Mareecham-Black pepper (Piper nigrum) Part 2

Mareecham-Black pepper (Piper nigrum) Part 2
Phytochemistry
Many investigators isolated different types of compounds viz. Phenolics, flavonoids, alkaloids, amides, steroids, lignans, neolignans, terpenes, chalcone, cardiac glycosides and many other compounds. As the chemicals are isolated from different parts of the plant, to avoid confusion I mention pharmacologically active compounds:
Alkaloids: Piperine, piperidine, chavicine,
Amides: Pitigrine and wisamine are insecticide 
Alkamides: Unsaturated fatty acids linked with amines by an amine bond
Benzodiaxole compounds: Dipiperamide D and Dipiperamide E,  
Oils: Oleoresin responsible for aromatic odor and the known pungent taste, a volatile oil, fatty oil
Bioactive compounds isolated recently are: Pellitorine, Guineensine, Pipnoohine, Trichostachine, Piperonal, trans-caryophyllene, cis- caryophyllene, 3-canene, Humulene, Limolene, Pinene, Brachyamide B, (2E,4E)-N-Eicosadienoyl-pereridine, N-trans-Feruloyltryamine, N-Formylpiperidine, Pentadienoyl as piperidine, (2E, 4E)-N-isobutyl-decadienamide, isobutyl-icosadeinamide, Tricholein, Trichostachine, isobutyl-eicosatrienamide, isobutyl-octadienamide, Piperamide, Piperamine, Piperretine, Pipericide, Piperolein B, Sarmentine Sarmentosine, Retrofractamide A 

Nutrient analysis per 100 g: energy 255 Kcal, proteins 10.95 g, carbohydrates 64.81 g, total fat 3.25 g, cholesterol 0 mg, dietary fiber 26.5 g.
Vitamins: choline 11.3 mg, folic acid 10 mcg, niacin 1.142 mg, pyridoxine 0.340 mg, riboflavin 0. 240 mg, thiamine 0. 109 mg, vitamin A 299 IU, vitamin C 21 mg, vitamin E 4.56 mg, vitamin K 163.7mcg.
Electrolytes: Sodium (Na) 44 mg, potassium (K) 1259 mg.
Minerals: Calcium 437 mg, copper (Cu) 1.127 mg, Iron (Fe) 28.86 mg, magnesium (Mg) 194 mg, manganese 5.625 mg, phosphorus (P) 173 mg, zinc 1.42mg
Phytonutrients: α carotene 0 mcg, β- carotene 156 mcg, crypto-xanthin- β 48 mcg, lutein-zeaxanthin 205 mcg, lycopene 6 mcg

Major chemical compounds responsible for aroma and pungency: α-terpineol (floral), acetophenone (irritant, sharp), hexonal (green apple), nerol (fresh, floral, herbal), nerolidol (mild spicy, rooty), 1, 8- cineol (camphory), dihydrocarveol (warm, wody), citral (citrusy), α-pinene (terperic, oxidized), piperolnol (sweet, floral) [24], [25], [26]

Identity, Purity and Strength   

Total Ash:                                                4.45 % w/w 

Water soluble ash:                                  4.21 % w/w

Acid insoluble ash:                                 0.45 % w/w

Water Soluble Extractive:                    25.04 % w/w

Methanol Soluble Extractive:              10.02 % w/w

Acetone Soluble Extractive:                 5.12 % w/w

Chloroform Soluble Extractive:          7.02 % w/w

Loss on Drying:                                 3.45%w/w  [27]

Heavy Metal Analysis of Piper nigrum

Determination of heavy metal residue in fruit extract of Marichi-Black Pepper (Piper nigrum) as per WHO guidelines. (Values in mg/kg)


   Element                  Observed Values               Permissible Values  

Cadmium (Cd)             0.22 +/- 0.04                            0.3
Lead (Pb)                     0. 45 +/- 0.05                         10.00
Arsenic (As)                  0.18 +/- 0.06                           0.5
Mercury (Hg)               0.27 +/- 0.06                            1.00                        [28]


Permissible maximum limits for heavy metals in extracts of herbal medicines

Metal                     Maximum Permissible Limit (μg/g)
Iron (Fe)                                   300
Manganese (Mn)                       100
Copper (Cu)                              20    
Zinc (Zn)                                   50
Lead (Pb)                                 10 
Cadmium (Cd)                         0.2

Source: WHO (2005)                         [29]

Aflatoxins in Piper nigrum fruit extract

The extracts of Piper nigrum fruit were subjected to aflatoxin analysis by HPLC method. The results showed that various aflatoxins e.g. B1, B2, G1 and G2 were absent in all the samples.  [30]

Permissible Microbial Load

Name of bacteria                               WHO Limit
Escherichia coli                                       102        
Salmonella species                              Absence
Shigella species                                  Absence         
Enterobacter species                                104
Total bacterial Count                              107
Yeast and Mould                                    104    

The high moisture content is an important cause for microbial contamination in the herbal extracts. Hence by various methods moisture is reduced from the herbs before processing them for the use of medicinal purposes. 

