International Journal of

Pharmaceutical Quality Assurance

ISSN: 0975 9506
Peer Review Journal

doi prefix: 10.25258/ijpqa

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1. The Study on Age of Menarche in Girls in the Buldana District of Maharashtra
Adhao V. S., Thenge R. R., Ajmire P. V.
The present study was designed to determine the age at which menarche occurs among school girls in Buldana district of Maharashtra state, India. A survey was conducted among 488 girls by writing the questionnaire from schools in the selected area. Respondents completed a questionnaire that recorded age at first menstruation by the recall, residential status, type of education, and diet/food habit. The mean age at menarche was 13.44 ± 0.75 years. Most girls (72.95%) of the respondents were found of normal age menarche (12–14 years); 27.05% of late-type menarche (> 14 years), and 0% were of early menarche (< 12 years). Our study suggests an influence of school education, residential area, and diet/food habit on menarcheal age.

2. The Correlation Between Anti-GAD65 And Coxsackievirus B-IgG (CVB-IgG) In Type 1 Diabetes-Coxsackievirus B (T1D-CVB) Patients
Ahmed S Kareem, Ghanim A. Almola, Oday J Alsalihi
Coxsackieviruses are one of the main causes of type 1 diabetes, due to the sequence similarity between the protein 2 C (P2-C) in the structure of the virus and the glutamic acid decarboxylase (GAD65) autoantigen in human beta-cells. The study aims to detect the anti-GAD65 and specific anti-CVB IgG in T1D patients to study the correlation between the levels of the GAD65 autoantigen and CVB- IgG autoantibody in T1D-CVB patients. A hospital-based cross-sectional study was carried out from November 2018 to July 2019 at Babylon Diabetic Center in Marjan Teaching city, Babylon teaching hospital for maternity and children and college of medicine at the University of Babylon. A total of 150 samples were obtained from diabetic patients and 50 samples from non-diabetic individuals as control. Diabetic mellitus (DM) patients diagnosed by clinical features, RBS test (above 200 mg/dL) and HbA1c test (above 6.5%). Anti-Gad and CVB-IgG detected by indirect ELISA assays. The study showed the age group A2 (1-5 years), the females group (B1), and rural group (C1) more susceptible to T1D-CVB infection. The study exhibited a positive correlation between anti-gad and anti-CVB-IgG (r =0.644**) in T1D-CVB patients.

3. Review of Elemental Impurities in Pharmaceuticals Arena
Alpa C. Sheth, Paresh U. Patel
Elemental impurities in drug products may arise from several sources; they may be residual catalysts that were added intentionally in the synthesis or may be present as impurities (e.g., through interactions with processing equipment or container/closure systems or by being present in components of the drug product). Because elemental impurities do not provide any therapeutic benefit to the patient, their levels in the drug product should be controlled within acceptable limits. The main objective of the ICH (Q3D) guideline applies to new finished drug products and new drug products containing existing drug substances. The drug products containing purified proteins and polypeptides. This guideline does not apply to herbal products, radiopharmaceuticals, vaccines, cell metabolites, DNA products, allergenic extracts, cells, whole blood, cellular blood components, or blood derivatives, including plasma and plasma derivatives. The evaluation of the toxicity data for potential elemental impurities; the establishment of a permitted daily exposure (PDE) for each element of toxicological concern; application of a risk-based approach to control elemental impurities in drug products. Different analytical techniques for elemental impurities detection: flame atomic absorption spectrometry (FAAS), graphite furnace atomic absorption spectrometry (GFAAS), atomic fluorescence spectrometry, X-ray fluorescence spectrometry (XRF), instrumental neutron activation analysis (INAA), inductively coupled plasma-atomic emission spectroscopy (optical emission spectroscopy, ICP-OES), inductively coupled plasma mass spectrometry (ICP-MS), and microwave plasma atomic emission spectrometry (MP-AES).

