Comparison of two methods for the extraction of fat from human milk

We compared two methods for the extraction of fat from human milk. Pure fat extraction techniques are necessary for qualitative and quantitative analysis of milk fat, of which triglycerides account for more than 98%. Method I was a conventional liquid-liquid system for the fat extraction while method II was a faster approach using a haematocrit technique. No significant differences were observed between both methods neither in the fat content determined gravimetrically, nor in qualitative and quantitative analysis of triglycerides by high-performance liquid chromatography (HPLC) with evaporative light-scattering detection (ELSD). We conclude that method II offers substantial advantages over the conventional method (method I). The former requires less reagents and material and is simpler and less time-consuming (approximately 30 min instead of 90 min). Therefore, a new method will make it possible to extract fat of more human milk in the same time.     

Chemical Fingerprint Analysis and Quantitative Analysis of Saccharides in Morindae Officinalis Radix by HPLC-ELSD

A method based on high performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD) was developed for the quantitative analysis of three active compounds and chemical fingerprint analyses of saccharides in Morindae officinalis radix. Ten batches of Morindae officinalis radix were collected from different plantations in the Guangdong region of China and used to establish the fingerprint. The samples were separated with a COSMOIL Sugar-D column (4.6 mm × 250 mm, 5 μm) by using gradient elution with water (A) and acetonitrile (B). In addition, Trapped-Ion-Mobility (tims) Time-Of-Flight (tims TOF) was used to identify saccharides of Morindae officinalis radix. Fingerprint chromatogram presented 26 common characteristic peaks in the roots of Morinda officinalis How, and the similarities were more than 0.926. In quantitative analysis, the three compounds showed good regression (r = 0.9995–0.9998) within the test ranges, and the recoveries of the method were in the range of 96.7–101.7%. The contents of sucrose, kestose and nystose in all samples were determined as 1.21–7.92%, 1.02–3.37%, and 2.38–6.55%, respectively. The developed HPLC fingerprint method is reliable and was validated for the quality control and identification of Morindae officinalis radix and can be successfully used to assess the quality of Morindae officinalis radix.     

HPLC-ELSD法测定绞股蓝中绞股蓝皂苷A的含量

采用HPLC-ELSD法测定绞股蓝中绞股蓝皂苷A的含量。色谱柱为HibarC18(250×4.6mm,5μm),流动相为乙腈-水梯度洗脱,流速0.5mL/min,柱温为20℃,绞股蓝皂苷A在0.0425—1.995mg/mL范围内线性关系良好(r2=0.9962),并通过正交法优化其提取工艺。该方法简便、有效,为绞股蓝的质量控制提供一种准确可行的检测方法。     

Detection of alkanolamines in liquid cement grinding aids by HPLC coupled with evaporative light scattering detector

Triethanolamine (TEA), triisopropanolamine (TIPA), diethanol isopropanolamine (DEIPA) are necessary cement additives, and knowing their contents is needed for quality control and also to understand final properties of the cement. Whether it is the production of grinding aids, technical research and development or application research all involve the detection of grinding aids. This work developed a simple analytical technique for the simultaneous analysis of these alkanolamines in liquid cement grinding aids using high-performance liquid chromatography (HPLC) combined with evaporative light scattering detection (ELSD). HPLC was conducted by an XBridge C18 column (with dimensions 4.6 × 250 mm and 5 µm particles) using methanol and 0.1% trichloroacetic acid as mobile elution phases. The ELSD sprayer and drift tube temperatures were 60 ºC and 90 ºC, respectively. HPLC-ELSD developed in this work demonstrated 1) high sensitivity with limits of detection for the three analytes are 0.15, 0.54, 1.04 µg/mL; 2) good linearity with correlation coefficients equal to 0.997–0.999 over the tested concentration range; 3) excellent repeatability with intra- and interday coefficient of variation (CV) below 2.71% and 4, satisfactory accuracy with recovery in the 95.5%–100.8% range. This novel method is a powerful, time- and costeffective tool for alkanolamine analyses and quality control.     

Correlations of Fat Content in Human Milk with Fat Droplet Size and Phospholipid Species

Fat globule size and phospholipid (PL) content in human milk (HM) were investigated. HM was classified into three groups depending on fat content (A < B < C). PL content (mg/100 g HM) was significantly higher in the C group (p < 0.05), indicating its positive relationship with HM fat content. When the PL content was normalized (mg/g fat), that of group A was significantly higher (p < 0.05) and fat droplet size in group C was slightly larger, suggesting that HM fat content is affected by fat droplet numbers to a larger extent than by fat droplet size. A correlation between PC and SM content in HM was observed regardless of fat content, while correlation between PE and either PC or SM increased in the order of C > B > A, hence the composition and content of PL species in HM varied according to its fat content.     

