Applicational possibilities of linear and non-linear (polynomial) regression and analysis of variance. III. Stability determination of pharmaceutical preparations: stability of diclofenac-sodium in Diclofen injections.

This paper presents the application of the regression analysis program and the program for comparing linear regressions (modified method for one-way, analysis of variance), writtens in BASIC program language, for instance, determination of content of Diclofenac-Sodium (active ingredient in DIKLOFEN injections, ampules á 75 mg/3 ml). Stability testing of Diclofenac-Sodium was done by isothermic method of accelerated aging at 4 different temperatures (30 degrees, 40 degrees, 50 degrees and 60 degrees C) as a function of time (4 different duration of treatment: (0-155, 0-145, 0-74 and 0-44 days). The decrease in stability (decrease in the mean value of the content of Diclofenac-Sodium (in %), at different temperatures as a function of time, is possible to describe by, linear dependance. According to the value for regression equation values, the times are assessed in which the content of Diclofenac-Sodium (in %) will decrease by 10%, of the initial value. The times are follows at 30 degrees C 761.02 days, at 40 degrees C 397.26 days, at 50 degrees C 201.96 days and at 60 degrees C 58.85 days. The estimated times (in days) in which the mean value for Diclofenac-Sodium content (in %) will by 10% of the initial values, as a junction of time, are most suitably described by 3rd order parabola. Based on the parameter values which describe the 3rd order parabola, the time was estimated in which Diclofenac-Sodium content mean value (in %) will fall by 10% of the initial one at average ambient temperatures of 20 degrees C and 25 degrees C. The times are: 1409.47 days (20 degrees C) and 1042.39 days (25 degrees C). Based on the value for Fischer's coefficien (F), the comparison of trenf of Diclofenac-Sodium content (in %) shows that, under the influence of different temperatures as a function of time, among them, depending on temperature value, there is: statistically very significant difference (P << .05) at 50 degrees C and lower toward 60 degrees C, i.e. statistically probably significant difference (P > 0.01) at 40 degrees C and lower towards 50 degrees C and there is no statistically significance difference (P >> 0.05) at 30 degrees C towards 40 degrees C.

Biological and chemical quality control of spermicidal raw material nonoxynol-9 and nonoxynol-9 containing vaginal contraceptive (ABF contraceptive film): determination of spermicidal activity and assay of nonoxynol-9.

The paper presents a routine biological and chemical quality control of nonoxynol-9 raw material and pharmaceutical formulation containing nonoxynol-9 (ABF contraceptive film). The biological control performed based on the determination and comparison of the spermicidal activities different dilutions of the standard raw materials solution and assay raw material or nonoxynol-9 pharmaceutical formulation solution on the human or breeding bull sperm using a modified Sander-Cramer method. The test preparations of the nonoxynol-9 raw materials or pharmaceutical formulation should show equal or higher spermicidal activity than the standard. The spermicidal activity should not be lower than dilution 1:90. The chemical control is performed based on the determination and comparison of nonoxynol-9 content in the standard raw material solution and assay raw material or nonoxynol-9 pharmaceutical formulation solution using UV-spectrophotometric method determining and comparing the absorbances at the maximum absorbance wavelength (275 nm). The content of nonoxynol-9 in the raw material assay solution or pharmaceutical formulation (ABF contraceptive film) should be within 90-110% of the declared value.

Possibilities of the application of regression analysis and analysis of variance. II. Assessment and comparison of acute toxicity: presentation and practical application of the interactive computer program "LD50-MORTALITY".

The paper presents the interactive computer program "LD50-MORTALITY", written in the program language BASIC and used for the assessment of the acute toxicity (calculation of LD50 and other LD values) based on the algorithm of the Finney's probit analysis method. The correctness of the program execution has been shown on the test-example of assessment the acute toxicity (LD50 an LD90) of insecticide rotenone on Macrosiphonelia sanborni (the example is taken from the literature on probit analysis presentation). The results obtained by the program execution have been identical or similar to those given in the example. The results of the assessment and comparison of LD50 have also been presented.

Applicational possibilities of linear and non-linear (polynomial) regressions and analysis of variance: kinetics of chloramphenicol.

The dependance of the content and microbiological activity of Chloramphenicol (active substance) at dissolution on time and on pH, as well as that of the content and microbiological activity of Chloramphenicol (250 mg capsules) at release, on time, in in-vitro conditions, was determined using linear and non-linear (polynomial and generalized dilution) regressions. Based on the square error value, the dependance of the content and microbiological activity of Chloramphenicol (active substance) at dissolution on time and different pH values, as well as the dependance of the content and microbiological activity of Chloramphenicol (capsules) at release on time were best described by polynomial function. The comparison of the content and microbiological activity of Chloramphenicol (active substance) at dissolution at different pH values, as well as of Chloramphenicol (capsules) at release showed the significant correlation between these parameters (r = 0.999, P << 0.001). The comparision of the content, on one hand, and microbiological activity of Chloramphenicol (active substance), on the other, at dissolution at different pH values, as a function of time, was done using a modified method of one-way analysis of variance for linear regression comparisons. Based on the value of Fischer's coefficient (F), there is a statistically very significant difference between the contents and between the microbiological activities of chloramphenicol (active substance) at dissolution and different pH as a function of time (P << 0.005).

Computerized calculation of biological potency: presentation of the interactive computer program "POTENCY" (assays: "2 + 2" doses, "3 + 3" doses, "twin cross-over assay" and "probit analysis" ("2 + 2" doses).

Paper presents the interactive computer program "POTENCY" written in BASIC program language, based on the algorithm for classifying biological assays using the assays: "2 + 2" doses, "3 + 3" doses, twin cross-over assay and probit analysis ("2 + 2" doses) described in Yugoslav Pharmacopeia (Ph. Jug. IV). The program execution is presented by assay examples of biological potency calculation described in European Pharmacopeia (Eur. Ph. 1997). The assay-examples have simultaneously been the measure of program execution correctness.