4. Development and Validation of Stability-Indicating Reverse Phase-High Performance Liquid Chromatography Method for Simultaneous Determination of Atenolol and Nifedipine in Bulk and Tablet Dosage Form
Ansari Yaasir Ahmed, Qazi Shoeb, Umme Rumana, Patel Afroza, Pathan Vahid Tajkhan, Makrani Shaharukh
The new stability-indicating high performance liquid chromatography (HPLC) method has been developed and validated with different parameters for atenolol and nifedipine in the combined dosage form. The chromatographic conditions were optimized using a mobile phase of MeOH:OPA (70:30) with a flow rate of 0.7 mL/min. Column (C18) of 4.6 × 250 mm dimension was used as a stationary phase; the particle size capacity of the column was 5 μm. The detection was carried out at 233 nm. The method was validated according to ICH guidelines for linearity, precision, repeatability, the limit of detection (LoD), and limit of quantitation (LoQ). The response was found to be linear in the concentration range of 20–100 mcg/mL for atenolol and 1–5 mcg/mL for nifedipine. The developed method shows the minimum quantity of drugs to be identified (LoD) and minimum drug to be quantified (LoQ). The LoD and LoQ were found to be 0.1415 and 0.4289, respectively, for atenolol, and 0.1834 and 0.5558, respectively, for nifedipine. The method was linear, simple, precise, and accurate and, therefore, suitable for routine analysis of drugs in tablet form. The forced degradation studies were also done through the exposure of analyte solution to four different stress conditions.

5. Prevalence of Some Gram-Negative Bacteria and Adenovirus in Breast Cancer Patients in Kirkuk City
Asmaa A Ali, Salah S Z Alabden, Najat A Zaman
Background: Breast cancer is one of the most prevalent types of malignant tumors in the world. Bacteria have been linked with cancer through two mechanisms the first stimulating chronic inflammation and the second production of carcinogenic bacterial metabolites. The most common types of adenovirus in cancer patients are serotypes 7 and 11 of type B, and serotypes 1, 2, and 5 are of type C. Methods: In this study (98 blood and 50 urine), samples were collected from women with breast cancer at the Center for Oncology and Hematology in Kirkuk city in addition to (30 blood and 20 urine) samples collected from healthy persons as control. Pathogenic gram-negative bacteria were isolated, identified in addition to checking their susceptibility to antibiotics. Antibodies (IgM, IgG) of adenovirus type-7 were involved in this study by the ELISA technique. Results: In this study, 22 gram-negative bacterial species were isolated from patients with breast cancer, including (31.81% K. pneumoniae, 45.45% E. coli, 18.18% P. mirabilis and 4.54% A. baumannii). Gram-negative bacteria showed high resistance to antibiotics (approximately absolute resistance) Amoxicillin/clavulanic acid, Ampicillin, Ceftazidime, Amoxicillin, Cephalothin, Vancomycin. Adenovirus Type-7 antibodies were detected by using the ELISA technique (3.84% IgM and 91.02% IgG). Viral and bacterial Co-infection were also included in this study caused by the involvement of E. coli + ADV-7, P. mirabilis + ADV-7, and K. pneumoniae + ADV-7.

6. Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of Ajwain (Trachyspermum ammi) Seed Extract
Basheer Mohammed Abdullah, Mansour Abdulnabi H. Mehdi, A. R. Khan, J. M. Pathan
The objectives of this research were to determine the chemical composition of the extract of Trachyspermum ammi L. seeds by using gas chromatography-mass spectrometry analysis (GC-MS). The GC-MS is a matchless method for the study and measuring quantity of organic volatile and semi-volatile compounds. Gas chromatography is employed to separates mixtures into individual components employing a temperature-controlled capillary column. Mass spectrometry is utilized to recognize a variety of components from their mass spectra. In the present study, volatile/ semi-volatile compounds present in Ajwain seed extract were analyzed. Ajwain seed extract is extracted by soxhlet extraction method and then analyzed by GC-MS. Total of 9 compounds were found and quantified in this study. The major bioactive compounds in Ajwain seed extract are 3,5-dimethylanisole (83.19%), 6-octadecenoic acid, methyl ester, (Z)-, 7-octadecenoic acid, methyl ester (7.42%), and 2-cyclohexyl-2,5-cyclohexadiene-1,4-dione, 4-oxime (3.01%).

