Densitometry Method Research Articles

The Effect of Roasting Temperature on Ferulic Acid Levels of Robusta Coffee Bean with Thin Layer Chromatography (TLC)-Densitometry

The purpose of this study was to quantitatively analyze ferulic acid levels of Robusta coffee bean extractsfrom two samples from different regions (Argopuro and Kaliwining, Jember District), with differentroasting temperatures (180 °C and 200 °C for 10 minutes) using Thin Layer Chromatography (TLC)-Densitometry method. The analytical method used was the TLC-Densitometry, which utilized methanolsolvent p.a; stationary phase: Merck silica Gel 60 F254 plate; chloroform mobile phase: methanol: formicacid (v/v/v) = (85:15:1) in a saturated chromatographic chamber; the test concentration of 50 ppm and theplates were analyzed with the Densitometry winCATS Camag scanner using a UV-Vis detector at 254 nmwavelength and blue fluorescence at 366 nm wavelength. The results showed that the standard correlationcoefficient (R) of ferulic acid was 0.9947. Quantitative analysis of ferulic acid levels obtained on theaverage results of Argupuro coffee bean extract at roasting temperatures 180 °C and 200 °C amounted to1.935598±0.020671 mg/g and 1.419963±0.051633 mg/g, while Robusta Kaliwining coffee bean extract at180°C and 200°C roasting temperature were 3.18087±0.06738 mg/g and 2.420237±0.1989 mg/g. Statisticalanalysis with One-Way Anova obtained p value <0.005. Least Significant Different Test (LSD) obtainedp value <0.005 in each group. Based on the statistical analysis it was concluded that there was an effectof roasting temperature on ferulic acid levels in Robusta coffee bean extract, with the highest ferulic acidcontent obtained in Robusta Kaliwining coffee bean extract in Jember Regency at roasting temperature of180 °C was 3.180875±0.020671 mg/g.

Quantification of the bioactive marker resveratrol in Morus alba Linn. fruits by high–performance thin–layer chromatography

The present study focuses on the development and validation of a high-performance thin-layer chromatography (HPTLC)‒densitometry method for the identification and quantification of resveratrol in the 70% methanolic extract of Morus alba fruits. The separation was achieved with the developer hexane‒ethyl acetate‒glacial acetic acid (4:6:0.1, V/V) on TLC aluminum plate pre-coated with silica gel 60 F254. Densitometric scanning was carried out at a wavelength of 302 nm in absorbance mode. The peak corresponding to reference standard resveratrol and that of the extract had the same RF of 0.556. The method was validated by following the International Conference of Harmonization (ICH) guidelines for precision, accuracy and reproducibility. The regression equation showed a polynomial function with a correlation coefficient of 0.994 ± 4.20% (R ± SD). The limit of detection and limit of quantification were 16 and 45 ng/band, respectively. The content of resveratrol was found to be 31.94 μg/100 mg of 70% methanolic extract of M. alba. The validated HPTLC method can be routinely used for the quality control and standardization of the bioactive marker resveratrol and formulations containing M. alba.

Determinants of bone health in adults Polish women: The influence of physical activity, nutrition, sun exposure and biological factors

