline Forms

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Example 21

The water content of each sample was measured on a Metrohm 874 Oven Sample Processor up to 150° C. with 851 Titrano Coulometer using Hydranal Coulomat AG oven reagent and nitrogen purge. Weighed solid samples were introduced into a sealed sample vial. Approximately 10 mg of sample was used per titration and duplicate determinations were made. An average of these results is presented unless otherwise stated. Data collection and analysis were performed using Tiamo software.

Results of the KF analysis for the crystalline forms are described in the following table:

Solid State FormKF
Form 10.7 wt % water
(0.21 mol equiv)
Form 219.1 wt % water (150° C.)
(7 mol equiv)
Pattern 91.7 wt %/2.3 wt % water (100° C.)
( ~0.6 mol equiv)
Form 30.5 wt % water (160° C.)
(0.15 mol equiv)
amorphous3.0 wt % water
(0.92 mol equiv)

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4

Water Content Analysis of Crystalline Forms

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Example 21

The water content of each sample was measured on a Metrohm 874 Oven Sample Processor up to 150° C. with 851 Titrano Coulometer using Hydranal Coulomat AG oven reagent and nitrogen purge. Weighed solid samples were introduced into a sealed sample vial. Approximately 10 mg of sample was used per titration and duplicate determinations were made. An average of these results is presented unless otherwise stated. Data collection and analysis were performed using Tiamo software.

Results of the KF analysis for the crystalline forms are described in the following table:

Solid State FormKF
Form 10.7 wt % water
(0.21 mol equiv)
Form 219.1 wt % water (150° C.)
(7 mol equiv)
Pattern 91.7 wt %/2.3 wt % water (100° C.)
(~0.6 mol equiv)
Form 30.5 wt % water (160° C.)
(0.15 mol equiv)
amorphous3.0 wt % water
(0.92 mol equiv)

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5

Water Content Determination of Hydrate Forms

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Example 9

The water content of each sample was measured on a Metrohm 874 Oven Sample Processor at 150° C. with 851 Titrano Coulometer using Hydranal Coulomat AG oven reagent and nitrogen purge. Weighed solid samples were introduced into a sealed sample vial. Approximately 10 mg of sample was used per titration and duplicate determinations were made. Data collection and analysis using Tiamo v2.2.

For Form D, a 7.6 wt. % loss was detected, corresponding to 2.0 mole equivalent water thus confirming the dihydrate form. For Form F, an 11.6 wt. % loss was detected, corresponding to 3.2 mole equivalents water, thus confirming the trihydrate form.

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6

Comprehensive Analytical Techniques for Sample Characterization

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An LC series 1200 instrument (Agilent, Waldbronn, Germany) consisting of a model G1379B degasser, G1312B binary pump SL, G1367C autosampler SL, and G1316B column thermostat SL was employed for LC-hrMS/MS analysis. The LC was coupled online to an LTQ-Orbitrap XL high-resolution tandem mass spectrometer (Thermo Electron, Bremen, Germany) fitted with a heated electrospray ionization (HESI) source. For AAA by IDMS, a microwave hydrolysis system (CEM, Saclay, France) and a rotational vacuum concentrator connected to a freeze-dryer system (Christ, Germany) were used to hydrolyse and concentrate samples, respectively. An Exion LC coupled online to a QTRAP 6500+ mass spectrometer (AB Sciex, Les Ulis, France) fitted to an electrospray ionisation (ESI) source was used for the detection of amino acids. A 930 compact IC flex and 863 compact autosampler (Metrohm AG, Switzerland) and a Metrohm Titrando 851 coupled to a Metrohm 874 Oven sample processor were used for IC and KF titration, respectively. Elemental analysis was performed using a Vario Micro Cube Elemental Analyser (Elementar, Lyon, France), whereas NMR measurements were performed on an ECS-400 MHz NMR instrument equipped with a direct type (Royal) automatic tuning probe (JEOL, Tokyo, Japan).
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7

Residual Moisture Determination by Karl Fischer

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Vials for residual moisture determination were tested by Karl Fischer using an 801 Coulometer equipped with an 874 Oven Sample Processor and 851 Titrando (Metrohm, Herisau, Switzerland). A 1% water standard was run to verify the instrument was operating properly (Apura, Burlington, MA). The DP (approximately 100 mg) was transferred to testing vials using a balance (Mettler Toledo, Columbus, OH). Vials were analyzed by heating to 125°C and the residual moisture determined by Tiamo software (version 2.5).
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