7. Novel Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) Method Development and Validation of Atorvastatin and Fenofibrate in Tablets
Chandrasekhar Kudupudi, Manikandan A
A novel, simple selective, precise, and accurate reverse-phase high-performance liquid chromatography (RP-HPLC) gradient method was developed for the simultaneous estimation of atorvastatin and fenofibrate in the combined formulation. The drugs atorvastatin calcium and fenofibrate were separated in the presence of their impurities atorvastatin related compound H, fenofibrate related compound A, and fenofibrate related compound B. The drugs and related compounds were separated on Kromasil C18 (250 x 4.6, 5μ) with reverse-phase gradient elution. Water adjusted pH 4.0 with phosphoric acid used as a buffer in pump A and acetonitrile used as a solvent in pump B as a mobile phase with gradient elution. The flow rate was 2.0 mL/min. 254 nm was the detection wavelength. The retention times were about 4.6 minutes for fenofibrate related compound A, 5.2 minutes for atorvastatin calcium, 5.7 minutes for fenofibrate related compound B, 8.7 minutes for atorvastatin related compound H, and 17.6 minutes for fenofibrate. The linearity ranges for atorvastatin calcium and fenofibrate were 5.00 to 15.00 and 80 to 240 mcg/mL, respectively, with correlation coefficient 0.999 for both. The proposed method validated statistically with respect to system suitability, specificity, linearity, precision, accuracy, range, robustness, and ruggedness. The method was accurate, linear, precise, specific, selective, and rapid suitable for the quantitative estimation of atorvastatin and fenofibrate in tablets.

8. Antileishmanial effect of Berberine and extracts of Berberis Vulgaris on the Growth of Leishmanial Species
Husain F. Hassan, Zina K. Najim
The present investigation was revealed that the Berberine (0.01,0.1,1mg/cm3), aqueous extracts and alcoholic extract (1,10,100 mg/cm3) of Berberis vulgaris roots were inhibitory on the growth of Leishmania Tropica, L.donovani and L.infantum promastigotes. High-pressure liquid chromatography (HPLC)analysis of the plant extracts proved to contain Berberine as an active compound. Berberine and B. vulgaris root extracts were caused a significant decrease in total nucleic acid contents of three leishmanial species, thus considering to be an excellent agents for antileishmanial chemotherapy.

9. Evaluation of Application of Effective vaccine management in Wassit Governorate, Iraq
Jalal A R Zayer, Ishraq A Chiad
Background: Immunization helps save a life, protect serious illness, and improve quality of life. It is recognized as one of the most cost-effective public health interventions available around the globe. However, the success of this program is heavily dependent on strong immunization supply chain practices. Proper immunization supply chain management ensures availing potent and live vaccines to the community. Objective: To evaluate the application of Effective vaccine management in Health facilities of Wassit governorate, Iraq. Subjects and Methods: It was a cross-sectional study involving a multistage sampling. A total of 45 health facilities sites were selected as follow: 1sub-national store (SN), six district store (LD), and 38 service point (SP) by using EVM Questionnaire, interview, reviewing the records, and observations for the agreed review period. Results: The overall scores of this assessment for all levels ( SN, LD and SP) of the supply chain 66.6% demonstrates a need for more improvement in most areas of the vaccine and supply management system as only two criteria (Storage Capacity E3 and Vaccine Management Practices E8) exceeds the WHO recommended minimum score of 80%. Performance levels of one criteria (Building, Equipment E4) were about 72% storage temperature (E2), maintenance(E5) and stock management (E6) were between 61% and 68%, while the vaccine distribution (E7), and information management (E9)were notably very weak with performance in each category less than 60%. Conclusions: The national average percentage at all levels was below the WHO recommended minimum score of 80%. Recommendation: The future vaccine storage capacity must be recalculated and stored to enter any new vaccine, receive large quantities of the influenza vaccine, and replace vaccine refrigerators at the sector level with cold rooms to accommodate current and future increases.

10. The Effect of Wounding, Indole Butyric Acid Levels, and Cutting Date on Rooting Ability of Ficus benghalensis
Mahmood S H Ahmed, Zeyad M Abdulrazzaq, Mohammed M Sharqi, Mustafa R Al-Shaheen
The experiment was conducted in the plastic house of the Department of Horticulture and Gardening Engineering at College of Agriculture / Anbar University during the period from March 2019 to September 2019 to study the effect of wounding, levels of indole butyric acid and date of cutting on the rotting ability of ficus. The experiment was carried out using complete randomized design (CRD) with a factorial experiment consisting of three factors; the first factor involved applying two levels of wounding (without wounding, two wounds at the base of the cutting); the second factor included the treatment of the cutting base with four levels of indole butyric acid (0, 1500 2000, and 3000 mg-liter -1), while the third factor was the date of cuttings (March and April). The results showed superiority of cutting during March on the percentage of rooting (99%), the number of days required for rooting (22 days), number of roots (89 per plant), a diameter of the roots (0.75 mm), and length of the roots (17.33 cm).

