Seabuckthorn (Hippophae rhamnoides, L.) pulp oil prevents ultraviolet-induced damage in human fibroblasts.

Seabuckthorn pulp oil (SBO) is used in beauty products because of its rich lipophilic substances with high nutraceutical and cosmeceutical potential. However, the mechanism through which SBO enhances skin elasticity remains unclear. Therefore, in this study, we examined the anti-photoaging activity of SBO in normal human dermal fibroblasts (NHDF) under ultraviolet (UV) irradiation. Pretreatment with SBO significantly suppressed UV-B-induced cell toxicity and collagen degradation, suggesting that SBO contains anti-photoaging substances. Further, palmitoleic acid, the main component of SBO, maintained cell viability and collagen levels in UV-B-irradiated NHDF by suppressing the expression of matrix metalloproteinase 1 and acted on the inhibition of p38 and JNK phosphorylation and nuclear translocation of nuclear factor-kappa B. These findings suggest the utility of SBO as an anti-photoaging agent.

Optimization of ultrasound-assisted aqueous two-phase extraction of polysaccharides from seabuckthorn fruits using response methodology, physicochemical characterization and bioactivities.

BACKGROUND: Seabuckthorn fruits contains many active subtances, among them, the seabuckthorn polysaccharide is one of the main active ingredients, and exhibits diverse bioactivities. The extraction of polysaccharides from seabuckthorn fruits is the most important step for their wide applications. Ultrasound-assisted aqueous two-phase extraction (UA-ATPE) is a promising green method for extracting polysaccharides. Additionally, physicochemical characterization and antioxidant activities can evaluate the potential functions and applications in the food and medicine industries. RESULTS: Based on the single-factor experiments, 20.70% (w/w) ammonium sulfate ((NH4 )2 SO4 ) and 27.56% (w/w) ethanol were determined as the suitable composition for aqueous two-phase. The optimum conditions of UA-ATPE obtained by response surface methodology were as follows: ultrasonic power (390 W), extraction time (41 min), liquid-to-material ratio (72: 1 mL/g), and the total yield of the polysaccharides reached 34.14 ± 0.10%, The molecular weights of the purified upper-phase seabuckthorn polysaccharide (PUSP) and the purified lower-phase seabuckthorn polysaccharide (PLSP) were 65 525 and 26 776 Da, respectively. PUSP and PLSP contained the same six monosaccharides (galacturonic acid, rhamnose, xylose, mannose, glucose and galactose), but with different molar ratios. Furthermore, PUSP and PLSP displayed certain viscoelastic property, had no triple helical structure, possessed different thermal stability, surface morphology and conformation in aqueous solution. PUSP and PLSP displayed strong antioxidant properties by the assays of scavenging ability of ABTS+ ·, the protection of DNA damage and erythrocyte hemolysis. CONCLUSION: UA-ATPE significantly increased the yield of seabuckthorn polysaccharides. PUSP and PLSP were different in many aspects, such as molar ratio, surface shape and antioxidant activities. Seabuckthornpolysaccharides possess great potential in medicine and functional foods. © 2022 Society of Chemical Industry.

Genome sequence and population genomics provide insights into chromosomal evolution and phytochemical innovation of Hippophae rhamnoides.

Plants of the Elaeagnaceae family are widely used to treat various health disorders owing to their natural phytochemicals. Seabuckthorn (Hippophae rhamnoides L.) is an economically and ecologically important species within the family with richness of biologically and pharmacologically active substances. Here, we present a chromosome-level genome assembly of seabuckthorn (http://hipp.shengxin.ren/), the first genome sequence of Elaeagnaceae, which has a total length of 849.04 Mb with scaffold N50 of 69.52 Mb and 30 864 annotated genes. Two sequential tetraploidizations with one occurring ~36-41 million years ago (Mya) and the last ~24-27 Mya were inferred, resulting in expansion of genes related to ascorbate and aldarate metabolism, lipid biosynthesis, and fatty acid elongation. Comparative genomic analysis reconstructed the evolutionary trajectories of the seabuckthorn genome with the predicted ancestral genome of 14 proto-chromosomes. Comparative transcriptomic and metabonomic analyses identified some key genes contributing to high content of polyunsaturated fatty acids and ascorbic acid (AsA). Additionally, we generated and analysed 55 whole-genome sequences of diverse accessions, and identified 9.80 million genetic variants in the seabuckthorn germplasms. Intriguingly, genes in selective sweep regions identified through population genomic analysis appeared to contribute to the richness of AsA and fatty acid in seabuckthorn fruits, among which GalLDH, GMPase and ACC, TER were the potentially major-effect causative genes controlling AsA and fatty acid content of the fruit, respectively. Our research offers novel insights into the molecular basis underlying phytochemical innovation of seabuckthorn, and provides valuable resources for exploring the evolution of the Elaeagnaceae family and molecular breeding.

