#Hydrogen Peroxide manufacturers in USA

[stellarexports]

Searching for Hydrogen Peroxide Manufacturer in USA?

Hydrogen Peroxide is highly flammable and exhibits excellent water absorption capacity and binding properties. Stellar Exports is a leading exporter of Hydrogen Peroxide and as a trusted and reliable Hydrogen Peroxide supplier in usa, our products are indispensable and appreciated by customers worldwide. For more information, visit our website.

[palvichemical]

Catalyzing Success with Hydrogen Peroxide Excellence | palvichemical

Elevate your chemical solutions with Palvichemical, your go-to partner for Hydrogen Peroxide brilliance. As a trusted Hydrogen Peroxide exporter and distributor in the USA, we transcend boundaries, delivering excellence in every drop. With precision manufacturing and unwavering commitment, Palvichemical stands as a leading hydrogen peroxide manufacturer and hydrogen peroxide supplier, setting new benchmarks in Hydrogen Peroxide innovation.

Choose us for purity, reliability, and a catalyst for success in your industrial endeavors.

[trendingreportz]

Merchant Hydrogen Market - Forecast(2024 - 2030)

Merchant Hydrogen Market Overview

The merchant hydrogen market size is forecast to reach US$124 billion by 2027 after growing at a CAGR of 7.2% during 2022-2027. Merchant hydrogen refers to the production of hydrogen in hydrogen production plants and selling it to several end-use industries for various industrial processes. Merchant hydrogen is extensively used in the oil and refinery industry in the manufacturing of gasoline, diesel, jet fuels, and various refining processes such as hydrocracking, hydrotreating, etc. The oil and refinery industry expanding globally and this will drive the growth of the market in the forecast period. For instance, according to the January 2020 data by the International Energy Agency, biofuel production is expected to increase fourfold from around 2 mboe/d in current times to almost 8 mboe/d by 2040. Furthermore, merchant hydrogen finds its broad uses in the production of various chemicals such as ammonia, methanol, cyclohexane, hydrogen peroxide, hydrochloric acid, etc. The chemical industry is booming globally and this will contribute to the growth of the market in the forecast period. For instance, according to the 2020 Chemical Industry Outlook Report by BASF, chemical production globally is expected to increase by 4.4% in 2021. The water electrolysis process is projected to witness the highest demand in the forecast period. Steam methane reforming will witness significant demand in the market. Lack of better infrastructure might hinder the growth of the market in the forecast period.

COVID-19 Impact

The merchant hydrogen market was moderately affected during the COVID-19 due to disruption in the supply chain and temporary shutdown of plants surfaced in the market. Market players implemented new work strategies to maintain a stable business operation. Despite executing new business strategy plans, businesses were affected due to the severity of the COVID-19 pandemic. As per the 2020 Annual Report by Air Products and Chemicals, Inc., operations were disrupted due to the pandemic, leading to reduced demand for industrial gas products in the company’s merchant business. The market witnessed decent demand towards the end of 2020. Going forward, the market is projected to have a positive growth rate owing to expansion in hydrogen production plants and increasing demand for hydrogen in end-use industries such as the oil and refinery industry, chemical industry, and automobile.

Request Sample

Merchant Hydrogen Market Report Coverage

The report: “Merchant Hydrogen Market Forecast (2022-2027)”, by IndustryARC, covers an in-depth analysis of the following segments of the Merchant Hydrogen Industry. 

By Process: Steam Methane Reforming, Water Electrolysis, Coal Gasification, Others

By End Use Industry: Oil and Refinery, Chemical, Food and Beverage, Construction, Residential, Commercial, Office, Hotels and Restaurants, Concert Halls and Museums, Educational Institutes, Automobile, Passenger Vehicle, Commercial Vehicle, Light Commercial Vehicle, Heavy Commercial Vehicle, Industrial, Electrical and Electronics, Agriculture, Paper, Others

By Geography: North America (USA, Canada, Mexico), Europe (UK, Germany, France, Italy, Netherlands, Spain, Russia, Belgium, Rest of Europe), Asia Pacific (China, Japan, India, South Korea, Australia, and New Zealand, Indonesia, Taiwan, Malaysia, Rest of Asia Pacific), South America (Brazil, Argentina, Colombia and Rest of South America), and RoW (Middle East and Africa).

Key Takeaways:

Water electrolysis is leading the merchant hydrogen market. This production process offers a robust option for carbon-free hydrogen production from renewable resources, making it a desirable choice among manufacturers.

The chemical industry will drive the growth of the market in the forecast period. According to the June 2020 data by the American Chemistry Council, net exports of chemicals will touch $37 billion by 2025.

The Asia-Pacific region will witness the highest demand for merchant hydrogen in the forecast period owing to the expanding oil and refinery industry in the region. According to the data by India Brand Equity Foundation, oil demand in India is expected to witness a 2x growth to reach 11 million barrels by 2045.

Inquiry Before Buying

Merchant Hydrogen Market - By Process

Water electrolysis dominated the merchant hydrogen market in 2021. This process helps in the production of carbon-free hydrogen from nuclear and renewable resources. Water electrolysis is done with the help of a unit called an electrolyzer. Proton exchange membrane (PEM) electrolyzer and alkaline electrolyzer are the two types of electrolyzers usually implemented. Water electrolysis involving electrolyzers is considered a mature technology compared to other processes. Owing to such diverse properties, market players are engaging in expanding the portfolio of water electrolysis in the market. For instance, in January 2021 Cummins Inc., installed a new 20-megawatt PEM electrolyzer system in Canada to generate green hydrogen. This is the world’s largest proton exchange membrane electrolyzer in operation. Such developments in the water electrolysis process will increase its demand in the forecast period. The steam methane reforming process is projected to witness significant demand in the market.

Merchant Hydrogen Market - By End Use Industry

Oil and refinery dominated the merchant hydrogen market in 2021 and is growing at a CAGR of 7.5% in the forecast period. Merchant hydrogen produced through processes such as steam methane reforming and water electrolysis is massively used in the production of high-quality lubricating oils and various refining processes in the oil and refinery industry. The oil and refinery industry expanding globally and this will lead to the growth of the market in the forecast period. For instance, as per the data by India Brand Equity Foundation, diesel demand in India is expected to double to 163 MT by 2029-30. Similarly, according to the December 2021 report by Energy Information Administration (EIA), total production of crude oil stood at 11.7 million b/d in November which is projected to touch an average of 12.1 million b/d in the fourth quarter of 2022. Such massive growth in the oil and refinery industry will increase the higher implementation of merchant hydrogen in the forecast period and this will contribute to the growth of the market in the forecast period. The chemical industry will drive the growth of the market significantly in the forecast period.

Schedule a Call

Merchant Hydrogen Market - By Geography

The Asia-Pacific region held the largest market share in the merchant hydrogen market in 2021 with a market share of up to 34%. The high demand for merchant hydrogen is attributed to the expanding oil and refinery industry in the region. For instance, as per the March 2021 report by Energy Information Administration (EIA), China’s refineries refined more crude oil for most of 2020 compared to US refineries. Similarly, as per the data by India Brand Equity Foundation, the Indian government has allowed 100% Foreign Direct Investment in upstream and private sector refining projects. Such increasing growth in the region’s oil and refinery industry will stimulate the higher uses of merchant hydrogen in the forecast period. The North American region is projected to witness significant demand for merchant hydrogen in the forecast period.

Merchant Hydrogen Market Drivers

The booming chemical industry will drive the growth of the market

Merchant hydrogen is deeply associated with the chemical industry as it is implemented in the production of several chemicals such as ammonia, methanol, cyclohexane, hydrogen peroxide, among others. The chemical industry is booming globally and this will contribute to the growth of the market in the forecast period. For instance, according to the June 2021 Mid-Year US Chemical Industry Outlook report, chemical volumes and shipments in the US are expected to increase by 3.2% and 8.2% respectively in 2022. Similarly, according to the report by India Brand Equity Foundation (IBEF), the domestic chemical sector's small and medium enterprises are projected to witness 18-23% revenue growth in FY22. This huge growth in the global chemical industry will increase the higher uses of merchant hydrogen and this, in turn, will contribute to the market’s growth in the forecast period.

Expanding oil and refinery industry will contribute to market’s growth

Merchant hydrogen is massively used in the oil and gas industry. Processes such as steam methane reforming and water electrolysis are implanted for the production of merchant hydrogen which is later used in the oil and refinery industry for the production of gasoline, diesel, jet fuels, and various high-quality lubricating oils. The oil and gas industry expanding globally and this will drive the growth of the market. For instance, as per the data by India Brand Equity Foundation, the oil and gas sector’s installed provisional refinery capacity accounted for 246.90 MMT as of September 2021 and Indian Oil Corporation was the largest domestic refiner with a capacity of 69.7 MMT. Similarly, according to the statistics by the International Energy Agency, global refining activity is expected to jump by 2.4 mb/d in 2022, and demand for oil will return to pre-pandemic levels by the end of 2022. Such massive expansion in the oil and refinery industry globally will augment the higher uses of merchant hydrogen and this will drive the growth in the forecast period.

Buy Now

Merchant Hydrogen Market Challenges

Lack of better infrastructure might hamper the market’s growth

The infrastructure barrier has been a key challenge in the merchant hydrogen and this might hinder the market’s growth in the forecast period. The distribution and storage of merchant hydrogen are limited and vary, restricting its widespread adoption. For instance, as per the June 2019 report by Energy Information Administration, better planning and coordination are required among government and industries for the proper delivery and storage of merchant hydrogen. Similarly, as per the August 2021 data by the Department for Business, Energy, and Industrial Strategy of the UK, better coordination is required for the wider rollout of hydrogen. Such infrastructure concerns associated with merchant hydrogen might limit the market’s growth.

Merchant Hydrogen Industry Outlook

Investment in R&D activities, acquisitions, product and technology launches are key strategies adopted by players in the merchant hydrogen market. Major players in the merchant hydrogen market are:

Airgas, Inc.