HPLC-ELSD与GC-MS法测定牛乳甘油三酯sn-2位脂肪酸组成

研究建立了快速、准确测定牛乳脂肪甘油三酯sn-2位脂肪酸组成的方法．利用胰脂酶专一水解甘油三酯sn-1和sn-3位置上的脂肪酸得到sn-2单甘油酯和游离脂肪酸,再通过蒸发光散射高效液相色谱分离出sn-2位单甘油酯,然后对其进行衍生,用气相色谱质谱联用仪对sn-2脂肪酸组成进行分析．结果显示用蒸发光散射高效液相色谱法分离sn-2位单甘油酯时方法的回收率达到83．3％～85．1％,该法省去了传统测定中费时费力的薄层色谱分离步骤．用气相色谱质谱联用法对产物进行分析,精密度高,结果可靠．分析结果表明,牛乳脂肪sn-2位脂肪酸由2．57％月桂酸、7．68％豆蔻酸、34．74％棕榈酸、11．56％亚油酸、22．53％油酸和15．21％硬酯酸组成．     

HPLC-ELSD测定茜芷胶囊中三七皂苷R1、人参皂苷Rg1及Rb1的含量

利用HPLC-ELSD对茜芷胶囊中的活性成分三七皂苷R1、人参皂苷Rg1和Rb1的含量进行测定.三七皂苷R1、人参皂苷Rg1及Rb1分别在0.13~2.60、0.30~6.00、0.38~7.60μg范围内呈现良好的线性关系,平均回收率分别为98.0%(RSD1.7%),97.1%(RSD1.8%),97.0%(RSD1.5%).该方法简便、准确、重现性好,可用于茜芷胶囊的质量控制.     

Determination of Triterpenoids and Phenolic Acids from Sanguisorba officinalis L. by HPLC-ELSD and Its Application

A novel analytical method involving high-performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD) was developed for simultaneous determination of 11 phenolic acids and 12 triterpenes in Sanguisorba officinalis L. Chromatographic separation was conducted with gradient elution mode by using a Diamonsil^TM C₁₈ column (250 mm × 4.6 mm, 5 μm) with the mobile phase of 0.1% acetic acid water (A) and methanol (B). The drift tube temperature of ELSD was set at 70 °C and the nitrogen cumulative flow rate was 1.6 L/min. The method was fully validated to be linear over a wide concentration range (R² ≥ 0.9991). The precisions (RSD) were less than 3.0% and the recoveries were between 97.7% and 101.4% for all compounds. The results indicated that this method is accurate and effective for the determination of 23 functional components in Sanguisorba officinalis L. and could also be successfully applied to study the influence of processing method on those functional components in Sanguisorba officinalis L.     

Evaluation of traditional Chinese herbal medicine: Chaihu (Bupleuri Radix) by both high-performance liquid chromatographic and high-performance thin-layer chromatographic fingerprint and chemometric analysis

Chaihu (Bupleuri Radix), roots of Bupleurum chinense and B. scorzonerifolium, is an authentic Chinese Materia Medica in the Chinese Pharmacopoeia. Some other species such as the roots of B. falcatum, B.bicaule and B. marginatum var. stenophyllum similar to Chaihu can also be occasionally found in local raw herb markets. The quality of 33 lots of authenticated Chaihu samples vs. 31 lots of commercial samples was evaluated by both high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) and high-performance thin-layer chromatography (HPTLC) analyses of its principal bioactive components (saikosaponins). The pre-treated data acquired from both HPLC fingerprints and HPTLC fluorescent images were processed by chemometrics for similarity and pattern recognition, including Artificial Neural Networks (ANNs), k-nearest neighbor (k-NN) and an expert’s panel. It was apparent that k-NN classifier exhibited good performance with sufficient flexibility for processing HPTLC fingerprint images which were otherwise not easily dealt with by other algorithms due to the shift of R_f values and varying hue/saturation of the band colours between different TLC plates. These two chromatographic fingerprint methods can be considered complementary measure of quality control. The roots of Chaihu from different species of the genus Bupleurum could readily be distinguished from each other so that commercial samples can easily be classified. Chaihu collected from several major herbal distribution centers was found to belong to B. chinense with great variation in the content of its major saikosaponins.     

Presence of Cholesterol in Non-Animal Organisms: Identification and Quantification of Cholesterol in Crude Seed Oil from Perilla frutescens and Dehydrated Pyropia tenera

Studies have reported that cholesterol, a molecule found mainly in animals, is also present in some plants and algae. This study aimed to determine whether cholesterol exists in three dehydrated algae species, namely, Pyropia tenera, Saccharina japonica, and Undaria pinnatifida, and in one plant species, namely, Perilla frutescens (four perilla seed oil samples were analyzed). These species were chosen for investigation because they are common ingredients in East Asian cuisine. Gas chromatography-flame ionization detection (GC-FID) analysis found that cholesterol was present in P. tenera (14.6 mg/100 g) and in all four perilla seed oil samples (0.3–0.5 mg/100 g). High-performance liquid chromatography with evaporative light-scattering detection (HPLC-ELSD) also demonstrated that cholesterol was present in P. tenera (14.2 mg/100 g) and allowed the separation of cholesterol from its isomer lathosterol. However, cholesterol could not be detected by HPLC-ELSD in the perilla seed oil samples, most likely because it is only present in trace amounts. Moreover, liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed the presence of cholesterol in both P. tenera and perilla seed oil. MRM results further suggested that lathosterol (a precursor of cholesterol) was present in P. tenera.