PurposeThe aim of this study was to assess the determinants of bone health in the group of women over 40 years old. Lifestyle factors such as past and present physical activity, past and present sun exposure, current dietary intake of calcium and vitamin D, nutritional status as measured by BMI, family history of osteoporosis and current hormonal status were analysed.MethodsThe study involved 500 women over 40 years old. All examined women was the same ethnicity- European origin. Methods used: densitometry method (DXA), bioelectrical impedance analysis, International Physical Activity Questionnaire, nutrition questionnaire, past and present sun exposure questionnaire. Past and present physical activity, past and present sun exposure and sufficient level of calcium in the diet proved to be the most important factors determining mineralization of bone tissue of women. In order to indicate an independent association of the correct bone tissue mineralization with individual factors, multivariate analysis was used—logistic regression.ResultsThe norm BMD in the distal part of the forearm was strongly influenced: recommended dietary calcium intake (OR = 5.95; p = 0.003), moderately (OR = 1.88; p = 0.053) and high (OR = 14.0; p<0.001) past physical activity, sufficient (OR = 4.97; p<0.001) and high (OR = 18.9; p = 0.004) level of present physical activity, sufficient past (OR = 5.15; p<0.001) and sufficient present sun exposure (OR = 10.0; p<0.001).The chance for the BMD prox norm was also increased several times: high past physical activity (OR = 68.4; p<0.0001) and sufficient past sun exposure (OR = 10.6; p<0.001), moderate past activity (OR = 4.20; p<0.001), sufficient (OR = 6.13; p<0.001) and high (OR = 10.0; p<0.001) present physical activity, sufficient present sun exposure (OR = 9.09; p<0.0001), recommended intake of calcium (OR = 9.57; p<0.001) and vitamin D (OR = 2.68; p = 0.052). Whereas e significantly lower likelihood for the BMD prox norm was found in women with the oldest hormonal status (postmenopausal period) (OR = 0.18; p<0.001), with osteoporosis in the family (OR = 0.37; p<0.001) and living in an agglomeration (OR = 0.68; p = 0.03).ConclusionInterventions to increase physical activity, especially outdoors, may help reduce risk of osteoporosis, fractures and subsequent healthcare costs.

Simultaneous Determination of 6-Shogaol and 6-Gingerol in Various Ginger (Zingiber officinale Roscoe) Extracts and Commercial Formulations Using a Green RP-HPTLC-Densitometry Method.

Various analytical methodologies have been reported for the determination of 6-shogaol (6-SHO) and 6-gingerol (6-GIN) in ginger extracts and commercial formulations. However, green analytical methods for the determination of 6-SHO and 6-GIN, either alone or in combination, have not yet been reported in literature. Hence, the present study was aimed to develop a rapid, simple, and cheaper green reversed phase high-performance thin-layer chromatography (RP-HPTLC) densitometry method for the simultaneous determination of 6-SHO and 6-GIN in the traditional and ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas. The simultaneous analysis of 6-SHO and 6-GIN was carried out via RP-18 silica gel 60 F254S HPTLC plates. The mixture of green solvents, i.e., ethanol:water (6.5:3.5 v/v) was utilized as a mobile phase for the simultaneous analysis of 6-SHO and 6-GIN. The analysis of 6-SHO and 6-GIN was performed at λmax = 200 nm for 6-SHO and 6-GIN. The densitograms of 6-SHO and 6-GIN from traditional and ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas were verified by obtaining their single band at Rf = 0.36 ± 0.01 for 6-SHO and Rf = 0.53 ± 0.01 for 6-GIN, compared to standard 6-SHO and 6-GIN. The green RP-HPTLC method was found to be linear, in the range of 100–700 ng/band with R2 = 0.9988 for 6-SHO and 50–600 ng/band with R2 = 0.9995 for 6-GIN. In addition, the method was recorded as “accurate, precise, robust and sensitive” for the simultaneous quantification of 6-SHO and 6-GIN in traditional and ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas. The amount of 6-SHO in traditional extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas was obtained as 12.1, 17.9, 10.5, and 9.6 mg/g of extract, respectively. However, the amount of 6-SHO in ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas were obtained as 14.6, 19.7, 11.6, and 10.7 mg/g of extract, respectively. The amount of 6-GIN in traditional extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas were found as 10.2, 15.1, 7.3, and 6.9 mg/g of extract, respectively. However, the amount of 6-GIN in ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas were obtained as 12.7, 17.8, 8.8, and 7.9 mg/g of extract, respectively. Overall, the results of this study indicated that the proposed analytical technique could be effectively used for the simultaneous quantification of 6-SHO and 6-GIN in a wide range of plant extracts and commercial formulations.

Design of Liver Functional Reserve Estimation Technique Based on Optical Densitometry.