11. Study of some virulence factors for Clostridium perfringens isolated from clinical samples and hospital environment and showing their sensitivity to antibiotics
Muhsin H Edham, Asma S Karomi, Zainab I Tahseen
The study included taking (100) samples from different clinical sources including (wounds and burns) and from the hospital environment, in Kirkuk General Hospital and Azadi Teaching Hospital in the city of Kirkuk for the period from November (2017) to August (2018). The results of isolation and diagnosis showed the growth of (30) isolates that are positive for Clostridium perfringens, distributed between (15) isolates (37.5%) from burns, (11) isolates (27.5%) from wounds and (4) isolates (20%) from the hospital environment. These isolates were diagnosed based on microscopical, cultural and biochemical tests, in addition to being diagnosed with the Api 20 A system. The sensitivity of isolates was tested toward a number of types of antibiotics, and all bacterial isolates showed a high sensitivity (100%) against Imipenem. As for the sensitivity to Vancomycin, Amikacin, Tetracycline was (96.66%, 90% and 66.66%) respectively. While all isolates showed a high resistance to Metronidazole and Colistin (100%). Some virulence factors of C.perfringens isolates have been studied , and showed that all isolates %100)) have the ability to produce Hemolysin, Lecithinase, capsule and Spore, while (70%) of the isolates produced DNAase.

12. GC-MS Profiling of Pimpinella anisum Oils and Its Antimicrobial and Antioxidant Activities
Muthanna J. Mohammed, Hiyam A. Ebraheem
Essential and fixed oils of anise plant (Pimpinella anisum (P. anisum)) growing in Iraq have been investigated regarding their chemical components, antioxidant and antimicrobial properties. Essential oils were extracted using the Clevenger-apparatus, while fixed oil was extracted using a Soxhlet apparatus. Gas chromatography-mass spectrometry (GC-MS) was used for the analysis of the oil components. Six strains of bacteria, namely S. epidermidis, S. aureus, E. coli, B. cereus, P. vulgaris and S. typhimurium were tested against the antimicrobial activity of each oil. Anise oil demonstrated a broad antibacterial property range, against Gram-positive and Gram-negative bacteria, through the inhibition zone. The antibiotic sensitivity test was performed by disk diffusion process against the test organisms. The agar dilution method was used at five different concentrations (12.5, 25, 50, 100, and 200 mg/mL) throughout the test. The Minimum Inhibitory Concentration (MIC) was determined for each volatile and fixed oil. The DPPH radical scavenging assay was used to test the antioxidant activities of essential and fixed oils. Anise oil showed excellent antioxidant activity, in comparison with the reference compounds. Anise oil has the potential to be used as a therapeutic, antimicrobial, and antioxidant agent.

13. Stability-indicating Simultaneous Method Development and Validation of Guaifenesin and Dextromethorphan HBr by Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC)
Nathi Rathnakar, D.G. Sankar
The developed method was validated according to ICH guidelines with respect to specificity, linearity, limits of detection, quantification, accuracy, precision, and robustness. The stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method is precise; it has been developed for the simultaneous estimation of assay of guaifenesin (GN) and dextromethorphan HBr (DN) in drug substance and drug product. The chromatographic separation was done in an isocratic mode using the Syncronus C8 (250 × 4.6 mm, 5 μ particle size) column with mobile phase containing a 10 mM ammonium acetate in water (modulated pH 4.30 with orthophosphoric acid) and acetonitrile in the ratio of 60:40 (% v/v) used for efficient chromatographic separation. The flow rate of the mobile phase was 1.0 mL/min with ambient column temperature and detection of wavelength at 279 nm; injection volume 10 μL was fixed for achieving good elution of eluents. The retention time for GN was found to 3.46 minutes and DN was found to 7.58 minutes. GN and DN were linear in the concentration range from 357 to 1428 and 19 to 75 μg/mL, respectively. Regression analysis showed that the r-value (correlation coefficient) greater than 0.999 for GN rvalue was found to be 0.999, DN r-value was found to be 0.999. Limit of detection (LoD) and limit of quantification (LoQ) of GN was found to be 0.151 and 0.904 μg/mL, DN was found to be 0.241 and 0.726 μg/mL. The developed method was validated and found to be accurate, specific, and robust. Both the drugs were subjected to the stress conditions like acidic, basic, oxidative, photolytic, and thermal conditions. The degradation results were found to be satisfactory. In peroxide stress condition, GN was found stable over DN, and DN was found to degrade significantly. The degradation products were well resolved from GN, DN, and their impurities. The peak purity test results confirmed that the GN and DN peak were homogenous and pure in all stress conditions, thus proving the stability-indicating nature of the method. This method could be applied for the simultaneous estimation of GN and DN in drug substance and drug product.

