Natural Remedies for Rheumatoid Arthritis

Rheumatoid arthritis is an autoimmune disease that greatly reduces a person’s quality of life, and can cause disability and premature death.

It affects an estimated 1.5 million people in the United States, and more women have it than men.

To understand the natural remedies for rheumatoid arthritis (RA), it is important to know what the disease is, what causes it, and what natural remedies can do to help.

Treating RA

The aims of any treatment for RA are to:

rheumatoid arthritic hands holding a mug
Rheumatoid arthritis is a progressive autoimmune disease that causes inflammation in the joints, often in the feet and hands first.
  • Educate people about the disease
  • Reduce pain and swelling
  • Help people stay active and feel better
  • Slow damage to the joints

Treatments may be medical or nonmedical, and they are often used in combination.

Since RA is a progressive disease that gets worse without intervention, treatment tends to be aggressive.

Disease-modifying antirheumatic drugs (DMARDs) are often prescribed within 3 months of diagnosis, to reduce disease activity and prevent the joints from deforming.

People with RA should work with a specialist to discuss medical treatment, and talk about other remedies that can reduce discomfort and improve mobility and quality of life.

Natural remedies for rheumatoid arthritis

According to clinical guidelines found in Orgão Oficial da Sociedade Portugue sa de Reumatologia, physical therapy may help people with RA.

Physical therapy can involve stretching, exercise, heat and cold, and balanced rest.

Stretching

Stretching the muscles surrounding the affected joints may provide relief from symptoms of RA.

A study posted to Health Technology Assessment found that simple stretches and strengthening exercises on the hands can bring relief to patients with RA.

Results suggested that a stretching and strengthening program to relieve symptoms in the hands and wrists may be an effective supplement to conventional care methods.

People with RA should talk to a doctor or physical therapist before beginning any stretching regimen to make sure it will not put the joints under any undue stress.

Exercise

In addition to stretching, a low-stress workout program may help. Dynamic, low-stress activities, such as swimming or cycling can strengthen the muscles around the affected joints, reduce the impact on joints, and slow the progression of the RA.

Heat and cold

There is conflicting medical evidence on the effect of applying heat or cold to areas affected by RA. However, some people may find temporary relief through heating or cool the sore areas of their bodies.

In the home, heat packs or ice packs can provide relief to sore wrists and feet. Ultrasound heat and cold sprays can also be used to deliver heat or cold to a deep level of tissue without changing the temperature of the skin too much.

Balanced rest

Rest is an important treatment for aching joints, but it must be balanced with exercise, as too much rest can make aching joints worse.

It is important to work closely with a doctor and physical therapist to ensure there is a balance between rest periods and strengthening exercise.

Diet

Since inflammation is one of the main symptoms of RA, an anti-inflammatory diet may help to reduce symptoms.

Fruit and vegetables
Studies suggest a diet of fresh fruit and vegetables alongside grains and nuts may help to reduce inflammation.

A study published in Complementary Therapies in Medicine looked at 600 participants who followed a plant-based diet rich in fresh fruits and vegetables, whole grains, legumes, nuts, and seeds.

The researchers found that most participants had a reduction in a specific protein, which is known to be active in causing inflammation.

People who followed this anti-inflammatory vegan diet appeared to have significantly reduced systemic inflammation.

People with RA should consider a healthy anti-inflammatory diet to reduce pain and support their overall wellbeing.

Supplements

Specific supplements may help promote a healthy lifestyle. Fish oil from cold water fish such as salmon, mackerel, tuna, cod, and herring contain high amounts of omega-3 fatty acids. Omega-3 fatty acids are anti-inflammatory and block inflammatory receptors in the body. These can help as RA is an inflammatory disease.

A meta-analysis posted to The Nutrition Society compiled research on fish oil and RA. Their findings indicate that people who used fish oil and nonsteroidal anti-inflammatory drugs (NSAIDs) for a long time had less tenderness in their joints. The researchers concluded that fish oil may be beneficial as a supplemental therapy for people with RA.

Boswellia, or frankincense, is another powerful anti-inflammatory supplement, which may help relieve symptoms of RA.

Turmeric shows promise as a natural anti-inflammatory. In a study posted to Arthritis and Rheumatology, researchers found that a specific extract of turmeric reduced joint inflammation in people with arthritis.

Increasing turmeric consumption by adding the spice to food could help without causing any side effects. Turmeric supplements should be used with care. People who also use blood-thinning medication such as Warfarin should avoid turmeric.

A recent study published in the journal Nutrition found that treating patients with a specific probiotic, L. casei 01, improved both disease activity and inflammation in patients with RA.

If further research confirms these results, probiotics could become part of the treatment for RA. Rather than using supplements, people with RA can get all the probiotics they need from foods, such as yogurt, pickles, and cheese.

Some supplements may help with RA, but it is important to discuss these with a doctor before taking any as they may have adverse side effects. The U.S. Food and Drug Administration (FDA) do not regulate herbs and supplements. As a result, the doses of supplements can be irregular; some could be high and others low.

Lifestyle changes

Reducing stress on the body and mind is likely to help people with RA.

Regular mindful meditation, Tai chi, yoga, and qigong are all gentle ways to bring balance and relaxation to both the body and mind.

Overview of RA

Rheumatoid arthritis is a systemic disease in which the immune system attacks the individual’s own body tissues.

three generations
Although the exact cause is unknown, rheumatoid arthritis may be hereditary.

Symptoms most commonly appear in the joints, where RA creates inflammation and causes the lining of the joints to thicken. It can affect other parts too, such as the lungs.

Unchecked, this inflammation can damage the cartilage and bones. Early diagnosis and treatment can help.

The joints most commonly affected are the small joints in the hands, and feet although other joints can be affected too, usually, both hands and or feet are affected similarly.

Causes and symptoms

The exact cause of RA is unknown, but it appears to involve genetic, environmental, and hormonal factors.