Hepatoprotective effect of seabuckthorn leaf-extract in lead acetate-intoxicated Wistar rats.

The present study investigated the hepatoprotective effects of seabuckthorn (Hippophae rhamnoides) leaf-extract (SLE) supplementation in lead acetate-intoxicated Wistar rats. Adult male Wistar rats (n = 35) were divided into five equal groups as per completely randomized design. Group I was kept as a control, group II received 250 ppm lead acetate in drinking water, and group III received SLE at 100 mg/kg body weight per os. Animals in group IV and group V received lead acetate at 250 ppm in drinking water for the first 45 days. Additionally, group IV received SLE at 100 mg/kg body weight per os throughout the experiment, whereas group V received SLE at 100 mg/kg body weight per os during the last 15 days of the trial. Blood samples were collected on day zero and at the 45th and 60th day to study the liver function enzymes. Lead exposure caused increase in aspartate aminotransferase (AST), alanine aminotransferase (ALT), acid phosphatase (ACP), and alkaline phosphatase (ALP) activity, and liver weight, and hepatic oxidative stress in lead acetate-intoxicated groups (groups II, IV, and V) as compared to group I. SLE on simultaneous supplementation with lead acetate (group IV) exerted a protective effect against lead toxicity. SLE supplemented after the establishment of lead acetate-toxicity (group V) also reduced the AST, ALT, and ALP activity and hepatic oxidative stress, indicating its ameliorative effect. SLE supplementation at 100 mg/kg body weight per os protects against hepatic damage caused by 250 ppm lead acetate in the drinking water of Wistar rats.

Morphometric and Microsatellite Diversity in Seabuckthorn (Hippophae rhamnoides L.) Natural Populations Originating from the Different Geographical Regions of the Indian Himalayas.

Seabuckthorn (Hippophae rhamnoides L.), an upcoming superfood plant, has attracted researchers' attention worldwide for its medicinal, nutritional, and socio-economic value, along with its characteristic features to sustain extreme climatic conditions. We have studied microsatellite marker-based genetic and morphometric diversity in 93 collections of H. rhamnoides from different geographic sites representing two regions, namely Leh and Lahaul of the Indian Himalayas. Microsatellite markers were isolated using two different approaches, including screening of microsatellite-enriched genomic library, and in silico screening of in-house developed seabuckthorn EST database and whole transcriptome assembly. In Leh and Lahaul collections, 32 and 30 microsatellite markers were found polymorphic, respectively. All the markers developed for H. rhamnoides showed cross-species transferability to H. salicifolia and H. tibetana. Two to six alleles were recorded in the two sets of collections with an average of 3.71 and 3.53 alleles per locus in Leh and Lahaul collections, respectively. Mean polymorphic information content (PIC) values for microsatellite markers were 0.39 and 0.41 for Leh and Lahaul collections, respectively. The average expected heterozygosity was less than the observed heterozygosity. Wright's fixation index (F IS) varied from (-)0.2045 to 1.0 and (-)0.1688 to 1.0 for Leh and Lahaul collections, respectively. Shannon's informative index (I) remained in the range of 0.6745 to 1.8621, and 0.6824 to 1.6308 for Leh and Lahaul collections, respectively. The UPGMA-based combined dendrogram showed clear demarcation between Leh and Lahaul collections, although a few ecotypes were regrouped with collections from the other region. No significant relationship was observed between the morphological distance matrix and molecular marker distance matrix. The findings of the present study may prove helpful in future breeding and conservation strategies aiming for seabuckthorn improvement. Supplementary Information: The online version contains supplementary material available at 10.1007/s11105-022-01338-6.

Gas chromatography-mass spectrometry (GC-MS) profiling reveals substantial metabolome diversity in seabuckthorn (Hippophae rhamnoides L.) berries originating from different geographical regions in the Indian Himalayas.