14. Hydrogen bond studies in interact 6-Mercaptopurine with it receptor HGPRT
Alaa A. Habeeb, Falah S A Suhail, Sami W Radhi
Objective: The aim of this study was to study some properties of the pharmaceutical compound (6-Mercaptopurine) a number of theoretical methods were carried out to calculate their molecular energy, the length of the bonds & angles, in addition to their chemical forms and the binding sites with the enzyme as important anti-cancer inhibitorsMethods: The intramolecular hydrogen bond, molecular structure, and vibrational frequencies of 6- Mercaptopurine have been investigated by means of density functional (DFT& AM1) methods with 6-311++G basis. Results: The nature of these interactions, known as resonance assisted hydrogen bonds, has been discussed. As a geometrical indicator of a local aromaticity, the geometry-based HOMA index has been applied. Additionally the linear correlation coefficients between substituent constants and selected parameters in R position have been calculated. Conclusion: The results show that the hydrogen bond strength is mainly governed by the resonance variations inside the chelate ring induced by the substituent groups. The topological properties of the electron density distributions for 6- MP H……N intra molecular bridges have been analyzed .Finally, the natural population analysis methods has been used to evaluate the hydrogen bonding interactions.

15. Novel First-Order Derivative UV Spectrophotometric Method for the Determination of Acarbose in Solid Dosage Forms
Santosh Karajgi, Gaviraj E. N., Sunayana Mali, C. C. Patil, Shripad Potdar, Kotnal R. B.
An easy, perfect, specific, and accurate process has been studied for the estimation of acarbose pure drug form as well as tablet dosage forms. Using methanol as a solvent, acarbose has absorbance maxima at 206 nm, and this drug shows a linear response according to Beer’s law in the concentration range of 24–40 μg/mL. The outcomes of the study were validated statistically, and recovery studies were satisfactory as per ICH guidelines. Thus, the projected method can be proficiently useful for the estimation of acarbose in regular analysis effort.

16. Biomarkers of allergic asthma and their Association with serum parameters
Suhayla H. Shareef, Kawa Amin
Background and objective: Asthma is not a single disease but, rather, a heterogeneous inflammatory disorder with various pathogenic mechanisms. A key element of severe asthma (SA) has been stated to be eosinophilic inflammation (EI). However, there is no serum biomarker that can reliably predict EI in SA. In this regard, the present study was carried out to evaluate the biomarkers of allergic asthma (AA) and their association with serum parameters. Methods: The present cohort study consisted of 66 AA and 61 healthy controls (HC) from Jan. 2017 to Nov. 2018. Data on variables like all types of leukocyte cells, total and specific serum IgE, ECP, Tryptase, IL-18, CD-20, and FEV1 were obtained. For comparison between the unpaired variables, the Mann-Whitney U test, χ2 test, and Kruskal Wallis H test was used. Results: Symptoms (cough, wheezes, dyspnea) and family history were found as significant indicators for AA (p <0.05). The AA and HC were significantly different in terms of blood and serum parameters [i.e., WBC, lymphocyte, monocyte, neutrophil, eosinophil, basophil, total IgE, ECP, Tryptase, IL-18, and CD-20] (p <0.05); therefore, these factors might be reliable indicators of AA. Also, most of the biomarkers were significantly correlated with the serum parameters, which reveal the reliability of these variables as indicators of AA. Conclusion: All types of leukocyte cells were found as reliable biomarkers to diagnose AA patients from HC. Also, FEV1%, Total WBC, and serum total IgE, ECP, Tryptase, IL-18, and CD-20 were found as reliable biomarkers for the severity of AA.