Symptoms often begin in middle age and are more common in older people. They include inflammation, stiffness, pain, and swelling around the affected areas. Fatigue and weight loss can also occur because of the inflammation in the body as a whole.

Symptoms vary from person to person and can come and go over time. If untreated, RA tends to damage the joints where there is inflammation.

When to see a doctor

It is always important to speak with a doctor before beginning any treatment, including natural remedies. If a person wants to use herbs and supplements as part of their treatment, they should discuss this with their doctor. This is particularly important because the FDA do regulate herbs and supplements.

If inflammation or other symptoms become worse, it is important to consult a doctor.

Anyone who has been diagnosed with RA should learn about the options and make appropriate lifestyle changes to support a healthy future and reduce pain as far as possible.

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Can Essential Oils Help Treat ADHD?

Attention deficit hyperactivity disorder is a brain disorder that involves inattention, hyperactivity, impulsive behavior, or all three.

The individual may find it hard to remain focused, and they may face challenges with organizational skills. They may find it hard to remember or to follow instructions, and this can lead to seemingly careless mistakes at work or school.

With attention deficit hyperactivity disorder (ADHD), the symptoms are chronic so they can last for a long time. They may be severe enough to interrupt daily functioning, learning ability, social habits, and even relationships.

This article takes a look at whether essential oils can be used as a form of treatment for people with ADHD.

What are essential oils?

Essential oils
Essential oils are natural oils from plants that are obtained by cold press extraction or distillation.

Essential oils are concentrated forms of compounds found in various plants. They are normally made through either cold press extraction or distillation.

In cold press extraction, a large amount of plant material is pressed using great pressure, until the natural compounds are released.

Distillation involves placing plant matter in a closed container and passing steam or water through it to extract the compounds from the plants. When the excess water is removed, the concentrated compounds that remain are called essential oils.

Essential oils aim to contain the highest possible concentration of beneficial compounds.

Many of these compounds are being used or studied for their potential in medicine.

Can essential oils help with ADHD?

Small studies and anecdotal evidence have suggested that some essential oils may benefit people with ADHD.

There is no evidence to suggest that essential oils can be harmful to people with ADHD. A healthcare provider may help a patient to experiment with some oils to see if they relieve symptoms.

Lavender essential oil

A person who is experiencing hyperactivity with ADHD may have trouble feeling relaxed enough to fall, or stay, asleep.

A recent study carried out at the University of Minnesota found that lavender essential oil may improve sleep.

The researchers studied the effectiveness of inhaling lavender essential oil for people with sleep issues.

Results indicated that those participants who inhaled lavender essential oil had a better quality of sleep than those who did not.

The use of inhaled lavender oil appeared to boost the well-being of the users over time.

Vetiver essential oil

If a person with ADHD is having difficulty staying focused, vetiver essential oil may help them to stay alerted and on task.

A study posted in the Journal of Intercultural Ethnopharmacology has measured the direct effects that inhaling vetiver essential oil has on the brain.

Those who inhaled vetiver essential oil showed an increase in their attention levels, and this was reflected in measurements of brain activity.

The researchers noted that the effects of vetiver essential oil may be beneficial for learning and memory processes. It appears to have a stimulating effect even at low inhaled doses, and it may improve alertness and task performance.

Rosemary essential oil

essential_oil_of_rosemary1,8-cineole is one of the main compounds in rosemary oil. Studies may suggest that its use can increase cognitive performance.

The fresh scent of the common cooking ingredient rosemary may help to keep the mind sharp.

A study posted to Therapeutic Advances in Psychopharmacology looked at the effects of one of the main compounds in rosemary essential oil, called 1,8-cineole.

The research showed that a higher concentration of this compound in the body resulted in better performance on cognitive tests.

The results were noted in both speed and accuracy. People with higher levels of the compound in their body described themselves as feeling more content.

More studies are necessary to confirm the findings.

Other essential oils

Other essential oils have been used by many people for many different symptoms of ADHD, with varying results.

Essential oils that may be useful include:

  • Frankincense
  • Ylang ylang
  • Bergamot
  • Eucalyptus
  • Lemon
  • Cedarwood

One study reports that hospice patients who received a hand massage using oils containing bergamot, frankincense, and lavender, for example, felt relief from symptoms of depression, although the effect on the separate components was not studied.

Bergamot is thought to have antidepressant qualities, and a combination of clary sage and ylang-ylang may also lift the mood.

Cedarwood oil is said to have sedative effects, while some claim that both lemon and frankincense oils can improve focus.

However, there is not enough scientific evidence to confirm the benefits.

How to use essential oils

Essential oils are highly concentrated and should, therefore, be used with care. Before starting to use any essential oil, a physician can advise about any possible interactions with other medications.

Here are three appropriate ways to use essential oils:

  • Inhaling by adding a few drops to an aromatherapy diffuser
  • Applying diluted oils topically, to be absorbed through the skin
  • Putting a few drops into a full bath or a hand or foot bath

It is important to dilute essential oils with a carrier oil before putting them on the body. Examples of carrier oils include olive, coconut, and grapeseed oil.

The high concentrations of compounds in the oils can cause reactions if applied undiluted anywhere on the body.

It is also important to perform an allergy test. This test can be carried out by applying a small amount of the diluted mixture to a small area of the body, like the back of the hand.

If any signs of allergy such as redness or burning sensations occur, do not use the oil.

Using essential oils for children

Since a child’s body and immune system are still developing, extra care is needed when using essential oils.

Children under 10 years of age are advised not to use essential oils containing 1,8-cineole, such as eucalyptus, rosemary, or peppermint.

The compound can cause reactions in young children and should, therefore, be avoided.

Making lifestyle changes to support ADHD

Women doing yoga
Steady lifestyle changes such as exercise, including yoga, may help to focus and balance the mind.