This work is aimed at creating a modified invasive technique for assessing the liver’s functional reserves. A study of the degree of hepatodepression is carried out by measuring the plasma elimination of indocyanine green using the method of optical densitometry. This paper presents test results for an aqueous solution and an albumin solution, as well as the results of measurements of plasma elimination of indocyanine green for patients with liver disease. Perfecting the proposed method will make an important scientific contribution to modern diagnostic medicine. Diagnosing the stages in the progression of the disease and its developing complications can make it possible to rapidly correct the patient’s treatment algorithm, achieving positive outcomes in medical practice.

Development and validation of a simple and rapid thin-layer chromatography–UV densitometry method for the determination of triazophos in human whole blood for forensic toxicological applications

No published studies reported triazophos (TZP) analysis in human whole blood following fatal intoxication. In this study, a method for the determination of TZP in human whole blood using liquid–liquid extraction (LLE) and thin-layer chromatography (TLC)–ultraviolet (UV) densitometry is described. Chromatography was performed on silica gel 60F254 TLC plates using mobile phase n-hexane:acetone (8:2, v/v) and UV densitometric detection at 248 nm. Better extractions were achieved by using ethyl acetate solvent at pH 5 with a recovery of 92.17%. Calibration curve for TZP in blood was linear from 2 to 100 μg/mL (0.04 to 2 μg/spot) and the sensitivity of the method (LLOQ) was 2 μg/mL (0.04 μg/spot). The method showed excellent within-day precision (0.37 to 0.82%, RSD) and between-day precision (0.39 to 1.47%, RSD) for spiked blood samples at 2, 10, and 50 μg/mL. The measured concentrations of TZP in blood did not deviate more than 2.69% under different storage conditions. TZP concentrations in two fatal cases of poisoning were reported. To the best of our knowledge, this is the first validated TLC–UV densitometric method developed to determine TZP in human whole blood.

Thin-layer chromatographic quantification of magnolol and honokiol in dietary supplements and selected biological properties of these preparations

Two isomeric biphenyl neolignans, magnolol and honokiol, are considered as constituents responsible for the healing effect of magnolia bark, a traditional Oriental medicine. To survey the increasing number of dietary supplements that contain magnolia bark or its extract, an affordable quantitative thin-layer chromatography (TLC) – densitometry method was developed. The methanol extracts were analyzed on the silica gel plates after manual sample application using n-hexane – ethyl acetate – ethanol (16:3:1, v/v/v) as a mobile phase. For quantitation, the chromatograms were scanned in the absorbance mode at the wavelength λ = 290 nm. The limits of detection and quantitation were 90 and 280 ng/zone for magnolol and 70 and 200 ng/zone for honokiol, respectively. None of the two targeted neolignans were detected in two of the six analyzed supplements. In the other four samples, the measured amounts were between 0.95-114.69 mg g−1 for magnolol and 4.88-84.86 mg g−1 for honokiol. Moreover, separations of these two neolignans on the TLC and high-performance TLC (HPTLC) layers were compared and HPTLC was combined with antioxidant (DPPH) and antibacterial (Bacillus subtilis and Aliivibrio fischeri) assays and mass spectrometry (MS), using the elution-based interface. Both magnolol and honokiol exhibited effects in all bioactivity assays. The HPTLC-MS tests confirmed purity of neolignan zones in the extracts of dietary supplements and supported tentative identification of the alkaloid piperine and the isoflavone daidzein as additional bioactive components of the investigated dietary supplements. Using the same mobile phase in the orthogonal directions 2D-HPTLC-MS experiments proved degradation, i.e., instability of magnolol and honokiol on the silica gel adsorbent.

