Immunofluorescent Staining of Mouse Pancreas for Islet Cell Mass Analysis v1

Author(s):  
Islet and Pancreas Analysis Core

This SOP defines the assay method used by the Vanderbilt Diabetes Center Islet and Pancreas Analysis (IPA) Core for quantitative determination of the islet cell composition and islet cell mass of mouse pancreas by immunofluorescent staining.

2009 ◽  
Vol 61 (4) ◽  
pp. 811-817 ◽  
Author(s):  
Jianbo Xiao ◽  
Xinlin Wei ◽  
Yuanfeng Wang

A reversed-phase liquid chromatographic method was developed for the quantitative determination of three major hypotensive compounds, namely geniposidic acid, chlorogenic acid, and geniposide in the bark of Eucommia ulmoides. Soxhlet extraction of GPA, GPS, and CA from E. ulmoides was optimized according to the Taguchi experimental design. Maximum global yields were obtained using the following conditions: extraction temperature, 80?C; extraction time, 1 h; number of extractions, three; solvent volume, 16 ml/g of sample; and 50% ethanol concentration in water. Optimal conditions of separation and detection were achieved on a Diamonsil ODS C18 column (150 mm ? 4.6 mm, 5 ?m) with a linear gradient of methanol and 0.04% aqueous phosphoric acid (v/v) at a flow rate of 1.0 ml/min and detection wavelength of 240 nm. All calibration curves showed good linearity (r2 > 0.999) within test ranges. The relative deviation of this method was less than 3% for intra- and inter-day assays, and the recovery percentage of the method was 95-104%, with a relative standard deviation (R.S.D.) of less than 5%. The current assay method was used for quantitative determination of geniposidic acid, chlorogenic acid, and geniposide in five samples of E. ulmoides with different age. The results indicate that the developed method could be readily utilized as a quality control method in working with E. ulmoides.


2021 ◽  
Author(s):  
IIDP-HIPP not provided

This Standard Operating Procedure (SOP) is based on the Human Islet Phenotyping Program of the IIDP Immunofluorescence Staining Procedure. This SOP provides the HIPP procedure for immunofluorescent staining, imaging, and analysis of islet preparations. This SOP defines the assay method used by the Human Islet Phenotyping Program (HIPP) for quantitative and qualitative determination of the Purified Human Pancreatic Islet product, post-shipment, manufactured for use in the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)-sponsored research in the Integrated Islet Distribution Program (IIDP).


Diabetes ◽  
1983 ◽  
Vol 32 (5) ◽  
pp. 460-465 ◽  
Author(s):  
A. H. Huen ◽  
M. Haneda ◽  
Z. Freedman ◽  
A. Lernmark ◽  
A. H. Rubenstein

Diabetes ◽  
1983 ◽  
Vol 32 (5) ◽  
pp. 460-465 ◽  
Author(s):  
A. H. -J. Huen ◽  
M. Haneda ◽  
Z. Freedman ◽  
A. Lernmark ◽  
A. H. Rubenstein

2020 ◽  
Vol 26 (4) ◽  
pp. 80-90
Author(s):  
I.V. Drapak ◽  

Aim. Validation of the method of quantitative determination of Cardiazole substance by high-performing liquid chromatography technique. Materials and Methods. The object of the study was the [3-allyl-4-(41-methoxyphenyl)-3H-thiazol-2-ylidene]-(32-trifluoromethylphenyl)-amine hydrobromide (Cardiazole) which possesses cardioprotective, anti-inflammatory, analgesic, hypolipidemic, antihypoxic, and antioxidant properties. The compound is patented and involved in the development plan of "Farmak" pharmaceutical company for further preclinical studies workflow. High-performance liquid chromatography was used for the quantitative determination of the Cardiazole substance. Validation of the proposed methodology was performed in accordance with the requirements of the State and European Pharmacopoeias requirements. The obtained data were analyzed using Analyst 1.5.2., as well as Statistica 10.0 and Microsoft Excel software. Results and Discussions. The high-performance liquid chromatography analytical method for the quantitative determination of Cardiazole substance was validated based on the main parameters according to the pharmacopeia requirements. The specificity of the technique was confirmed by comparing the chromatograms of the comparison solution, the test solution, and the blank solution. The linearity parameters are set over the entire range of the analyzed Cardiazole concentrations. The parameters of the correctness of the 9 prepared test solutions within the range of the analytical method application have met the following criteria: requirements for statistical and practical insignificance. The study of the parameters of intra-laboratory precision of 3 tests of the same sample was carried out by two analysts on different days during one week using different measuring ware. Thus, compliance with the criteria was confirmed. The obtained results of the experimental determining of the validation characteristics confirmed the correctness of the technique while reproduced in other laboratories. The results of the stability study showed: for optimal chromatography conditions it is necessary to use a freshly prepared solution of Cardiazole (within 24 hours). Conclusions. The evaluation of the validation parameters of the method of quantitative determination of Cardiazole by high-performance liquid chromatography was carried out. It was shown that the presented method of analysis of Cardiazole meets the requirements for specificity, linearity, accuracy, precision, and stability. The validated analytical method for the quantitative determination of Cardiazole by high-performance liquid chromatography can be used for standardization of the Cardiazole substance, as well as for studying the pharmacokinetics and pharmacodynamics, bioequivalence, and pharmaceutical development of dosage forms. Keywords: validation parameters, Cardiazole, high-performance liquid chromatography


Author(s):  
L. S. Lohoyda

The aim of this study was the validation of methods of quantitative determination of nifedipine in  tablets by liquid chromatography. The chromatographic analysis was performed on nifedipine liquid chromatograph Agilent 1290 Infinity II LC System. A validation of methods of quantitative determination of nifedipine by high performance liquid chromatography tablets was performed. It was established that the method proves the requirements of the State Pharmacopoeia of Ukraine for the main validation parameters: specificity, accuracy, linearity, robasnist. The results obtained in this study clearly indicate that the developed HPLC method is fast, economical, simple, accurate and suitable for determination of nifedipine in medicines.


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