Many people also find relief from ADHD symptoms by making steady lifestyle changes.

For example, drinking herbal teas such as chamomile may promote a calm state of mind.

Likewise, a diet that is rich in omega-3 fatty acids could lead to a decrease in ADHD symptoms.

Many people find relief from restlessness by exercising daily. Practices including meditation, yoga, and Tai chi may help to provide a more balanced state of mind.

Medical treatments for ADHD

Anyone who has been diagnosed with ADHD should discuss treatment options closely with a healthcare provider.

A physician will often prescribe stimulant drugs to treat ADHD. Stimulant drugs such as Ritalin, Adderall, Concerta, Vyvanse, and Dexedrine have an effect on brain chemistry, which calms the patient’s mind. Nonstimulant drugs include Strattera.

There are side effects associated with most drugs used for ADHD.

It is important to work with a doctor in order to find the best treatment and to discuss any alternative treatment options with the physician.

Quantitative Determination of 3-O-Acetyl-11-Keto-β-Boswellic Acid (AKBA) and Other Boswellic Acids in Boswellia sacra Flueck (syn. B. carteri Birdw) and Boswellia serrata Roxb

Giuseppe Mannino 1, Andrea Occhipinti 1,2 and Massimo E. Maffei 1,2,*
1
Biosfered S.r.l., Innovation Centre, Academic Spin-Off of the University of Turin, Via Quarello 15/A, Turin 10135, Italy
2
Department of Life Sciences and Systems Biology, Innovation Centre, University of Turin, Via Quarello 15/A, Turin 10135, Italy
*
Correspondence: Tel.: +39-011-670-5967
Academic Editor: Vassilios Roussis
Received: 29 July 2016 / Accepted: 1 October 2016 / Published: 6 October 2016

Abstract

:

Boswellia serrata and Boswellia sacra (syn. B. carteri) are important medicinal plants widely used for their content of bioactive lipophilic triterpenes. The qualitative and quantitative determination of boswellic acids (BAs) is important for their use in dietary supplements aimed to provide a support for osteoarthritic and inflammatory diseases. We used High Performance Liquid Chromatography (HPLC)-Diode Array Detector (DAD) coupled to ElectroSpray Ionization and tandem Mass Spectrometry (ESI-MS/MS) for the qualitative and quantitative determination of BAs extracted from the gum resins of B. sacra and B. serrata. Limit of detection (LOD), limit of quantification (LOQ), and Matrix Effect were assessed in order to validate quantitative data. Here we show that the BAs quantitative determination was 491.20 g·kg−1 d. wt (49%) in B. sacra and 295.25 g·kg−1 d. wt (30%) in B. serrata. Lower percentages of BAs content were obtained when BAs were expressed on the gum resin weight (29% and 16% for B. sacra and B. serrata, respectively). The content of Acetyl-11-Keto-β-Boswellic Acid (AKBA) was higher in B. sacra(70.81 g·kg−1 d. wt; 7%) than in B. serrata (7.35 g·kg−1 d. wt; 0.7%). Our results show that any claim of BAs content in either B. sacra or B. serrata gum resins equal to or higher than 70% or AKBA contents of 30% are simply unrealistic or based on a wrong quantitative determination.
Keywords:

Acetyl-11-Keto-β-Boswellic Acid; 11-Keto-beta-Boswellic Acid; boswellic acids; standardization; Boswellia serrata; Boswellia sacra; HPLC-DAD-ESI-MS/MS

1. Introduction

The genus Boswellia (Burseraceae), comprises 25 species of trees and shrubs which are widely spread in Arabia, the north-eastern coast of Africa and India [1]. Since ancient times, the natural resin of Boswellia trees has been collected and used to produce the oleo gum resin, frankincense (olibanum). The gum resin is harvested from incisions made on the trunk of the tree and the darkening of resin droplets is an index of oxidation [2]. Among Boswellia species, only a few are of economic importance as a natural source of phytopharmaceutical compounds, including B. serrata Roxb. and B. sacra Flueck (syn. B. carteri Birdw, syn. B. undulatocrenata Engl.) [3,4,5].
B. serrata is used for the treatment of oxidative and inflammatory damage [2], rhinitis [6] asthma [7], age-related disorders [8], neurorecovery [9], arthritis [10], skin disorder [11], cancer [12], and against several human pathogenic and plant pathogenic fungi [13]. Recently, the pharmacological properties and clinical effectiveness of B. serrata have been studied systematically [3].
B. sacra oleo gum resin is used in the treatment of gastric and hepatic disorders [14], skin disorders [15], for its hepatoprotective activity [16], analgesic effect [4], antiglycation and antioxidant activities [17], tumor suppression [18], anticoagulation effects [19], antinflammatory activity [20], and cardioprotective effects [21].
The main constituents of B. serrata and B. sacra are volatile oils, composed of monoterpenes and sesquiterpenes [22,23], diterpenes including incensole, incensole acetate and cembrenol (serratol) [24], lipophilic pentacyclic triterpene acids of the oleanane-(α-boswellic acids), ursane-(β-boswellic acids) and lupane-type (lupeolic acids), as well as an ether-insoluble fraction containing polysaccharides (arabinose, galactose, xylose) [25]. Among triterpenoids, bioactive boswellic acids are of particular interest, particularly 3-O-Acetyl-11-Keto-β-Boswellic Acid (AKBA), 11-Keto-beta-Boswellic Acid (KBA), and the various β-boswellic acids (βBAs), and α-boswellic acids (αBAs) and their esters. The analysis of these triterpenes is performed by different analytical methods including High Performance Thin Layer Chromatography (HPTLC) [26], although the most used methods are based on HPLC coupled to both photodiode array detection [27] and mass spectrometry detection [28]. In accordance with the spectral properties of the boswellic acids, their analysis is performed at three different wavelengths, 210 nm for αBAs, βBAs as well as lupeolic acid, 250 nm for AKBA and KBA, and 280 nm for 9,11-dehydro-α- and -β-boswellic acids [27]. However, a precise identification and quantification of boswellic acids is usually obtained with liquid chromatography electrospray ionisation tandem mass spectrometry (LC-ESI-MS/MS) by using selected ion monitoring (SIM) detection [28].
In general, the total organic acids from B. serrata and B. sacra constitute approximately 65%–70% by weight of the total alcoholic extract. Of this fraction, approximately 25% is made of triterpenes. In market products, these percentages are often misinterpreted and it is not unusual to find claims of 70% boswellic acids content or 30% AKBA content, which is obviously misleading because the highest amounts so far reported of boswellic acids in B. serrata is about 140 mg/g (i.e., 14%) and in B. sacra is about 190 mg/g (i.e., 19%) [27]. Since both Boswellia extracts are used in several formulations, it is important to express unequivocally the “real content” of boswellic acids in both B. serrata and B. sacra. Therefore, the aim of this work is to provide a guideline for the accurate identification and quantification of boswellic acids in these two important Boswellia species by using HPLC-DAD-ESI-MS/MS.