The Effect of Vitamin C Addition on Epigallocatechin Gallate (EGCG) Stability in Green Tea Solution

Background: Epigallocatechin gallate (EGCG) is the most abundant green tea catechin with a powerful antioxidant effect to prevent cancer cells. EGCG in green tea solution is highly susceptible to degradation, this it is urgent to increase the stability of EGCG by vitamin C addition. Vitamin C can regenerate radical EGCG to be normal EGCG. Objective: This research aim was to determine percent of decreased level of EGCG before and after vitamin C addition. Methods: This paper was focused on enhancing the stability of EGCG by 1 mg (GTVC1), 1.5 mg (GTVC2), 2 mg (GTVC3), 2.5 mg (GTVC4) and 3 mg (GTVC5) of vitamin C addition to 10 g/L of green tea solution concentration. Evaluations of EGCG were conducted at 0 days, 1 day, 2 days, 3 days, and 4 days of storage time. EGCG was analyzed using thin layer chromatography (TLC) densitometry methods. The stability of EGCG was determined by % of decreased EGCG. Results: Percent of loss of EGCG in GT, GTVC1, GTVC2, GTVC3, GTVC4 and GTVC5 after 4 days storage were 19.93%, 10.89%, 21.08%, 18.18%, 28.56%, and 9.76%, respectively. Conclusion: This study showed that in 4 days storage, the decreased level of EGCG in green tea solution with 3 mg of vitamin C addition (GTVC5) was 9.76% which was smaller than green tea solution without vitamin C addition (GT) which EGCG decreasing 19.93%.

Spectrophotometric and spectrodensitometric quantification of a new antiviral combination

Accurate, simple, and selective spectrophotometric and spectrodensitometric methods were developed and adopted to quantify velpatasvir (VPS) and sofosbuvir (SFV) concurrently in their pure forms and tablets. The spectrophotometric technique was based on the first derivative of ratio spectra (1DD) technique and developed to determine VPS and SFV simultaneously in table formulation. However, the spectrodensitometric technique was based on thin-layer chromatography (TLC) and densitometry and developed to determine VPS and SFV simultaneously in tablet dosage form. Chromatographic separation was performed using chloroform:methanol 9.5:0.5 (%, v/v) as the mobile phase on glass-coated TLC plates. Detection was achieved using a 265-nm deuterium lamp in absorbance mode. Both analytical methods were validated according to the International Conference on Harmonization (ICH)-Q2B guidelines. The linearity in the range of concentration ranges of 1–50 μg/mL and 5–80 μg/mL were obtained for VPS and SFV, respectively, using 1DD spectrometric method. However, the linearity in the range of 5–50 and 10–70 μg/band for VPS and SFV, respectively, were recorded using TLC–densitometric method. Accuracy was recorded ˃100% for VPS and SFV using both methods. This is the first TLC–densitometry method that can separate and quantify the studied mixture of the drugs. The proposed analytical methods were found to be accurate, precise, selective, robust and sensitive for simultaneous analysis of VPS and SFV in tablet dosage forms.

PROFIL BIOAUTOGRAFI DAN UJI PENANGKAP RADIKAL 2,2-DIFENIL-1-PIKRIHIDRAZIL OLEH EKSTRAK ETANOL DAUN BINAHONG (Anredera scandens (L.) Moq.) DAN FRAKSI-FRAKSINYA

Free radicals are atoms or molecules that have one or more unpaired electrons and are highly reactive. Antioxidants can inhibit the excessive oxidation reaction. The leaves of binahong (Anredera scandens (L.) Moq.) are one part of plants which contain flavonoid compounds and tannins that have scientific activity as antioxidants.This study aims to determine the bioautographic profile and to test the antioxidant activity of the ethanol extract of the leaves of Anredera scandens (L.) Moq. useing 2.2-diphenyl-1-pikrilhidrazil radical capture method. The extraction process produced 19.22% yield, 14.68% (14.684 grams) chloroform fraction, 11.22% (11.224 grams) n-hexane fraction, 3.09% (3.094 grams) ethyl acetate fraction, and 23.90% (23.904 grams) n-butanol fraction. The results of bioautographic profile test using KLT densitometry method proved that ethanol extract, chloroform fraction and ethyl acetate fraction have DPPH radical capture activities. The determination of total flavonoid content found that ethyl acetate fraction contained higher total flavonoids than other fractions. In the ethyl acetate fraction, high total flavonoid level and low IC50 values were obtained, while the n-hexane fraction had low flavonoid level and high IC50 values.&#x0D; &#x0D; Keywords: antioxidants, binahong leaves, free radicals, flavonoids, IC50.

