( 1 2 )

United S ta tes Pa tent

Whittle et a l.
US 007344736B2
US 7, 344, 736 B2
Ma r. 1 8 , 2 008
( 1 0) Pa tent N 0. :
( 45 ) Da te o f Pa tent:
( 5 4)
( 75 )
( 73)
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EXTRACTION OF PHARMACEUTICALLY
ACTIVE COMPON EN TS FROM PLAN T
MATERIALS
Inv ento rs : Bria n Whittle, S a lis b u ry ( G B) ; Co lin
A. Hill, Cha tha m l ( G B) ; Ia n R.
Flo c k ha rt, Wo o dm a ns ey ( G B) ; Da v id
Vic to r Do w ns , Ca nterb u ry ( G B) ; Peter
G ib s o n, S a lis b u ry ( G B) ; G a ry Willia m
Whea tley , S a lis b u ry ( G B)
As s ig nee: G W Pha rm a Lim ited, S a lis b u ry ( G B)
N o tic e: S u b j ec t to a ny dis c la im er, the term o f this
p a tent is ex tended o r a dj u s ted u nder 35
U. S . C. 1 5 4( b ) b y 749 da y s .
Ap p l. N o . : 1 0/ 2 1 8 , 9 72
Filed: Au g . 1 4, 2 002
Prio r Pu b lic a tio n Da ta
US 2 004/ 00332 8 0 A1 Feb . 1 9 , 2 004
Int. Cl.
A61 K 36/ 00 ( 2 006. 01 )
A61 K 9 / 00 ( 2 006. 01 )
U. S . Cl. . . . . . . . . . . . . . . . . . . . . . . . 42 4/ 72 5 ; 42 4/ 774; 42 4/ 400
Field o f Cla s s i?c a tio n S ea rc h . . . . . . . . . . . . . . . . 42 4/ 774;
5 49 / 39 2
S ee a p p lic a tio n ?le f o r c o m p lete s ea rc h his to ry .
Ref erenc es Cited
U. S . PATEN T DOCUMEN TS
4, 2 79 , 8 2 4 A * 7/ 1 9 8 1 Mc Kinney . . . . . . . . . . . . . . . . . . 5 49 / 39 0
5 , 2 2 7, 5 37 A 7/ 1 9 9 3 S to s s et a l.
5 , 478 , 736 A * 1 2 / 1 9 9 5 N a ir . . . . . . . . . . . . . . . . . . . . . . . . . . . 435 / 1 2 3
FOREIG N PATEN T DOCUMEN TS
1 2 1 2 2 73 3/ 1 9 9 9
1 9 8 00 330 C2 9 / 2 002
2 1 032 38 9 / 1 9 9 7
WO 00/ 2 5 1 2 7 5 / 2 000
WO 01 / 2 8 649 A1 4/ 2 001
WO02 / 32 42 0 * 4/ 2 002
WO 02 / 0641 09 A2 8 / 2 002
OTHER PUBLICATION S
T. Veres s , S a m p le p rep a ra tio n b y s u p erc ritic a l ?u id ex tra c tio n f o r
q u a nti?c a tio n. A m o del b a s ed o n the dif f u s io n- la y er theo ry f o r
determ ina tio n o f ex tra c tio n tim e, J o u rna l o f Chro m a to g ra p hy A,
668 ( 1 9 9 4) 2 8 5 - 2 9 1 .
[ N o a u tho r lis ted] Va s c u la r Pla nts o f Ru s s ia a nd Adj a c ent
Co u ntries iHu m u lu s ( Oc t. 2 6, 1 9 9 6) .
[ N o a u tho r lis ted] Va s c u la r Pla nts o f Ru s s ia a nd Adj a c ent
Co u ntries iCa nna b is L. ( Oc t. 2 6, 1 9 9 6) .
R. A. N els o n, Hem p Hu s b a ndry , Cha p ter 6iCa nna b ino id Chem
is try , http : / / WWW. rex res ea rc h. c o m / hhu s b / hh6thc htm , 2 000.
D. P. Wes t, Hem p a nd Ma rij u a na : My ths & Rea lities , http : / / WWW.
na ihc . o rg / hem p iinf o rm a tio n/ c o ntent/ hem p . m j . htrnl, 1 9 9 8 .
A. Aw a s thi et a l. , A Rev iew o n S u p erc ritic a l Ca rb o n Dio x ide
Ex tra c tio n o f N a tu ra l Pro du c ts , Chem ic a l Eng ineering Wo rld, v o l.
XXXII, N o . 1 0, Oc t. 1 9 9 7, p p . 65 - 71 .
Hendrik s , H. et a l. , Mo no - a nd S es q u i- terp ene Hy dro c a rb o ns o f the
Es s entia l Oil o f Ca nna b is s a tiv a , Ps y c ho c hem is try ( 1 9 75 ) 5 1 4
8 1 5 .
Tu rner, C. E. , Co ns titu ents o f Ca nna b is s a tiv a L. , Bio m edic a l
Ma s s S p ec tro m etry , v o l. 7, N o . 6 ( 1 9 9 0) 2 47- 2 5 6.
Tu rner, C. E. , Co ns titu ents o f Ca nna b is s a tiv a L. XVII. Arev iew o f
the N a tu ra l Co ns titu ents , J o u rna l o f N a tu ra l Pro du c ts , v o l. 43, N o .
2 ( Ma r- Ap r. 1 9 8 0) 1 69 - 2 34.
Tu rner, C. E. , Chem is try a nd Meta b o lis m , S u p p lied b y The Britis h
Lib ra ry , p p . 8 1 - 9 7.
The United King do m Pa rlia m ent, S elec t Co m m ittee o n S c ienc e a nd
Tec hno lo g y N inth Rep o rt ( 1 9 9 8 ) a t http : / / WWW. p a rlia m ent. the- s ta
tio nery - o f ?c e. c o . u ldp a / Id1 9 9 79 8 / Ids elec t/ Ids c tec h/ l5 1 / 1 5 1 01 . htm .
The United King do m Pa rlia m ent, S elec t Co m m ittee o n S c ienc e a nd
Tec hno lo g y N inth Rep o rt ( Ma r. 1 4, 2 001 ) a t http : / / WWW. p u b lic a
tio ns . p a rlia m ent. u 1 dp a / Id2 00001 / Ids elec t/ Ids c tec h/ 5 0/ 5 001 . htm .
* c ited b y ex a m iner
Prim a ry Ex a m ineriTerry Mc Kelv ey
As s is ta nt Ex a m ineriRa nda ll Wins to n
( 74) Atto rney , Ag ent, o r Firm iWo lf , G reen?eld & S a c k s ,
RC.
( 5 7) ABS TRACT
The inv entio n rela tes to the ex tra c tio n o f p ha rrna c eu tic a lly
a c tiv e c o m p o nents f ro m p la nt m a teria ls , a nd m o re p a rtic u
la rly to the p rep a ra tio n o f a b o ta nic a l dru g s u b s ta nc e ( BDS )
f o r inc o rp o ra tio n in to a m edic a m ent. It a ls o rela tes to a BDS
o f g iv en p u rity , f o r u s e in p ha rm a c eu tic a l f o rm u la tio ns . In
p a rtic u la r it rela tes to BDS c o m p ris ing c a nna b ino ids
o b ta ined b y ex tra c tio n f ro m c a nna b is .
2 1 Cla im s , 3 Dra w ing S heets
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US 7, 344, 736 B2
1
EXTRACTION OF PHARMACEUTICALLY
ACTIVE COMPON EN TS FROM PLAN T
MATERIALS
FIELD OF THE IN VEN TION
The p res ent inv entio n rela tes to the ex tra c tio n o f p ha r
m a c eu tic a lly a c tiv e c o m p o nents f ro m p la nt m a teria ls , a nd
m o re p a rtic u la rly to the p rep a ra tio n o f a b o ta nic a l dru g
s u b s ta nc e ( BDS ) f o r inc o rp o ra tio n in to a m edic a m ent. It
a ls o rela tes to a BDS o f g iv en p u rity , f o r u s e in p ha rm a c eu
tic a l f o rm u la tio ns . In p a rtic u la r it rela tes to BDS c o m p ris ing
c a nna b ino ids o b ta ined b y ex tra c tio n f ro m c a nna b is .
BACKG ROUN D OF THE IN VEN TION
In PCT/ G B02 / 0062 0 the a p p lic a nt dis c lo s es a m etho d o f
p rep a ring a herb a l dru g ex tra c t ( b o ta nic a l dru g s u b s ta nc e)
f ro m m edic ina l c a nna b is . The p ro c es s c o m p ris es :
1 . a hea ting s tep to dec a rb o x y la te the a c id f o rm o f the
c a nna b ino ids to their neu tra l f o rm ;
2 . a ?rs t ex tra c tio n With a s p ec i?ed v o lu m e o f liq u id c a rb o n
dio x ide f o r 648 ho u rs ; a nd
3. a s tep to redu c e the p ro p o rtio n o f no n- ta rg et m a teria ls ,
ref erred to a s Winteris a tio n, Whic h s tep p rec ip ita tes o u t
Wa x es .
Mo re s p ec i?c a lly , PCT/ G B02 / 0062 0 des c rib es a p ro c es s
Wherein:
s tep 1 c o m p ris es hea ting c ho p p ed c a nna b is ( 2 43 m m ) a t
1 0041 5 0o C. f o r s u f ?c ient tim e to a llo W dec a rb o x y la tio n;
s tep 2 c o m p ris es CO2 ex tra c tio n u s ing :
a ) a c o a rs e p o Wder ( the p a rtic les a re p a s s ed thro u g h a 3 m m
m es h) ;
b ) a p a c k ing dens ity o f 0. 3; a nd
c ) s u p erc ritic a l c o nditio ns o f 600 b a r a t 35 C. f o r 4 ho u rs ,
a ltho u g h o ther c o m b ina tio ns o f tem p a nd p res s u re ra ng ing
f ro m 1 ( L35 o C. a nd 604600 b a r ( b o th s u p er c ritic a l a nd
s u b c ritic a l c o nditio ns ) c o u ld, it is a c k no w ledg ed, b e
u s ed; a nd
s tep 3 c o m p ris es c o ndu c ting a n etha no lic p rec ip ita tio n a t
2 00 C. f o r 2 4 ho u rs a nd rem o v ing the Wa x y m a teria l b y
?ltra tio n.
The s u p erc ritic a l m etho d dis c lo s ed in PCT/ G B02 / 0062 0
p ro du c ed:
a ) a hig h THC ex tra c t c o nta ining :
60% THC ( Ag - tetra hy dro c a nna b ino l)
1 42 % CBD ( c a nna b idio l)
445 % o ther m ino r c a nna b ino ids inc lu ding CBN ( c a nna b
ino l)
( Q u a nta tiv e y ields Were 9 % Wt/ Wt b a s ed o n dry Weig ht o f
m edic ina l c a nna b is ) ; a nd
b ) a hig h CBD ex tra c t c o nta ining :
60% CBD
4% THC
2 % o ther c a nna b ino ids
( Q u a nta tiv e y ields Were 9 % Wt/ Wt b a s ed o n dry Weig ht o f
m edic ina l c a nna b is ) .
Clea rly a s the res u lting BDS is to b e u s ed in a p ha rm a
c eu tic a l p ro du c t it is es s entia l tha t the p ro c es s is s a f e,
s c a la b le to G MP a nd g iv es hig h deg rees o f p ro du c t c o ns is
tenc y a nd, p ref era b ly a ls o g o o d y ields .
The p rinc ip les o f s u p erc ritic a l ?u id ex tra c tio n ( S FE) ha v e
b een k no Wn s inc e the Wo rk o f Ba ro n Ca g nia rd de le To u r in
1 8 2 2 When it Wa s no ted tha t the g a s - liq u id b o u nda ry dis a p
p ea red When the tem p era tu re o f c erta in m a teria ls Wa s
inc rea s ed b y hea ting them in a c lo s ed g la s s c o nta iner. Fro m
this ea rly Wo rk the c ritic a l p o int o f a s u b s ta nc e Wa s ?rs t
2 0
2 5
30
35
40
45
5 0
5 5
60
65
2
dis c o v ered. The c ritic a l p o int is the tem p era tu re a b o v e
Whic h a s u b s ta nc e c a n c o - ex is t in g a s , liq u id a nd s o lid
p ha s es . It Wa s la ter f o u nd tha t b y ta k ing s u b s ta nc es to o r
a b o v e their c ritic a l tem p era tu re a nd p res s u re they c o u ld b e
u s ed a s s o p his tic a ted s o lv ents f o r ex tra c tio n a nd f ra c tio n
a tio n o f c o m p lex m ix tu res .
The tec hniq u e is Widely u s ed in the f u el o il p ro c es s ing
b u s ines s a nd ha s b een a p p lied to , f o r ex a m p le, the p u ri?
c a tio n a nd s ep a ra tio n o f v eg eta b le a nd ?s h o ils .