Air Products and Chemicals, Inc.

Praxair, Inc.

Linde plc

Air Liquide

Uniper

Engie SA

FuelCell Energy, Inc.

Cummins Inc.

Others

Recent Developments

In May 2020, Engie collaborated with Neste to produce renewable hydrogen on a large scale for the production of high-quality biofuels. Such collaborations will contribute to the growth of the market in the forecast period.

[chemanalystdata]

Hydrogen Peroxide Prices Trend, Monitor, News, Analytics and Forecast | ChemAnalyst

Hydrogen Peroxide Prices: During the Quarter Ending December 2023

North America:

During Q4 2023, the Hydrogen Peroxide market in North America underwent significant shifts influenced by several factors. Notably, a bearish trend prevailed due to reduced demand from key sectors like textiles and paper industries, resulting in decreased purchasing activity and overall market dynamics.

Moreover, the market experienced high supply levels, leading to an oversupply situation and necessitating inventory management by suppliers. Notably, no specific plant shutdown incidents were reported by market participants during this period. Specifically in the United States, significant price changes occurred as the market followed a descending trajectory. This was largely attributed to shifting demand patterns and decreased interest from industries such as textiles and paper.

Furthermore, lowered costs of essential feedstocks like sulfuric acid and natural gas impacted pricing strategies, ensuring competitiveness and viability for producers. Price-wise, there was a notable -13% decrease compared to the same quarter last year and a -5% decrease from the previous quarter. However, specific price comparisons between the first and second halves of the quarter were not provided. The latest Hydrogen Peroxide price FOB Illinois in the USA for the current quarter stands at USD 585/MT, influenced by reduced demand, oversupply, and lowered feedstock costs.

Get Real Time Prices of Hydrogen Peroxide: https://www.chemanalyst.com/Pricing-data/hydrogen-peroxide-1169

APAC:

In Q4 2023, the Hydrogen Peroxide market in the APAC region navigated through several influential factors. Despite stable supply conditions with a moderate product influx from manufacturing units, a bearish trend prevailed due to high supply levels, particularly in domestic ports, leading to an oversupply situation. Continuous material inflows from the Chinese market contributed significantly to this oversupply.

Despite low demand and subdued purchasing activities, the region maintained adequate Hydrogen Peroxide supplies. In Japan, prices experienced a bearish trend, with a 2% decrease compared to the previous month, attributed to high supply levels and low domestic demand. Price-wise, there was a significant 16% decrease compared to the same quarter of the previous year and a slight 6% decrease from the previous quarter. The Hydrogen Peroxide price CFR Tokyo in Japan for the current quarter is USD 480/MT, reflecting stable supply conditions amidst a bearish market trend.

Europe:

During Q4 2023, the European Hydrogen Peroxide market faced various factors influencing prices and market dynamics. An oversupply situation prevailed due to high availability from manufacturing units in countries like the Netherlands, Germany, and Belgium, resulting in surplus material in the market. Businesses reported sufficient inventory to meet demand, further contributing to the bearish market sentiment.

Significant price changes were observed in Belgium, with a 1.4% decrease in FOB Antwerp prices throughout the quarter. This decline was influenced by satisfactory supply and reduced purchasing activities, leading to lower bidding costs. Overall, there was an -8% decrease in prices from the previous quarter, indicating a slight downward trend.

The Hydrogen Peroxide market in Europe witnessed a bearish scenario characterized by high supply and low demand. Belgium experienced price declines due to satisfactory supply and reduced purchasing activities. Prices significantly decreased compared to the same quarter of the previous year, with a slight decline from the previous quarter. The latest Hydrogen Peroxide price FOB Antwerp in Belgium for the current quarter is USD 319/MT.

Get Real Time Prices of Hydrogen Peroxide: https://www.chemanalyst.com/Pricing-data/hydrogen-peroxide-1169

Contact Us:

ChemAnalyst

GmbH - S-01, 2.floor, Subbelrather Straße,

15a Cologne, 50823, Germany

Call: +49-221-6505-8833

Email: sales@chemanalyst.com

Website: https://www.chemanalyst.com

[nurseshannansreviews]

💙 My favorite place to get tp is @bumroll_ because it's a 100% sustainable, recyclable toilet paper that reinvests in the environment! Love how it's chlorine-free and plastic wrap-free! It's also a healthy choice and better for your #skin because it's free of perfume-free and hydrogen peroxide whitening! Cute packaging and in each roll you get 400 soft, strong sheets, while staying eco-friendly.

💙 Bumroll is ethically made in the USA supporting local jobs and communities. They also have an Eco Commitment: by planting a tree in the USA for every box sold. Try bumroll as a subscription or a one time purchase. Get $3 off your first shipment with coupon code: INSTAGRAM. https://joinbumroll.com

[wxwlchem]

Sodium Percarbonate Manufacturer in China

Wuxi Wanli Chemical Co; Ltd is one of the leading manufacturer of Sodium Percarbonate in China. Sodium Percarbonate is an additional compound of sodium carbonate and hydrogen peroxide. It is frequently used in washing bleaching, detergent powder for clothes, kitchen, bathroom, Carpet stains. We export our material to Japan, south korea, Israel, USA, etc.

[diamondbeautycompany]

Shop teeth whitening gel pens online

Teeth whitening gel pens are a popular at-home teeth whitening option that comes in a convenient pen-like applicator. These teeth whitening gel pens typically contain a bleaching gel that is applied directly to the teeth using the built-in brush or pen tip. The gel is usually formulated with hydrogen peroxide or carbamide peroxide, which are active ingredients that help break down stains and whiten the teeth. Gently brush the gel onto the front surface of each tooth you wish to whiten. Be careful to avoid applying the gel to your gums or any areas other than the teeth. Once the gel is applied, it's generally recommended to avoid eating or drinking for about 30 minutes to allow the gel to work effectively. This professional teeth whitening pen offers an easy and effective way to brighten smiles. Order teeth whitening gel in bulk. Diamond Beauty Company USA is a proud manufacturer and distributor of dental whitening gel options. Enjoy our extra strength formula, safe for sensitive teeth. Shop today!

[speediipl]

Sodium Permanganate Manufacturers in India

Sodium permanganate is the inorganic compound with the formula NaMnO4. It is closely related to the more commonly encountered potassium permanganate, but it is generally less desirable, because it is more expensive to produce. It is mainly available as the monohydrate. This salt absorbs water from the atmosphere and has a low melting point. Being about 15 times more soluble than KMnO4, sodium permanganate finds some applications where very high concentrations of MnO4− are sought. Because of its high solubility, its aqueous solutions are used as etchants in printed circuitry. It is gaining popularity in water treatment for taste, odor, and zebra mussel control. The V-2 rocket used it in combination with hydrogen peroxide to drive a steam turbopump.

[go-redgirl]

Coronavirus: Distilleries Around the Country Divert Operations to Produce Hand Sanitizer

Distilleries nationwide are changing their operations to produce more hand sanitizer to combat coronavirus, the president announced at the White House on Saturday.

President Donald Trump used Pernod Ricard USA as an example of this development:

This is really an example where we are repurposing alcohol. They went out and repurposed their alcohol production capabilities in Arkansas, Kentucky, and West Virginia to make hand sanitizer, and it’s a big difference.

“And they’ve been unbelievable,” Trump said.

“Their first delivery will go out on Tuesday,” Trump said. “It’s going to go to various states.”

“They’re going to start I think in New York and they’re going to work their way around,” Trump said. “They’re making a tremendous amount of hand sanitizer.”

The Centers for Disease Control and Prevention requires hand sanitizers to be at least 60 percent alcohol to make them effective.

Two distillers located in the nation’s capitol are also stepping up to the plate.

“We’ve got all this high-proof booze around, why not put it to use?” Cotton & Reed co-founder Jordan Cotton said in a DCist article.

“You can’t get [hand sanitizer], so therefore you have to make it. In order to make it, you have to have good, high-proof spirit,” Republic Restoratives founder and CEO Pia Carusone said in the same article. “These sort of crises demand innovation.”

The DCist reported on how that innovation is taking place:

Republic Restoratives’ cleanser is made from 140-proof ethyl alcohol, vegetable glycerine, and hydrogen peroxide. Cotton & Reed’s is made with 66 percent ABV rum (the only spirit they manufacture) vegetable glycerine, and essential oils like bergamot, lemon, and orange.

Cotton & Reed is also handing out free groceries and hand sanitizer to service and hospitality workers affected by the coronavirus pandemic, as part of the Friends and Family Meal initiative—a nonprofit that provides food from local farmers to those in need.

“The oils take over [the aroma], but there’s a certain rum-iness to it,” Cotton said.

An order from D.C. Mayor Muriel Bowser resulted in both distilleries shuttering their tasting rooms and bars, but alcohol retailers can still sell their products through delivery services or at their retail counters.

U.S. News and World Report also reported on this trend:

Distilleries must follow strict rules for the products they sell under the Alcohol and Tobacco Tax and Trade Bureau, but many are working with state officials to legally make the switch to producing hand sanitizer.

The recipe “starts with ethanol, which is what we have plenty of in the distillery, then you add glycerin, hydrogen peroxide water and you mix it up,” Scott Jendrek, owner of Patapsco Distilling Co. in Sykesville, Maryland, told a local NBC News affiliate.

In Vermont, Smugglers’ Notch Distillery plans to release a hand sanitizer this week at its Waterbury and Jeffersonville locations. According to NBC News, part of the proceeds will be donated to the state’s efforts to combat the outbreak.

“I know I have a unique opportunity to help out a little bit and keep my staff employed,” Jeremy Elliott, the company’s co-owner, said in the article.

And in Colorado, Spirit Hound Distillers is storing its hand sanitizer solution in spray bottles donated from a local skin and body care company. Spirit Hound has donated its hand sanitizer to its local fire department, businesses, and a local home health care nurse.

“Within two hours, we filled 1,000 four-ounce bottles and a bunch of gallon jugs and we are giving them out,” Craig Engelhorn, the company’s head distiller, told the Denver Post.