MINERAL DENSITY OF BONE TISSUE AND INDICATORS OF BONE REMODELING IN HIGHLY SKILLED ATHLETES IN THE ANNUAL TRAINING CYCLE

Aim. The article aims at studying the relationship between the mineral density of heel bone and bone remodeling in cyclistduring the annual training cycle. Materials and methods. The study involved 14 highly skilled male cyclists (cycle track, MS, MSIC) aged 20–25 years with a sports experience of 5 to 16 years. The measurements were performed in the preparatory, pre-competitive and competitive periods of the annual training cycle. Bone mineral density (BMD) was measured by the method of quantitative ultrasound densitometry (QUS) using the Achilles Express ultrasonometer (Lunar, USA). BMD was estimated as a percentage of the peak norm for a given section of the skeleton (reference norms for 20–29 years). Results. The dynamics of BMD in the annual cycle correlates with a change in bone remodeling and phosphorus-calcium metabolism. A decrease in BMD during the competitive period is associated with an increase in β-Cross Laps and a decrease in osteocalcin, alkaline phosphatase, ionized calcium and serum phosphorus. Conclusion. A decrease in BMD is associated with an increase in the activity of osteoclasts and a decrease in osteoblastic activity. Indicators of bone remodeling and phosphorus-calcium metabolism reflect the status of bone tissue and the processes of general adaptation to training loads, and can be used as markers of risk of decreased BMD. The heel bone QUS method can be used to identify athletes at risk for hypotrophic conditions.

Tibial tray thickness does not significantly increase medial tibial bone resorption: Using tibial bone density as an objective measurement method

BackgroundRecent studies have reported the occurrence of medial tibial bone resorption following total knee replacement. One study proposed that a thick tibial tray results in stress shielding and increases the risk of bone resorption, but its findings were based on subjective radiological assessment. This study aimed to verify this hypothesis and to objectively quantify medial tibial bone density by using serial measurements with digital radiological densitometry. MethodsThis was a retrospective cohort study involving 140 patients (70 thick tray vs. 70 thin tray) with cobalt–chromium implants with at least 24 months of follow-up. Standard radiographs were reviewed to look for incidence of medial tibial bone loss. Serial measurement of medial tibial bone density was also performed using the method of digital radiological densitometry. ResultsThere was no significant difference in the incidence of medial tibial bone loss. Both groups showed a significant drop in medial tibial bone density after operation (P < 0.01). Medial tibial bone density of the thick-tray cohort was significantly higher than the thin-tray cohort at one year (93.3 vs. 83.1 Greyscale; P = 0.04), but not at two and three years. Clinical outcomes in terms of postoperative range of motion, Knee Society score and complication rates were similar. ConclusionsMedial tibial bone resorption is a common phenomenon. Implants with thicker tibial trays suffer less than those with thinner trays at one year, but the difference is transient and does not affect clinical outcome.

Growth Rate and Histamine Production of Citrobacter freundii CK01 in Various Incubation Temperatures

Improper handling and temperature fluctuations during postharvest of fish commonly allow the growth of histamine-forming bacteria (HFB) that may cause histamine poisoning. Citrobacter freundii CK01 is one of the HFB isolated from Skipjack landed on Sadeng, Yogyakarta. This study aimed to determine the temperature effect on the growth rate and histamine production by C. freundii CK01. Bacterial growth and histamine production was tested in tuna fish infusion broth (TFIB) at 5, 15, 30 and 40°C. The bacterial growth was tested using Total Plate Count method, while histamine was determined using Thin Layer Chromatography and densitometry method. The primary model for bacterial growth was plotted with incubation time using DMFit referred to Baranyi &amp; Roberts model. The secondary model was converted from growth rate and modeled using Ratkowsky Square Root Model. The equation for growth rate of C. freundii CK01 was μmax=[0.0105 (T + 13.886)]2 (Root Mean Square Error &lt;10%). Histamine production reached the highest concentration at 15°C in 96 hours up to 117,13 ppm. This study shows that temperature affected the growth rate and histamine production of C. freundii, indicating the importance of maintaining the low temperature stability during handling of skipjack.