An a ttra c tiv e f ea tu re o f S FE o v er the u s e o f c o nv entio na l
s o lv ents is tha t the s o lv ent p o Wer ( E ) c a n b e v a ried b y
m a nip u la tio n o f tem p era tu re a nd p res s u re a b o v e the c ritic a l
p o int.
In a ty p ic a l p res s u re- tem p era tu re dia g ra m f o r a s u b s ta nc e
there a re three lines Whic h de?ne the eq u ilib riu m b etWeen
tWo o f the p ha s es . Thes e lines m eet a t the trip le p o int. The
lines de?ne the interf a c e b etWeen g a s , liq u id a nd s o lid s ta tes ,
a nd p o ints a lo ng the line de?ne the eq u ilib riu m b etWeen
p a irs o f p ha s es . Fo r ex a m p le, the v a p o u r p res s u re ( b o iling
p o int) c u rv e s ta rts a t the trip le p o int a nd ends a t the c ritic a l
p o int. The c ritic a l reg io n s ta rts a t this p o int a nd a s u p er
c ritic a l ?u id is a ny s u b s ta nc e tha t is a b o v e its c ritic a l
tem p era tu re ( Tc ) a nd c ritic a l p res s u re ( Pc ) . The c ritic a l
tem p era tu re is thu s the hig hes t tem p era tu re a t Whic h a g a s
c a n b e c o nv erted to a liq u id b y a n inc rea s e in p res s u re a nd
the c ritic a l p res s u re is the hig hes t p res s u re a t Whic h a liq u id
c a n b e c o nv erted into a tra ditio na l g a s b y inc rea s ing the
tem p era tu re. In the s o - c a lled c ritic a l reg io n, there is o nly o ne
p ha s e a nd it p o s s es s es s o m e o f the p ro p erties o f b o th a g a s
a nd a liq u id.
S UMMARY OF THE IN VEN TION
There a re a nu m b er o f s o lv ents , Whic h c a n b e u s ed f o r
ex tra c tio n o f a c tiv e s u b s ta nc es f ro m p la nt m a teria ls , a nd
Ta b le 1 s ho Ws the c ritic a l tem p era tu re a nd p res s u res f o r
s o m e o f thes e s o lv ents .
TABLE 1
Critic a l Co nditio ns f o r S o lv ents
Critic a l
S o lv ents Critic a l Tem p era tu re ( 0 C. ) Pres s u re ( b a r)
Ca rb o n dio x ide 31 . 1 73. 8
Etha ne 32 . 2 48 . 8
Ethy lene 9 . 3 5 0. 4
Pro p a ne 9 6. 7 42 . 5
Pro p y lene 9 1 . 9 46. 2
Cy c lo hex a ne 2 8 0. 3 40. 7
Is o p ro p a no l 2 35 . 2 47. 6
Benzene 2 8 9 . 0 48 . 9
To lu ene 31 8 . 6 41 . 1
p - Xy lene 343. 1 35 . 2
Chlo ro tri?u o ro m etha ne 2 8 . 9 39 . 2
Tric hlo ro ?u o ro rnetha ne 1 9 8 . 1 44. 1
Am m o nia 1 32 . 5 1 1 2 . 8
Wa ter 374. 2 2 2 0. 5
The a p p lic a nt ha s s elec ted a s a p ref erred s o lv ent c a rb o n
dio x ide, Whic h ha s a c ritic a l tem p era tu re o f 31 . 1 0 C. a nd a
c ritic a l p res s u re o f 73. 8 b a r.
Ca rb o n dio x ide is p a rtic u la rly a dv a nta g eo u s b ec a u s e it is
a v a ila b le in p lentif u l s u p p ly , a t lo W c o s t, a nd c a n if nec es
s a ry b e rec y c led. Any lo s s es o f CO2 a re a ls o ec o lo g ic a lly
neu tra l. Fu rtherm o re, CO2 ex tra c tio n is a c o ns erv a tiv e
m etho d o f p rep a ra tio n a nd q u ite f ra g ile m o lec u les c a n b e
ex tra c ted With p rec is io n.
A k ey c o ns idera tio n in the initia l s elec tio n o f liq u id CO2
a s the s o lv ent f o r the p ro du c tio n o f a hig h p o tenc y s ta nda r
US 7, 344, 736 B2
3
dis ed ex tra c t o f c a nna b is herb Wa s the hig h deg ree o f
s elec tiv ity Whic h c a n b e a c hiev ed. In the CO2 s y s tem it ha s
b een determ ined tha t s o lv a ting p o w er c a n p rim a rily b e
reg a rded a s b eing a f u nc tio n o f dens ity a nd tem p era tu re,
With the s o lv ent dens ity b eing the m o re im p o rta nt f a c to r.
Co ntra ry to ex p ec ta tio n the a p p lic a nt ha s determ ined tha t
c a nna b ino ids a re b es t o b ta ined u nder s u b - c ritic a l ra ther tha n
s u p er- c ritic a l c o nditio ns
By c a ref u lly c o ntro lling tem p era tu re a nd p res s u re b elo W
the s u p er- c ritic a l tem p era tu re a nd p res s u re the a p p lic a nt ha s
b een a b le to s ep a ra te o u t s p ec i?c lip o p hilic o r hy dro p hilic
f ra c tio ns ric h in c a nna b ino ids With o ther c o m p o nents Whic h
c a n b e s ep a ra ted rela tiv ely ea s ily to o b ta in a b o ta nic a l dru g
s u b s ta nc e ( BDS ) Whic h c o nta ins the des ira b le c o m p o nents
in a f o rm Whic h is p ha rm a c eu tic a lly a c c ep ta b le. Thu s c o m
p o u nds Whic h a re k no Wn to b e a c tiv e s u b s ta nc es c a n b e
s ep a ra ted f ro m c o m p lex m ix tu res Whic h o c c u r in b o ta nic a l
ra W m a teria l.
Fu rtherm o re, v ery g o o d b a tc h- to - b a tc h rep ro du c ib ility
c a n b e o b ta ined b etWeen b a tc hes a nd u nWa nted c o ns titu ents ,
s u c h a s hea v y m eta ls , Whic h m a y b e p res ent to v a ry ing
ex tents in the b o ta nic a l ra W m a teria l, c a n b e lef t b ehind in
the ex ha u s ted m a teria l.
Ex tra c tio n c o nditio ns c a n a ls o b e m o di?ed to rej ec t
p es tic ide res idu es Whic h m a y b e p res ent in the o rig ina l
m a teria l.
The b ene?ts o f u s ing s u b - c ritic a l c o nditio ns inc lu de the
s elec tiv e na tu re o f the ex tra c tio n. In c o ntra s t, the a p p lic a nt
f o u nd tha t With S FE the s o lv ent, a s Well a s s o lu b ilis ing the
des ira b le c a nna b ino ids , dis a dv a nta g eo u s ly s o lu b ilis ed o ther
no n ta rg et m a teria ls Whic h p ro v ed dif ?c u lt to s ep a ra te o u t in
a s u b s eq u ent c lea n- u p s tep .
To ex p la in, the dens ity o f s u b - c ritic a l CO2 is lo W, a nd
rem a ins lo W ev en a s p res s u re is inc rea s ed u ntil the c ritic a l
p o int o f the s y s tem is rea c hed. Thu s , Whils t the s o lv a ting
p o Wer o f s u b - c ritic a l CO2 is redu c ed a hig h deg ree o f
s elec tiv ity c a n b e a c hiev ed, a s o nly the m o s t s o lu b le c o m
p o nents a re e?ic iently dis s o lv ed b y the CO2 ; in this c a s e the
c a nna b ino id f ra c tio n. The res u lt is the p ro du c tio n o f a
rela tiv ely s im p le ex tra c t c o nta ining , a s Well a s the c a nna b
ino ids , o nly a lim ited nu m b er o f no n- ta rg et c o m p o u nds ,
m a ny o f Whic h c a n b e rem o v ed rela tiv ely ea s ily in a s im p le
s tep . Fu rtherm o re, the c o s t s a v ing s m a de b y o p era ting a t
rela tiv ely lo W p res s u res a nd tem p era tu res a re a f u rther
b ene?t.
In c o ntra s t, a b o v e the c ritic a l tem p era tu re o f 3 1 C. , there
is a s ig ni?c a nt inc rea s e in the dens ity o f the CO2 a s it no W
ex is ts in a s u p erc ritic a l ?u id s ta te. This ha s the ef f ec t o f
g rea tly inc rea s ing the s o lv a ting p o Wer o f the s o lv ent, Whic h
Whils t g enera lly a dv a nta g eo u s in tha t m o re c a nna b ino ids a re
s o lu b ilis ed thereb y g iv ing hig h y ields , in f a c t p ro v es dis a d
v a nta g eo u s b ec a u s e the dec rea s ed s elec tiv ity o f the m o re
p o Werf u l s o lv ent res u lts in inc rea s ed s o lu b ility o f a ra ng e o f
no n- ta rg et c o m p o u nds Whic h m a k es the res u lting ex tra c t
ha rder to p u rif y . In o ther Wo rds , it res u lts in the p ro du c tio n
o f m o re c o m p lex ex tra c ts in Whic h the c o nc entra tio n o f the
ta rg et c o m p o u nd m a y b e s ig ni?c a ntly dilu ted ( i. e. the
p o tenc y o f the ex tra c t is dec rea s ed) .
In a ?rs t a s p ec t the inv entio n p ro v ides a m etho d o f
ex tra c ting c a nna b ino ids f ro m p la nt m a teria l c o m p ris ing a
dec a rb o x y la tio n s tep , a n ex tra c tio n With liq u id c a rb o n dio x
ide ( CO2 ) , a nd a s tep to redu c e the p ro p o rtio n o f no n- ta rg et
m a teria ls in the ex tra c t, c ha ra c teris ed in tha t the ex tra c tio n
With liq u id CO2 is c o ndu c ted u nder s u b - c ritic a l c o nditio ns a t
a tem p era tu re o f b etWeen 5 * l5 C. a nd a p res s u re o f
b etWeen 5 0* 70 b a r.
2 0
35
40
45
5 0
5 5
60
65
4
The m etho d o f the inv entio n m a y b e u s ed to p rep a re a
c a nna b ino id- ric h ex tra c t f ro m c a nna b is p la nt m a teria l. In a
p ref erred em b o dim ent, the m etho d m a y b e u s ed to p ro du c e
a c a nna b is ex tra c t Whic h is a b o ta nic a l dru g s u b s ta nc e.
In the c o ntex t o f this a p p lic a tio n a b o ta nic a l dru g s u b
s ta nc e is a n ex tra c t deriv ed f ro m c a nna b is p la nt m a teria l,
Whic h ex tra c t f u l?ls the de?nitio n o f b o ta nic a l dru g s u b
s ta nc e p ro v ided in the G u ida nc e f o r Indu s try Bo ta nic a l
Dru g Pro du c ts Dra f t G u ida nc e, Au g u s t 2 000, US Dep a rt
m ent o f Hea lth a nd Hu m a n S erv ic es , Fo o d a nd Dru g Adm in
is tra tio n Center f o r Dru g Ev a lu a tio n a nd Res ea rc h o f : A
dru g s u b s ta nc e deriv ed f ro m o ne o r m o re p la nts , a lg a e, o r
m a c ro s c o p ic f u ng i. It is p rep a red f ro m b o ta nic a l ra W m a te
ria ls b y o ne o r m o re o f the f o llo Wing p ro c es s es : p u lv eris a
tio n, dec o c tio n, ex p res s io n, a q u eo u s ex tra c tio n, etha no lic
ex tra c tio n, o r o ther s im ila r p ro c es s es .
Pla nt m a teria l is de?ned a s a p la nt o r p la nt p a rt ( e. g .
b a rk , Wo o d, lea v es , s tem s , ro o ts , ?o Wers , f ru its , s eeds ,
b erries o r p a rts thereo f ) a s Well a s ex u da tes , a nd inc lu des
m a teria l f a lling Within the de?nitio n o f b o ta nic a l ra W
m a teria l in the G u ida nc e f o r Indu s try Bo ta nic a l Dru g
Pro du c ts Dra f t G u ida nc e, Au g u s t 2 000, US Dep a rtm ent o f
Hea lth a nd Hu m a n S erv ic es , Fo o d a nd Dru g Adm inis tra tio n
Center f o r Dru g Ev a lu a tio n a nd Res ea rc h.
The m etho d o f the inv entio n m a y b e u s ed to ex tra c t
c a nna b ino ids f ro m a ny p la nt m a teria l k no Wn to c o nta in s u c h
c a nna b ino ids . Mo s t ty p ic a lly , b u t no t nec es s a rily , the p la nt
m a teria l Will b e p la nt m a teria l o r b o ta nic a l ra W m a te
ria l deriv ed f ro m o ne o r m o re c a nna b is p la nts .