2. Results and Discussion

2.1. Identification of Boswellic Acids

Gum resin of B. sacra and B. serrata were extracted using methanol in order to evaluate the total BAs content. The total yield for B. sacra methanolic extract was 598.88 g·kg−1 (±11.40 g·kg−1) gum resin dry weight whereas the yield for B. serratawas 549.78 g·kg−1 (±29.31 g·kg−1) gum resin dry weight. The total recovery of methanolic extracts from B. sacra was 99.17% (±1.93%), whereas the total recovery from B. serrata was 98.96% (±1.97%). Our findings indicate that the total content of the lipophilic extracts (excluding the polysaccharide moiety of both species) never exceeded 60%, in agreement with previous works [27,29,30]. By considering that the methanolic fraction contains several lipophilic compounds, including mono-, di- and triterpenes, it is evident that a claim of 70% BAs is not sustainable, being the BAs only a portion of the total methanolic extract.
In order to define the content and to identify the BAs present in the two Boswellia methanolic extracts, we performed HPLC-DAD-ESI-MS/MS analyses. Several BAs were present in both methanolic extracts. Table 1 reports the molecular mass and the fragmentation pattern of compounds identified in the methanolic extracts, whereas Figure 1 shows the chemical structure of the identified BAs. The identification of these compounds was achieved by both mass spectrometry and comparison with pure standards (see Supplementary Figure S1 for mass spectra of identified compounds and Supplementary Figure S2 for UV chromatograms). In both species, the main BAs were represented by AKBA, KBA, αBA, βBA, acetyl-αBA (A-αBA) and acetyl-βBA (A-βBA), in accordance with the literature data [2,31,32].

Figure 1. Chemical structure of the boswellic acids identified in Boswellia serrata and Boswellia sacra. Numbers correspond to compounds listed in Table 1.
Table 1. Molecular mass and fragmentation pattern of compounds identified in the methanolic extracts of B. sacra and B. serrata.

2.2. Quantification of Boswellic Acids

Table 2 shows for both species the quantitative determination of the main BAs. The total amount of the main BAs was statistically (p < 0.05) higher in B. sacra than in B. serrata. In B. sacra, the total amount of the main BAs in the methanolic extract was lower than 50% and this value was reduced to about 29% when the amount was considered in terms of the total gum resin dry weight (Table 2). In B. serrata the total content of the main BAs in the methanolic extract was lower than 30% and the value dropped to 16% when BAs were calculated in terms of the gum resin dry weight (Table 2). In both species, the major BAs were represented by αBA and βBA, in agreement with literature data [32]. A direct comparison between the two species shows that the contents of B. sacra AKBA (about 10 fold), αBA (1.5 fold) and βBA (1.6 fold) were statistically (p < 0.05) higher than in B. serrata.

Table 2. Quantitative determination of boswellic acids in Boswellia sacra and Boswellia serrata by High Performance Liquid Chromatography-Diode Array Detector coupled to ElectroSpray Ionization and tandem Mass Spectrometry (HPLC-DAD-ESI-MS/MS), by using calibration curves from pure standards. Data are expressed as g·kg−1 gum resin dry weight. (Standard deviation), in the same row, different letters indicate significant (p < 0.05) differences.
In order to validate the quantitative analyses reported in Table 2, we calculated the linearity and precision of the identified BAs standard curves, the detection limit (LOD), the quantification limit (LOQ) and the Matrix Effect (ME). Table 3shows the validation results for the main identified BAs. All compounds showed a high R2 value, which indicates a high linearity in the calibration curves. The lowest LOD and LOQ values were found for A-βBA, followed by equal values for KBA and AKBA. The highest LOD and LOQ values were found for A-αBA. In order to complete the validation process, we assessed the ME, to assure that precision, selectivity and sensitivity were not compromised during HPLC-ESI-MS/MS analyses. The absolute ME was calculated by comparing the slope of matrix-matched standard curve with the slope of the standard calibration curve, according to [33]. Table 3 reports, for each species, the ME accuracy (expressed as percent values) of the main identified BAs. In B. sacra extracts, KBA (CV = 14.06) and A-βBA (CV = 11.61) showed the highest percentage of accuracy, followed by AKBA (CV = 7.53) and A-αBA (CV = 1.90). βBA (CV = 8.72) showed the lowest ME accuracy percentages. In B. serrata, the highest percentages of accuracy were found for αBA (CV = 0.22), followed by KBA (CV = 14.13), A-αBA (CV = 2.66) and AKBA (CV = 7.74).