INTRODUCTION: Seabuckthorn (Hippophae rhamnoides L.) is a high-altitude plant with immense medicinal, nutritional, and therapeutic value. Earlier studies have documented the presence of various useful bioactive substances in this species; however, comprehensive metabolome profiling of seabuckthorn berries originating from different regions of the Indian Himalayas has not been undertaken. OBJECTIVE: Metabolomic profiling of seabuckthorn berries originating from different geographical sites in the Himachal Pradesh and Jammu & Kashmir regions of the Indian Himalayas was performed by using gas chromatography-mass spectrometry. MATERIALS AND METHODS: The GC-MS metabolome profiles of seabuckthorn berries collected from different sites (altitude 1,400-4,270 m; average temperature 8°C-27°C) were subjected to multivariate analysis following principal component analysis and hierarchical clustering analysis. RESULTS: The GC-MS results showed substantial variability for berry metabolites, including fatty acids, alkyl ethers, and alkyl esters. Fatty acids and their esters were mainly responsible for the variation in the berry metabolome. The metabolite expression profile heat map revealed two distinct groups of seabuckthorn berries originating from Himachal Pradesh (Lahaul and Spiti) and Jammu & Kashmir (Leh, Nubra, and Kargil), the former showing higher expression of metabolites. Interestingly, a strong negative association existed between altitude and the amounts of metabolites such as amides, alkyl esters, alcohols, sugars, and sugar esters. In contrast, temperature showed a strong positive association with ketone and alkyl ether levels. CONCLUSION: GC-MS profiling provides important phytochemical indicators to distinguish between seabuckthorn berries from different geographical sites. Our metabolome profiling analysis generated valuable information that will be useful in the formulation of various seabuckthorn products, benefiting farmers and industries.

Feasibility of Defatted Juice from Sea-Buckthorn Berries (Hippophae rhamnoides L.) as a Wheat Beer Enhancer.

Juice made from sea-buckthorn berries (Hippophae rhamnoides L.) is a valuable source of bioactive compounds, vitamins, as well as micro- and macronutrients. By applying defatted sea-buckthorn juice, it is possible to enhance wheat beer and change its sensory properties and the contents of bioactive compounds in the finished product. A sensory assessment showed that wheat beers with a 5% v/v addition of sea-buckthorn juice were characterised by a balanced taste and aroma (overall impression). Physicochemical analyses showed that, compared to the control samples, wheat beers enhanced with defatted sea-buckthorn juice at a rate of 5% v/v or 10% v/v had high total acidity with respective mean values of 5.30 and 6.88 (0.1 M NaOH/100 mL), energy values lower on average by 4.04% and 8.35%, respective polyphenol contents of 274.1 mg GAE/L and 249.7 mg GAE/L, as well as higher antioxidant activity (measured using DPPH, FRAP, and ABTS assays). The findings show that the samples of wheat beer enhanced with sea-buckthorn juice had average ascorbic acid contents of 2.5 and 4.5 mg/100 mL (in samples with 5% v/v and 10% v/v additions, respectively) and contained flavone glycosides, e.g., kaempferol-3-O-glucuronide-7-O-hexoside. Based on the current findings, it can be concluded that wheat beer enhanced with sea-buckthorn juice could emerge as a new trend in the brewing industry.

Improving the color and functional properties of seabuckthorn seed protein with phytase treatment combined with alkaline solubilization and isoelectric precipitation.

BACKGROUND: Reducing anti-nutritional factors like phytates in seed protein products requires an ongoing effort. This study was the first to investigate the phytic acid content in seabuckthorn seed protein (SSP) and its reduction by an exogenous phytase during protein isolation from seabuckthorn seed meal through the common alkaline solubilization-isoelectric precipitation process. RESULTS: The additional phytase treatment could reduce the content of phytic acid from 22.46 to 13.27 g kg-1 , leading to SSP products with lighter color (lower ΔE* ), higher protein solubility, higher in vitro digestibility, but lower phenolic antioxidant content (including flavonoids and procyanidins) and some beneficial ions like Ca, Fe, Mg, and Zn. The Fourier transform infrared (FTIR) results indicated that the secondary structure of protein changed under the treatment with phytase. Correlation analysis showed that L* was significantly negatively correlated with TP, TPC and TF (P < 0.001), while a* and b* were significantly positively correlated with them (P < 0.001). CONCLUSIONS: There may be a trade-off between protein functionalities and other health-promoting components when a phytase treatment is included in SSP isolation. © 2021 Society of Chemical Industry.