The term Ca nna b is p la nt( s ) enc o m p a s s es Wild ty p e
Ca nna b is s a liv a a nd a ls o v a ria nts thereo f , inc lu ding c a n
na b is c hem o v a rs Whic h na tu ra lly c o nta in dif f erent a m o u nts
o f the indiv idu a l c a nna b ino ids , Ca nna b is s a liv a s u b s p ec ies
indic a inc lu ding the v a ria nts v a r. indic a a nd v a r. k a ?ris
Z a nic a , Ca nna b is indic a a nd a ls o p la nts Whic h a re the res u lt
o f g enetic c ro s s es , s elf - c ro s s es o r hy b rids thereo f . The term
Ca nna b is p la nt m a teria l is to b e interp reted a c c o rding ly a s
enc o m p a s s ing p la nt m a teria l deriv ed f ro m o ne o r m o re
c a nna b is p la nts . Fo r the a v o ida nc e o f do u b t it is hereb y
s ta ted tha t c a nna b is p la nt m a teria l inc lu des dried c a nna b is
b io m a s s .
The ex tra c tio n With liq u id CO2 is p ref era b ly c a rried o u t a t
a tem p era tu re b etWeen 8 * l2 C. , m o s t p ref era b ly a t a
tem p era tu re o f a b o u t 1 00 C.
The ex tra c tio n With liq u id CO2 is p ref era b ly c a rried o u t a t
a p res s u re b etWeen 5 5 * 65 b a r, m o s t p ref era b ly a t a p res s u re
o f s u b s ta ntia lly 60 b a r.
Mo s t p ref era b ly the CO2 ha s a m a s s Ho w o f f ro m
l000il5 00 Kg / h, m o re p ref era b ly a m a s s Ho w o f s u b s ta n
tia lly 1 2 5 0 Kg / h.
Pref era b ly the liq u id CO2 ex tra c tio n is ru n f o r u p to 1 0
ho u rs , m o s t p ref era b ly a b o u t 8 ho u rs .
In a p ref erred em b o dim ent liq u id CO2 is rem o v ed b y
dep res s u ris a tio n a nd the rec o v ered ex tra c t held a t a tem
p era tu re in the ra ng e f ro m l5 C. to 2 0 C.
The s tep to redu c e the p ro p o rtio n o f no n- ta rg et m a teria ls
in the b o ta nic a l dru g s u b s ta nc e m a y b e es s entia lly a ny
trea tm ent tha t res u lts in s elec tiv e rem o v a l o f u ndes ira b le
c o m p o nents ( a s o p p o s ed to c a nna b ino ids ) , s u c h tha t the
a m o u nt o f the u ndes ira b le c o m p o nents p res ent in the ?na l
b o ta nic a l dru g s u b s ta nc e p ro du c t is redu c ed. N o n- ta rg et
m a teria ls a re a ny m a teria ls deriv ed f ro m the s ta rting p la nt
m a teria l tha t a re no t des ired to b e p res ent in the ?na l
b o ta nic a l dru g s u b s ta nc e. In a p ref erred em b o dim ent this
s tep m a y c o m p ris e a p rec ip ita tio n With a CliC5 a lc o ho l,
Wherein the m a teria l to b e trea ted in the a lc o ho l p rec ip ita tio n
s tep is Wa rm ed to a b o v e ro o m tem p era tu re b ef o re the CliC5
US 7, 344, 736 B2
5
a lc o ho l is a dded. Ty p ic a lly , the s tep to redu c e the p ro p o rtio n
o f no n- ta rg et m a teria ls in the b o ta nic a l dru g s u b s ta nc e is
c a rried o u t a f ter ex tra c tio n With liq u id CO2 , in Whic h c a s e
the m a teria l to b e trea ted in the a lc o ho lic p rec ip ita tio n is
the p ro du c t o f the liq u id CO2 ex tra c tio n. This ex tra c t is its elf
a b o ta nic a l dru g s u b s ta nc e Within the de?nitio n g iv en
a b o v e.
The C1 4C5 a lc o ho l is p ref era b ly etha no l. The ex tra c t is
p ref era b ly Wa rm ed to a tem p era tu re in the ra ng e f ro m 36 C.
to 44 C. , m o s t p ref era b ly a b o u t 40 C. Wa rm ing o f the
m a teria l to b e trea ted p rio r to a dditio n o f the C1 5 a lc o ho l
ha s the ef f ec t o f im p ro v ing m ix ing o f this m a teria l With the
C1 5 a lc o ho l, a nd henc e im p ro v es the p erf o rm a nc e o f the
a lc o ho l p rec ip ita tio n s tep .
The C1 5 a lc o ho l is p ref era b ly a dded in a n a m o u nt o f
f ro m 3: 1 to 1 : 1 C1 4C5 a lc o ho l v o lu m e to Weig ht o f the
m a teria l to b e trea ted, m o re p ref era b ly a n a m o u nt o f a b o u t
2 : 1 C1 4C5 a lc o ho l v o lu m e to Weig ht o f the m a teria l to b e
trea ted.
The s o lu tio n res u lting f ro m a dditio n o f C1 4C5 a lc o ho l to
the m a teria l to b e trea ted is c hilled a nd ins o lu b le m a teria ls
a re a llo Wed to p rec ip ita te o u t. Pref era b ly the s o lu tio n is
c hilled to a tem p era tu re in the ra ng e f ro m 1 5 C. to 2 5
C. , a nd p ref era b ly the s o lu tio n is c hilled f o r u p to 5 2 ho u rs .
The p rec ip ita te o f ins o lu b le m a teria ls is then rem o v ed,
ty p ic a lly b y ?ltra tio n. Pref era b ly ?ltra tio n is c a rried o u t
thro u g h a 2 0 u m m em b ra ne.
In a f u rther p ref erred em b o dim ent the m etho d m a y f u rther
c o m p ris e a m u lti- s tep ev a p o ra tio n u nder redu c ed p res s u re.
This m a y b e b y ro ta ry ev a p o ra tio n o r o ther k no Wn tec h
niq u es .
Ty p ic a lly the m u lti- s tep ev a p o ra tio n is c a rried o u t o n the
p ro du c t o f the C1 5 a lc o ho l p rec ip ita tio n s tep in o rder to
rem o v e s u b s ta ntia lly a ll o f the C1 5 a lc o ho l a nd Wa ter.
Pref era b ly , the C1 4C5 a lc o ho l is rem o v ed ?rs t a nd then the
Wa ter.
The C1 5 a lc o ho l is p ref era b ly rem o v ed b y hea ting to a
tem p era tu re in the ra ng e o f 5 8 462 C. to g iv e a v a p o u r
tem p era tu re in the ra ng e o f 38 442 C. u nder a v a c u u m in the
ra ng e o f 1 68 41 72 m b a r u ntil there is little o r no v is ib le
c o ndens a te.
Wa ter is then a dditio na lly rem o v ed, p ref era b ly b y a
s tep Wis e redu c tio n o f the v a c u u m in s ta g es to a b o u t 5 0 m b a r.
The dec a rb o x y la tio n s tep m a y b e c a rried o u t p rio r to o r
a f ter ex tra c tio n With liq u id CO2 .
In a p ref erred em b o dim ent the dec a rb o x y la tio n s tep is
c a rried o u t p rio r to ex tra c tio n With liq u id CO2 a nd is
c o ndu c ted b y hea ting the p la nt m a teria l to tem p era tu res a nd
f o r tim es Whic h ens u re a t lea s t 9 5 % c o nv ers io n o f the a c id
c a nna b ino ids f ro m the a c id f o rm to their neu tra l f o rm Whils t
ens u ring therm a l deg ra da tio n o f THC to CBN is les s tha n
1 0% .
Dec a rb o x y la tio n o f c a nna b ino id a c ids is a f u nc tio n o f
tim e a nd tem p era tu re, thu s a t hig her tem p era tu res a s ho rter
p erio d o f tim e Will b e ta k en f o r c o m p lete dec a rb o x y la tio n o f
a g iv en a m o u nt o f c a nna b ino id a c id. In s elec ting a p p ro p ria te
c o nditio ns f o r dec a rb o x y la tio n c o ns idera tio n m u s t, ho W
ev er, b e g iv en to m inim is ing therm a l deg ra da tio n o f the
des ira b le, p ha rm a c o lo g ic a l c a nna b ino ids into u ndes ira b le
deg ra da tio n p ro du c ts , p a rtic u la rly therm a l deg ra da tio n o f
THC to c a nna b ino l ( CBN ) .
Pref era b ly , dec a rb o x y la tio n is c a rried o u t in a m u lti- s tep
hea ting p ro c es s in Whic h the p la nt m a teria l is :
i) hea ted to a ?rs t tem p era tu re f o r a ?rs t ( rela tiv ely s ho rt)
tim e p erio d to ev a p o ra te o if reta ined Wa ter a nd a llo W f o r
u nif o rm hea ting o f the p la nt m a teria l; a nd
2 0
2 5
30
35
40
45
5 0
5 5
60
65
6
ii) the tem p era tu re is inc rea s ed to a s ec o nd tem p era tu re f o r
a s ec o nd tim e p erio d ( ty p ic a lly lo ng er tha n the ?rs t tim e
p erio d) u ntil a t lea s t 9 5 % c o nv ers io n o f the a c id c a nna b
ino ids to their neu tra l f o rm ha s o c c u rred.
Pref era b ly the ?rs t s tep is c o ndu c ted a t a tem p era tu re in
the ra ng e o f 1 00 C. to 1 1 0 C. f o r 1 042 0 m inu tes . Mo re
p ref era b ly the ?rs t tem p era tu re is a b o u t 1 05 C. a nd the ?rs t
tim e p erio d is a b o u t 1 5 m inu tes .
If the p la nt m a teria l is deriv ed f ro m c a nna b is p la nts
ha v ing a hig h CBD c o ntent ( de?ned a s >9 0% CBD a s a
p erc enta g e o f to ta l c a nna b ino id c o ntent) , the s ec o nd tem
p era tu re is p ref era b ly in the ra ng e f ro m 1 1 5 C. to 1 2 5 C. ,
p ref era b ly a b o u t 1 2 0 C. a nd the s ec o nd tim e p erio d is in the
ra ng e f ro m 45 to 75 m inu tes , p ref era b ly a b o u t 60 m inu tes .
Mo re p ref era b ly the s ec o nd tem p era tu re is in the ra ng e f ro m
1 35 C. to 1 45 C. , p ref era b ly 1 40 C. a nd the s ec o nd tim e
p erio d is in the ra ng e f ro m 1 5 to 45 m inu tes , p ref era b ly
a b o u t 30 m inu tes . In a no ther em b o dim ent, m o s t p ref erred
f o r a m a s s o f p la nt m a teria l g rea ter tha n 4 k g , the s ec o nd
tem p era tu re is in the ra ng e f ro m 1 40 C. to 1 5 0 C. ,
p ref era b ly 1 45 C. a nd the s ec o nd tim e p erio d is in the ra ng e
f ro m 5 49 0 m inu tes . The la tter c o nditio ns a re p ref erred f o r
p ro c es s ing a m o u nts o f , f o r ex a m p le, 446 k g o f s ta rting p la nt
m a teria l a nd the ex a c t ?g u res , p a rtic u la rly tim e, m a y v a ry
s lig htly With inc rea s ed m a s s .
If the p la nt m a teria l is deriv ed f ro m c a nna b is p la nts
ha v ing a hig h THC c o ntent ( de?ned a s >9 0% THC a s a
p erc enta g e o f to ta l c a nna b ino id c o ntent) , the s ec o nd tem
p era tu re is p ref era b ly in the ra ng e o f 1 1 5 C. to 1 2 5 C. ,
ty p ic a lly 1 2 0 C. , a nd the s ec o nd tim e p erio d is p ref era b ly
in the ra ng e o f 45 m inu tes to 75 m inu tes , ty p ic a lly a b o u t 60
m inu tes . Mo re p ref era b ly the s ec o nd tem p era tu re is in the
ra ng e o f 1 00 C. to 1 1 0 C. , ty p ic a lly 1 05 C. , a nd the
s ec o nd tim e p erio d is in the ra ng e o f 60 to 1 2 0 m inu tes . In
a no ther em b o dim ent, m o s t p ref erred f o r a m a s s o f p la nt
m a teria l g rea ter tha n 4 k g , the s ec o nd tem p era tu re is in the
ra ng e o f 1 40 C. to 1 5 0 C. , p ref era b ly 1 45 C. , a nd the
s ec o nd tim e p erio d is in the ra ng e o f 45 to 5 5 m inu tes .
Mo s t p ref era b ly the dec a rb o x y la tio n s tep is c o ndu c ted a t
tem p era tu res a nd f o r tim es Whic h ens u re a t lea s t 9 7%
c o nv ers io n o f the a c id c a nna b ino ids to their neu tra l f o rm ,
Whils t ens u ring therm a l deg ra da tio n o f THC to CBN is les s
tha n 5 % .
S ta nda rd c o nditio ns f o r c a nna b ino id a s s a y s , a nd m etho ds
o f c a lc u la ting c a nna b ino id c o ntent ( a s % ) a re g iv en in the
a c c o m p a ny ing Ex a m p les .
The p la nt m a teria l u s ed a s the s ta rting m a teria l f o r the
ex tra c tio n p ro c es s is p ref era b ly g ro u nd, m illed o r o therWis e
p ro c es s ed to g iv e a p a rtic le s iZ e o f les s tha n 2 m m , b u t
p ref era b ly g rea ter tha n 1 m m . S u c h trea tm ent g enera lly
res u lts in im p ro v ed ex tra c tio n o f c a nna b ino ids f ro m the
p la nt m a teria l, a s p a c k a g ing dens ity is im p ro v ed.