Table 3. Validation of boswellic acids (BAs) quantitative analyses of methanolic extracts from B. sacra and B. serrata. (Standard deviation).
Finally, Table 4 shows the recovery of the identified BAs from B. sacra and B. serrata methanolic extracts. In both species, the total recovery was higher than 98%. In B. sacra, the highest recovery was found for AKBA and αBA, whereas in B. serrata the highest recovery was found for A-βBA and A-αBA (Table 4).

Table 4. Percentage of recovery of identified BAs from B. sacra and B. serratamethanolic extracts. Values are expressed as percentage of recovery.
The quantitative determination of B. sacra and B. serrata BAs content and the validation of the quantitative chemical analysis show that any claim of BAs content in either B. sacra or B. serrata gum resins equal to or higher than 70% or 30% AKBA are simply unrealistic or based on a wrong quantitative determination. The same is true when the percentage of BAs is calculated in the methanolic extract.

3. Materials and Methods

3.1. Plant Material and Chemicals

Boswellia serrata Roxb. and Boswellia sacra Flueck gum resins were purchased from Bauer S.r.l. (Udine, Italy). The origin of B. sacra samples was from Ethiopia, whereas B. serrata samples originated from India. The gum resins were milled to coarse powder and used for all extractions. All chemicals were of analytical reagent-grade unless stated otherwise. Pure standards were purchased for the quantification by external calibration curves: 11-Keto-β-boswellic acid, α-Boswellic acid and β-Boswellic acid (ExtraSynthese, Lyon, France), 3-O-Acetyl-11-keto-β-Boswellic Acid (Merck, Darmstadt, Germany), 3-O-Acetyl-α-boswellic acid, 3-O-Acetyl-β-boswellic acid (Sigma-Aldrich, St. Louis, MO. USA).

3.2. Solvent Extraction of Boswellia serrata and Boswellia sacra oleo Gum Resins

One hundred grams of ground B. serrata and B. sacra oleo gum-resins were extracted with 1 L methanol (VWR International, Radnor, PA, USA) (extraction ratio 1:10 w/v). Samples were then placed on an orbital shaker for 5 days in the dark. Extracts were then filtered and the resin was rinsed with 400 mL of methanol. To evaluate the recovery of analyzed compounds, the exhaust gum resin was re-extracted with methanol as previously described. Samples were then concentrated by vacuum evaporation (Rotavapor, Büchi, Flawil, Switzerland). Concentrated extracts were then dried in a ventilated oven at 70 °C for 4 h. The powdered extracts were stored at room temperature in the dark until chemical analysis. Extractions were performed in triplicate.

3.3. Isolation and Quantification of Boswellic Acids by HPLC-DAD-ESI-MS/MS

Boswellic acids were identified and quantified by liquid chromatography (1200 HPLC, Agilent Technologies, Santa Clara, CA, USA) equipped with a reverse phase column, Luna C18 (3 µm, 150 mm × 3.0 mm, Phenomenex, Torrance, CA, USA). B. serrata and B. sacra powdered extracts were dissolved (30 mg·mL−1) in HPLC-grade methanol and properly diluted. The binary solvent system was: (A) MilliQ H2O (Millipore, Billerica, MA, USA):Methanol 50:50 containing 5 mM ammonium acetate (Sigma-Aldrich, USA); and (B) Methanol:1-Propanol (VWR International, Radnor, PA, USA) 80:20 containing 5 mM ammonium acetate. The chromatographic separation was carried out at constant flow rate (200 µL·min−1) with the following conditions: linear gradient from 30% to 50% of B in 2 min, then 80% of B in 35 min, then at 47 min B concentration was raised to 98%. The concentration of solvent B was maintained at 98% for 6 min. The initial mobile phase was re-established for 10 min before the next injection. The temperature of wellplate autosempler G1377A was set 4 °C while chromatography was carried out at constant temperature (30 °C) controlled by an Agilent 1100 HPLC G1316A Column Compartment.
Tandem mass spectrometry analyses were performed with a 6330 Series Ion Trap LC-MS System (Agilent Technologies, USA) equipped with an electrospray ionization source (ESI) operating in negative mode. The flow rate of nitrogen was set 325 °C and 5.0 L·min−1, while the Capillary Voltage was 1.5 kV. Helium was used as a collision gas.
Identification of Boswellia oleo gum resin compounds was performed by scan analyses with a 50–750 m/z scan range and by monitoring the absorption at 210, 250 and 280 nm. Quantitative analyses were performed by Multiple Reaction Monitoring (MRM) by monitoring the fragmentation of quasi-molecular ions for αBA and βBA and KBA (Table 1) and by Diode Array Detector (DAD) at 250 nm for AKBA and 210 nm for A-αBA and A-βBA. Quantification was performed by external calibration curves with pure standards dissolved in HPLC grade Methanol. Limit of Detections (LOD) and Limit of Quantifications (LOQ) for each compounds were determined as described in [34].
To evaluate the ME in the quantification of target compounds, B. serrata and B. sacra powdered extracts were dissolved (30 mg·mL−1) in HPLC-grade methanol and properly diluted. These sample solutions were used to prepare the calibration curves in the presence of other extracted gum resin compounds [33]. The slope of standard curves obtained with the solvent (methanol) and in the extracts were used to compare the ME percentage (%ME = Calibration Slope(sample)/CalibrationSlope(standard) × 100). 100% ME percentage indicates no ME, a ME% < 100% indicates ionization suppression and a ME% > 100% indicates ionization enhancement.

4. Conclusions

Boswellia sacra and Boswellia serrata extracts are widely used in pharmaceutical and nutraceutical preparations. The bioactivity of these Boswellia extracts is based on the content of BAs. Clearly, the dose of bioavailable BAs is central to the issue of Boswellia efficacy. Claims of 70% BAs or even 30%–40% AKBA are currently found, but this work confirms that the BAs content never exceeds 50% of the methanolic extract, whereas lower percentages are obtained when BAs are expressed in terms of the gum resin weight. Moreover, the highest percentage of AKBA found in B. sacra was below 8%. Only analytical methods based on HPLC coupled to mass spectrometry allow the precise quantification and identification of BAs in Boswellia extracts, whereas other methods based only on HPLC or spectrophotometric methods do not sufficiently allow an accurate quantification of BAs. Therefore, we recommend LC-MS technology for BAs determination and quantification.