Protective Effect of Two Alkaloids from Hippophae rhamnoides Linn. against Doxorubicin-Induced Toxicity in H9c2 Cardiomyoblasts.

BACKGROUND: Doxorubicin (Dox) is one of the most frequently prescribed anti-cancer drugs. However, clinical application with Dox is limited due to its potentially fatal cumulative cardiotoxicity. N-p-coumaroyl-4-aminobutan-1-ol (alk-A), an organic amide alkaloid and hippophamide (alk-B), a rare pyridoindole alkaloid were successfully obtained by purification and separation of seabuckthorn seed residue in our previous research. This study was undertaken to investigate the protective effect of alk-A and alk-B against Dox-induced embryonic rat cardiac cells (H9c2 cells) apoptosis. METHODS: H9c2 cells were treated with Dox (2.5 µM) in the presence of alk-A and alk-B (10, 20, and 40 µM) and incubated for 24 h. RESULTS: It was shown that pretreatment of the H9c2 cells with alk-A and alk-B significantly reduced Dox-induced apoptosis. Alk-A and alk-B both inhibited reactive oxygen species (ROS) production and suppressed cleaved-caspase-3 protein expression and the activation of JNK (Jun N-terminal kinases), as well as increasing ATP levels, favoring mitochondrial mitofusin protein expression, and relieving damage to mitochondrial DNA. CONCLUSIONS: These results suggest that alk-A and alk-B can inhibit Dox-induced apoptosis in H9C2 cardiac muscle cells via inhibition of cell apoptosis and improvement of mitochondrial function, while alk-B showed more protection. Alk-B could be a potential candidate agent for protecting against cardiotoxicity in Dox-exposed patients.

[Research progress on chemical constituents and pharmacological activities of seabuckthorn and prediction of its Q-markers].

Seabuckthorn contains flavonoids, tannins, terpenoids, polysaccharides, and vitamins, which have anti-inflammation,anti-oxidation, liver protection, anti-cardiovascular disease, anti-aging, immune enhancing, anti-tumor, and anti-bacterial activities.We reviewed the papers focusing on the chemical constituents, pharmacological activities, and utilization of seabuckthorn. The quality markers(Q-markers) of seabuckthorn were predicted and analyzed based on original plant phylogeny, chemical composition correlation, traditional medicinal properties, pharmacodynamic correlation, traditional and extended efficacy, pharmacokinetics, metabolic processes, and measurable components. With this review, we aim to provide theoretical reference for the quality control and quality standard establishment of seabuckthorn, so as to promote the rational exploitation and utilization of seabuckthorn resources, and improve the healthy and sustainable development of seabuckthorn industry.

Supplementation with Seabuckthorn Oil Augmented in 16:1n-7t Increases Serum Trans-Palmitoleic Acid in Metabolically Healthy Adults: A Randomized Crossover Dose-Escalation Study.

BACKGROUND: In animal models cis-palmitoleic acid (9-hexadecenoic acid; 16:1n-7c), a lipokine, improves insulin sensitivity, inflammation, and lipoprotein profiles; in humans trans-palmitoleic acid (16:1n-7t) has been associated with lower incidence of type 2 diabetes. The response to dose-escalation of supplements containing cis- and trans-palmitoleic acid has not been evaluated. OBJECTIVES: We examined dose-escalation effects of oral supplementation with seabuckthorn oil and seabuckthorn oil augmented in 16:1n-7t on serum phospholipid fatty acids (PLFAs). METHODS: Thirteen participants (7 women and 6 men; age 48 ± 16 y, BMI 30.4 ± 3.7 kg/m2) participated in a randomized, double-blind, crossover, dose-escalation trial of unmodified seabuckthorn oils relatively high in 16:1n-7c (380, 760, and 1520 mg 16:1n-7c/d) and seabuckthorn oils augmented in 16:1n-7t (120, 240, and 480 mg 16:1n-7t/d). Each of the 3 escalation doses was provided for 3 wk, with a 4-wk washout period between the 2 supplements. At the end of each dose period, fasting blood samples were used to determine the primary outcomes (serum concentrations of the PLFAs 16:1n-7t and 16:1n-7c) and the secondary outcomes (glucose homeostasis, serum lipids, and clinical measures). Trends across doses were evaluated using linear regression. RESULTS: Compared with baseline, supplementation with seabuckthorn oil augmented in 16:1n-7t increased phospholipid 16:1n-7t by 26.6% at the highest dose (P = 0.0343). Supplementation with unmodified seabuckthorn oil resulted in a positive trend across the dose-escalations (P-trend = 0.0199). No significant effects of either supplement were identified on blood glucose, insulin, lipids, or other clinical measures, although this dosing study was not powered to detect such effects. No carryover or adverse effects were observed. CONCLUSIONS: Supplementation with seabuckthorn oil augmented in 16:1n-7t and unmodified seabuckthorn oil moderately increased concentrations of their corresponding PLFAs in metabolically healthy adults, supporting the use of supplementation with these fatty acids to test potential clinical effects in humans.This trial was registered at clinicaltrials.gov as NCT02311790.