In a p ref erred em b o dim ent the m etho d o f the inv entio n
m a y f u rther c o m p ris e a s tep o f trea ting a n ex tra c t ( o r
b o ta nic a l dru g s u b s ta nc e m a teria l) deriv ed f ro m the p la nt
m a teria l With a c tiv a ted c ha rc o a l.
Ty p ic a lly , this s tep Will b e c a rried o u t o n the p ro du c t o f a
p rec ip ita tio n With C1 4C5 a lc o ho l, u s u a lly im m edia tely f o l
lo Wing ?ltra tio n to rem o v e the p rec ip ita te. The liq u id p ro d
u c t o f the a lc o ho lic p rec ip ita tio n is c la s s i?ed a s a b o ta nic a l
dru g s u b s ta nc e a c c o rding to the de?nitio n g iv en a b o v e.
Co nv eniently , trea tm ent With a c tiv a ted c ha rc o a l m a y b e
c a rried o u t b y p a s s ing liq u id m a teria l to b e trea ted do Wn a n
a c tiv a ted c ha rc o a l c o lu m n.
As illu s tra ted in the a c c o m p a ny ing ex a m p les , trea tm ent
With a c tiv a ted c ha rc o a l s ig ni?c a ntly im p ro v es the s ta b ility
o f b o ta nic a l dru g s u b s ta nc es deriv ed f ro m c a nna b is p la nt
US 7, 344, 736 B2
7
m a teria l, s ig ni?c a ntly im p ro v ing res is ta nc e to therm a l deg
ra da tio n o f the a c tiv e c a nna b ino ids .
In a p ref erred em b o dim ent the m etho d o f the inv entio n
Will c o m p ris e the f o llo w ing s tep s , p ref era b ly c a rried o u t in
the s ta ted o rder s ta rting f ro m c a nna b is p la nt m a teria l:
i) dec a rb o x y la tio n,
ii) ex tra c tio n With liq u id CO2 , to p ro du c e a c ru de b o ta nic a l
dru g s u b s ta nc e,
iii) p rec ip ita tio n With C1 * C5 a lc o ho l to redu c e the p ro p o r
tio n o f no n- ta rg et m a teria ls ,
iv ) ?ltra tio n to rem o v e the p rec ip ita te,
v ) ev a p o ra tio n to rem o v e C1 * C5 a lc o ho l a nd Wa ter, to
p ro du c e a ?na l b o ta nic a l dru g s u b s ta nc e ( BDS ) .
A s tep o f trea tm ent With a c tiv a ted c ha rc o a l m a y b e
inc lu ded b etWeen s tep iv ) a nd s tep v ) , res u lting in im p ro v ed
s ta b ility o f the ?na l BDS .
The a p p lic a nt ha s f u rther determ ined tha t the a dditio n o f
a p ro p o rtio n o f m o di?er o r p o la r s o lv ent, f o r ex a m p le a C1
to C5 a lc o ho l, a s ex em p li?ed b y etha no l, to liq u id c a rb o n
dio x ide s o lv ent m a y f u rther inc rea s e s elec tiv ity o f the
ex tra c tio n p ro c es s .
Ac c o rding ly , the inv entio n f u rther p ro v ides a m etho d o f
ex tra c ting c a nna b ino ids f ro m p la nt m a teria l c o m p ris ing a n
ex tra c tio n With liq u id CO2 , c ha ra c teris ed in tha t a n o rg a nic
m o di?er o r p o la r s o lv ent is a dded to the c a rb o n dio x ide.
Pref era b ly the m o di?er o r p o la r s o lv ent is a dded in a n
a m o u nt o f u p to 1 0% b y Weig ht.
Pref era b ly the m o di?er is a C1 * C5 a lc o ho l, m o s t p ref er
a b ly etha no l.
In a f u rther a s p ec t the inv entio n f u rther rela tes to b o ta ni
c a l dru g s u b s ta nc es deriv ed f ro m c a nna b is p la nt m a teria l.
Theref o re, the inv entio n p ro v ides a b o ta nic a l dru g s u b s ta nc e
o b ta ina b le f ro m b o ta nic a l ra W m a teria l f ro m a hig h THC
c o nta ining c a nna b is p la nt ha v ing a THC c o ntent o f a t lea s t
9 0% W/ W o f to ta l c a nna b ino id c o ntent, Wherein s a id b o ta ni
c a l dru g s u b s ta nc e is a n ex tra c t deriv ed f ro m the hig h THC
c a nna b is p la nt c o m p ris ing a t lea s t 5 0% THC W/ W o f ex tra c t,
no m o re tha n 5 % CBD W/ W o f the THC c o ntent, a nd no
m o re tha n 5 % c a nna b ino ids o ther tha n THC a nd CBD W/ W
o f the THC c o ntent.
The % THC Wt/ Wt o f ex tra c t is m o re p ref era b ly a t lea s t
5 5 % , a nd m o re p ref era b ly s till a t lea s t 60% . The o ther
c a nna b ino ids a nd the a s s a y m etho do lo g y f o r determ ining the
a m o u nts a re g iv en la ter.
The inv entio n a ls o p ro v ides a b o ta nic a l dru g s u b s ta nc e
o b ta ina b le f ro m b o ta nic a l ra W m a teria l f ro m a hig h CBD
c o nta ining c a nna b is p la nt ha v ing a CBD c o ntent o f a t lea s t
9 0% W/ W o f to ta l c a nna b ino id c o ntent, Wherein s a id b o ta ni
c a l dru g s u b s ta nc e is a n ex tra c t deriv ed f ro m a hig h CBD
c a nna b is p la nt, Whic h ex tra c t c o m p ris es a t lea s t 5 0% CBD
W/ W o f ex tra c t, no m o re tha n 7. 5 % THC W/ W o f the CBD
c o ntent, a nd no m o re tha n 5 % c a nna b ino ids o ther tha n CBD
a nd THC ex p res s ed a s % W/ W o f the CBD c o ntent.
The s k illed m a n Will a p p rec ia te tha t hig h THC p la nts s u c h
a s , f o r ex a m p le, S k u nk ha v e b een b red, a lb eit f o r rec re
a tio na l u s e, u s ing tra ditio na l b reeding tec hniq u es Whic h c a n
lik eWis e b e u s ed to dev elo p p la nts ric h in o ther c a nna b ino ids
e. g CBD b y na tu ra l s elec tio n o r b y g enetic tec hniq u es a s the
g enes f o r c a nna b idio la te s y ntha s e a nd THC s y ntha s e ha v e
b een identi?ed, s ee J P 2 001 02 9 08 2 a nd J P2 000078 9 79 .
CPRO 9 2 1 01 8 La nd ra c e Tu rk ey is a n ex a m p le o f hig h CBD
p la nt.
The b o ta nic a l dru g s u b s ta nc es m a y b e o b ta ined s ta rting
f ro m c a nna b is p la nt m a teria l ( b o ta nic a l ra W m a teria l) u s ing
the ex tra c tio n m etho d a c c o rding to the inv entio n.
In a p ref erred em b o dim ent the b o ta nic a l dru g s u b s ta nc e
c o m p ris es no m o re tha n 4 p p b a ?a to x in.
2 0
2 5
30
35
40
45
5 0
5 5
60
65
8
In a f u rther p ref erred em b o dim ent the b o ta nic a l dru g
s u b s ta nc e c o m p ris es no m o re tha n 2 0 p p m to ta l hea v y
m eta ls .
In a f u rther p ref erred em b o dim ent the b o ta nic a l dru g
s u b s ta nc e c o m p ris es no m o re tha n 1 5 % W/ W res idu a l s o l
v ents , m o re s p ec i?c a lly no m o re tha t 1 5 % W/ W etha no l.
In a f u rther p ref erred em b o dim ent the b o ta nic a l dru g
s u b s ta nc e c o m p ris es no m o re tha n 1 05 c f u / g TVC ( To ta l
Via b le Co u nt) , no m o re tha n 1 04 c f u / g f u ng i, no m o re tha n
1 03 c f u / g entero b a c teria a nd o ther no n g ra m neg a tiv e o rg a n
is m s , a nd no detec ta b le E. c o li, S a lm o nella o r S . a u reu s .
The a b o v e- lis ted p a ra m eters rela te to p u rity o f the b o ta ni
c a l dru g s u b s ta nc e a nd de?ne a lev el o f p u rity Whic h is
p ref erred if the b o ta nic a l dru g s u b s ta nc e is to b e inc o rp o
ra ted into a p ha rm a c eu tic a l p ro du c t. Bo ta nic a l dru g s u b
s ta nc es ha v ing the req u ired lev el o f p u rity m a y b e o b ta ined
u s ing the ex tra c tio n p ro c es s a c c o rding to the inv entio n,
p a rtic u la rly u s ing the o p era ting c o nditio ns a nd q u a lity c o n
tro l p ro c edu res des c rib ed in the a c c o m p a ny ing ex a m p les .
S ta nda rd a s s a y tec hniq u es f o r u s e in determ ining the lev els
o f a ?a to x in, hea v y m eta ls , res idu a l s o lv ents a nd b a c teria l
c o nta m ina nts in a b o ta nic a l dru g s u b s ta nc e a re k no Wn in the
a rt ( e. g . Eu ro p ea n Pha rm a c o p o eia ( Ph. Eu r) s ta nda rd p ro c e
du res ) a nd f u rther deta ils a re p ro v ided in the a c c o m p a ny ing
Ex a m p les .
Bo ta nic a l dru g s u b s ta nc es p rep a red f ro m c a nna b is p la nt
m a teria l a c c o rding to the m etho ds o f the inv entio n m a y b e
f o rm u la ted With o ne o r m o re p ha rm a c eu tic a lly a c c ep ta b le
c a rriers , dilu ents o r ex c ip ients o r dep o s ited o n a p ha rm a
c eu tic a lly a c c ep ta b le s u rf a c e f o r v a p o ris a tio n in o rder to
p ro du c e p ha rm a c eu tic a l f o rm u la tio ns c o nta ining c a nna b
ino ids a s the p ha rm a c eu tic a lly a c tiv e a g ents .
Theref o re, in a f u rther a s p ec t the inv entio n p ro v ides a
m etho d o f m a k ing a p ha rm a c eu tic a l c o m p o s itio n c o m p ris
ing , a s a n a c tiv e a g ent, a b o ta nic a l dru g s u b s ta nc e Whic h is
a n ex tra c t f ro m a t lea s t o ne c a nna b is p la nt v a riety , Whic h
m etho d c o m p ris es p rep a ring a b o ta nic a l dru g s u b s ta nc e
c o nta ining c a nna b ino ids f ro m the a t lea s t o ne c a nna b is p la nt
v a riety u s ing the ex tra c tio n m etho d a c c o rding to the inv en
tio n, a nd f o rm u la ting the b o ta nic a l dru g s u b s ta nc e With o ne
o r m o re p ha rm a c eu tic a lly a c c ep ta b le dilu ents , c a rriers o r
ex c ip ients o r dep o s iting the b o ta nic a l dru g s u b s ta nc e o n a
p ha rm a c eu tic a lly a c c ep ta b le s u rf a c e f o r v a p o ris a tio n to p ro
du c e a p ha rm a c eu tic a l c o m p o s itio n.
S ep a ra te b o ta nic a l dru g s u b s ta nc es m a y b e p rep a red f ro m
s ing le c a nna b is p la nt v a rieties ha v ing dif f ering c a nna b ino id
c o ntent ( eg hig h THC a nd hig h CBD p la nts ) a nd then
m ix ed o r b lended to g ether p rio r to f o rm u la tio n to p ro du c e
the ?na l p ha rm a c eu tic a l c o m p o s itio n. This a p p ro a c h is p re
f erred if , f o r ex a m p le, it is des ired to a c hiev e a de?ned ra tio
b y Weig ht o f indiv idu a l c a nna b ino ids in the ?na l f o rm u la
tio n. Alterna tiv ely , b o ta nic a l ra W m a teria l f ro m o ne o r m o re
c a nna b is p la nt v a rieties o f de?ned c a nna b ino id c o ntent m a y
b e m ix ed to g ether p rio r to ex tra c tio n o f a s ing le b o ta nic a l
dru g s u b s ta nc e ha v ing the des ired c a nna b ino id c o ntent,
Whic h m a y then b e f o rm u la ted into a ?na l p ha rm a c eu tic a l
c o m p o s itio n.