“Nearby in Boulder, J&L Distilling has also delivered handmade hand sanitizer to its local fire department and some senior care organizations,” World Report reported.

“There’s a need in the community and I’m uniquely positioned to fill it,” co-founder Seth Johnson said.

READ MORE STORIES ABOUT:

Health Politics coronavirus distilleries Donald Trump Hand Sanitizer Pernod Ricard USA

[jobssok]

GLO Brilliant Teeth Whitening Device Kit with Patented Heat

GLO Brilliant Teeth Whitening Device Kit with Patented Heat

Price: (as of – Details) From the manufacturer Whitening Gel Treatment Vials Sensitivity Free Refreshing Mint Flavor Made in the USA Recyclable Vegan, Cruelty-Free, Gluten-Free Patented technology Vials have (6%) hydrogen peroxide GLO Good Foundation Established by Dr. Jonathan B. Levine and co-founder Stacey Levine. GLO Good provides free dental care and life-changing smile transformations to…

[palvichemical]

Industry Insights:  Navigating Challenges as a Hydrogen Peroxide Professional

Welcome to Palvi Industries Ltd., your trusted partner in the dynamic world of hydrogen peroxide.  As a leading Hydrogen Peroxide Manufacturer, Exporter, Distributor, and Supplier, we understand the challenges and triumphs that come with being a part of this essential industry.  Join us as we delve into the nuances of navigating challenges as a Hydrogen Peroxide professional, exploring the critical role we play as a Hydrogen Peroxide Exporter in the USA, Distributor, Manufacturer, and Supplier.

Introduction:  The Essence of Hydrogen Peroxide

Hydrogen peroxide, a versatile chemical compound, is a staple in numerous industries, serving as a powerful oxidizer, bleaching agent, and disinfectant.  As a Hydrogen Peroxide Manufacturer, we recognize the pivotal role this compound plays in various applications, from healthcare and cosmetics to wastewater treatment and beyond.

The Challenges of Manufacturing Hydrogen Peroxide

Being a Hydrogen Peroxide Manufacturer entails overcoming a myriad of challenges.  From ensuring optimal production processes to adhering to stringent quality standards, the journey from raw materials to the final product is intricate.  As a Hydrogen Peroxide Supplier, Palvi Industries Ltd. places a premium on precision and excellence in manufacturing, addressing challenges head-on to deliver top-notch products.

Global Outreach:  Navigating the Export Landscape

Palvi Industries Ltd. takes pride in being a Hydrogen Peroxide Exporter in the USA.  Exporting hydrogen peroxide globally involves tackling logistical complexities, compliance with international regulations, and maintaining product integrity during transit.  Our commitment to quality and reliability positions us as a trusted partner for businesses seeking a Hydrogen Peroxide Supplier with a global reach.

Distributor Dynamics:  Challenges in the Distribution Network

As a Hydrogen Peroxide Distributor in the USA, we understand the intricacies of the distribution network.  Timely deliveries, inventory management, and ensuring customer satisfaction are paramount.  Palvi Industries Ltd.  employs state-of-the-art distribution strategies to overcome these challenges, ensuring that our clients receive their hydrogen peroxide products efficiently and seamlessly.

Sustainable Solutions:  Meeting Environmental Challenges

The hydrogen peroxide industry faces increasing scrutiny regarding environmental impact.  As a responsible Hydrogen Peroxide Supplier and Manufacturer, Palvi Industries Ltd. is dedicated to implementing sustainable practices.  We continuously invest in research and development to explore eco-friendly alternatives and reduce our carbon footprint, contributing to a cleaner, greener future.

Innovations in Hydrogen Peroxide:  Staying Ahead of the Curve

Remaining at the forefront of the industry requires a commitment to innovation.  Palvi Industries Ltd. embraces cutting-edge technologies and explores novel applications for hydrogen peroxide.  As a Hydrogen Peroxide Distributor and Exporter, we take pride in providing our clients with the latest advancements, ensuring that they stay ahead in their respective fields.

Hydrogen Peroxide in Healthcare:  A Critical Perspective

The healthcare industry relies on hydrogen peroxide for various applications, from wound care to surface disinfection.  Palvi Industries Ltd. acknowledges the critical role we play as a Hydrogen Peroxide Supplier in supporting the healthcare sector, especially during challenging times.  Our commitment to quality and reliability ensures that healthcare professionals can trust our products for their disinfection needs.

The Palvi Advantage:  Reliability, Quality, and Customer Satisfaction

Navigating challenges as a Hydrogen Peroxide professional requires a reliable partner.  Palvi Industries Ltd. distinguishes itself through unwavering commitment to quality, adherence to international standards, and a customer-centric approach.  Whether you are seeking a Hydrogen Peroxide Manufacturer, Exporter, Distributor, or Supplier, Palvi Industries Ltd. is your trusted ally in the industry.

Conclusion:  Charting the Course for a Hydrogen Peroxide Future

In conclusion, being a Hydrogen Peroxide professional is a journey filled with challenges and opportunities.  Palvi Industries Ltd. embraces these challenges, leveraging our expertise as a Hydrogen Peroxide Manufacturer, Exporter, Distributor, and Supplier to contribute to the success of our clients worldwide.  As we navigate the dynamic landscape of the hydrogen peroxide industry, we remain dedicated to innovation, sustainability, and excellence.  Partner with Palvi Industries Ltd. for a reliable and rewarding hydrogen peroxide experience.

[agexpharma-123]

GLYCIDOL: MEANING AND ITS APPLICATIONS

I.                   INTRODUCTION:

Glycidol is an epoxide and an alcohol, and as such is a highly reactive compound. It is miscible with water but also reacts with it. It decomposes when distilled at atmospheric pressure. In 1909, chemist Nikolai Prilezhaev at the Warsaw University of Technology prepared glycidol by epoxidizing allyl alcohol with peroxybenzoic acid. This reaction was a breakthrough at the time and became known as the Prilezhaev oxidation. Glycidol is still manufactured in much the same way; but in 2018, a group of companies built a pilot plant in Teesside, UK, to make it via a “green” process.

Glycidol has a chiral center, but it is generally produced and used as the racemic mixture. Glycidol (556-52-5 ) manufacturer USA supplies only the best and high Glycidol which is also used as a chemical intermediate and used in the production of detergents, healthcare products, and industrial paints and coatings.

 II.                CHEMICAL AND PHYSICAL DATA

 1.      Nomenclature

Chem. Abstr. Serv. Reg. No.: 556-52-5 Deleted CAS Reg. Nos: 61915-27-3; 98913-54-3 Chem. Abstr. Name: Oxiranemethanol

IUPAC Systematic Name: 2,3-Epoxypropan-1-ol

Synonyms: Allyl alcohol oxide; epihydrin alcohol; 1,2-epoxy-3-hydroxypropane; 2,3-epoxy-1-propanol; (±)-2,3-epoxy-1-propanol; glycide; (±)-glycidol; (RS)- glycidol; dl-glycidol; glycidyl alcohol; hydroxy-1,2-epoxypropane; 1-hydroxy-2,3- epoxypropane; 2-(hydroxymethyl)oxirane; 3-hydroxypropylene oxide; oxiranylmethanol; racemic glycidol

   2.      Structural and molecular formulae and relative molecular mass

                                                                      O              

                                                               H2C         C    CH2  OH      

                                                                            H

                                                                                                                        C3H6O2                                                                                Relative molecular mass: 74.08

 3. Chemical and physical properties of the pure substance

(a) Description: Colourless, odourless liquid (Sienel et al., 1987)

(b) Boiling-point: 162 °C (decomposes) (Verschueren, 1996)

(c) Melting-point: –54 °C (Verschueren, 1996)

(d) Density: 1.143 g/cm3 at 25 °C (Lide & Milne, 1996)

(e) Spectroscopy data: Infrared (prism [15765]; grating [28381]), nuclear magnetic resonance (proton [18790]) and mass spectral data have been reported (Sadtler Research Laboratories, 1980; Lide & Milne, 1996)

(f) Solubility: Miscible in all proportions in water, alcohols, ketones, esters, ethers and aromatics; almost insoluble in aliphatic hydrocarbons (Sienel et al., 1987)

(g) Volatility: Vapour pressure, 120 Pa at 25 °C (American Conference of Governmental Industrial Hygienists, 1999); relative vapour density (air = 1), 2.15 (Verschueren, 1996)

(h) Stability: Flash-point, 71 °C (Sienel et al., 1987); reacts vigorously with strong caustic soda, strong sulfuric acid and with anhydrous metal halides, such as stannic and ferric chlorides (Dixie Chemical Co., 1995)

(i) Octanol/water partition coefficient (P): log P, –0.95 (Hansch et al., 1995)

(j) Conversion factor1: mg/m3 = 3.03 × ppm

 4.      Technical products and impurities

Glycidol is commercially available with a minimum purity of 95% and a maximum water content of 1% (Dixie Chemical Co., 1999). Trade names for glycidol include: Epiol OH.

5.      Analysis

Glycidol can be determined in workplace air by adsorbing the air sample on charcoal, desorbing with tetrahydrofuran and analysing by gas chromatography with flame ionization detection (Eller, 1994)

III.             PRODUCTION

Glycidol  is commercially produced by two methods:

(1) epoxidation of allyl alcohol with hydrogen peroxide and a catalyst (tungsten or vanadium); and

(2) reaction of epichlorohydrin with caustic .

Information available in 1999 indicated that glycidol was manufactured by two companies in Japan, and one company each in Germany and the United States (Chemical Information Services, 1999).