Genotoxicity and teratogenicity of seabuckthorn (Hippophae rhamnoides L.) berry oil.

As botanicals and dietary supplements are used increasingly in many countries, the issue of safety is particularly critical for regulation of food products containing these substances. Seabuckthorn (Hippophae rhamnoides L.) has been used for centuries as a medicine and nutritional supplement in Asia and Europe. However, data regarding to the safety assessment of the plant and its extracts is still rare. This study was to evaluate the potential toxicity of seabuckthorn berry (SB) oil conducted in three genotoxicity studies and a teratogenicity study. Results of the genotoxicity studies indicated that SB oil has no genotoxicity under the experimental conditions of this study. Specifically, SB oil did not display any mutagenic activity on histidine dependent strains of Salmonella typhimurium under exposure concentrations of 8, 40, 200, 1000, and 5000 µg/plate; SB oil did not have significant effect on sperm morphology and have no influence on micronucleus rate of polychromatic erythrocytes in mice at doses of 9.36, 4.68, and 2.34 g/kg body weight. In the teratogenicity study, pregnant rats were treated with 4.68, 2.34, and 1.17 g/kg SB oil from gestation day 7 to 16 and no treatment-related maternal toxicity or embryo toxicity was observed. Taken together, these results support the safe use of seabuckthorn berry oil for potential dietary consumption in food or as a dietary supplement.

Adjuvant activity of ethanol extract of Hippophae rhamnoides leaves with inactivated rabies virus antigen.

CONTEXT: Hippophae rhamnoides L. (Elaeagnaceae), commonly known as seabuckthorn (SBT), is known for its medicinal and nutritional properties. OBJECTIVE: Evaluation of in vivo adjuvant activity of SBT leaf extract (SBTE) with inactivated rabies virus antigen (Rb). MATERIALS AND METHODS: Swiss albino mice were immunized with aqueous-alcoholic SBTE (100 mg/kg body weight) or algel (aluminium hydroxide gel) with or without Rb (5% v/v). After priming, booster was administered on day 14. Rabies virus neutralizing antibody (RVNA) titers were estimated by rapid fluorescent focus inhibition test in sera samples collected on days 7, 14, 21, 28 and 35. Effect of adjuvant administration on cytotoxic T lymphocytes (CTLs), memory T cells, plasma and CD11c+ cells was studied by flow cytometry. In vitro hemolysis was assayed in human RBC. RESULTS: RVNA titers were significantly enhanced (p < 0.05) after booster administration in mice immunized with SBTE + Rb as compared to the controls. In combination, SBTE, algel and Rb, enhanced the RVNA titers. CTLs significantly increased (p < 0.05) in SBTE + Rb immunized mice. Memory T cells and plasma cells were 27.9 and 15.9%, respectively, in SBTE + Rb immunized mice as compared to that of 20.3 and 11.3%, respectively, in Rb immunized group. SBTE + Rb enhanced peritoneal CD11c+ cells (25.8%) as compared to 9.4% cells in Rb immunized mice, showed 3.2-fold increment in LPS induced IL-1ß. No RBC hemolysis was observed with SBTE. CONCLUSIONS: This study demonstrates the potential adjuvant activity of SBTE with Rb by increasing RVNA titers and CTL response.

Acute and subchronic toxicity studies of seabuckthorn (Hippophae rhamnoides L.) oil in rodents.