The b o ta nic a l dru g s u b s ta nc e m a y b e f o rm u la ted With a ny
c o nv enient p ha rm a c eu tic a lly a c c ep ta b le dilu ents , c a rriers o r
ex c ip ients to p ro du c e a p ha rm a c eu tic a l c o m p o s itio n. The
c ho ic e o f dilu ents , c a rriers o r ex c ip ients Will dep end o n the
des ired do s a g e f o rm , Whic h m a y in tu rn b e dep endent o n the
intended ro u te o f a dm inis tra tio n to a p a tient. Pref erred
do s a g e f o rm s inc lu de, inter a lia , liq u id do s a g e f o rm s f o r
a dm inis tra tio n v ia p u m p - a c tio n o r a ero s o l s p ra y s , ta b lets ,
p a s tilles , g els , c a p s u les , s u p p o s ito ries , p o Wders , etc a nd
v a p o u ris ers . S u c h do s a g e f o rm s m a y b e p rep a red in a c c o r
US 7, 344, 736 B2
da nc e With s ta nda rd p rinc ip les o f p ha rm a c eu tic a l f o rm u la
tio n, k no w n to tho s e s k illed in the a rt. Pref erred do s a g e
f o rm s , a nd m etho ds o f p rep a ring s u c h do s a g e f o rm s , a re
des c rib ed in the a p p lic a nt s c o - p ending Interna tio na l a p p li
c a tio n PCT/ G B02 / 0062 0.
Liq u id f o rm u la tio ns a re p a rtic u la rly p ref erred. A p a rtic u
la rly p ref erred f o rm u la tio n f o r a dm inis tra tio n o f c a nna b
ino ids , tho u g h no t intended to b e lim iting to the inv entio n,
is a liq u id f o rm u la tio n c o m p ris ing the b o ta nic a l dru g s u b
s ta nc e, etha no l a nd p ro p y lene g ly c o l, a nd o p tio na lly a
?a v o u ring , s u c h a s p ep p erm int o il. This f o rm u la tio n m a y b e
c o nv eniently a dm inis tered to the b u c c a l o r s u b ling u a l m u c o
s a e v ia a p u m p - a c tio n s p ra y , a nd p ro v ides f o r ef ?c ient
a b s o rp tio n o f the a c tiv e c a nna b ino ids .
The v a rio u s a s p ec ts o f the inv entio ns a re f u rther illu s
tra ted, b y Wa y o f ex a m p le o nly , b y the f o llo Wing ex a m p les ,
to g ether With the a c c o m p a ny ing Fig u res .
BRIEF DES CRIPTION OF THE DRAWIN G S
FIG . 1 illu s tra tes lo s s o f THC o v er tim e a t 400 C. f o r
s ta nda rd THC b o ta nic a l dru g s u b s ta nc e ( BDS ) a nd a c tiv a ted
c ha rc o a l- trea ted THC BDS ( p u ri?ed BDS ) . Y- a x is : a m o u nt
o f THC ( ex p res s ed a s p erc enta g e o f to v a lu e) , x - a x is : tim e
in m o nths .
FIG . 2 illu s tra tes lo s s o f CBD o v er tim e a t 400 C. f o r
s ta nda rd CBD b o ta nic a l dru g s u b s ta nc e ( BDS ) a nd a c tiv a ted
c ha rc o a l- trea ted CBD BDS ( p u ri?ed BDS ) . Y- a x is : a m o u nt
o f CBD ( ex p res s ed a s p erc enta g e o f to v a lu e) , x - a x is : tim e
in m o nths .
FIG . 3 illu s tra tes f o rm a tio n o f c a nna b ino l ( CBN ) o v er
tim e a t 400 C. f o r s ta nda rd THC b o ta nic a l dru g s u b s ta nc e
( BDS ) a nd a c tiv a ted c ha rc o a l- trea ted THC BDS ( p u ri?ed
BDS ) . Y- a x is : a m o u nt o f CBN ( ex p res s ed a s p erc enta g e o f
t0 v a lu e) , x - a x is : tim e in m o nths .
EXAMPLES
Ex a m p le 1
Dev elo p m ent o f a Pro c es s f o r Ex tra c tio n o f
Ca nna b ino ids f ro m Ca nna b is Pla nts
S elec tio n o f Ca nna b is Chem o v a rs
G W Pha rm a Ltd ha s dev elo p ed dis tinc t v a rieties o f Ca n
na b is p la nt hy b rids to m a x im is e the o u tp u t o f the s p ec i?c
c hem ic a l c o ns titu ents , c a nna b ino ids . TWo ty p es o f p la nt a re
u s ed; o ne c hem o v a r p ro du c es p rim a rily THC a nd a f u rther
c hem o v a r p ro du c es p redo m ina tely CBD. Ho Wev er a lterna
tiv e v a rieties c a n b e o b ta inedis ee f o r ex a m p le, Co m m o n
c a nna b ino ids p heno ty p es in 35 0 s to c k s o f c a nna b is , S m a ll
a nd Bec k s tea d, LLo y dia v o l 36b , 1 9 73 p l44il 5 6ia nd b red
u s ing tec hniq u es Well k no Wn to the s k illed m a n to m a x im is e
c a nna b ino id c o ntent.
Chem ic a l a nd s tru c tu ra l s im ila rities ex is t b etWeen THC
a nd CBD. Du e to thes e s im ila rities to g ether With the b o ta nic
o rig in o f the s ta rting m a teria ls , ea c h c a n b e c o ns idered to b e
interc ha ng ea b le With res p ec t to the dev elo p m ent o f p ro
c es s es f o r ex tra c tio n o f c a nna b ino ids .
Pref era b ly , ea c h Ca nna b is c hem o v a r is p ro c es s ed a nd
c o ntro lled s ep a ra tely to y ield tWo dis tinc t BDS s . Ho Wev er,
it is p o s s ib le to m ix p la nt m a teria l f ro m tWo o r m o re
c hem o v a rs o r u s e a v a riety Whic h Will p ro du c e the des ired
ra tio o f g iv en c a nna b ino ids p rio r to ex tra c tio n, a nd thu s
p rep a re a s ing le BDS .
2 0
2 5
30
35
40
45
5 0
5 5
60
65
1 0
Pro du c tio n o f Bo ta nic a l Ra W Ma teria l
BDS is p rep a red f ro m ex tra c ts o f Ca nna b is s a liv a L.
( f a m ily Ca nna b ida c ea e) . Ca nna b is s a liv a Wa s des c rib ed in
the 1 9 34 Britis h Pha rm a c o p o eia . Ca nna b is is g ro Wn u nder
United King do m Ho m e Of ?c e lic enc e u nder the c o ntro l o f
G W Pha rm a Ltd in the United King do m . G ro Wing f a c ilities
a re eq u ip p ed With s ha des a nd f u ll c lim a tic c o ntro l ( tem p era
tu re, hu m idity a nd hig h intens ity lig hting ) s o tha t s ev era l
c ro p s p er y ea r c a n b e p ro du c ed in a lm o s t identic a l g ro Wing
c o nditio ns thu s ens u ring c o ntinu ity o f s u p p ly .
Cu ltiv a tio n:
Ca nna b is p la nts a re p ro p a g a ted f ro m c u tting s ta k en f ro m
the m o ther p la nts , o rig ina ting f ro m a s ing le s eed s o u rc e.
Theref o re a c ro p is p ro du c ed thro u g h a s ex u a l p ro p a g a tio n
Where the p la nts a re a ll f em a le. Pro p a g a tio n u s ing c u tting s
c o ntro ls g eno ty p e c o ns is tenc y .
The c u tting s a re ro o ted in c o m p o s t s u p p lied a s p es tic ide
f ree. The p la nts a re Wa tered a nd s u s ta ined relea s e f ertiliZ er
is a p p lied du ring the g ro Wing c y c le. Thro u g h c o ntro lled
g ro Wing c o nditio ns the p la nts ta k e a p p ro x im a tely 1 2 Week s
to rea c h m a tu rity .
The p la nts a re irrig a ted thro u g ho u t their g ro Wing c y c le
With p o ta b le q u a lity Wa ter.
N o s y nthetic herb ic ides o r p es tic ides a re u s ed in the
c u ltiv a tio n o f Ca nna b is p la nts .
Co m p o s t:
Ef ?c ient c u ltiv a tio n o f Ca nna b is nec es s ita tes the s u p p ly
o f a relia b ly u nif o rm g ro Wing m edia .
The c o m p o s t p ro v ides a s o f t tex tu re, hig h a ir p o ro s ity ,
rea dy Wetting , lo W c o ndu c tiv ity a nd b a la nc ed nu trient s u p
p ly . The c o m p o s t c o ns is ts o f p ea t a nd a dded na tu ra l m inera ls
inc lu ding lim e ( m a g nes iu m a nd c a lc iu m c a rb o na tes ) to
p ro v ide p H c o ntro l o f the c o m p o s t du ring the g ro Wing c y c le
o f the Ca nna b is p la nts .
The c o m p o s t c o nta ins a n a deq u a te s u p p ly o f es s entia l
m inera ls a nd a m inim u m o f m inera ls With k no Wn a dv ers e
ef f ec ts o n the p la nts . S o m e m inera ls inc lu ding m a ng a nes e
c a n b e p res ent in a n ins o lu b le f o rm in c o m p o s t a nd b e
relea s ed in a f reely s o lu b le f o rm o v er tim e. Co ntro lling
c o m p o s t p H a nd m o nito ring irrig a tio n to a v o id Wa terlo g g ing
Will c o ntro l s o lu b le m a ng a nes e lev els . Co m p o s t p H is m a in
ta ined a b o v e 5 . 5 .
The c o m p o s t is dec la red a s p es tic ide f ree, a s no p es tic ides
o r herb ic ides a re a dded.
Fertilis er:
The c o m p o s t c o nta ins f ertilis er identi?a b le in tWo dis c rete
f o rm s , a b a s e f ertilis er a nd a s lo W relea s e f ertilis er. Addi
tio na l s lo W relea s e f ertilis er is a p p lied to the p la nts du ring
g ro Wing .
Dis ea s e a nd Pes t Co ntro l:
N o a rti?c ia l herb ic ides o r p es tic ides a re u s ed du ring
c u ltiv a tio n. S tring ent hy g iene c o nditio ns redu c e ing res s o f
p es ts a nd dis ea s es .
By c o ntro lling the g ro Wing c o nditio ns , env iro nm enta l
s tres s es s u c h a s dro u g ht, ins u f ?c ient lig ht a nd u nf a v o u ra b le
tem p era tu res redu c es the ris k o f dis ea s e.
Reg u la r ins p ec tio n o f the p la nts du ring the g ro Wing c y c le
a llo Ws f o r the detec tio n o f a ny ro g u e p la nts a nd p es ts . Ro g u e
m a le p la nts m a y a ris e, tho u g h Weeds s ho u ld b e a b s ent du e
to the c o ntro lled g ro Wing c o nditio ns a nd m edia . Freq u ent
ins p ec tio ns a nd b io lo g ic a l c o ntro l m etho ds a re u s ed to
m a na g e a ny p es ts a nd dis ea s es tha t m a y o c c u r.
Pla nt Co llec tio n:
Thro u g h s tric t c o ntro l o f g ro Wing c o nditio ns the Ca n
na b is p la nts rea c h m a tu rity in a p p ro x im a tely 1 2 Week s . In
US 7, 344, 736 B2
1 1
the la s t w eek s o f g ro w th, dens e res ino u s ?o w ers dev elo p . By
the end o f a p p ro x im a tely w eek 1 1 the c a nna b ino id b io s y n
thes is ha s s lo w ed m a rk edly , a nd the p la nts a re rea dy f o r
ha rv es t.
The entire p la nt is c u t a nd dried in a tem p era tu re a nd
hu m idity c o ntro lled env iro nm ent.
Ap p ro x im a tely 2 1 0 C.
Ap p ro x im a tely 38 * 45 % RH.
Dried p la nt is p hy s ic a lly a s s es s ed f o r end- p o int.
THC a nd CBD a re the p rinc ip le b io a c tiv e c o ns titu ents in
the BDS . Ho w ev er, thes e c o ns titu ents a re p res ent a s b io
lo g ic a lly ina c tiv e c a rb o x y lic a c ids in the BRM.
THCA
CBDA
The a c id f o rm s s lo w ly dec a rb o x y la te o v er tim e du ring
dry ing . The lea v es a nd ?o w ers a re s trip p ed f ro m the la rg er
s tem s to p ro v ide the Bo ta nic a l Ra w Ma teria l ( BRM) .
S to ra g e o f BRM:
Under c o nditio ns o f s to ra g e the lo s s o n dry ing rea c hes
eq u ilib riu m o f a p p ro x im a tely 1 0% . The s to ra g e c o nditio ns
f o r the dried BRM w ill b e dep endent o n the p hy s ic a l s ta tu s
o f the BRM.
G enera l S to ra g e Co nditio ns f o r BRM:
Pro tec ted f ro m lig ht.
Ap p ro x im a tely 1 5 * 2 5 o C. o r 2 00 C.
Ap p ro x im a tely 38 * 42 % RH.