IV.             USE

In 1956, glycidol was only used for research purposes, but by 1978 it was used in the preparation of glycerol, glycidyl ethers, esters and amines in the pharmaceutical industry (Proctor & Hughes, 1978) and as a sterilant in pharmaceuticals (Ivashkiv & Dunham, 1973). Calculated from: mg/m3 = (relative molecular mass/24.45) × ppm, assuming a temperature of  25 °C and a pressure of 101 kPa Glycidol has become an important intermediate for the production of functional epoxides. For example, reaction of phosgene with glycidol yields 2,3-epoxypropyl chloroformate. Reaction of glycidol with isocyanates affords the commercially important glycidyl urethanes (Sienel et al., 1987). It is used as an intermediate in the production of pharmaceuticals, as an additive for synthetic hydraulic fluids and as a reactive diluent in some epoxy resin systems (Hooper et al., 1992; American Conference of Governmental Industrial Hygienists, 1999). It is a stabilizer for natural oils and vinyl polymers, a dye-levelling agent and a demulsifier (American Conference of Governmental Industrial Hygienists, 1986).

 There were presented the applications of glycidol as an intermediate in the syntheses of many biologically active compounds used in medicine, cosmetology, and in economic chemistry. Glycidol is half-finished product in the synthesis of surfactants. These agents are used as components of cosmetic products for moistening and cleansing of skin, hair shampoo, toothpastes, washing detergents and disinfectants. Another applications of glycidol are plasticizers, fabric dyes, photochemical compounds, caoutchouc, varnish and plastics. Glycidol is also used in the synthesis of many biologically active compounds, which originally were obtained from living organisms (algae, fungi). One of the most important applications of glycidol is the synthesis of antiviral and analgesic drugs. The especially important group of antivirial drugs comprise the active compounds against the HIV virus.

V.                ENVIRONMENTAL OCCURRENCE

Production of glycidol and its broad applications as an intermediate, as a reactive diluent in epoxy resins and as a stabilizer and a sterilant may result in its release into the environment through various waste streams .

VI.             HAZARDS IDENTIFICATION

Emergency Overview

1.      OSHA Hazards

Combustible Liquid, Carcinogen, Target Organ Effect, Toxic by inhalation., Toxic by ingestion, Toxic by skin absorption, Irritant, Teratogen, Mutagen

2.      Target Organs

Nerves.

3.      Other hazards which do not result in classification

Rapidly absorbed through skin.

GHS Classification

Flammable liquids (Category 4)

Acute toxicity, Oral (Category 4)

Acute toxicity, Inhalation (Category 3)

Acute toxicity, Dermal (Category 4)

Skin irritation (Category 2)

Serious eye damage (Category 1)

Germ cell mutagenicity (Category 2)

Carcinogenicity (Category 1B)

Reproductive toxicity (Category 1B)

Specific target organ toxicity - single exposure (Category 3)

VII.          POTENTIAL HEALTH EFFECTS

Inhalation                                   Toxic if inhaled. Causes respiratory tract irritation.

Skin                                               Toxic if absorbed through skin. Causes skin irritation.

Eyes                                             Causes eye irritation.

Ingestion                                      Toxic if swallowed.

  VIII.       RESPIRATOR RECOMMENDATIONS

 NIOSH

Upto150ppm: (APF=10)Anysupplied-airrespirator* (APF = 50) Any self-contained breathing apparatus with a full facepiece

Emergency or planned entry into unknown concentrations or IDLH conditions: (APF = 10,000) Any self-contained breathing apparatus that has a full facepiece and is operated in a pressure-demand or other positive-pressure mode (APF = 10,000) Any supplied-air respirator that has a full facepiece and is operated in a pressure-demand or other positive-pressure mode in combination with an auxiliary self-contained positive-pressure breathing apparatus

Escape: (APF = 50) Any air-purifying, full-facepiece respirator (gas mask) with a chin-style, front- or back-mounted organic vapor canister Any appropriate escape-type, self-contained breathing apparatus

[daisyri-me]

Sodium Chlorite Market to Account for USD 256.4 million by 2025

Global Sodium Chlorite Market size is expected to reach USD 256.4 million by 2025. Sodium chlorite is a chemical compound with a formula “NaClO2”. It is a white crystalline and odor less powder, which is used in the production of chlorine dioxide. It is commonly known as “sodium salt”. It completely dissolves in water and slightly soluble in methanol & ethanol. Sodium chlorite is used for water disinfection and purification, bleaching agent, weed control, and others. 

Also, combination of sodium chlorite and zinc chloride is used as a component in toothpaste, mouthwash, sprays and gels. The compound is hard to burn, but accelerates the burning of organic constituents. It is available in two forms like liquid and pellets large crystals. The sodium chlorite industry is expected to register a CAGR of 6.0% over the forecast period as the scope, product types, and its applications are increasing across the world.

The increasing demand for sodium chlorite from various industries like healthcare and sanitation due to its excellent disinfectant, antimicrobial, and bleaching properties and growing urban population that increases use of sodium chlorite for water purification are major factors expected to boost the sodium chlorite market growth in the future period. However, increasing cost of raw material, stringent government rules & regulations regarding the use of sodium chlorite, and availability of alternatives such as hydrogen peroxide & ozone used for pulp bleaching are expected to restrain the sodium chlorite industry in the forecast period. The sodium chlorite market is categorized based on application, end-use and geography.

Application that could be explored in sodium chlorite industry includes antimicrobial agent, disinfectant, bleaching agent, and others. The demand for sodium chlorite as a bleaching agent is expected to grow at a CAGR of 4.6% in the forthcoming years due to its superior oxidizing properties. Also, growing demand for sodium chlorite as antimicrobial agents in the food processing industry to enhance the shelf life of packaged products is anticipated to have a positive impact on market growth in the forecast period.

The market may be categorized based on end-users such as paper & pulp, water treatment, textile, medical, and others. In 2016, water treatment sector accounted for 53.2% market share and is expected to lead the overall market in the coming years. This is due to superior antimicrobial & biocidal properties associated the sodium chlorite. However, textile industry is anticipated to grow at a CAGR of 6.3% in the years to come due to its emergence as an excellent source for chlorine dioxide.

Enquire About This Report: https://www.millioninsights.com/industry-reports/sodium-chlorite-market/request-sample

Asia Pacific is accounted for 38.7% of total market share in 2016 and is anticipated to lead the overall market growth in the future period. The reason behind overall market growth may be growing pulp & paper and water treatment industries and presence of key manufactures in this region. Moreover, China is a leading consumer of sodium chlorite in Asia Pacific region. Further, North America and Europe are expected to follow the market growth in the coming years.

Europe is the second largest region with significant market share. However, North America is expected to witness a growth of 5.4% in the forecast period due to rising government initiative to improve water and wastewater treatment processes. In this region, sodium chlorite is used to de-ice the highways and streets during heavy snowfall. DuPont, Occidental Petroleum Corporation (OxyChem) and American Elements are the major producer of sodium chlorite in North American region. Headline and Adox are the two major brands of sodium chlorite manufactured by DuPont.

The key players operating in the sodium chlorite market are DuPont, Occidental Petroleum Corporation (OxyChem), ERCO Worldwide, American Elements, Alfa Aesar, AngeneChembo Pharma, ABI Chemicals, AOK Chem, Erco Worldwide, J and K Industry, Fintech Industry, Santa Cruz Biotechnology, Sigma Aldrich, Shanghai IS Chemical Technology, Tractus Co. Ltd., JalorChem, and OxyChem. These market players strongly invest in the expansion of their business and development to maintain a top position in the market. Also, these players concentrating on new joint ventures, collaborations, agreements, and strategies to improve their production facilities and gain a larger share in the market.

Get Full Details On: https://www.millioninsights.com/industry-reports/sodium-chlorite-market

Market Segment:

Application Outlook (Volume, Tons; Revenue, USD Thousands, 2012 - 2025)     • Disinfectant     • Antimicrobial agent     • Bleaching agent     • Others

Get in touch

At Million Insights, we work with the aim to reach the highest levels of customer satisfaction. Our representatives strive to understand diverse client requirements and cater to the same with the most innovative and functional solutions.

Contact Person:

Ryan Manuel

Research Support Specialist, USA

Email:ryan@millioninsights.com

[wahid007posts]

Electronic Wet Chemicals Market Share Will Take a Big Hit in the Coming Years

According to the new market research report "Electronic Wet Chemicals Market By Type, Form (Solid Liquid, Gas), Application (Semiconductor, IC Packaging, PCB), End-Use Industry (Consumer Goods, Automotive, Aerospace & Defence, Medical), And Region - Global Forecast To 2025" published by MarketsandMarkets™, the global Electronic Wet Chemicals Market size is expected to grow from USD 3.3 billion in 2020 to USD 3.9 billion by 2025, at a CAGR of 3.5%, during the forecast period.

Get PDF brochure of the report: https://www.marketsandmarkets.com/pdfdownloadNew.asp?id=265979069

Browse in-depth TOC on "Electronic Wet Chemicals Market"

164- Market Data Tables

39 - Figures

185 – Pages

The major factors driving the market include growing consumption of electronic wet chemicals in the emerging clusters of APAC is mainly because of the rising per capita expenditure on consumer electronics products such as smartphones, laptops and others and abundant availability of raw materials and cheap labor force, and growing demand from various end-use industries such as automotive and consumer goods.

Acetic Acid to dominate the global Electronic Wet Chemicals Market during the forecast period

The electronic wet chemicals industry has been segmented based on type as acetic acid, isopropyl alcohol hydrogen peroxide, hydrochloric acid, ammonium hydroxide, hydrofluoric acid, nitric acid, phosphoric acid, sulfuric acid and others. Acetic acid segment accounted for the larger share of the market in 2019. The growth in this segment is attributed to the increase in use in integrated circuits and it serve as one of the main substrates for microelectronics. These factors are expected to drive the demand for electronic wet chemicals during the forecast period.

