[vc_row lg_spacing=”padding_top:60;padding_bottom:60″ md_spacing=”padding_top:40;padding_bottom:40″][vc_column width=”1\/6″][\/vc_column][vc_column width=”2\/3″][tm_spacer size=”lg:40″][tm_heading custom_google_font=”1″ google_fonts=”font_family:Playfair%20Display%3Aregular%2Citalic%2C700%2C700italic%2C900%2C900italic|font_style:700%20bold%20regular%3A700%3Anormal” font_weight=”700″ align=”center” text=”COA Tool” max_width=”1000px” font_size=”lg:36″ letter_spacing=”2″][tm_spacer size=”lg:10″][tm_image align=”center” image=”4510″][tm_spacer size=”lg:20″][tm_spacer size=”lg:20″][tm_image align=”center” image=”4870″][tm_spacer size=”lg:20″][\/vc_column][vc_column width=”1\/6″][\/vc_column][\/vc_row][vc_row full_width=”stretch_row” background_color=”custom” custom_background_color=”#f7f7f7″ lg_spacing=”padding_top:60;padding_bottom:40″][vc_column width=”1\/6″][\/vc_column][vc_column width=”2\/3″][tm_heading custom_google_font=”” font_weight=”700″ text=”Analysis of cannabinoids”][tm_spacer size=”lg:10″][tm_heading tag=”p” custom_google_font=”” text=”0.01 g (\u00b1.0001) of crystals was dissolved in 1 ml of methanol (HPLC grade). Solution was sonicated for 2 min and vortexing for 10 sec. Samples before HPLC analysis were further diluted with methanol to the final concentration of 0.01 mg\/ml.”][tm_spacer size=”lg:30″][tm_heading custom_google_font=”” font_weight=”700″ text=”Chromatographic Analysis”][tm_spacer size=”lg:10″][tm_heading tag=”p” custom_google_font=”” text=”Analysis of cannabinoids content was performed using Waters 2695 (Milford, MA, USA) separation module equipped with auto injector, sample cooler, vacuum degasser and column heater units. Separation of all cannabinoids was accomplished on YMC PRO C18 (150 x 4 mm I.D., S-3\u03bcm) RP column coupled with C18 precolumn maintained at 30 \u00b0C by a CTO-20AC column oven. Isocratic elution consisted of acetonitrile:water (FA 0.1%) (4:1) was done in 30 min. The flow rate was maintained at 0.8 ml\/min. The cannabinoid CBD and CBD-A were monitored at 225 and 306 nm wave length respectively using dual absorbance detector Waters 2487 (Milford, MA, USA). The injection volume of 20 \u03bcl was injected using auto sampler at 10 \u00b0C. Data evaluation was performed using Empower 2 software.<\/p>\n
Quantification of CBD and CBD-A were obtained from linear regression equation of calibration curve of individual reference standards by plotting concentration versus the area ratio. The calibration range for CBD and CBD-A were linear from 5 to 500 \u03bcg\/ml. Samples which contain CBD or CBD-A concentration higher than 40% were weaken diluting 10 times beforeinjection. Retention time of CBD-A was at 7.1 min and CBD at 8.1 min.”][tm_spacer size=”lg:30″][tm_heading custom_google_font=”” font_weight=”700″ text=”Analysis of terpenes”][tm_spacer size=”lg:10″][tm_heading tag=”p” custom_google_font=”” text=”10 mg of homogenous sample was scaled and diluted with 1 ml of pentane containing 0.04 % of decane as internal standard. The tube containing the sample solution was placed in ultrasonic bath for 5 min and then mixed. 200 \u00b5L of prepared solution was diluted with 800 \u00b5L of pure pentane mixed and individually analysed by GC-FID.<\/p>\n
An Agilent HP 6890 gas chromatograph equipped with FID was used for the analysis of terpenoids. Separation was accomplished on a Rtx-5 w\/Integra-Guard capillary column (30 m length, 0.25 mm i.d. and 0.25 \u00b5m df). Injections were carried out in split mode using a general purpose split\/splitless liner packed with glass wool. The program started at 50 \u00b0C, increased to 280 \u00b0C (at 15 \u00b0C\/min) and held for 15 min for a total of 31 min. 2 \u00b5L of each sample was injected with helium as the carrier gas (constant flow mode, 1 ml\/min) using a split ratio 1:10. Temperatures were applied 280 \u00b0C for the injector, 260 \u00b0C for the transfer line. Data was analysed using Chemstation v.D.02.00.275 (Agilent Technologies).<\/p>\n
List of the target Terpenes:<\/p>\n
\u2022 Compound RT
\n\u2022 Decane (IS) 5,165
\n\u2022 \u03b1-pinene 4,635
\n\u2022 Myrcene 5,095
\n\u2022 Limonene 5,538
\n\u2022 Linalool 6,176
\n\u2022 E-Caryophyllene 9,328<\/p>\n
1. Decane concentration in pentane 0,04% = 1 ml pentane contain 0.292 mg decane.
\n2. 0.01g of sample dissolved with 1000 \u00b5L pentane containing 0.04 % decane.
\n3. Extract with concentration of 10 mg\/ml dissolved 5 times. Final concentration of the extract in solution 2.5 mg\/ml (0.0025 g\/ml).
\n4. Final concentration of decane in solution 0.073 mg\/ml.
\n5. 0.073 mg of decane \/ 0.0025 g of extract = 29.2 mg\/g extract
\n6. Calculations<\/p>\n
Peak Area of decane – 29.2 mg\/g extract
\nPeak area of target compound – x mg\/g extract
\nx= 29.2 \u00d7 Peak area of target compound \/ Peak area of decane = amount of target compound mg\/ g extract.”][tm_spacer size=”lg:30″][tm_heading custom_google_font=”” font_weight=”700″ text=”COA & Lot Numbers:”][tm_spacer size=”lg:10″][vc_column_text]<\/p>\n
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