Determining Caffeine in Fat-Burning Supplements Using Thin Layer Chromatography-Densitometry and UV-Vis Spectrophotometer

There are so many ways to lose weight, such as consuming fat burner supplements to burn fat faster. A fat burner supplement consists of various ingredients, such as caffeine. Fat burner supplements are usually not registered in BPOM RI. This study aims to evaluate caffeine in fat burner supplements qualitatively and quantitatively. These supplements were analyzed in the pharmaceutical laboratory in Universitas Muhammadiyah Yogyakarta using Thin Layer Chromatography (TLC)–Densitometry and UV–Vis Spectrophotometer. Sample preparation was conducted by separating caffeine from supplements by a separatory funnel with chloroform as an organic solvent. Qualitative analysis was carried out by TLC and analyzing standard spectrum, and sampling technique was carried out with UV–Vis Spectrophotometer. The first quantitative analysis used densitometry by measuring the spot on the TLC plate. Meanwhile, the second quantitative analysis used UV–Vis Spectrophotometer by observing absorbency value on samples with λ 273.5 nm. The result of the qualitative test using TLC was analyzed by comparing the Rf value of the standard and the sample. The Rf value of caffeine standard was 0.63, and every sample showed similar value with caffeine standard, indicating that all samples contain caffeine. The result of quantitative test with TLC - Densitometry method revealed 1 st sample 5.68 mg/ml, 2 nd sample 5.74 mg/ml, 3 rd sample 3.43 mg/ml, 4 th sample 8.90 mg/ml, and 5 th sample 1.88 mg/ml. The qualitative test result was analyzed using the UV–Vis Spectrophotometer method, and all of the caffeine standard spectra can be read at wavelength 273.5 nm, which means all samples contain caffeine. The second quantitative test result analyzed by using UV–Vis Spectrophotometer method showed 1 st sample 3.22 mg/g, 2 nd sample 4.56 mg/g, 3 rd sample 2.23 mg/g, 4 th sample 11.22 mg/g, and 5 th sample 0.26 mg/g. It can be concluded that all samples (fat burners) contained caffeine.

Association between body composition and aerobic capacity in karate athletes

Abstract The aim of this study was to analyze the association between body composition and performance in the karate specific aerobic test (KSAT). This is a study carried out with thirteen athletes (6 females) with a mean age of 20.7 ± 4.2 years, affiliated to the Pernambuco Federation of Karate Associations, were developing this training routine during the 2018 season. Body mass and height were measured. To measure body composition, the body densitometry method was used by the x-ray double-ray absorptiometry (DEXA) technique. Karate Specific Aerobic Test was used to verify aerobic performance. Association analyzes were performed between body composition variables and KSAT performance using Pearson's correlation test and linear regression. All analyzes adopted a significance of p &lt;0.05. KSAT performance was negatively correlated with total fat mass (r = -0.797; p = 0.001) and fat percentage (r = -0.757; p = 0.003). The linear regression model with the highest explanatory power included total fat and KSAT (adjusted R² = 0.732; p &lt;0.001) exhibited a negative association with total fat (? = -0.21; p &lt;0.001), and the model that included % body fat and KSAT (adjusted R² = 0.708; p = 0.003) were negatively associated with% body fat (? = -22.937; p = 0.001), both adjusted for gender and age. There is a negative association between total fat mass, body fat percentage and karate specific aerobic test performance.