Semiconductor segment to lead the global Electronic Wet Chemicals Market during the forecast period

The market has been segmented based on application into semiconductor, IC Packaging, PCB and others. The semiconductor segment accounted for the largest share of the market in 2019 and is expected to witness significant growth during the forecast period. The growth in this segment is attributed to increasing consumption in the advanced electronics and consumer goods such as smartphones, laptops and others in emerging economies such as APAC. Furthermore, the demand for semiconductor is increasing in flat panel display and automotive industry. These factors are expected to drive demand during the forecast period.

https://www.prnewswire.com/news-releases/electronic-wet-chemicals-market-worth-3-9-billion-by-2025---exclusive-report-by-marketsandmarkets-301133095.html

APAC is expected to have the largest market size in the global Electronic Wet Chemicals Market during the forecast period

APAC is projected to be the leading electronic wet chemicals industry globally during the forecast period. The growth in the APAC region can be attributed to the rising demand from consumer goods industry. The presence of several microelectronic devices manufacturers, favorable government policies, and low labor costs along with growing demand for electronic wet chemicals in the region are further strengthening the market and attracting major players to invest in the region.

The Avantor Inc (US), BASF SE (Germany), Cabot Microelctronics (US), Honeywell International Inc. (US), Kanto Chemical Co. Inc (Japan), Eastman (US), Solvay (Belgium), Fujifilm Holding (Japan), Technic Inc. (US), and Linde Plc. (Ireland) among others are the key players operating in the Electronic Wet Chemicals Market.

Early buyers will receive 10% free customization on this report.

Don't miss out on business opportunities in Electronic Wet Chemicals Market.

Speak to Our Analyst and gain crucial industry insights that will help your business grow.

https://www.marketsandmarkets.com/speaktoanalystNew.asp?id=265979069

About MarketsandMarkets

MarketsandMarkets is the largest market research firm worldwide in terms of annually published premium market research reports. Serving 1700 global fortune enterprises with more than 1200 premium studies in a year, M&M is catering to a multitude of clients across 8 different industrial verticals. We specialize in consulting assignments and business research across high growth markets, cutting edge technologies and newer applications.

Contact: Mr. Ashish Mehra MarketsandMarkets™ INC.  630 Dundee Road  Suite 430  Northbrook, IL 60062  USA: 1-888-600-6441  newsletter@marketsandmarkets.com

[halocantik]

How honey kills bacteria

The FASEB Journal

•

Research Communication

How honey kills bacteria

Paulus H. S. Kwakman,* Anje A. te Velde,† Leonie de Boer,* Dave Speijer,‡ Christina M. J. E. Vandenbroucke-Grauls,*,§ and Sebastian A. J. Zaat*,1

*Department of Medical Microbiology, Center for Infection and Immunity Amsterdam, †Laboratory of Experimental Gastroenterology and Hepatology, and ‡Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, Amsterdam; and §Department of Medical Microbiology and Infectious Diseases, Vrije Universiteit Medical Center, Amsterdam,

The Netherlands

  ABSTRACT         With the rise in prevalence of antibiotic- resistant bacteria,

honey

is increasingly valued for its antibacterial activity. To characterize all bactericidal factors in a medical-grade honey, we used a novel approach of successive neutralization of individual honey bactericidal factors. All bacteria tested, including

Bacillus subtilis

, methicillin-resistant

Staphylococcus au-

reus

, extended-spectrum J3-lactamase producing

Esche-

richia coli

, ciprofloxacin-resistant

Pseudomonas aerugi-

nosa

, and vancomycin-resistant

Enterococcus faecium

, were killed by 10 –20% (v/v) honey, whereas >40% (v/v) of a honey-equivalent sugar solution was required for similar activity. Honey accumulated up to 5.62 ±

0.54 mM H2O2 and contained 0.25 ± 0.01 mM methyl-

glyoxal (MGO). After enzymatic neutralization of these two compounds, honey retained substantial activity. Using

B. subtilis

for activity-guided isolation of the additional antimicrobial factors, we discovered bee defensin-1 in honey. After combined neutralization of H

2

O

2

, MGO, and bee defensin-1, 20% honey had only minimal activity left, and subsequent adjustment of the pH of this honey from 3.3 to 7.0 reduced the activity to that of sugar alone. Activity against all other bacteria tested depended on sugar, H

2

O

2

, MGO, and bee defensin-1. Thus, we fully characterized the antibacterial activity of medical-grade honey.—Kwak- man, P. H. S., te Velde, A. A., de Boer, L., Speijer, D., Vandenbroucke-Grauls, C. M. J. E., Zaat, S. A. J. How honey kills bacteria.

FASEB J.

24, 2576 –2582 (2010).

www.fasebj.org

 Key Words: antibacterial agents · drug resistance · isolation and purification · methicillin-resistant Staphylococcus aureus

· peptides

 Honey has been renowned for its wound-healing properties since ancient times (1). At least part of its positive influence is attributed to antibacterial proper- ties (2, 3). With the advent of antibiotics, clinical application of honey was abandoned in modern West-

The potent in vitro activity of honey against antibiotic- resistant bacteria (6, 7) and its successful application in treatment of chronic wound infections not re- sponding to antibiotic therapy (3) have attracted considerable attention (8 –10).

The broad spectrum antibacterial activity of honey is multifactorial in nature. Hydrogen peroxide and high osmolarity— honey consists of ~80% (w/v) of sugars— are the only well-characterized antibacterial factors in

honey (11). Recently, high concentrations of the anti- bacterial compound methylglyoxal (MGO) were found specifically in Manuka honey, derived from the Manuka tree (Leptospermum scoparium) (12, 13). Until now, no honey has ever been fully characterized, which ham- pers clinical application of honey.

Recently, we determined that Revamil medical-grade honey, produced under standardized conditions in greenhouses, has potent, reproducible bactericidal ac- tivity (14). In the current study, we identified all bactericidal factors in the honey used as source for this product and assessed their contribution to honey bac- tericidal activity.

To accomplish this, we used a novel approach of successive neutralization of individual honey bacteri- cidal factors combined with activity-guided identifica- tion of unknown factors.

  MATERIALS AND METHODS

 Honey

 Unprocessed Revamil source (RS) honey was kindly provided by Bfactory Health Products (Rhenen, The Netherlands). RS honey has a density of 1.4 kg/L and contains 333 g/kg glucose, 385 g/kg fructose, 73 g/kg sucrose, and 62 g/kg maltose. To study the contribution of the sugars to the bactericidal activity of honey, a solution with a sugar compo- sition identical to that of the honey was prepared.

ern medicine, although in many cultures, it is still used                                  

(4). These days, however, abundant use of antibiotics has resulted in widespread resistance. With the devel- opment of novel antibiotics lagging behind (5), alter- native antimicrobial strategies are urgently needed.

1 Correspondence: Department of Medical Microbiology, Academic Medical Center, Meibergdreef 15, 1105 AZ Amster- dam, The Netherlands, E-mail: s.a.zaat@amc.uva.nl

doi: 10.1096/fj.09-150789

Microorganisms

 Bactericidal activity of honey was assessed against the labora- tory strains Bacillus subtilis ATCC6633, Staphylococcus aureus 42D, Escherichia coli ML-35p (15), and Pseudomonas aeruginosa PAO-1 (ATCC 15692), and against clinical isolates of methi-

cillin-resistant S. aureus (MRSA), vancomycin-resistant Entero- coccus faecium (VREF), extended-spectrum [3-lactamase-pro- ducing E. coli (E. coli ESBL) and ciprofloxacin-resistant

P. aeruginosa (CRPA).

aliquots of undiluted and 10-fold serially diluted incubations were plated on blood agar. Bacterial survival was quantified after overnight incubation at 37°C. The detection level of this assay is 100 CFU/ml.

To assess the contribution of H2O2 to the bactericidal activity of honey, bovine liver catalase (Sigma) was added to a final concentration of 600 U/ml. A catalase stock solution was prepared according to the manufacturers’ instructions in 50 mM phosphate buffer (pH 7.0). The addition of 0.25% (v/v) of this catalase stock solution reduced the amount of H2O2 to undetectable levels at all honey concentrations tested and did

Determination of H O

concentration in honey

not affect bacterial viability.

2     2                                                                                                                     Sodium polyanetholsulfonate (SPS) (Sigma) was added to neutralize cationic bactericidal components (19) at a final

Hydrogen peroxide concentrations in honey were deter-

mined quantitatively using a modification of a method de- scribed previously (16). Undiluted and 10-fold diluted sam-

ples of honey (40 µl) were mixed in wells of microtiter plates with 135 µl reagent, consisting of 50 µg/ml O-dianisidine (Sigma, St. Louis, MO, USA) and 20 µg/ml horseradish peroxidase type IV (Sigma) in 10 mM phosphate buffer (pH

6.5). O-dianisidine and peroxidase solutions were freshly prepared from a 1 mg/ml stock in demineralized water and from a 10 mg/ml stock in 10 mM phosphate buffer (pH 6.5), respectively. After 5-min incubations at room temperature,

reactions were stopped by addition of 120 µl6MH SO , and

concentration of 0.025% (w/v). The incubation buffer did not affect the pH of the concentrations of honey used in our experiments.A1M NaOH solution was used to titrate honey solutions to pH 7.0.

 Agar diffusion assay

 To assess antibacterial activity of fractionated honey, an agar diffusion assay was used (20). In brief, a B. subtilis inoculum suspension was prepared as described for the liquid bacteri- cidal assay. Bacteria (107 CFU) were mixed with 20 ml

2    4             nutrient-poor agar [0.03% (w/v) TSB in 10 mM sodium

absorption at 540 nm was measured. Hydrogen peroxide concentrations were calculated using a calibration curve of 2-fold serial dilutions of H2O2 ranging from 2200 to 2.1 µM.

 MGO neutralization assay

 Reduced glutathione (Sigma) was added to diluted honey to a final concentration of 15 mM, and conversion of MGO to S-d-lactoyl-glutathione (SLG) was initiated by addition of 0.5 U/ml glyoxalase I (Sigma). The amount of MGO converted was determined using the extinction coefficient of SLG of

3.37 mM-1 at 240 nm (17). Thus, we determined that up to 10 mM of exogenous MGO added to 40% honey was com-

pletely converted, and that undiluted RS honey contained

0.25 0.01 mM of MGO.