Seabuckthorn (Hippophae rhamnoides L.) has been traditionally used as medicine and nutritional supplement for a long period of time. However, information on the systemic toxicity and safety evaluation of seabuckthorn and its extracts is still scarce. The purpose of this study was to evaluate the potential toxicity of seabuckthorn oil by an acute oral toxicity study in mice and a 90-day repeated oral toxicity study in rats. No mortality or signs of toxicity was observed in mice treated with 20 mL/kg body weight seabuckthorn oil in the acute toxicity study. In the subchronic toxicity study, 80 Sprague-Dawley rats (10 animals per sex per treatment group) were administrated with 10, 5, 2.5 and 0 (control) mL/kg body weight of seabuckthorn oil daily for 90 days by gavage. There were no signs of toxicity and treatment-related changes in rats treated with seabuckthorn oil on mortality, body and organ weights, food consumption, blood biochemistry and hematology, gross necropsy and histopathological examinations. Based on the finding of this study, the maximum tolerated dose of seabuckthorn oil was >20 mL/kg for mice for acute toxicity study, and the no-observed-adverse-effect level was 10 mL/kg body weight for both male and female rats for 90-day toxicity study.

Effect of Sea-Buckthorn (Hippophaë rhamnoides L.) Pulp Oil Consumption on Fatty Acids and Vitamin A and E Accumulation in Adipose Tissue and Liver of Rats.

An in vivo experiment was conducted to determine the effect of sea-buckthorn pulp oil feeding on the fatty acid composition of liver and adipose tissue of Wistar rats and the liver accumulation of retinol, its esters and α-tocopherol. For a period of 28 days, rats were given a modified casein diet (AIN-93) in which sea-buckthorn pulp oil, soybean oil and pork lard were used as sources of fat. Compared to the other fat sources, sea-buckthorn pulp oil was the most abundant in C16 fatty acids, carotenoids (mainly ß-carotene) and tocopherols (mainly α-tocopherol). Its consumption was reflected in an increased share of palmitoleic acid in adipose tissue and the liver and an increased level of retinol in liver tissues (this was not observed for its esters). Although the type of fat did not have a significant effect on the average content of α-tocopherol in the liver, the variation of saturation of this tissue with α-tocopherol was the lowest when rats were fed a diet containing sea-buckthorn oil. This experiment indicates the possibility of affecting adipose tissue and liver by a diet.

Phytochemical characterization and evaluation of anticataract potential of seabuckthorn leaf extract.

OBJECTIVE: This study was undertaken to carry out phytochemical characterization of aqueous extract of Seabuckthorn (SBT, Hippophae rhamnoides L.) leaves and evaluation of its therapeutic role in oxidative stress-induced cataract in isolated goat lenses using Vit. E as reference compound. ANIMAL STUDIED: A total of 42 goat eye lenses were used in the present study. PROCEDURE: Seabuckthorn leaf extract was characterized by total phenol content estimation and HPLC analysis of quercetin and catechin. Further, cataract was induced in goat lenses using hydrogen peroxide (H2O2) and anticataract activity was evaluated using the extract in the dose range of 100, 200, 500, and 1000 µg/mL concentrations through estimation of biochemical markers such as superoxide dismutase (SOD), reduced glutathione (GSH), and malonaldehyde (MDA). RESULTS: The results of the phytochemical characterization showed the total phenol content of the extract to be 365 mg/g in terms of gallic acid equivalents. Quercetin and catechin were estimated to be 0.01 and 0.12% w/w, respectively. In biochemical analysis, H2O2 introduction resulted in a decrease in SOD (approximately 85%) and GSH (approximately 63%) contents and an increase in MDA content (approximately 300%). The decreased levels of SOD and GSH were significantly restored in experimental groups receiving 500 and 1000 1g/mL of SBT extract. All the experimental groups showed significantly reduced MDA level in all the doses. CONCLUSION: Aqueous extract of SBT leaves showed the potential to delay onset and/or progression of cataract, at least during in vitro conditions. Results indicate the possibilities of evaluating this extract for its use as anticataract agent during in vivo conditions.

[Rapid analysis of compounds in leaves of Chinese seabuckthorn and Tibetan seabuckthorn by UPLC/Q-TOF-MS].

