A Novel RP-HPLC Method for Estimating Fulvestrant, Benzoyl Alcohol, and Benzyl Benzoate in Injection Formulation

For the quantitative determination of Fulvestrant, Benzyl alcohol, and Benzyl benzoate in Fulvestrant injection, an original RP-HPLC approach was developed. The gradient method was developed using HPLC and a Phenomenex Luna C8, 150 × 4.6 mm, i.d 3.0 μm particle size column with a gradient programme of mobile phases A and B. With a flow rate of 1.5 mL/minute, injection volume of 10 μL, and column temperature of 35°C, UV wavelength detection at 254 nm for Benzyl alcohol and Benzoyl Benzoate and 280 nm for Fulvestrant, mobile phase-A consists of DI water and mobile phase-B consists of Acetonitrile. The current study describes a single HPLC method for developing a Fulvestrant (Active), Benzyl alcohol (Cosolvent), and Benzyl Benzoate (Cosolvent) assay for Fulvestrant injection. The assay method was determined to be suitable for quantifying three components in the pharmaceutical product and was verified according to ICH guidelines.

Benzyl Alcohol-Benzyl Benzoate Clearing Reveals the Dose-Dependent Effect of Cyclophosphamide on Follicle Damage in Mice

Objective:Cyclophosphamide(CTX),which is commonly used in clinical chemotherapy,has a damaging effect on ovarian follicles.This study aimed to establish a new method to count the number of follicles in mouse ovaries using benzyl alcohol-benzyl benzoate(BABB)-based tissue-clearing technology and evaluate the follicle-damaging effects of different doses of CTX.Methods:C57BL/6 mice were divided into four groups and administered intraperitoneal injections of 0,40,80,or 120 mg/kg CTX.The serum levels of estradiol(E 2)and follicle-stimulating hormone(FSH)were detected using an ELISA kit.Mouse ovaries were subjected to BABB clearing and labeled with DAPI,β-actin,and DDX4 to observe the ovarian structure and follicles.A three-dimensional software,Imaris,was used to reconstruct the ovarian structure and automatically count the number of follicles.The effects of different CTX doses on the total follicle number and estrous cycle were determined.Results:As the CTX dose increased,E 2 levels in CTX mice declined from 212.3 to 57.7 pg/mL;the FSH levels increased from 3.2 to 29 ng/mL.Mouse ovaries became transparent after BABB treatment.After fixation,microscopy,and Imaris processing,immunofluorescence signals ofβ-actin and DAPI from all levels in intact ovaries could be obtained and follicle number in half ovaries could be automatically counted using anti-DDX4 antibody labeling.In the NC,CTX40,CTX80,and CTX120 groups,the proportion of mice in the diestrus phase was 26.67%,51.67%,73.33%,and 95.00%,respectively,and the total follicle number was 2,603,1,761,1,043,and 262,respectively.E 2 levels were positively correlated with follicle number and FSH levels were negatively correlated with follicle number,indicating that the damaging effect of CTX on follicle number may be dose dependent.Conclusion:BABB can be used to clear intact ovaries from adult mice,and follicle number in half ovaries can be automatically counted.The damaging effect of CTX on follicles and the endocrine system is dose dependent.

Intraspecific Chemical Variability of Essential Oil of Hoslundia opposita

Chemical investigation on leaves oil of Hoslundia opposita, aromatic plant used in traditional medicine, harvested from 20 localities of C?te d’Ivoire was realized by GC and GC/MS. In total, 101 compounds were identified accounting to 92% - 99%, aromatic compounds such benzyl benzoate and sesquiterpenes namely germacrene D, β-caryophyllene and α-humulene hydroxyl were found as the major constituents. As regard, the chemical profiles of our samples, it was found a great chemical variability suggesting a high polymorphism degree. In order to study this chemical variability, the results were submitted to the principal components and cluster analysis which allowed distinguishing two main groups of essential oils with respect to sampling site. The cluster I was dominated by benzyl benzoate (15%) and thymol (7.4%) comprising 6 localities, cluster II was characterized by phytol (8.5%) and α-humulene (7%) and constituted by 14 localities.

Kinetic Evidence for the Solvent Intervention in the Solvolysis of Tertiary Benzylic Benzoates

Rates of solvolyses of 2-（4-methylphenyl）-2-propyl benzoate （2b）, 2-（4-methylthiopheny1）-2-propyl benzoate （3b） and 2,2-dimethy1-1-（4-methoxypheny1）-1-phenyl-1-propy1 benzoate （4b） were measured, respectively, in ten or more different solvent systems. Additional YBnOPXB and Y, BnOPNB values in binary trifluoroethanol-water systems were determined, Grunwald-Winstein type correlation analysis showed that YBaIOBB values were applicable to benzylic benzoates. The rate ratio, koPNB/KOBZ, was found to be solvent-dependent in the range of 29 for 4 in ethanol to 4 for 2 in trifluoroethanol-water. Non-limiting mechanism, including both nucleophilic and electrophilic solvent assistance, in the solvolysis of 2b was discussed,