Supplementary Materials

The following are available online at http://www.mdpi.com/1420-3049/21/10/1329/s1, Figure S1: mass spectra of boswellic acids isolated in this study; Figure S2: UV chromatograms of boswellic acids isolated in this study.

Acknowledgments

This work was supported by the PhD Program in Pharmaceutical and Biomolecular Sciences of the University of Turin and by the Academic Spin-Off Biosfered S.r.l.

Author Contributions

M.E.M and A.O. conceived and designed the experiments; A.O. and G.M. performed the experiments; M.E.M. and A.O. analyzed the data; M.E.M. contributed reagents/materials/analysis tools; M.E.M. wrote the paper.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Atta, U.R.; Naz, H.; Makhmoor, F.T.; Yasin, A.; Fatima, N.; Ngounou, F.N.; Kimbu, S.F.; Sondengam, B.L.; Choudhary, M.I. Bioactive constituents from Boswellia papyrifera. J. Nat. Prod. 2005, 68, 189–193. [Google Scholar] [CrossRef] [PubMed]
  2. Siddiqui, M.Z. Boswellia serrata, a potential antiinflammatory agent: An overview. Indian J. Pharm. Sci. 2011, 73, 255–261. [Google Scholar] [PubMed]
  3. Ernst, E. Frankincense: Systematic review. Brit. Med. J. 2008, 337, 1439–1441. [Google Scholar] [CrossRef] [PubMed]
  4. Al-Harrasi, A.; Ali, L.; Hussain, J.; Rehman, N.U.; Mehjabeen; Ahmed, M.; Al-Rawahi, A. Analgesic effects of crude extracts and fractions of Omani frankincense obtained from traditional medicinal plant Boswellia sacra on animal models. Asian Pac. J. Trop. Med. 2014, 7, S485–S490. [Google Scholar] [CrossRef]
  5. ThePlantList, Boswellia sacra Flueck. is an accepted name. Available online: http://www.theplantlist.org/tpl1.1/record/kew-2680579 (accessed on July 20 2016).
  6. Marogna, M.; Braidi, C.; Colombo, C.; Colombo, F.; Palumbo, L. A randomized controlled trial of a phytotherapic compound containing Boswellia serrata and bromeline for seasonal allergic rhinitis complicated by upper airways recurrent respiratory infections. J. Allergy Clin. Immun. 2015, 135, Ab271–Ab271. [Google Scholar] [CrossRef]
  7. Soni, K.K.; Lawal, T.; Wicks, S.; Patel, U.; Mahady, G.B. Boswellia serrata and Ocimum sanctum extracts reduce inflammation in an ova-induced asthma model of BALB/c mice. Planta Med. 2015, 81, 879–879. [Google Scholar] [CrossRef]
  8. Hosseini-Sharifabad, M.; Esfandiari, E. Effect of Boswellia serrata gum resin on the morphology of hippocampal CA1 pyramidal cells in aged rat. Anat Sci. Int. 2015, 90, 47–53. [Google Scholar] [CrossRef] [PubMed]
  9. Moein, P.; Fard, S.A.; Asnaashari, A.; Baratian, H.; Barekatain, M.; Tavakoli, N.; Moein, H. The effect of Boswellia serrata on neurorecovery following diffuse axonal injury. Brain Injury 2013, 27, 1454–1460. [Google Scholar] [CrossRef] [PubMed]
  10. Wang, H.; Zhang, C.N.; Wu, Y.; Ai, Y.; Lee, D.Y.W.; Dai, R.H. Comparative pharmacokinetic study of two boswellic acids in normal and arthritic rat plasma after oral administration of Boswellia serrata extract or Huo Luo Xiao Ling Dan by LC-MS. Biomed. Chromatogr. 2014, 28, 1402–1408. [Google Scholar] [CrossRef] [PubMed]
  11. Togni, S.; Maramaldi, G.; Bonetta, A.; Giacomelli, L.; Pierro, F. Clinical evaluation of safety and efficacy of Boswellia-based cream for prevention of adjuvant radiotherapy skin damage in mammary carcinoma: A randomized placebo controlled trial. Eur. Rev. Med. Pharm. 2015, 19, 1338–1344. [Google Scholar]
  12. Ahmed, H.; Abdel-Rahman, M.; Salem, F.E.; Shalby, A.; Lokman, M.S. Boswellia serrata and colon cancer: Evidence based approach for treatment. Planta Med. 2014, 80, 1545–1545. [Google Scholar] [CrossRef]
  13. Gangwal, M.L.; Vardhan, D.K. Antifungal Studies of Volatile Constituents of Boswellia serrata. Asian J. Chem. 1995, 7, 675–676. [Google Scholar]
  14. Asad, M.; Alhomoud, M. Proulcerogenic effect of water extract of Boswellia sacra oleo gum resin in rats. Pharm. Biol.2016, 54, 225–230. [Google Scholar] [CrossRef] [PubMed]
  15. Nardoni, S.; Giovanelli, S.; Pistelli, L.; Mugnaini, L.; Profili, G.; Pisseri, F.; Mancianti, F. In Vitro Activity of Twenty Commercially Available, Plant-Derived Essential Oils against Selected Dermatophyte Species. Nat. Prod. Commun.2015, 10, 1473–1478. [Google Scholar] [PubMed]
  16. Asad, M.; Alhumoud, M. Hepatoprotective effect and GC-MS analysis of traditionally used Boswellia sacra oleo gum resin (Frankincense) extract in rats. Afr. J. Tradit. Complement. 2015, 12, 1–5. [Google Scholar] [CrossRef]
  17. Al-Harrasi, A.; Ali, L.; Ceniviva, E.; Al-Rawahi, A.; Hussain, J.; Hussain, H.; Rehman, N.U.; Abbas, G.; Al-Harrasi, R. Antiglycation and Antioxidant Activities and HPTLC Analysis of Boswellia sacra Oleogum Resin: The Sacred Frankincense. Trop. J. Pharm. Res. 2013, 12, 597–602. [Google Scholar] [CrossRef]