S u m m a ry - p ro du c tio n o f a BRM:
Ha rv es t o f p la nts
Dry ing
( lig ht ex c lu s io n)
BRM
( c o nta ins : THCA + CBDA)
Milling to les s tha n 2 000 u m to redu c e p a rtic le s ize
Dec a rb o x y la tio n o f a c id f o rm o f c a nna b ino ids ( THCA + CBDA)
to p ro du c e neu tra l c a nna b ino ids ( THC + CBD)
Ty p ic a l BRM S p ec i?c a tio n Deriv ed f ro m a Hig h CBD
Va riety is Illu s tra ted in Ta b le 2 :
Tes t Metho d S p ec i?c a tio n
Identi?c a tio n:
A Vis u a l Co m p lies
B TLC Co rres p o nds to s ta nda rd ( f o r CBD
& CBDA)
C HPLC/ U V Po s itiv e f o r CBDA
As s a y : In- ho u s e N o t les s tha n ( N LT) 9 0% o f
CBDA + CBD ( HPLC/ UV) a s s a y ed c a nna b ino ids b y p ea k a rea
Lo s s o n Dry ing : Ph. Eu r. N o t m o re tha n ( N MT) 1 5 %
5
1 0
2 0
2 5
30
35
40
45
5 0
5 5
60
65
1 2
- c o ntinu ed
Tes t Metho d S p ec i?c a tio n
A?a to x in: UKAS * m etho d N MT 4 p p b
Mic ro b ia l: Ph. Eu r.
TVC N MT 1 07 c f u / g
Fu ng i N MT 1 O5 c f u / g
E. c a Z i N MT 1 02 c f u / g
Fo reig n Ma tter: Ph. Eu r. N MT 2 %
Res idu a l Ph. Eu r. Co m p lies
Herb ic ides a nd
Pes tic ides :
* United King do m Ac c redita tio n S erv ic e
Ana ly tic a l Metho ds :
Identi?c a tio n b y Vis u a l:
Ma c ro s c o p ic c ha ra c teris tic s a llo w the f ea tu res o f the
Ca nna b is p la nt to b e dis ting u is hed f ro m p o tentia l a du lter
a nts a nd s u b s titu tes . It is a v is u a l identi?c a tio n a g a ins t a
p ho to g ra p hic s ta nda rd.
Identi?c a tio n b y TLC:
TLC u s es b o th retentio n tim e a nd c ha ra c teris tic s p o t
c o lo u r to ef f ec tiv ely identif y the v a riety o f Ca nna b is . La b o
ra to ry s a m p les a re p rep a red f o r TLC a na ly s is b y ex tra c ting
the dried herb . An a liq u o t is s p o tted o nto a TLC p la te,
a lo ng s ide ref erenc e s a m p les f o r THC a nd CBD. Fo llo w ing
ex p o s u re to Fa s t Blu e B rea g ent, THC a nd THCA p res ent a s
p ink s p o ts , w hile CBD a nd CBDA a re o ra ng e in c o lo u r.
N eu tra ls c a n b e dis ting u is hed f ro m the a c ids b y c o m p a ris o n
o f the Rf v a lu e to tha t o b ta ined f o r the s ta nda rds . Identity is
c o n?rm ed b y c o m p a ris o n o f Rf a nd c o lo u r o f the s a m p le
s p o t, to tha t o b ta ined f o r the a p p ro p ria te s ta nda rd.
Identi?c a tio n b y HPLC:
HPLC u s es retentio n tim e c o m p a ris o n o f c a nna b ino ids to
ef f ec tiv ely identif y the v a riety o f Ca nna b is . The rev ers ed
p ha s e HPLC m etho d is s p ec i?c f o r CBD a nd CBDA, a nd
theref o re m a y b e u s ed a s a n identity tes t. S a m p les o f
b io m a s s a re ex tra c ted a nd c entrif u g ed. Detec tio n o f a ll
a na ly tes is a c c o m p lis hed a t 2 2 0 nm w ith a dditio na l c o n?r
m a tio n o f a c idic a na ly tes a t 31 0 nm .
As s a y ( CBD+CBDA) :
This a s s a y is u s ed to m o nito r the CBD a nd CBDA c o ntent
in the p la nt. CBD a nd CBDA a s s a y a re determ ined u s ing a n
HPLC m etho d.
The ef ?c ienc y o f the dec a rb o x y la tio n p ro c es s is deter
m ined b y div iding the % c o ntent in term s o f w / w o f CBD b y
the to ta l CBD+CBDA c o ntent.
Lo s s o n Dry ing :
Lo s s o n Dry ing is ev a lu a ted u s ing Ph. Eu r. tes t m etho d.
A?a to x in is a na ly s ed u s ing a United King do m Ac c redi
ta tio n S erv ic e ( UKAS ) a c c redited m etho d.
Mic ro b ia l :
Mic ro b io lo g ic a l q u a lity is determ ined u s ing Ph. Eu r.
m etho do lo g y .
Fo reig n Ma tter:
Fo reig n Ma tter is ev a lu a ted u s ing the Ph. Eu r. tes t m etho d.
Flo w ers , lea v es a nd s ide s tem s a re s p rea d o u t in a thin la y er
o n a c lea n la b o ra to ry s u rf a c e. Fo reig n Ma tter is s ep a ra ted b y
ha nd a s c o m p letely a s p o s s ib le, a nd is w eig hed. Res u lts a re
US 7, 344, 736 B2
1 3
ex p res s ed a s % W/ W o f Fo reig n Ma tter in the herb a l b io m a s s
s a m p le. Fo reig n Ma tter m a y c o m p ris e no m o re tha n 2 % o f
the b io m a s s .
Res idu a l Herb ic ides a nd Pes tic ides :
The Ca nna b is p la nts a re g ro Wn in a Well c o ntro lled
env iro nm ent. N o a rti?c ia l herb ic ides o r p es tic ides a re u s ed
o r needed du ring c u ltiv a tio n.
An eq u iv a lent BRM s p ec i?c a tio n ( c o m p a re ta b le 2 ) is
deriv ed f o r a hig h THC v a riety a nd identic a l a na ly tic a l
m etho ds f o llo Wed, ex c ep t tha t THC/ THCA rep la c es CBD/
CBDA.
Dec a rb o x y la tio n
THC a nd CBD a re the p rinc ip le b io a c tiv e c o ns titu ents in
Ca nna b is . Ho Wev er, thes e c o ns titu ents a re p res ent a s the
b io lo g ic a lly ina c tiv e c a rb o x y lic a c ids in Ca nna b is p la nts . In
o rder to ex tra c t THC o r CBD f ro m c a nna b is p la nt m a teria l,
it is nec es s a ry to c o nv ert the s to ra g e p rec u rs o r c o m p o u nds
o f THCA a nd CBDA into their m o re rea dily ex tra c ta b le a nd
p ha rm a c o lo g ic a lly a c tiv e f o rm s . THC a nd CBD a c ids
s lo Wly dec a rb o x y la te na tu ra lly o v er tim e. The tra ditio na l
Wa y to inc rea s e ra te o f dec a rb o x y la tio n is b y the a p p lic a tio n
o f hea t. Ho Wev er, THCA is c o nv erted no t o nly to THC, b u t
a ls o to a no ther c a nna b ino id, c a nna b ino l ( CBN ) .
o
1 .
THCA o r CBDA ( c 2 2 rr30o 4) > THC o rCBD( C2 1 H30O2 )
The dec a rb o x y la tio n p ro c edu re is g enera lly c a rried o u t
Within the p rep a ra tio n o f the s ta rting m a teria l o r b o ta nic a l
ra W m a teria l ( BRM) , p rio r to the initia tio n o f the ex tra c tio n
p ro c es s .
La b o ra to ry S tu dies - dec a rb o x y la tio n
Po rtio ns o f m illed dried p la nt m a teria l Were s u b j ec ted to
hea t ( a p p ro x im a tely 0. 2 5 g With p a rtic le s iZ e 1 42 m m ) . A
p ilo t s c a le ex p erim enta l s y s tem Wa s s et u p , With the o b j ec
tiv e o f determ ining p a ra m eters f o r the o p tim a l c o nv ers io n o f
THCA o r CBDA into THC a nd CBD res p ec tiv ely , With
c o nc o m ita nt m inim a l lo s s o f thes e ens u ing c o m p o u nds into
their therm a l deg ra da tio n p ro du c ts , in the c a s e o f THC the
f o rm a tio n o f CBN .
Brief Des c rip tio n o f Ma teria ls a nd Metho ds :
Po rtio ns ( 0. 2 5 g ) o f m illed ( a p p ro x im a tely 1 42 m m p a r
tic le s iZ e) o f b o th THCA a nd CBDA herb a l m a teria ls Were
p la c ed in 2 0- m l g la s s hea ds p a c e v ia ls a nd the v ia ls s ea led
tig htly With c rim p c a p p ed Te?o n- f a c ed b u ty l ru b b er s ea ls .
S ea led v ia ls Were hea ted a t o ne o f three tem p era tu res , f o r
p erio ds o f u p to 4 hrs a s f o llo Ws :
1 05 C. , 1 2 00 C. , 1 400 C. f o r 0. 5 , 1 . 0, 2 . 0 a nd 4. 0 ho u rs .
The hea ting Wa s p erf o rm ed in a n o v en With f o rc ed a ir
c irc u la tio n. Ov en c o nditio ns Were s ho Wn to b e a c c u ra te to
Within 05 41 . 0 deg ree a t the three tem p era tu res u s ed.
Af ter the hea ting p ro c es s Wa s c o m p lete rep res enta tiv e
s a m p les o f the dec a rb o x y la ted herb Were a s s a y ed u s ing
HPLC, G C a nd TLC tec hniq u es . S ta nda rds o f THC, CBD
a nd CBN Were inc lu de in the HPLC a nd G C s eq u enc es .
Res u lts a nd Dis c u s s io ns :
HPLC a na ly s is o f the s o lv ent ex tra c ts Wa s a b le to dem
o ns tra te the dis a p p ea ra nc e o f either CBDA o r THCA a s a
f u nc tio n o f tim e a t the tWo lo Wer tem p era tu res . At 1 40 C. ,
the ea rlies t tim e p o int s a m p les a t 0. 5 ho u r c o nta ined o nly
v ery m o des t lev els o f a p ea k elu ting a t the retentio n tim es o f
CBDA o r THCA.
2 0
2 5
30
35
40
45
5 0
5 5
60
65
1 4
Ta b les 3 a nd 4 p res ent HPLC da ta q u a ntif y ing the c o n
v ers io n o f CBDA o r THCA into the f ree c o m p o u nds ; a ls o
p res ented is da ta s ho Wing the c o ntent o f CBD o r THC a nd
the ra tio o f CBD/ CBDA+CBD o r THC/ THCA+THC. The
c o nv ers io n o f the c a rb o x y lic a c id f o rm s to the c o rres p o nding
dec a rb o x y la ted f o rm c a n b e m o nito red b y c o m p a ring the
dec a rb o x y la ted/ dec a rb o x y la ted p lu s u n- dec a rb o x y la ted
ra tio With the a b s o lu te c o ntent o f the dec a rb o x y la ted c o m
p o u nds . Thu s , When the ra tio rea c hes a m a x im u m v a lu e
( >0. 9 5 ) , the ea rlies t tim e/ tem p era tu re p o int a t Whic h the
c o ntent o f THC o r CBD is a ls o m a x im a l, s ho u ld b e o p tim a l
f o r the c o nv ers io n p ro c es s .
Thu s , f o r CBD c o nta ining herb , 1 ho u r a t 1 2 0 C. o r 0. 5
ho u r a t 1 40 C. , Wa s a p p ro p ria te.
This is c o n?rm ed b y ex a m ina tio n o f the TLC c hro m a to
g ra m f o r the s o lv ent ex tra c ts , CBDA is a b s ent a f ter 1 ho u r
a t 1 2 0 C. o r a t a ny tim e p o int a t 1 40 C.
Fo r THC there is a third c riterio n, f o rm a tio n o f CBN ,
Where it is des ira b le to m inim is e the f o rm a tio n o f this
c o m p o u nd du ring the therm a l dec a rb o x y la tio n p ro c es s .
Ta b le 5 p ro v ides G a s Chro m a to g ra p hy ( G C) da ta Where a
CBN / THC ra tio c a n b e deriv ed. Ta k en into c o ns idera tio n,
a lo ng s ide the THC/ THCA+THC ra tio a nd the m a x im u m
THC c o ntent, m inim a l CBN f o rm a tio n o c c u rs a f ter 0. 5 o r
1 . 0 ho u r a t 1 2 0 C. At 1 40 C. , ev en 0. 5 ho u r g iv es a hig her
c o ntent o f CBN tha n either o f the tWo lo Wer tim e/ tem p era
tu re p o ints .
Theref o re la b o ra to ry s tu dies dem o ns tra te the o p tim u m
c o nditio ns f o r the dec a rb o x y la tio n o f :
Chem o v a r p ro du c ing p rim a rily CBD is 1 ho u r a t 1 2 0 C.
o r 0. 5 ho u r a t 1 40 C.
Chem o v a r p ro du c ing p rim a rily THC to m inim is e CBN
f o rm a tio n, is 1 to 2 ho u rs a t 1 05 C. o r 1 ho u r a t 1 2 0
C.