The exposure of Maricham-Black pepper (Piper nigrum) to Microwaves at 663 Watts for 12.5 minutes was found to be sufficient to reduce the microbial load to permissible level suggested by the International Commission on Microbial Specifications for the Foods and the European Spice Association. After this treatment the moisture content reduced from initial 260+/-3 g/kg to 100 +/- 1 g/kg. After the treatment the retention of volatile oil was 91.3 +/- 0.03 %, piperine 87.6 +/- 0.02 % and resin 90.7 +/- 0.05 percent. [31]

Muraz B and Chaigneau M used ethylene oxide for decontamination of spices. But the process had some toxicity. So some researchers started using ozone as a substitute for ethylene oxide. The black peppercorns were immersed in water and sparged with ozonised air (ozone concentration, 6.7 mg/litre) for 10 minutes at an air flow rate of 6 litres/min. By this process the microbial population reduced markedly. [32]   
    
Genetic Identity

To avoid adulteration, it is absolutely mandatory to obtain genetically pure form of Maricham-Black pepper (Piper nigrum) for medicinal purposes. The genetic identity of Maricham-Black pepper (Piper nigrum) was determined by analyzing the meristem (rapidly growing top) of the flowers. This helps to avoid adulteration while processing the plant for medicinal purpose. The first ever floral gene identified was APETAL1. [33] 


PCR Analysis and SSR Sequencing

Maricham-Black pepper (Piper nigrum) is an autogamous (self pollinating, self reproducing) plant with a small percentage of cross pollination despite its fertile seeds. This gives rise to many “look-alike” plants which do not give expected pharmacological effects. To overcome these difficulties researchers have developed PCR analysis, RAPD and SSR (Simple Sequence Repeats, or microsatellites) markers for accurate identification of Maricham-Black pepper (Piper nigrum).  [34]  


Chromosomal Identity

Various research workers have reported variable numbers of somatic chromosome numbers Maricham-Black pepper (Piper nigrum). Here I mention all the numbers:

In cultivated and in many wild varieties, Mathew 1958, Martin and Gregory 1962, Samuel and Bavappa 1981, Jose and Sharma 1984, Okada 1986, Samuel and Morawetz reported chromosome numbers 2n=52.
Dasgupta and Datta 1976:  2n=36 and 2n=60
Jose and Sharma 1984:  2n=104.
R. R. Nair et al 1993: 2n=78
Progenies of the cultivar showed a range of chromosomal variation from2n=52 to 2n=104. Out of 20 progenies examined, six showed chromosome number of 2n=78, 2n=65, three showed 2n=82 and the other six showed chromosome number of 2n=52, 55, 72, 73, 76 and 104.  [35]

Recently many researchers reported chromosome number in Maricham-Black pepper (Piper nigrum) as 2 N= 52. [36]

                                        
HPTLC Analysis

Recently researchers found that rapid HPTLC method provided a new and powerful approach to estimate phytochemicals in the extract as well as its commercial formulations for routine quality control. [37]         

Properties and Pharmacology

Ayurvedic Properties

Rasa: Katu (Acrid, Pungent)
Weerya/ Virya: Ushna (Hot),   According to Sushruta: Naati ushna-Naati sheeta (not very hot not very cool)
Wipaaka: Katu (Acrid, Pungent)

Guna:

Ushna (hot), teekshna (penetrating) [38]

According to Sushruta: Naati ushna-Naati sheeta (not very hot not very cool) [39]

Ayurvedic properties:

Jwarahara: (antipyretic)
Chakshushya/Netrya: (beneficial for vision, relieves diseases of the eye)
Rochana: (improves taste),
Kaphahara: (reduces phlegm, expectorant),
Kaasahara: (antitussive),
Shwasahara:  (antiasthmatic),
Shoolahara:  (anticolic, relieves abdominal pain)  
Deepana: (improves appetite, digestion)
Pittakrut/ Pittakara: (increases gastric secretion, increases biliary secretion and improves bile flow)
Krimighna/ Krimihara: (anthelmintic, de-worming, expelling intestinal worms)
Waatahara: (improves neuromuscular activity)
Hridya:  (beneficial for heart diseases)
Pramaathi: (removes all pathological matter from tissues)
Awrishyam:  (spermicidal, contraceptive, anti-aphrodisiac)
Rasaayana: (adaptogen)
Chhedana: expels tenacious phlegm
Medohara: Antiobesity          [40], [41], [42]

Ayurvedic Ganas (class of drugs)

Charaka:
Deepaneeya: appetizer, improving digestion
Shoolanut, Shoolaprashamana: anticolic, relieving abdominal pain
Krimighna: anthelmintic, de-worming
Shirowirechana: expelling doshas (defects) from head and neck

Sushrut:
Pippalyadi, tri-ushna: (three hot herbs)

Waagbhata:
Pippalyadi, tri-ushna: (three hot herbs)

Dhanwantari Nighantu: Shatapushpaadi warga

Sodhala Nighantu: Shatapushpaadi warga  [43], [44]

  
                
References:

[24] http://www.stuartxchange.org/Paminta.html

[25] Zoheir A Damanhouril and Aftab Ahmad, A Review on Therapeutic Potential of Piper nigrum L. (Black Pepper): The King of Spices, Medicinal and Aromatic Plants, 2014, 3:3

[26] Shilika Shetty and K. K. Vijayalaxmi, Phytochemical Investigation of Extract/ Solvent Fractions of Piper nigrum Linn. Seeds and Piper betle Linn. Leaves, International Journal of Pharma and Biosciences Volume 3 Issue 2 April-June 2012
[27] Manisha N Trivedi et al, Pharmacognostic, Phytochemical Analysis and Antimicrobial Activity of Two Piper Species, Pharmacie Globale, (IJCP), 2011, 7 (05)
[28] Aftab Ahmad et al, Quantification of total phenol, flavonoid content and pharmacognostical evaluation in HPTLC fingerprinting for the standardization of Piper nigrum Linn fruits, Asian Pacific Journal of Tropical Biomedicine, Volume 5, Issue 2, February 2015, Pages 101-107

[29] P. T. Sandanayake et al, Analysis of Metal Content in Black Pepper Powder Available in the Sri Lankan Market, International Journal of Engineering Sciences & Research Technology, 6(10): October 2017, 

[30] Aftab Ahmad et al, Quantification of total phenol, flavonoid content and pharmacognostical evaluation in HPTLC fingerprinting for the standardization of Piper nigrum Linn fruits, Asian Pacific Journal of Tropical Biomedicine, Volume 5, Issue 2, February 2015, Pages 101-107

[31] G Chengaiyan et al, Application of microwaves for microbial load reduction in black pepper (Piper longum L) Journal of the science of food and agriculture, 19 January 2016

[32] Jian Zhao et al, Microbial decontamination of black pepper and the effect of the treatment on volatile oil constituents of the spice. Journal of the science of food and agriculture May 1995   

[33] S Lekha, Molecular analysis of floral meristem identity genes in black pepper, Krishikosh.egranth.ac.in/handle/1/5810041675; 2014,


[34] Remmia Raghavan et al, Molecular Characterization of Black Pepper (Piper nigrum) Using RAPD and SSR Markers, Biosciences, Biotechnology Research Asia 2010, 7(2)

[35] R. R. Nair et al, Polyploidy in a Cultivar of Black Pepper (Piper nigrum L) and its Open Pollinated Progenies, Japan Mendel Society, Cytologia 58: 27-31, 1993

[36] Nisha Joy et al, A preliminary assessment of genetic relationships among agronomically important cultivars of black pepper, Biomed Central, Genet, 2007 Jun 29 8: 42.

[37] Anagha Rajopadhye et al, Rapid validated HPTLC method for estimation of piperine and piperlongumine in root of Piper longum extract and its commercial formulation, Revista Brasileira de Farmacognosia 22(6): 1355-1361, December 2012 

[38] Charaka Samhitaa, Sootrasthaana 27 

[39] Sushruta Samhita, Soortasthana 46

[40] Charaka Samhitaa, Sootrasthaana 27 

[41] Sushruta Samhita, Soortasthana 46

[42] https:/www.bimbima.com/ayurveda/medicinal-use-of-black-pepper-kali-mirch/1572?

[43] Charaka Samhitaa, Sootrasthaana 27 

[44] Sushruta Samhita, Soortasthana 46




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