  18. Suhail, M.M.; Wu, W.J.; Cao, A.; Mondalek, F.G.; Fung, K.M.; Shih, P.T.; Fang, Y.T.; Woolley, C.; Young, G.; Lin, H.K. Boswellia sacra essential oil induces tumor cell-specific apoptosis and suppresses tumor aggressiveness in cultured human breast cancer cells. BMC Complement. Altern. Med. 2011, 11. [Google Scholar] [CrossRef] [PubMed]
  19. Pan, Y.N.; Liang, X.X.; Niu, L.Y.; Wang, Y.N.; Tong, X.; Hua, H.M.; Zheng, J.; Meng, D.Y.; Liu, X.Q. Comparative studies of pharmacokinetics and anticoagulatory effect in rats after oral administration of Frankincense and its processed products. J. Ethnopharmacol. 2015, 172, 118–123. [Google Scholar] [CrossRef] [PubMed]
  20. Banno, N.; Akihisa, T.; Yasukawa, K.; Tokuda, H.; Tabata, K.; Nakamura, Y.; Nishimura, R.; Kimura, Y.; Suzuki, T. Anti-inflammatory activities of the triterpene acids from the resin of Boswellia carteri. J. Ethnopharmacol. 2006, 107, 249–253. [Google Scholar] [CrossRef] [PubMed]
  21. Zaki, A.A.; Hashish, N.E.; Amer, M.A.; Lahloub, M.F. Cardioprotective and antioxidant effects of oleogum resin “Olibanum” from Boswellia carteri Birdw. (Bursearceae). Chin. J. Nat. Med. 2014, 12, 345–350. [Google Scholar] [CrossRef]
  22. Niebler, J.; Buttner, A. Unraveling the smell of frankincense: Identification of potent odorants in Boswellia sacra resin and pyrolysate. Abs. Pap. Am. Chem. Soc. 2014, 248, 159. [Google Scholar]
  23. Singh, B.; Kumar, R.; Bhandari, S.; Pathania, S.; Lal, B. Volatile constituents of natural Boswellia serrata oleo-gum-resin and commercial samples. Flav. Fragr. J. 2007, 22, 145–147. [Google Scholar] [CrossRef]
  24. Pollastro, F.; Golin, S.; Chianese, G.; Putra, M.Y.; Moriello, A.S.; de Petrocellis, L.; Garcia, V.; Munoz, E.; Taglialatela-Scafati, O.; Appendino, G. Neuractive and anti-inflammatory Frankincense cembranes: A structure-activity study. J. Nat. Prod. 2016, 79, 1762–1768. [Google Scholar] [CrossRef] [PubMed]
  25. Herrmann, A.; Lechtenberg, M.; Hensel, A. Comparative isolation and structural investigations of polysaccharides from Boswellia serrata ROXB and Boswellia carteri BIRDW. Planta Med. 2007, 73, 815–815. [Google Scholar] [CrossRef]
  26. Krohn, K.; Rao, M.S.; Raman, N.V.; Khalilullah, M. High-performance thin layer chromatographic analysis of anti-inflammatory triterpenoids from Boswellia serrata Roxb. Phytochem. Anal. 2001, 12, 374–376. [Google Scholar] [CrossRef] [PubMed]
  27. Buchele, B.; Zugmaier, W.; Simmet, T. Analysis of pentacyclic triterpenic acids from frankincense gum resins and related phytopharmaceuticals by high-performance liquid chromatography. Identification of lupeolic acid, a novel pentacyclic triterpene. J. Chromatogr. B-Anal. Technol. Biomed. Life Sci. 2003, 791, 21–30. [Google Scholar] [CrossRef]
  28. Frank, A.; Unger, M. Analysis of frankincense from various Boswellia species with inhibitory activity on human drug metabolising cytochrome P450 enzymes using liquid chromatography mass spectrometry after automated on-line extraction. J. Chromatogr. A 2006, 1112, 255–262. [Google Scholar] [CrossRef] [PubMed]
  29. Buchele, B.; Zugmaier, W.; Genze, F.; Simmet, T. High-performance liquid chromatographic determination of acetyl-11-keto-alpha-boswellic acid, a novel pentacyclic triterpenoid, in plasma using a fluorinated stationary phase and photodiode array detection: Application in pharmacokinetic studies. J. Chromatogr. B 2005, 829, 144–148. [Google Scholar] [CrossRef] [PubMed]
  30. Buchele, B.; Simmet, T. Analysis of 12 different pentacyclic triterpenic acids from frankincense in human plasma by high-performance liquid chromatography and photodiode array detection. J. Chromatogr. B 2003, 795, 355–362. [Google Scholar] [CrossRef]
  31. Paul, M.; Bruning, G.; Bergmann, J.; Jauch, J. A Thin-layer Chromatography Method for the Identification of Three Different Olibanum Resins (Boswellia serrata, Boswellia papyrifera and Boswellia carterii, respectively, Boswellia sacra). Phytochem. Anal. 2012, 23, 184–189. [Google Scholar] [CrossRef] [PubMed]
  32. Shah, B.A.; Qazi, G.N.; Taneja, S.C. Boswellic acids: a group of medicinally important compounds. Nat. Prod. Rep.2009, 26, 72–89. [Google Scholar] [CrossRef]
  33. Villagrasa, M.; Guillamon, M.; Eljarrat, E.; Barcelo, D. Matrix effect in liquid chromatography-electrospray ionization mass spectrometry analysis of benzoxazinoid derivatives in plant material. J. Chromatogr. A 2007, 1157, 108–114. [Google Scholar] [CrossRef] [PubMed]
  34. Validation of analytical procedures: Text and Methodology Q2(R1). Available online: http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf (accessed on 27 July 2016).
  • Sample Availability: Samples of the compounds are not available.