Thin la y er c hro m a to g ra p hy rev ea ls tha t v irtu a lly a ll o f the
THCA ha s dis a p p ea red a f ter 4 ho u rs a t 1 05 C. a nd a f ter 1
ho u r a t 1 2 0 C. N o THCA is v is ib le a t a ny tim e p o int When
the herb is hea ted a t 1 40 C. A s m a ll a m o u nt o f res idu a l
s ta ining a t this retentio n v a lu e o n TLC a nd the p res enc e a t
lo W lev els o f a p ea k c o inc ident With THCA o n HPLC
a na ly s is m a y indic a te the p res enc e o f a m ino r c a nna b ino id
ra ther tha n res idu a l THCA.
TABLE 3
HPLC Da ta f ro m Dec a rb o x y la tio n o f CBDA Herb a l Ma teria l
CBD/ CBD + CBD p ea k a rea / 0. 1 g
Tem p era tu re Tim e ( ho u rs ) CBDA o f herb
1 05 C. Z ero 0. 1 5 4769
0. 5 0. 2 2 5 2 62
1 . 0 0. 8 6 5 5 9 8
2 . 0 0. 9 3 5 2 5 1
4. 0 0. 9 8 5 2 42
1 2 0 C. 0. 5 0. 9 1 5 1 2 9
1 . 0 0. 9 7 5 2 1 7
2 . 0 0. 9 9 5 037
4. 0 1 . 00 5 2 00
1 40 C. 0. 5 0. 9 6 5 440
1 . 0 1 . 00 5 1 05
2 . 0 1 . 00 5 1 5 7
4. 0 1 . 00 5 005
US 7, 344, 736 B2
1 5
TABLE 4
HPLC Da ta f ro m Dec a rb o x y la tio n o f THCA Herb a l Ma teria l
THC p ea k
THC/ THC + a rea / 0. 1 g
Tem p era tu re Tim e ( ho u rs ) THCA o f herb
1 05 C. Z ero 0. 1 7 9 9 2 . 9
0. 5 0. 8 7 5 749
1 . 0 0. 9 3 5 2 73
2 . 0 0. 9 8 7734
4. 0 0. 9 9 7068
1 2 0 C. 0. 5 0. 9 7 71 8 9
1 . 0 0. 9 9 639 1
2 . 0 0. 9 9 65 00
4. 0 1 . 00 5 8 70
1 40 C. 0. 5 1 . 00 672 4
1 . 0 1 . 00 5 9 8 1
2 . 0 1 . 00 5 361
4. 0 1 . 00 478 7
TABLE 5
G C Da ta f ro m Dec a rb o x y la tio n o f THC Herb a l Ma teria l
Tem p era tu re Tim e ( ho u rs ) CBN / THC ( % )
1 05 C. Z ero 2 . 4
0. 5 3. 5
1 . 0 4. 2
2 . 0 3. 7
4. 0 5 . 6
1 2 0 0. 5 3. 2
1 . 0 4. 1
2 . 0 6. 7
4. 0 1 1 . 3
1 40 C. 0. 5 5 . 7
1 . 0 1 3. 0
2 . 0 1 7. 5
4. 0 2 3. 8
The dec a rb o x y la tio n c o nditio ns f o r a b a tc h s c a le o f a b o u t
4 k g o f b o ta nic a l ra w m a teria l ( BRM) a re a s f o llo w s :
Ap p ro x im a tely 4 k g o f m illed BRM ( either THCA o r
CBDA) to b e dec a rb o x y la ted w a s initia lly hea ted to 1 05 C.
a nd held a t this tem p era tu re f o r a b o u t 1 5 m inu tes to ev a p o
ra te o f f a ny reta ined w a ter a nd to a llo w u nif o rm hea ting o f
the BRM. The b a tc h w a s then f u rther hea ted to 1 45 C. a nd
held a t this tem p era tu re f o r 45 m inu tes to a llo w dec a rb o x y
la tio n to b e c o m p leted to g rea ter tha n 9 5 % ef f ic ienc y .
The hea ting tim e f o r CBDA BRM w a s ex tended to 5 5
m inu tes a t 1 45 0 C. a s it b ec a m e a p p a rent f ro m res u lts tha t
CBDA w a s s lig htly m o re res is ta nt to dec a rb o x y la tio n tha n
THCA. This dif f erenc e b etw een CBD a nd THC w o u ld b e
ev en m o re p ro no u nc ed a t c o m m erc ia l s c a le b a tc hes . The
THC BRM hea ting tim e w a s reta ined a t 1 45 0 C. f o r 45
m inu tes .
The c o nditio ns u s ed in p ilo t s c a le c lo s ely re?ec t tho s e
c o nditio ns determ ined a s o p tim a l f ro m the la b o ra to ry s tu d
ies . The dif f erenc es c a n b e ex p la ined b y s lo w er a nd les s
ef ?c ient hea t tra ns f er v ia the c o nta iners a nd thro u g h the
BRM a t the inc rea s ed b a tc h s ize f o r the p ilo t s c a le.
Ta b les 6 a nd 7 p ro v ide da ta to dem o ns tra te the ef ?c ienc y
o f dec a rb o x y la tio n m ea s u red in term s o f c o ntent o f the
b io lo g ic a lly a c tiv e c a nna b ino id, THC o r CBD.
2 0
2 5
30
35
40
45
5 0
5 5
60
65
1 6
TABLE 6
Dec a rb o x y la tio n Ef f ic ienc y f o r CBD BRM
Ba tc h % Elf ic ienc y o f
N u m b er Dec a rb o x y la tio n
CBD S p ec i?c a tio n >9 5 %
A 9 8 . 8
B 9 9 . 5
C 9 8 . 3
D 1 00. 0
E 1 00. 0
F 1 00
G 9 6. 9
H 1 00. 0
Inc rea s e in b a tc h s ize o f CBD BRM f ro m a p p ro x im a tely
4 k g to 6 k g res u lted in a need to inc rea s e dec a rb o x y la tio n
tim e. The dec a rb o x y la tio n tim e a t 1 45 0 C. w a s inc rea s ed
f ro m 5 5 m inu tes to 9 0 m inu tes .
TABLE 7
% Elf ic ienc y o f
Dec a rb o x y la tio n
S p ec i?c a tio n >9 5 %
B a tc h
N u m b er THC
9 9 . 4
9 7. 3
9 8 . 5
1 00. 0
9 7. 8
9 9 . 9
1 00. 0 OZ Z HWHH
Ov erv iew o f Ex tra c tio n Pro c es s :
The BDS is ex tra c ted f ro m dec a rb o x y la ted BRM u s ing
liq u id c a rb o n dio x ide m etho do lo g y . This inv o lv es c o ntinu
o u s ly p a s s ing liq u e?ed c a rb o n dio x ide thro u g h the c ho p p ed
b io m a s s , w hic h is c o nta ined in a hig h- p res s u re v es s el. The
c ru de ex tra c t is dis s o lv ed in etha no l, c o o led to a lo w
tem p era tu re then ?ltered to rem o v e p rec ip ita ted c o ns titu ents
s u c h a s w a x es . Rem o v ing etha no l a nd w a ter in v a c u o
p ro du c es BDS c o nta ining either hig h c o nc entra tio ns o f CBD
o r THC, dep ending o n the b io m a s s u s ed.
Flo w Dia g ra m o f Ty p ic a l Ex tra c tio n Pro c es s :
BRM is dec a rb o x y la ted b y hea ting to a p p ro x im a tely 1 05 0 C.
f o r 1 5 m inu tes , f o llo w ed b y a p p ro x im a tely 1 45 0 C. f o r m inim u m
o f 5 5 m inu tes f o r THCA a nd 9 0 m inu tes f o r CBDA.
l
Ex tra c tio n w ith liq u id c a rb o n dio x ide ( CO2 ) [ Fo o d G ra de]
f o r u p to 1 0 ho u rs
Co nditio ns : Ap p ro x im a tely 60 b a r i 1 0 b a r p res s u re a nd 1 0 C. i 5 C.
l
Rem o v a l o f CO2 b y dep res s u ris a tio n to rec o v er c ru de ex tra c t
l
Winteris a tio n" Dis s o lu tio n o f c ru de ex tra c t in etha no l f o llo w ed b y
c hilling s o lu tio n ( 2 0 C. i 5 C. / u p to 5 2 ho u rs )
to p rec ip ita te u nw a nted w a x es
l
US 7, 344, 736 B2
1 7
c o ntinu ed
Rem o v a l o f u nw a nted w a x y m a teria l b y c o ld ?ltra tio n
( 2 0 u m ?lter)
l
Rem o v a l o f etha no l a nd w a ter f ro m the ?ltra te b y
thin ?lm ev a p o ra tio n u nder redu c ed p res s u re
( 60 C. i 2 C. , w ith v a p o u r a t 40 C. i 2 C. / 1 72 m b a r
a nd 72 m b a r i 4 m b a r)
1
BDS
( S to red a t - 2 00 c . i 5 0 c . )
Ex tra c tio n N o . 1
The ?rs t s ta g e in the m a nu f a c tu ring p ro c es s is Ex tra c tio n
u s ing liq u id CO2 u nder s u b - c ritic a l c o nditio ns .
Ex p erim ents indic a ted tha t b o th THC a nd CBD c o u ld b e
ex tra c ted f ro m Ca nna b is p la nt m a teria l in hig h ef f ic ienc y
u s ing s u b - c ritic a l CO2 a t lo w tem p era tu re, o f a p p ro x im a tely
1 0 C1 5 o C. u s ing a p res s u re o f a p p ro x im a tely 60 b a rzlO
b a r.
the Ta b le 8 b elo w s ho w s c o m p a ra tiv e da ta g enera ted f o r a
BDS rc h in THC
Pres s u re Tem p % w / w Wa x % thc w / w p o s t
Cha rg e N o b a r C. rem o v ed w interis a tio n
Ac l2 02 400 60 8 . 2 67. 2
Ac l2 05 400 60 6. 1 67. 0
Ac l2 06 400 60 6. 1 68 . 0
Three ru ns 60 1 0 2 . 2 4. 8 5 9 . 9 473]
Av e a b o u t 3 Av e 65 %
Fro m the res u lts it c a n b e s een tha t there is lo s s o f
s elec tiv ity , a s indic a ted b y the hig h w a x b u rden u nder s u p er
c ritic a l c o nditio ns . Whils t Winteris a tio n c a n rem o v e la rg er
a m o u nts o f w a x , p ro c es s ing is dif ?c u lt a s , f o r ex a m p le,
?lters b lo c k .
S im ila r res u lts w ere o b ta ined w ith CBD.
Pref erred c o nditio ns f o r liq u id CO2 ex tra c tio n a re a s
f o llo w s :
Dec a rb o x y la ted b o ta nic a l ra w m a teria l is p a c k ed into a
s ing le c o lu m n a nd ex p o s ed to liq u id CO2 u nder p res s u re.
Ba tc h s ize: Ap p ro x im a tely 60 k g
Pres s u re: 60 b a rzlO b a r
Tem p era tu re: 1 00 C1 5 0 C.
Tim e: Ap p ro x im a tely 8 ho u rs
CO2 m a s s ?o w 1 2 5 0 k g / hr 1 2 0% .
Pref erred p ro c es s p a ra m eters f o r p ro du c tio n o f BDS a re:
ex tra c tio n tim e >1 0 ho u rs , CO2 p res s u re 5 0470 b a r, ex tra c
tio n tem p 5 41 5 0 C. , CO2 m a s s 1 67 k g / k g BRM.
Fo llo w ing dep res s u ris a tio n a nd v enting o f f o f the CO2 the
c ru de BDS ex tra c t is c o llec ted into s ea led v es s els . The
o rig ina l BRM redu c es to a p p ro x im a tely 1 0% w / w o f c ru de
BDS ex tra c t. The c ru de BDS ex tra c t is held a t 2 00 C1 5 0
C.
The c ru de BDS ex tra c t c o nta ins w a x es a nd lo ng c ha in
m o lec u les . Rem o v a l is b y Winteris a tio n p ro c edu re ( ex tra c
tio n 2 ) , w hereb y the c ru de BDS ex tra c t is w a rm ed to eg 400
C1 40 C. to liq u ef y the m a teria l. Etha no l is a dded in the ra tio
o f 2 : 1 etha no l v o lu m e to w eig ht o f c ru de BDS ex tra c t. The
2 0
2 5
30
35
40
45
5 0
5 5
60
65
1 8
etha no lic s o lu tio n is then c o o led to 2 00 C1 5 0 C. a nd held
a t this tem p era tu re f o r a p p ro x im a tely 48 ho u rs .
On c o m p letio n o f the Winteris a tio n the p rec ip ita te is
rem o v ed b y c o ld ?ltra tio n thro u g h a 2 0 u m ?lter.
Ex tra c tio n N o . 2
The s ec o nd s ta g e in the m a nu f a c tu ring p ro c es s is Ex tra c
tio n N o . 2 , ref erred to a s Winteris a tio n u s ing etha no l.
Cru de BDS ex tra c t is p ro du c ed f ro m Ex tra c tio n N o . 1 tha t
c o nta ins c o ns titu ents , s u c h a s w a x es . Etha no l ef f ec tiv ely
ex tra c ts lo ng c ha ined m o lec u les f ro m the c ru de ex tra c t.