The study is aimed to analyze the chemical components in leaves of Chinese seabuckthorn and Tibetan seabuckthorn qualitatively and compare the differences between them by using ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS).The chromatographic separation of the components was achieved ona Waters ACQUITY UPLC-T3 column (2.1 mm×100 mm, 1.7 µm）using gradient mobile phase consisting of acetonitrile (A) and aqueous solution (B). The identification of the separated compounds was performed on atandem mass spectrometry (MS/MS)by fragmentation patterns under the negative electrospray ionization. The parameters of ion source were as follows：capillary voltage, 2 000 V; Cone voltage, 40 V. The ion source temperature, 100 ℃; collision gas argon; sheath gas flow rate, 900 L•h⁻¹; sheath gas temperature, 450 ℃. Through the analysis of mass spectrometry data and with the help of literature data, a total of 35 compounds were detected and most of them were flavonoids. Among these compounds, 29 were common components for the two species, two components were unique to Chinese seabuckthorn and 4 were characteristic components of Tibetan seabuckthorn. The results indicated that the compositions of the two kinds of seabuckthorn leaves were quite similar. It is also demonstrated that UPLC/Q-TOF-MS method could be applied to rapidly and effectively analyze and speculate the compounds in leaves of Chinese seabuckthorn and Tibetan seabuckthorn.

[Study on HPLC fingerprint and quantitative analysis of 3 components of Tibetan medicine Gyantse Seabuckthorn].

This study is to establish an HPLC fingerprint and quantitative analysis of 3 components of Gyantse Seabuckthorn from different producing areas.The separation was developed on Shimadzu InertSustain C18column (4.6 mm × 250 mm,5 µm) by gradient elution with acetonitrile and 0.2% phosphoric acid water as mobile phase at a flow rate of 1.0 mL•min ⁻¹; the detection wavelength was set at 360 nm and column temperature was set at 30 ℃. The data calculation was performed with similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine(Version 2004A).The fingerprints of 10 batches of Gyantse Seabuckthorn were carried out by similarity comparison, and 12 chromatographic peaks were extracted as the common peaks of fingerprint, of which three main active ingredients were successfully determined. This is the first established fingerprint and multi-component quantitative determination of Gyantse Seabuckthorn by using HPLC. This method has good precision stability and repeatability that could provide basis for quality control and evaluation of Gyantse Seabuckthorn.

[Genetic and chemical discrimination of traditional Tibetan medicine seabuckthorn based on DNA barcode and ¹H-NMR metabolic method].

To differentiate three medicinal Hippopahe species of seabuckthorn, a combined genetic and chemical identification method was established in this study. ITS2 and psbA-trnH were tested for identification of 3 species of seabuckthorn. Detection of the kimura 2-parameter (K2P) distance, the neighbor-joining (NJ) tree and the barcoding gap were used to assess the identification efficiency. ¹H-NMR based metabolic method was applied to acquire the profile of metabolites. PCA was used to analysis the metabolite data. The results indicated that DNA barcode combined ¹H-NMR based metabolic method is a powerful tool for the identification of 3 medicinal Hippopahe species of seabuckthorn. The finding demonstrated that different genetic variation and chemical constituents existed among 3 medicinal Hippopahe species of seabuckthorn. The combined identification method will improve the reliability of species discrimination and could be applicable to much other ethnic medicine which has various origins in China.

Optimization of seabuckthorn fruit yogurt formulation using response surface methodology.

The response surface methodology was used to optimize the formulation of seabuckthorn fruit yogurt. The independent variables were proportions of seabuckthorn fruit syrup and skimmed milk powder. The responses were counts of Streptococcus thermophilus and Lactobacillus bulgaricus, taste and viscosity of the product. Statistical analysis revealed that fruit syrup and skimmed milk powder significantly affected all the responses. Contour plots for each response were used to generate an optimum area by superimposition. Optimum formulation conditions of fruit syrup (15 %) and skimmed milk powder (12.5 %) are recommended for the blend formulation yielding an acceptable and good quality seabuckthorn fruit yogurt. Model validation was conducted using separate experiments at optimum conditions. The experimental values were found to be in close agreement to the predicted values and were within the acceptable limits indicating the suitability of the model in predicting quality attributes of seabuckthorn fruit yogurt. The resultant product also exhibited more amounts of fat, protein, carbohydrate and antioxidants viz., vitamin C, E, carotenoids, phenols and anthocyanins when compared to a commercial one.