 Antibee defensin-1 polyclonal antibody

 An affinity-purified polyclonal antibee defensin-1 antibody was purchased from Eurogentec (Seraing, Belgium). The N-terminal part of bee defensin-1 is hydrophobic and con- tains 3 disulfide bonds, whereas the hydrophilic C-terminal region lacks cysteine residues (18). Therefore, rabbits were immunized with a synthetic peptide corresponding to the C terminus of bee defensin-1 (CRKTSFKDLWDKRF), and anti- bodies were subsequently affinity-purified using this peptide coupled to AF-Amino Toyopearl 650 M resin (Toso, Tokyo, Japan).

 Liquid bactericidal assay

 Bactericidal activity of honey was quantified in 100-µl volume liquid tests, in polypropylene microtiter plates (Costar Corn- ing, New York, NY, USA). For each experiment, a 50% (v/v)

stock solution of honey was freshly prepared in incubation buffer containing 10 mM phosphate buffer (pH 7.0) supple- mented with 0.03% (w/v) trypticase soy broth (TSB; BD Difco, Detroit, MI, USA). Bacteria from logarithmic phase cultures in TSB were washed twice with incubation buffer and suspended at a final concentration of 1 X 106 CFU/ml, based

on optical density. Plates were incubated at 37°C on a rotary

shaker at 150 rpm. At indicated time points, duplicate 10-µl

phosphate buffer (pH 7.0) with 1% low EEO agarose (Sigma)] of 45°C, and immediately poured into 10- X 10-cm culture plates. Wells of 1 mm diameter were punched into the agarose, and 2.5-µl samples were added to the wells and allowed to diffuse into the agarose for 3 h at 37°C. Subse- quently, the agarose was overlaid with 20 ml of double- strength nutrient agarose [6% TSB and 1% Bacto-agar (BD Difco), 45°C], and plates were incubated overnight at 37°C. Clear zones around the wells indicated antibacterial activity.

 Ultrafiltration of honey components

 Fifteen milliliters of 20% honey was centrifuged in a 5-kDa molecular weight cutoff Amicon Ultra-15 tube (Millipore, Bedford, MA, USA) at 4000 g for 45 min at room tempera- ture. The <5-kDa filtrate was collected, and the >5-kDa reten- tate was subsequently washed 3 times in the filter tube with 15 ml of demineralized water and concentrated to 0.4 ml.

 Bacterial overlay assay

 Native cationic proteins were separated by acid urea polyacryl- amide gel electrophoresis (AU-PAGE) (21). Gels were either stained with PAGE-Blue (Fermentas, St. Leon-Rot, Germany) or washed 3 X 8 min in 10 mM phosphate buffer (pH 7.0) for a bacterial overlay assay. After washing, the gel was incubated for 3 h on B. subtilis-inoculated nutrient-poor agarose (see

Agar Diffusion Assay). After removal of the gel, the agarose was overlaid with double-strength nutrient agarose and treated as described for the agar diffusion assay.

 Immunoblotting

 Proteins were separated by tris-tricine SDS-PAGE, as de- scribed previously (22), and transferred onto nitrocellulose membranes (Schleicher and Schuell, Keene, NH, USA). Membranes were subsequently blocked with 5% nonfat dry milk (Bio-Rad, Veenendaal, The Netherlands) plus 0.5 M NaCl and 0.5% (v/v) Tween-20 in 10 mM Tris-HCl, pH 7.5 (rinse buffer), for 1 h. Blocked membranes were incubated with affinity-purified antibee defensin-1 antibody at 1.4

µg/ml in rinse buffer for 2 h. After incubation with primary antibody, membranes were washed 2X for 15 min in rinse buffer, incubated with horseradish peroxidase-labeled goat-

anti-rabbit secondary antibody (Jackson ImmunoResearch West Grove, PA, USA) at 0.4 µg/ml in rinse buffer for 1 h, and washed again for 10 min. in rinse buffer and 5 min in PBS, respectively. The membrane was developed using a DAB liquid substrate kit (Sigma).

 Purification of antibacterial peptide from honey

 An amount of >5-kDa honey retentate equivalent to 13 ml of honey was dissolved in loading buffer (3M urea in 5% acetic acid with methyl green as tracking dye) and loaded on a preparative acid-urea PAGE, as described previously (21) with

slight modifications. A cylindrical gel (3.7 cm diameter, 6 cm height) in a model 491 Prep Cell (Bio-Rad) was prepared, prerun at reversed polarity for3h at 150V in 5% acetic acid at 4°C, and protein was electrophoresed at 40 mA with reversed polarity. Protein was eluted in 5% acetic acid at 0.5 ml/min and collected in fractions of 2 ml. Fractions were assessed for protein composition by tris-tricine SDS-PAGE and for antibacterial activity by bacterial overlay assay. Frac- tions containing purified antibacterial protein were pooled, concentrated, dialyzed against 0.01% acetic acid in a 3.5-kDa molecular weight cutoff MINI Slide-A-Lyzer tube (Pierce, Rockford, IL, USA), freeze-dried, and dissolved in deminer- alized water.

 Protein identification by V8 digestion with subsequent mass analysis

 Duplicate fractions (estimated to contain ~2 µg of protein each) were adjusted to 50 mM sodium phosphate (pH 7.9)

and 5% (v/v) acetonitrile. Approximately 0.5 µg of endopro- teinase Glu-C (Fluka) was added per fraction and incubated at 25°C overnight. The resulting peptide mixtures were purified and concentrated with the aid of C18 ziptips (Milli- pore) and eluted in 10 µl 90% (v/v) acetonitrile and 1% (v/v) formic acid. The samples were checked for the presence of nonautodigest peptides with a reflectron MALDI-TOF mass spectrometer (MALDI; Waters, Milford, MA, USA). Next,

samples were analyzed with ESI-tandem mass spectrometry (MS/MS). Data were acquired with a QT of 1 (Waters) coupled to an Ultimate nano-LC system (LC Packings Di- onex, Sunnyvale, CA, USA). One microliter of peptide mix- ture was diluted in 10 µl of 0.1% TFA. The peptides of both

samples were separated on a nanoanalytical column (75 µm

i.d. X 15 cm C18 PepMap; LC Packings Dionex) using a standard gradient of acetonitrile in 0.1% formic acid. The

flow of 300 nl/min was directly electrosprayed in the QT of 1 operating in data-dependent MS and MS/MS mode. The resulting MS/MS spectra were analyzed with Mascot software (Matrix Science, Boston, MA, USA). In both fractions, a doubly charged ion (VTCDLLSFKGQVND, mass 1537.8) with a sequence corresponding to the mature N terminus of bee defensin-1 could be identified (MOWSE scores >73).

  RESULTS

 Hydrogen peroxide is produced by the Apis mellifera (honeybee) glucose oxidase enzyme on dilution of honey. RS honey diluted to 40 to 20% accumulated high levels of H2O2 24 h after dilution, with a maximum of

5.62 0.54 mM H2O2 formed in 30% honey (Fig. 1A). The addition of catalase reduced H2O2 to negligible

   Figure 1. Contribution of H2O2, sugars, and MGO to the bactericidal activity of honey after 24 h. A) Mean se hydrogen peroxide accumulation in different concentrations of honey, without catalase (squares) or with catalase added (asterisks).

B) Bactericidal activity against indicated laboratory strains (top row) and against clinical isolates of vancomycin-resistant

E. faecium (VREF), methicillin-resistant S. aureus (MRSA), extended-spectrum [3-lactamase-producing E. coli (E. coli ESBL), and ciprofloxacin-resistant P. aeruginosa (CRPA) (bottom row). Bacteria were exposed to various concentrations of honey (squares), honey with catalase added (asterisks), or to honey-equivalent sugar solutions (circles). C) Killing of B. subtilis by honey in incubation buffer without addition (squares), with catalase (asterisk), with glyoxalase (small solid circles), or with catalase and glyoxalase I (inverted triangles), added to neutralize H2O2 and MGO, respectively, or by a honey-equivalent sugar solution

(circles). Data are mean se log-transformed bacterial concentration (CFU/ml).

levels (Fig. 1A) and markedly reduced the bactericidal activity against all bacteria tested, except B. subtilis (Fig. 1B). However, H2O2-neutralized honey exerted stron- ger bactericidal activity than equivalent sugar solutions (Fig. 1B). This indicates that H2O2 is important for the bactericidal activity of honey, but that additional factors must also be present. As B. subtilis was the most susceptible bacterium for nonperoxide bactericidal activity, we used it for identification of additional bactericidal factors.

The honey bactericidal compound MGO can be converted into S-lactoylglutathione (SLG) by glyoxalase I, and this product can be measured spectrophoto- metrically. RS honey contained 0.25 0.01 mM MGO. We aimed to apply glyoxalase I to neutralize the bactericidal activity of MGO in honey. This required that SLG, the reaction product of MGO, would be nonbactericidal. Indeed, the activity of up to 20 mM MGO was neutralized by conversion into SLG (Supple- mental Fig. 1), indicating that SLG up to high concen- trations did not kill the bacteria. Neutralization of MGO or H2O2 alone did not alter bactericidal activity of RS honey, but simultaneous neutralization of MGO and H2O2 in 10% honey reduced the killing of B. subtilis by 4-logs (Fig. 1C). At higher concentrations of honey, the bactericidal activity was not affected by neutralization of H2O2 and MGO (Fig. 1C), indicating that still more factors were involved.

As a first step to characterize the unknown bacteri- cidal factors, we size-fractionated honey by ultrafiltra- tion with a 5-kDa molecular weight cutoff membrane. Unfractionated honey produced a small zone of com- plete bacterial growth inhibition and a larger zone with partial growth inhibition in an agar diffusion assay with

B. subtilis (Fig. 2A). After ultrafiltration, the factors that caused complete and partial bacterial growth inhibition were separated and were present in the >5-kDa reten-

tate and the <5-kDa filtrate, respectively (Fig. 2A).

Ion exchange chromatography of the retentate indi-

cated a cationic nature of the antibacterial factors. Indeed, the polyanionic compound SPS abolished the antibacterial activity of the retentate (Fig. 2B). More- over, pepsin treatment also abolished this activity (Fig. 2B). Together, this implies that cationic antibacterial proteins were present.