Transfer of TLC screening methods to quantitative HPTLC–densitometry methods for pharmaceutical products containing amlodipine besylate, cefpodoxime proxetil, cetirizine 2HCl, diclofenac sodium, efavirenz, mefenamic acid, and atovaquone + proguanil HCl

Transfer of seven thin-layer chromatography (TLC) Global Pharma Health Fund E.V. Minilab protocols for screening counterfeit pharmaceutical products in the field to quantitative high-performance TLC (HPTLC)–densitometry methods was performed using a model process published previously. The developed and validated methods for tablets containing amlodipine besylate, cefpodoxime proxetil, cetirizine 2HCl, diclofenac sodium, efavirenz, mefenamic acid, and atovaquone + proguanil HCl involved the use of only relatively inexpensive and nontoxic solvents, Merck KGaA Premium Purity HPTLC silica gel 60 F254 plates, semi-automated sample and standard solution application with a CAMAG Linomat 4, and automated densitometry with a CAMAG Scanner 3 for detection, identification, and quantification. In addition, previously transferred HPTLC–densitometry methods for azithromycin and for cephalexin were used to analyze a new product of each drug to demonstrate the applicability of the methods.

No Association Between Bone Mineral Density and Breast Arterial Calcification Among Postmenopausal Women

ContextThe association between bone mineral density (BMD) and breast arterial calcification (BAC) remains poorly understood and controversial.ObjectiveThe objective of this article is to examine the association between BMD and BAC in a large cohort of postmenopausal women undergoing routine mammography.DesignA cross-sectional analysis of baseline data from a multiethnic cohort was performed.SettingThe setting for this analysis is an integrated health care delivery system in Northern California in the United States.PatientsA total of 1273 women age 60 to 79 years (mean age, 67 years) were recruited within 12 months of screening mammography.Main outcome measureA BAC score (mg) was obtained from digital mammograms using a novel densitometry method. BAC presence was defined as a BAC score greater than 0 mg, and severe BAC as a BAC score greater than 20 mg.ResultsOverall, 53% of women had osteopenia and 21% had osteoporosis. The prevalence of BAC greater than 0 mg was 29%, 30%, and 29% among women with normal BMD, osteopenia, and osteoporosis, respectively (P = 0.98). The prevalence of BAC greater than 20 mg was 5%, 3%, and 5% among women with normal BMD, osteopenia and osteoporosis, respectively (P = .65). The odds ratios (ORs) of BAC greater than 0 mg vs BAC = 0 mg after multivariable adjustment were 1.09 (95% CI, 0.81-1.48; P = .54) for osteopenia and 0.99 (95% CI, 0.69-1.48; P = .98) for osteoporosis. The adjusted ORs for BAC greater than 20 mg vs BAC 20 mg or less were 1.03 (95% CI, 0.52-2.01; P = .93) for osteopenia and 1.89 (95 CI, 0.81-4.47; P = .14) for osteoporosis.ConclusionOur findings do not support an association of either osteopenia or osteoporosis with BAC presence or severity among postmenopausal women.

Development and Validation of a High-Performance Thin-Layer Chromatography Method for the Quantification of Rutin in the Fruit Pulp of Benincasa hispida (Thunb.) Cogniaux

The fruits of Benincasa hispida (Thunb.) Cogniaux, a ‘rasayana’ in ‘Ayurveda’, are enriched with several secondary metabolites, and rutin is one of them. Fruits are used for their anabolic, brain tonic, carminative, diuretic, memory enhancer, refrigerant, and vitalizer properties. In view of the fact that herbal medicines and/or products are facing challenges towards global acceptance due to the lack of universally accepted standardization method(s), the aim of the current investigation was to develop and validate a high-performance thin-layer chromatography (HPTLC)—densitometry method for the quantification of rutin in the hydroalcoholic extracts of the fruit pulp of B. hispida (HABH). The separation was achieved in a solvent system consisting of ethyl acetate—formic acid—acetic acid—water at a ratio of 7.2:0.7:0.7:1.4 by volume on a TLC aluminum plate pre-coated with silica gel 60 F254. Quantifications were performed by densitometric scanning under a deuterium lamp at a wavelength of 268 nm in the absorbance mode. The precision, accuracy, and reproducibility of the HPTLC method were validated by the International Conference on Harmonization (ICH) guidelines. The mobile phase employed for HPTLC/TLC resulted in good separation for rutin (RF = 0.357). The limit of detection and limit of quantification of the analysis were found to be 0.1 and 0.3 μg per band, respectively. The rutin concentration in the HABH was found to be 178.28 ± 3.62 μg in 10 mg of the extract. The method developed here is simple, fast, reliable, and sensitive and can be implemented in the analysis and routine quality control of B. hispida formulations containing rutin.