Can Frankincense Treat Cancer?

Frankincense oil is derived from the Boswellia tree. It has a long history in myth and folk medicine. In the Bible, it is one of three gifts offered to Jesus by the wise men, possibly because of its apparent healing powers.

Some supporters of herbal medicine argue that frankincense offers numerous health benefits. These supposed benefits include controlling bleeding, speeding up the wound-healing process, improving oral health, fighting inflammatory conditions such as arthritis, and improving uterine health.

Its most promising use may be as a cancer treatment. Cancer is a leading cause of death, killing 8.2 million people worldwide in 2012. Current research on the effectiveness of frankincense is limited, but early results are promising.

How might frankincense reduce inflammation?

Frankincense contains boswellic acid, which may help fight inflammation.

Inflammation is one of the key processes through which the body fights infection. When tissue becomes inflamed, white blood cells arrive to fight infection. Local inflammation causes redness, swelling, and heat. It can occur with injuries ranging from mild to life-threatening.

three-wise-men

Frankincense is steeped in history, but will it work as a modern-day cure?

 

Pimples, for example, are pockets of inflammation. Serious skin infections such as cellulitis can cause inflammation to spread and become life-threatening.

Long-term inflammation, especially when it occurs in multiple areas of the body, is associated with a wide range of health issues. Arthritis is an inflammatory disease, and evidence increasingly points to inflammation as a factor in depression.

A 2006 study published in Planta Medica uncovered a number of ways the boswellic acid in frankincense might fight infection. Boswellic acid inhibited 5-lipoxygenase, a chemical involved in inflammatory processes. Researchers also found that boswellic acid might target free radicals and cytokines. Both of these play a role in inflammation.

This has important implications for the fight against cancer. Numerous studies have linked inflammation to cancer. By disrupting inflammatory processes, frankincense could stop cancer before it starts.

The anti-inflammatory properties of frankincense suggest that it might also be effective in the treatment of diseases such as:

  • Rheumatoid arthritis
  • Crohn’s disease
  • Bronchial asthma
  • Ulcerative colitis.

Could frankincense fight cancer directly?

Frankincense might not just reduce inflammation. It may also directly attack cancer cells.

[frankincense boswellia tree]
Frankincense comes from the Boswellia tree.

 

One of the challenges of cancer treatment is that, unlike bacteria or viruses, cancer cells are not foreign invaders. Instead, cancer occurs when the body’s cells grow out of control, attacking healthy tissue.

This process makes it difficult to fight cancer without also killing healthy cells. In fact, most cancer treatments do kill healthy cells.

Chemotherapy, for example, kills many healthy cells as it fights cancer. This is why cancer patients undergoing chemotherapy often lose their hair, experience nausea, and become more vulnerable to infection.

Some evidence suggests that frankincense might target cancer cells without harming healthy cells.

A 2009 study of bladder cancer studied how frankincense affected cultures of normal and cancerous bladder cells. The oil targeted cancerous cells, but it did not destroy healthy cells.

A 2015 study found similar effects in breast cancer. The researchers found that frankincense could kill breast cancer cells and disrupt the growth of future cancer cells.

The results of these studies are preliminary. However, they offer hope that frankincense might one day fight some forms of cancer without the potentially life-threatening effects of chemotherapy.

Future research: Is a cancer cure on the horizon?

Research on frankincense as a cancer treatment has only looked at cell cultures and not cancer growing in a living, human being. Before frankincense can be used to treat cancer, researchers must perform human trials to prove that it works and is reasonably safe.

A number of studies have uncovered substances that can kill cancer cells in a petri dish. Substances ranging from bleach to antiparasite drugs have killed cancer in a lab, but not yet in a human.

Human bodies are complicated systems. Before using frankincense on people, scientists must work out a safe dosage, explore potential side effects, and decide how best to deliver treatment. As research is still in its early stages, it is unlikely that frankincense will become a mainstream cancer treatment in the near future.

Safe use of frankincense

People should talk to a doctor before trying frankincense or any other essential oil.

Frankincense is not an alternative to mainstream cancer treatments. No research currently supports using the oil in place of other cancer treatments. Frankincense may, however, be used as a supplement to medical treatment.

Frankincense has not been approved as a drug for any specific disease, and there are no scientifically proven guidelines for its use.

These include:

  • Using frankincense in skin care products, for example, adding a drop or two of frankincense oil to a favorite lotion.
  • Soaking in frankincense in the bath tub. A few drops create an aromatic soak, and the body may absorb some of the oil.
  • Using frankincense on pulse points during meditation or yoga, or applying a few drops of oil to a hot compress.
  • Ingesting frankincense, but remembering to dilute the oil first. One popular recommendation advises a 10 to 1 ratio of water to frankincense.

People can also add frankincense to honey or another sweetener. They can conceal its pungent taste by adding the sweetener to coffee or tea. It is recommended to limit intake to just a few drops per day and to speak with a doctor for approval first.

Users should watch carefully for side effects, and they should stop use immediately if any ill effects develop. If anyone plans to use frankincense on their skin, they should try testing a diluted version of the oil on a small patch of skin first.

Essential oils can be diluted with olive oil. If someone intends to ingest the oil, they should start with a heavily diluted drop, then gradually increase the dosing over several days.

Frankincense is natural, but like many other natural substances, it can be poisonous. People who are pregnant, lactating, have a history of allergic reactions, or have a weakened immune system, should avoid using frankincense unless their doctor says otherwise.