17. Antibiotic Resistance Patterns and Horizontal Gene Transferring of Some Bacterial Species Isolated from Tonsillitis in Babylon Province
Yusra A Radeef, Ahmed A Mhawesh
Tonsillitis or throat infection is one of the most frequent health problems worldwide. In the current study, 100 tonsil swabs were collected from patients who were suffering from tonsillitis and tonsillectomy and 10 control whose did not take any drugs for both genera (male and female) with age ranging from 3-82 years admitted in Al-Hilla Teaching Hospital and Babylon Hospital for birth and children and people, who are suffering from tonsillitis and living in Hilla during a period from December 2018 to May 2019. The total numbers of gram-positive (G+ve) bacteria were 75 (71.5%), while the number of gram-negative (G-ve) was 30 (28.5%). Antibiotics susceptibility was tested, and results were showed significant differences between antibiotics at (p <0.05). Antibiotic resistance genes (ARGs), in addition associated genetic elements, were distinguished by polymerase chain reaction (PCR) test with definite primers. Conjugation was involved gene transfer, where the PCR test was exhibited only the successful transferring of blaTEM1A gene beginning K. pneumoniae to standard strain (E. coli Hb101) with the range of conjugation frequency was (2.5˟10-3-1.59˟10-4), while the rest bacteria reveal negative results. In relevant, we observed by transformation, the successful transferring of erm B gene from bacteria S. aureus to S. pyogenes, tet M gene from bacteria S. pyogenes to S. aureus and blaTEM1A gene from bacteria P. aeruginosea to K. pneumoniae except blaIMP-1 gene was not transferred from K. pneumoniae to P. aeruginosea, where the results reveal the transformation frequency of bacteria S. aureus was 1.036˟10-1, S. pyogenes was 1.7, P. aeruginosea was 1.43 and K. pneumoniae was 2.25˟10-1, respectively.

18. Reverse-Phase High-Performance Liquid Chromatography Method Development and Validation for Estimation of Glimepiride in Bulk and Tablet Dosage Form
Aseem Kumar, Anil Kumar Sharma, Rohit Dutt
The present work demonstrates a simple, rapid, precise, specific, and sensitive reverse-phase high-performance liquid chromatography (RP-HPLC) method for analyzing glimepiride in pure and tablet forms. The present method was developed using a C18 column 150 × 4.6 mm, with 5 μm, and packing L1 maintained at a temperature of 30°C. The mobile phase was prepared by dissolving 0.5 gram of monobasic sodium phosphate in 500 mL of distilled water, pH of the solution adjusted to 2.1 to 2.7 with 10% phosphoric acid, and added 500 mL of acetonitrile. The mobile phase was pumped in the high-performance liquid chromatography (HPLC) system at a flow rate of 1 mL/min, and separation was carried out at 228 nm, using an ultraviolet (UV) detector. The chromatographic separation was achieved with peak retention time (RT) at about 9.30 minutes, and the method was found to be linear over a concentration range of 40 to 140 μg/mL. The specificity of the method represented no interference of the excipients during the analysis, and stability testing after 24 hours also showed that the method is suitable and specific. The accuracy was between 99.93 to 99.96%, with limit of detection (LOD) and limit of quantitation (LOQ) being 0.354 μg/mL, 1.18 μg/mL, respectively. Satisfactory results were found for precision and robustness parameters during the development and validation stage for the analytical method. The proposed method was also adopted for the analysis of glimepiride tablets to improve the overall quality control. Using this method, symmetric peak shape was obtained with reasonable retention time. The retention time of glimepiride for six repetitions is 9.3 ± 0.1 minutes; the run time is 21 minutes. The proposed RP-HPLC method is a modification of the United States Pharmacopeia (USP) method, and it was found to be valid for glimepiride within concentration ranges 40 to 140 μg/mL, using C18 analytical columns, and isocratic elution with UV detection, and at 1 mL/min of flow rate.

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