Development of quantitative HPTLC methods for dolutegravir, lamivudine, and tenofovir disproxil fumarate in a combination pharmaceutical product using a model process published earlier for transfer of minilab TLC screening methods to HPTLC-densitometry

High-performance thin-layer chromatography (HPLTC)–densitometry methods are described for the analysis of the anti(retro)virals dolutegravir (D), lamivudine (L), and tenofovir disoproxil fumarate (TDF) in a pharmaceutical tablet product. To the best of our knowledge, no previous quantitative planar chromatography method has been reported in the literature for this combination formulation. The method for L was transferred from a thin-layer chromatography (TLC) screening method published in the Global Pharma Health Fund (GPHF) Minilab Manual designed for identification of counterfeit and substandard drug products using a model process published earlier. D and TDF are not included in the list of drugs for which TLC screening methods are published for the Minilab, but HPTLC–densitometry procedures were developed for them using the transfer process guidelines. L was analyzed simultaneously with TDF on Merck Premium Purity silica gel 60 F plates using the mobile phase ethyl acetate–methanol–acetone–concentrated ammonium hydroxide (30:7:3:1) and densitometric scanning at 254 nm. D was analyzed on a second plate by scanning at 366 nm after chromatography with the chloroform–methanol–formic acid (32:8:2) mobile phase. Data for all three drugs are shown to meet the requirements of the model transfer process for calibration curve r values, assay of tablets relative to their label values, peak purity/peak identity tests, and validation by standard addition analysis of samples spiked at 50%, 100%, and 150% of the label value of active ingredients. A TLC screening method for TDF in the combination product was developed and published online with open access.

Factor Optimization Study to Develop and Validate a Reversed-Phase Thin-Layer Chromatography Method for the Determination of Trimetazidine Dihydrochloride and Its Reported Impurities in Pharmaceuticals

A validated reversed-phase thin-layer chromatography (RP-TLC)—densitometry method was developed and optimized for the determination of trimetazidine dihydrochloride (TMZ) and its potential impurities listed in the British Pharmacopoeia, namely, piperazinecarboxaldehyde (Y-145), trimethoxybenzyl alcohol (Y-235), and trimethoxybenzaldehyde (Y-234). Chromatographic separation was performed on aluminum plates pre-coated with silica gel 60 RP-18F using a mixture of acetonitrile—methanol—0.1% aqueous ortho-phosphoric acid pH 6.2 (4.5:4.5:1, V/V) as the developing system. Different factors affecting resolution were studied and optimized. Successful resolution was observed with significant difference in the RF values of 0.21 ± 0.02, 0.35 ± 0.02, 0.5 ± 0.02, and 0.85± 0.02 for TMZ and its impurities, respectively. Densitometric measurement was done at 215 nm over the range of 0.05–10, 0.05–1.1, 0.05–1.2, and 0.04–1.0 µg per spot with lower limits of detection (LOD) and quantification (LOQ) of 20 and 50 ng per spot for TMZ, Y-145, and Y-235, respectively, and 15 and 40 ng per spot for Y-234 impurity. Good accuracy was obtained with mean percentage recoveries of 99.55 ± 1.06, 100.50 ± 1.16, 100.07 ± 1.24, and 99.41 ± 1.11 for TMZ and its impurities, respectively. The developed method was used to investigate the impurity profile of TMZ in drug substance and different products and it was validated as per the International Conference on Harmonisation (ICH) guidelines.