We separated cationic proteins in the retentate using a native acid-urea PAGE gel, and allowed the separated components to diffuse from this gel into a B. subtilis- inoculated agar to identify antibacterial proteins. This yielded a single zone of bacterial growth inhibition that corresponded to a protein band in a Coomassie-stained gel run in parallel (Fig. 2C). This protein was purified from a larger amount of retentate using preparative acid-urea PAGE (Fig. 2D), and identified by peptide mass analysis as bee defensin-1.

To specifically assess the contribution of bee defen- sin-1 to the bactericidal activity of honey, an antibee defensin-1 antibody was raised (Fig. 2E). Like SPS, this antibody negated all bactericidal activity of the >5-kDa

retentate against B. subtilis (Fig. 3A). The <5-kDa

filtrate had only minor bactericidal activity (Fig. 3A), but this was not due to cationic compounds, since SPS

failed to neutralize this activity (Fig. 3A). Thus, bee defensin-1 was the only cationic bactericidal compound present in RS honey.

Next, we assessed the contribution of bee defensin-1 to the bactericidal activity of nonfractionated honey

               Figure 2. Identification of bee defensin-1 in honey. A) Honey was fractionated by ultrafiltration using a 5-kDa molecular weight cutoff filter tube; antibacterial activity of 2.5 µl of 80% honey, and equivalent amounts of the <5-kDa filtrate and >5-kDa retentate, were tested in an agar diffusion assay. B) Retentate equivalent to 7.5 µl of undiluted honey was tested for the presence of cationic and proteinaceous antibacterial components. Activity of cationic components was neutralized by

adding SPS, and protein was digested with pepsin, followed by 5-min inactivation at 100°C. As control, incubation for 5 min at 100°C without pepsin was performed. Activity in retentate (ret.)

was compared with that of 0.2 µg hen egg white lysozyme (lys.). C) To identify cationic antibacterial proteins in retentate, amounts of this fraction equivalent to 750 µl honey, and 3 µg lysozyme as a reference, were run in duplicate sets on a single native acid-urea PAGE gel. One half

of the gel was Coomassie-stained (left); other was used for a bacterial overlay assay with B. subtilis

(right). D) Silverstained tris-tricine SDS-PAGE of different amounts of lysozyme and preparative acid-urea PAGE-purified bee defensin-1, separated by an empty lane. E) Retentate separated on tris-tricine SDS-PAGE, blotted to nitrocellulose, stained with either Ponceau S (Pon. S, left) or immunostained with antibee defensin-1 (right).

  Figure 3. Roles of bee defensin-1 and pH in bactericidal activity of honey against B. subtilis. A) Contribution to bacte- ricidal activity of cationic components in general and of bee defensin-1 specifically was tested by neutralization with SPS or with antibee defensin-1 antibody (C-bd), respectively, at con- centrations of retentate equivalent to 20% honey (open bars) and 40% honey (solid bars); ctrl. indicates survival without

neutralization. B) To assess the contribution of bee defen- sin-1 to bactericidal activity of unfractionated honey, B. subtilis was incubated in various concentrations of honey in incuba- tion buffer (squares), or with catalase and glyoxalase I added either without (triangles) or with SPS (diamonds), or in a honey-equivalent sugar solution (circles). C) To assess the contribution of the low pH to the bactericidal activity of honey, B. subtilis was incubated in various concentrations of honey in incubation buffer (squares), or with catalase, glyox- alase I, and SPS added either without (triangles) or with neutralization to pH 7 (diamonds), or in a honey-equivalent sugar solution (circles). After 24 h, numbers of surviving bacteria were determined. Data are mean se log-trans- formed bacterial concentration (CFU/ml).

 against B. subtilis. As previously observed, >20% honey retained bactericidal activity when H2O2 and MGO were neutralized. Additional neutralization of bee de- fensin-1 strongly reduced the bactericidal activity of 20% honey but did not affect the activity of 30 and 40% honey (Fig. 3B). So, bee defensin-1 contributed to the bactericidal activity of honey, but still other bactericidal factors were involved.

Honey has a low pH, mainly because of the conver- sion of glucose into hydrogen peroxide and gluconic acid by glucose oxidase. This low pH might also con- tribute to the bactericidal activity of honey (23). Titra- tion of the pH of 40 –10% RS honey from 3.4 –3.5 to 7.0, combined with neutralization of H2O2, MGO and bee defensin-1, reduced the bactericidal activity of honey to a level identical to that of a honey-equivalent sugar solution (Fig. 3C). Thus, with this experiment, we

succeeded in identifying all bactericidal factors in RS honey responsible for killing of B. subtilis.

The contribution of the identified bactericidal fac- tors to activity against antibiotic-susceptible and -resis- tant strains of various species was tested with honey diluted to 20%, since this killed the entire inocula of all bacteria tested independent of sugar (Fig. 1). Simulta- neous neutralization of H2O2, MGO and bee defensin-1 negated all activity (Fig. 4), showing that these were the major factors responsible for broad spectrum bacteri- cidal activity of honey.

We studied the contribution of the honey bacteri- cidal factors in more detail by neutralizing the factors individually or combined. Neutralization of H2O2 alone strongly reduced the bactericidal activity against all bacteria tested except B. subtilis (Fig. 4). Neutralization of MGO alone strongly reduced killing of E. coli and

P. aeruginosa strains (Fig. 4). Neutralization of bee defensin-1 alone reduced killing of VREF, but not of the other bacteria tested (Fig. 4). When compared to neutralization of MGO alone, the additional neutraliza- tion of bee defensin-1 reduced killing of all bacteria tested, except E. coli ESBL (Fig. 4). In summary, H2O2, MGO, and bee defensin-1 differentially contributed to the activity of honey against specific bacteria, and their combined presence was required for the broad-spec- trum activity.

   DISCUSSION

 All bacterial species tested were susceptible to different combinations of bactericidal factors in honey, indicat- ing that these bacteria were killed via distinct mecha- nisms. This clearly demonstrates the importance of the multifactorial nature of honey for its potent, broad- spectrum bactericidal activity.

Some factors had overlapping activity. For instance, the activity of bee defensin-1 against most bacteria was only revealed after neutralization of MGO. This clearly demonstrates the importance of neutralizing known bactericidal factors in honey to reveal the presence of additional factors. Similarly, the contribution of the low pH for activity of honey against B. subtilis was only revealed when H2O2, MGO, and bee defensin-1 were simultaneously neutralized.

In other situations, bactericidal activity depended on the combined presence of different factors. Thus, the activity of honey against E. coli and P. aeruginosa was markedly reduced by neutralization of either H2O2 or MGO. Alternatively, the activity of certain bactericidal factors likely is more potent in the context of honey than as pure substances. This is most clearly illustrated by the activity of MGO. When tested in a buffer, >0.3 mM MGO was required for activity against B. subtilis (Supplemental Fig. 1). In contrast, as little as 0.05 mM MGO, the concentration in 20% RS honey, was suffi- cient to substantially contribute to the bactericidal activity. This suggests that the presence of the other bactericidal factors in honey enhanced the effect of

 Figure 4. Effect of neutralization of H2O2, MGO, and bee defensin-1 on bactericidal activity of honey. Hydrogen peroxide, MGO, and bee defensin-1 were neutralized in 20% honey by adding catalase (cat.), glyoxalase I (gly I) and SPS, respectively. Bactericidal activity was tested against indicated laboratory strains (left 4 panels) and against clinical isolates of VREF, MRSA,

E. coli ESBL, and CRPA (right 4 panels). A sugar solution equivalent to 20% honey was used as a reference. After 24 h, numbers of surviving bacteria were determined. Data are mean se log-transformed bacterial concentration (CFU/ml).

 MGO. It is not possible to quantify the contribution of the different factors to honey bactericidal activity since, as we have shown, these factors may have redundant activity, be mutually dependent, or have additive or synergistic activity depending on the bacterial species targeted.

We have demonstrated for the first time that honey contains an antimicrobial peptide, bee defensin-1, and that this peptide substantially contributes to the bacte- ricidal activity. Bee defensin-1 was previously isolated from royal jelly (24), the major food source for bee queen larvae (and then referred to as “royalisin”), and was identified in honeybee hemolymph (18). Royal jelly is produced by young worker bees and contains their hypopharyngeal and mandibular gland secretions (25, 26). Bee defensin-1 mRNA has been identified in the hypopharyngeal gland of young worker bees (18), suggesting this gland is involved in production of bee defensin-1 found in royal jelly (24). When worker bees age, they become the major producers of honey. Major differences develop in morphology and protein expres- sion of their hypopharyngeal glands (27, 28), e.g., several important carbohydrate-metabolizing enzymes, including glucose oxidase are expressed (29). The bees add the secretion from their hypopharyngeal glands to the collected nectar. The carbohydrate-metabolizing enzymes then convert sucrose to glucose and fructose, and glucose oxidase converts the glucose to hydrogen peroxide and gluconic acid. These latter compounds presumably are involved in prevention of microbial spoilage of unripe honey (11). Since we have found bee defensin-1 in honey, this suggests that after the transi- tion in hypopharyngeal gland function of the worker bees with age, the gland still produces bee defensin-1. This peptide, therefore, likely contributes to protection of both royal jelly and honey against microbial spoilage. It remains to be established whether bee defensin-1 is also present in other honeys. In Manuka honey, no evidence was found for the presence of antimicrobial peptides (30). For several other honeys, proteins were reported to contribute to the antibacterial activity (31, 32), but their identity remains unknown. Using our antibee defensin-1 antibody, we aim to assess the role of

bee defensin-1 for the antibacterial activity of other honeys.

Previous studies regarding the effect of low pH to antibacterial activity of honey have yielded conflicting results (11). In our study, the contribution of the low pH for activity against B. subtilis was only revealed on inactivation of all other bactericidal factors. So, in other studies, which did not employ an approach of neutral- ization of bactericidal factors in honey, the contribu- tion of the low pH of honey may easily have been overlooked.

