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.

Development on the Technique of Total Recovery of Benzoic Acid Residue

Benzoic acid residue is solid waste produced from the production of benzoic acid by oxidizing toluene.Because it contained important chemical raw materials such as benzoic acid,benzyl benzoate and fluorenone,it is necessary to recover them from the residue.In this work the technique featured with high efficiency evaporation and vacuum distillation was developed to obtain total recovery and utilization of the benzoic acid residue.By controlling the operation temperature at 260℃ and the pressure of 16kPa in the rising and falling film evaporators,heavy components separated efficiently from the residue can be polymerized and the light components consisting of 63% of the residue entered into a benzoic acid vacuum distillation column.Keeping the temperature of polymerization at(280±10)℃,coumarone resin was produced after adjusting the softening point according to the market requirements.Vacuum distillation was operated under the following conditions:top temperature at 186℃,top pressure of 16kPa,bottom temperature at 240-250℃,reflux ratio being 3︰1.Benzoic acid of 98% purity was distilled out from the column as a side stream and the bottom product was crude benzyl benzoate.By the developed technique,the benzoic acid residue was splitted into three products,benzoic acid,crude benzyl benzoate and coumarone resin without any surplus waste.

Separation of Benzoic Acid Residue by Flash-Vacuum Distillation-Melt Crystallization

A large amount of residue including benzoic acid, benzyl benzoate and fluorenone can form in the production of benzoic acid by oxidizing toluene. To recycle the resources and reduce secondary pollution, the treatment of benzoic acid residue was carried out to obtain the three purified materials by flash-vacuum distillation combination method, and the influence of the operating parameters, such as the top pressure, reflux ratio and top and bottom temperatures was investigated to obtain the best operating conditions. The experimental results show that the benzoic acid purity can reach 97% through distillation under the following conditions： the top pressure is 1 600 Pa, the bottom temperature is 190--200℃, the top temperature is 130--135℃, and reflux ratio is 5：1. The best operating conditions for benzyl benzoate distillation column are： the top pressure is 400 Pa, bottom temperature is 250-- 260 ℃, the top temperature is 150--160 , and reflux ratio is 5：1. The benzyl benzoate purity can reach 95%, and the fiuorenone purity can reach 92 %. When refined by melt crystallization, the benzoic acid purity can be improved up to 99.6%, and the purities of benzyl benzoate and fluorenone are both above 95%.

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,