Much effort has been put into identification of phenolic antibacterial components in honey (11). Sev- eral of these compounds have been isolated from honey, but as they were tested at concentrations far exceeding those in honey, no conclusions can be drawn regarding their contribution to honey bactericidal ac- tivity (11). Our data do not show a role of phenolic compounds in RS honey bactericidal activity.

Our approach of selectively neutralizing individual bactericidal factors present in a medical-grade honey allowed us to unravel the multifactorial bactericidal activity of a honey for the first time. We presently use the same approach to assess the contribution of these factors to activity of other honeys, and simultaneously to screen for novel bactericidal factors. Such honeys, or isolated components thereof, may serve as novel agents to prevent or treat infections, in particular those caused by antibiotic-resistant bacteria.                                                                            

 The authors thank Jorn Blom and Sadira Thomas for their help with purification of bee defensin-1; Henk Dekker for expert nano ESI-ms/ms experiments; and Ton Bisseling, Ben Berkhout, Mark van Passel, and Brendan McMorran for critically reviewing the manuscript.

   REFERENCES

 1.     Majno, G. (1975) Man and Wound in the Ancient World. Harvard University Press, Cambridge, MA, USA

2.     Molan, P. C. (2001) Why honey is effective as a medicine: its use in modern medicine. In Honey and Healing (Munn, P., and Jones, R., eds.) pp. 5–14, International Bee Research Associa- tion, Cardiff, UK

3.     Efem, S. E. E. (1988) Clinical observations on the wound- healing properties of honey. Brit. J. Surg. 75, 679 – 681

4.     Jones, R. (2001) Honey and healing through the ages. In Honey and Healing (Munn, P., and Jones, R., eds.) pp. 1– 4, Interna- tional Bee Research Association: Cardiff, UK

5.    Nathan, C. (2004) Antibiotics at the crossroads. Nature 431,

899 –902

6.     Cooper, R. A., Halas, E., and Molan, P. C. (2002) The efficacy of honey in inhibiting strains of Pseudomonas aeruginosa from infected burns. J. Burn Care Rehabil. 23, 366 –370

7.     Cooper, R. A., Molan, P. C., and Harding, K. G. (2002) The sensitivity to honey of gram-positive cocci of clinical significance isolated from wounds. J. Appl. Microbiol. 93, 857– 863

8.     Bonn, D. (2003) Sweet solution to superbug infections? Lancet Infect. Dis. 3, 608

9.    Dixon, B. (2003) Bacteria can’t resist honey. Lancet Infect. Dis. 3,

116

10.     Lusby, P. E., Coombes, A., and Wilkinson, J. M. (2002) Honey: a potent agent for wound healing? J. Wound Ostomy Continence Nurs. 29, 295–300

11.     Molan, P. C. (1992) The antibacterial activity of honey. 1. The nature of the antibacterial activity. Bee World 73, 5–28

12.     Adams, C. J., Boult, C. H., Deadman, B. J., Farr, J. M., Grainger,

M. N., Manley-Harris, M., and Snow, M. J. (2008) Isolation by HPLC and characterisation of the bioactive fraction of New Zealand manuka (Leptospermum scoparium) honey. Carbohydr. Res. 343, 651– 659.

13.     Mavric, E., Wittmann, S., Barth, G., and Henle, T. (2008) Identification and quantification of methylglyoxal as the domi- nant antibacterial constituent of Manuka (Leptospermum scopa- rium) honeys from New Zealand. Mol. Nutr. Food Res. 52, 483– 489

14.     Kwakman, P. H. S., Van den Akker, J. P. C., Guclu, A., Aslami, H., Binnekade, J. M., de Boer, L., Boszhard, L., Paulus, F., Middelhoek, P., Te Velde, A. A., Vandenbroucke-Grauls,

C. M. J. E., Schultz, M. J., and Zaat, S. A. J. (2008) Medical-grade honey kills antibiotic-resistant bacteria in vitro and eradicates skin colonization. Clin Infect Dis. 46, 1677–1682

15.     Lehrer, R. I., Barton, A., Daher, K. A., Harwig, S. S., Ganz, T., and Selsted, M. E. (1989) Interaction of human defensins with Escherichia coli. Mechanism of bactericidal activity. J. Clin. Invest. 84, 553–561

16.     White, J. W. Jr., and Subers, M. H. (1963) Studies on honey inhibine. 2. A chemical assay. J. Apicul. Res. 2, 93–100

17.     Racker, E. (1951) The mechanism of action of glyoxalase. J. Biol. Chem. 190, 685– 696

18.     Klaudiny, J., Albert, S., Bachanova, K., Kopernicky, J., and Simuth, J. (2005) Two structurally different defensin genes, one of them encoding a novel defensin isoform, are expressed in honeybee Apis mellifera. Insect Biochem. Mol. Biol. 35, 11–22

19.     Dankert, J., Van der Werff, J., Zaat, S. A., Joldersma, W., Klein, D., and Hess, J. (1995) Involvement of bactericidal factors from

thrombin-stimulated platelets in clearance of adherent viridans streptococci in experimental infective endocarditis. Infect. Im- mun. 63, 663– 671

20.     Lehrer, R. I., Rosenman, M., Harwig, S. S., Jackson, R., and Eisenhauer, P. (1991) Ultrasensitive assays for endogenous antimicrobial polypeptides. J. Immunol. Methods 137, 167–173

21.     Harwig, S. S., Chen, N. P., Park, A. S., and Lehrer, R. I. (1993) Purification of cysteine-rich bioactive peptides from leukocytes by continuous acid-urea-polyacrylamide gel electrophoresis. Anal. Biochem. 208, 382–386

22.     Schagger, H., and von Jagow, G. (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem. 166, 368 –379

23.     Yatsunami, K., and Echigo, T. (1984) Antibacterial activity of honey and royal jelly. Honeybee Sci. 5, 125–130

24.     Fujiwara, S., Imai, J., Fujiwara, M., Yaeshima, T., Kawashima, T., and Kobayashi, K. (1990) A potent antibacterial protein in royal jelly. Purification and determination of the primary structure of royalisin. J. Biol. Chem. 265, 11333–11337

25.     Lensky, Y., and Rakover, Y. (1983) Separate protein body compartments of the worker honeybee (Apis mellifera L). Comp. Biochem. Physiol. B Biochem. Mol. Biol. 75, 607– 615

26.     Knecht, D., and Kaatz, H. H. (1990) Patterns of larval food- production by hypopharyngeal glands in adult worker honey- bees. Apidologie 21, 457– 468

27.     Ohashi, K., Natori, S., and Kubo, T. (1997) Change in the mode of gene expression of the hypopharyngeal gland cells with an age-dependent role change of the worker honeybee Apis mellif- era L. Eur. J. Biochem. 249, 797– 802

28.     Sasagawa, H., Sasaki, M., and Okada, I. (1989) Hormonal-control

of the division of labor in adult honeybees (Apis-Mellifera L). 1. Effect of methoprene on corpora allata and hypopharyngeal gland, and its alpha-glucosidase activity. Appl. Entomol. Zool. 24, 66 –77

29.     Ohashi, K., Natori, S., and Kubo, T. (1999) Expression of amylase and glucose oxidase in the hypopharyngeal gland with an age-dependent role change of the worker honeybee (Apis mellifera L.). Eur. J. Biochem. 265, 127–133

30.     Weston, R. J., Brocklebank, L. K., and Lu, Y. R. (2000) Identi-

fication and quantitative levels of antibacterial components of some New Zealand honeys. Food Chem. 70, 427– 435

31.     Mundo, M. A., Padilla-Zakour, O. I., and Worobo, R. W. (2004) Growth inhibition of foodborne pathogens and food spoilage organisms by select raw honeys. Int. J. Food Microbiol. 97, 1– 8

32.     Gallardo-Chacon, J. J., Casellies, M., Izquierdo-Pulido, M., and Rius, N. (2008) Inhibitory activity of monofloral and multifloral honeys against bacterial pathogens. J. Apicul. Res. 47, 131–136

33.   https://cloverhoney.web.id/

34.   https://cloverhoney.web.id/clover-honey-madu-hdi/

35.   https://cloverhoney.web.id/propoelix/

36.   https://cloverhoney.web.id/royal-jelly-hdi/

37.   https://cloverhoney.web.id/clover-honey-harga/

38.   https://cloverhoney.web.id/propoelix-harga/

39.   https://cloverhoney.web.id/hdi-propoelix-adalah/

40.   https://cloverhoney.web.id/manfaat-propoelix/

41.   https://cloverhoney.web.id/madu-hdi-harga/

42.   https://cloverhoney.web.id/propoelix-plus/

43.   https://cloverhoney.web.id/madu-hdi-manfaatnya/

44.   https://cloverhoney.web.id/clover-honey-manfaatnya/

45.      

Received for publication November 18, 2009.

Accepted for publication February 4, 2010.

[jobssok]

Twice Oral Wellness Extra Strength Tooth Whitening Pen - Gentle, Sensitive Teeth Whitening Pens - Professional, on-The-go, or at Home use! Easy Teeth Stain Removal and Teeth Brightening. (2 Pack)

Twice Oral Wellness Extra Strength Tooth Whitening Pen – Gentle, Sensitive Teeth Whitening Pens – Professional, on-The-go, or at Home use! Easy Teeth Stain Removal and Teeth Brightening. (2 Pack)

Price: (as of – Details) Product Description Product Dimensions ‏ : ‎ 2.12 x 1.75 x 5.75 inches; 3.03 Ounces Item model number ‏ : ‎ 202 Date First Available ‏ : ‎ April 21, 2022 Manufacturer ‏ : ‎ Smiletwice, Inc. ASIN ‏ : ‎ B09YL4VP7J Country of Origin ‏ : ‎ USA A TEETH WHITENING PEN FOR SENSITIVE TEETH | Clinically proven hydrogen peroxide gel combined with desensitizing and plant based…