S tu dies :
It w a s f o u nd b y w a rm ing the c ru de BDS ex tra c t to
a p p ro x im a tely 400 C. the m ix ing a b ility o f the c ru de ex tra c t
w ith s o lv ent w a s im p ro v ed.
It w a s p ref erred to c hill the Winteris a tio n s o lu tio n to
2 00 C. f o r a b o u t 48 ho u rs .
Pref erred p ro c es s p a ra m eters f o r p ro du c tio n o f BDS a re:
ex tra c tio n tem p 3644 C. , ra tio etha no l: p ro du c t a p p ro x .
2 : 1 , f reezer tem p 2 5 C. to 1 5 C. , tim e 48 5 4 ho u rs .
Filtra tio n
The etha no lic s o lu tio n p ro du c ed in the s ec o nd ex tra c tio n
s ta g e req u ires ?ltra tio n to rem o v e the res u lting p rec ip ita tio n.
Filter s ize is p ref era b ly 2 0 u m .
Pref erred p ro c es s p a ra m eters f o r p ro du c tio n o f BDS a re:
to ta l ?ltra tio n tim e >6 ho u rs .
Ev a p o ra tio n
The ?na l s ta g e o f the m a nu f a c tu ring p ro c es s is the
rem o v a l o f etha no l a nd a ny w a ter tha t m a y b e p res ent.
Pref era b ly this is c a rried o u t b y hea ting a t 60 C. : 2 o C. to
g iv e a v a p o u r tem p era tu re o f 400 C1 2 0 C. u nder a v a c u u m
o f 1 72 m b a r1 4 m b a r. The dis tilla tio n u nder thes e c o nditio ns
c o ntinu es u ntil there is little o r no v is ib le c o ndens a te.
Redu c ing the v a c u u m f u rther, in s ta g es , do w n to a p p ro x i
m a tely 5 0 m b a r, c o m p letes w a ter rem o v a l. On c o m p letio n
the BDS is tra ns f erred into s ea led s ta inles s s teel c o nta iners
a nd s to red in a f reezer a t 2 00 C1 5 0 C.
Pref erred p ro c es s p a ra m eters f o r p ro du c tio n o f BDS a re:
ev a p o ra tio n v a p o u r tem p era tu re 38 442 0 C. , v a c u u m p res
s u re rem o v a l o f etha no l 1 6741 77 m b a r, v a c u u m p res s u re
rem o v a l o f w a ter 7( L75 m b a r 62 45 8 m b a r 5 2 448 m b a r, tim e
<8 ho u rs .
Cha ra c teris a tio n o f BDS
The THC BDS is a b ro w n, v is c o u s , s em i- s o lid ex tra c t
c o ns is ting o f a t lea s t 60% c a nna b ino ids c o ns titu ents . The
c a nna b ino id c o ns titu ents inc lu de a t lea s t 9 0% THC, a b o u t
1 . 5 % CBD w ith the rem a inder b eing m a de u p o f o ther m ino r
c a nna b ino ids .
The c hem ic a l c o m p o s itio n o f Ca nna b is ha s b een tho r
o u g hly s tu died w ith o v er 400 c o m p o u nds identi?ed ( Hen
dric k s et a l. , 1 9 75 ; Tu rner et a l. , 1 9 8 0) . Mo re tha n 60
c a nna b ino ids ha v e b een identi?ed, w ith CBDA a nd THCA
( the CBD a nd THC p re- c u rs o rs ) b eing the m o s t a b u nda nt.
G enera lly , the no n- c a nna b ino id c o ns titu ents c o m p ris e u p to
5 0% o f ex tra c ts , dep ending o n the ex tra c tio n p ro c es s .
Chem ic a l c la s s es identi?ed inc lu de a lk a nes ( 2 5 430 c a rb o n
c ha in) , nitro g eno u s c o m p o u nds , a m ino a c ids , s u g a rs , a lde
hy des , a lc o ho ls a nd k eto nes , ?a v a no ids , g ly c o s ides , v ita
m ins , p ig m ents a nd terp enes . Ab o u t 9 5 m o no - a nd s es q u i
terp enes ha v e b een identi?ed in Ca nna b is a nd a re
res p o ns ib le f o r the c ha ra c teris tic o do u r.
US 7, 344, 736 B2
1 9
Co ns idera b le Wo rk ha s b een c a rried o u t to c o m p letely
elu c ida te the s tru c tu re o f b o th CBD a nd THC ( s u m m a ris ed
in the a b o v e p a p ers ) a nd b o th ha v e b een p rep a red s y ntheti
c a lly . Pu re THC ha s b een s u c c es s f u lly is o la ted in s u f ?c ient
q u a ntity f ro m the BDS to b e u s ed a s ref erenc e m a teria l f o r
identi?c a tio n a nd q u a nti?c a tio n.
Im p u rities :
The BDS s u b s ta nc e is a s elec tiv e ex tra c t f ro m dried
dec a rb o x y la ted lea v es a nd ?o w ering hea ds o f s p ec i?c
c hem o v a rs o f Ca nna b is s a liv a . A ra ng e o f o v er 400 c o m
p o u nds , inc lu ding o v er 60 c a nna b ino ids , ha v e b een f o u nd in
Ca nna b is p la nts ( Tu rner 1 9 8 0) . As thes e a re na tu ra lly o c c u r
ring it is no t c o ns idered nec es s a ry to deem a ny o f thes e
c o m p o nents a s im p u rities . The m a j o r im p u rities theref o re
o c c u r in f o u r a rea s , p es tic ides intro du c ed du ring the g ro Wing
p ro c es s , a ?a to x ins , a ny neW p ro du c ts f o rm ed b y dec a rb o x y
la tio n a nd the m a teria ls o ther tha n the c a nna b ino ids , Whic h
m a k e u p the BDS .
The g ro Wing p ro c es s is c lo s ely c o ntro lled u s ing G AP
g u idelines a nd ta k es p la c e in a c lim a te c o ntro lled indo o r
g ro Wing env iro nm ent. N o p es tic ides a re a p p lied to the c ro p s
du ring g ro Wth, a ll p es t c o ntro l b eing m a na g ed b y b io lo g ic a l
m ea ns . N o p es tic ides a re inc o rp o ra ted in the g ro Wing
m ediu m .
To ens u re tha t no p es tic ide res idu es a re intro du c ed into the
p ro du c t the g ro Wing m ediu m is p erio dic a lly tes ted f o r
p es tic ides k no Wn to b e u s ed b y the g ro Wing m ediu m s u p
p lier.
Onc e the p la nt m a teria l ha s b een ha rv es ted a nd dried
f u rther s a m p les a re p erio dic a lly tes ted u s ing a g enera l
p es tic ide s c reen to ens u re no c o nta m ina tio n o f the c ro p ha s
o c c u rred Po tentia l im p u rities a re a deq u a tely c o ntro lled a t
the BRM s ta g e.
Altho u g h the g ro Wing c o nditio ns a re c a ref u lly c o ntro lled
to p rev ent this , the ra W m a teria l ha s the p o tentia l f o r
m ic ro b io lo g ic a l c o nta m ina tio n res u lting in a ?a to x ins in the
p ro du c t. The BRM a nd the BDS a re theref o re tes ted p eri
o dic a lly f o r a ?a to x ins c o ntent.
The na tu ra lly o c c u rring f o rm o f THC in the f res hly g ro Wn
p la nt is the a c id THCA, a ltho u g h s m a ll q u a ntities o f the
neu tra l THC do o c c u r. Bef o re ex tra c tio n the THCA is
dec a rb o x y la ted b y hea ting to y ield the neu tra l THC. The
p ro c es s is ef ?c ient b u t a s m a ll a m o u nt o f THCA rem a ins a nd
this is m o nito red du ring the ?na l tes ting o f the BDS .
Therm a l deg ra da tio n o f the THCA a nd THC du ring the
dec a rb o x y la tio n p ro c es s is p o s s ib le to y ield CBN A a nd
CBN . Thes e a re m o nito red in the BDS .
The no n- c a nna b ino id c o m p o nents tha t m a k e u p the b a lla s t
p o rtio n o f the BDS inc lu de hy dro c a rb o n a nd trig ly c eride
Wa x es , p la nt p ig m ents a nd terp enes . Thes e a re c o m m o n
c o m p o nents o f m a ny o ther ex tra c ts o f m edic ina l p la nts a nd
a re c o ns idered to b e o f little to x ic o lo g ic a l a nd p ha rm a c o
lo g ic a l s ig ni?c a nc e. The ra ng e o f o ther c o m p o nents p res ent
is Wide b u t they a re g enera lly p res ent in o nly s m a ll q u a n
tities
The q u a ntity o f b a lla s t is redu c ed b y the Winteris a tio n
p ro c es s Whic h p rec ip ita tes the Wa x es . The b a lla s t m a teria ls
a re c o ns idered to b e a dilu ent o f the a c tiv e c o ns titu ents a nd
a re no t a s s a y ed o r c o ntro lled.
2 0
2 5
30
35
40
45
5 0
5 5
60
65
2 0
TABLE 9
S p ec i?c a tio n f o r the c o ntro l o f BDS hig h in CBD:
Tes t Tes t Metho d Lim its
Ap p ea ra nc e In- Ho u s e Bro Wn v is c o u s s em i- s o lid
Identi?c a tio n:
A TLC S p o ts ha v e c ha ra c teris tic R1 a nd
c o lo u rs , c o m p a red With CBD
s ta nda rd
B HPLC/ UV Po s itiv e f o r CBD
CBD c o ntent In- ho u s e N LT 5 5 % W/ W o f ex tra c t
( HPLC- UV)
Rela ted In- ho u s e
c a nna b ino ids : ( HPLC/ UV)
THC c o ntent N MT 7. 5 % o f the CBD c o ntent
Others ( to ta l) N MT 5 % o f the CBD c o ntent
A?a to x in: TBA N MT 4 p p b
To ta l Hea v y Meta ls : Ph. Eu r. N MT 2 0 p p m
Res idu a l s o lv ents : In- ho u s e
Etha no l N MT 5 % W/ W
Mic ro b ia l: Ph. Eu r.
TVC N MT 1 05 c f u / g
Fu ng i N MT 1 04 c f u / g
Other N MT 1 03 c f u / g
entero b a c teria &
c erta in o ther g ra m
neg a tiv e o rg a nis m s
E. c a li Ab s ent in 1 g
S a lm o nella Ab s ent in 1 0 g
S . a a rea s Ab s ent in 1 g
Ana ly tic a l Pro c edu res
Identi?c a tio n, As s a y a nd Rela ted Ca nna b ino ids :
The c o ntent o f THC, CBD a nd Ca nna b ino l ( CBN ) in the
BRM a nd BDS , a re q u a ntita tiv ely determ ined b y ex tra c tio n
With m etha no l o r m etha no l/ c hlo ro f o rm ( 9 : 1 ) . Rev ers e- p ha s e
Hig h Perf o rm a nc e Liq u id Chro m a to g ra p hy ( HPLC) With
UV detec tio n a t 2 2 0 nm is the m etho d o f q u a nti?c a tio n. All
a na ly s is m u s t b e p erf o rm ed u nder a m b er lig ht b ec a u s e the
c o m p o u nds o f interes t a re k no Wn to b e lig ht s ens itiv e.
Chro m o to g ra p hy Eq u ip m ent a nd c o nditio ns :
Eq u ip m ent Ag ilent ( HP) 1 1 00 HPLC s y s tem With v a ria b le Wa v e
leng th UV detec to r o r dio de a rra y detec to r.
HPLC Co lu m n Dis c o v ery C8 5 p m 1 5 c m x 0. 46 c m
Pre- Co lu m n King s o rb C1 8 5 p m 3 c m x 0. 46 c m
Mo b ile Pha s e Ac eto nitrile: Metha no l: 0. 2 5 % W/ v a c etic a c id
( 1 6: 7: 6 b y v o lu m e)
Co lu m n Tem p 2 5 0 C.
Flo W Ra te 1 . 0 m l m in 1
Detec tio n 2 2 0 nm 600 m A f . s . d. S ec o nd Wa v eleng th 31 0 nm
Inj ec tio n 1 0 p l
Vo lu m e
Ru n Tim e 2 0* 2 5 m inu tes ( m a y b e ex tended f o r s a m p les c o nta ining
s m a ll a m o u nt o f la te- elu ting p ea k s )
Elu tio n Order CBD, CBDA, A9 THCV, CBN , A9 THC, CBC, A9 THCA
S ta nda rd Prep a ra tio n:
S to c k s ta nda rd s o lu tio ns o f CBD, CBN a nd A9 THC in
m etha no l a t a p p ro x im a tely 1 m g m l 1 a re s to red a t 2 00 C.
Dilu ted Wo rk ing s ta nda rds ( 0. 1 m g / m l f o r A9 THC a nd
CBD a nd 0. 01 m g / m l f o r CBN ) a re p rep a red in m etha no l
f ro m the s to c k s ta nda rds a nd s to red a t 2 00 C. ( m a x im u m
p erio d o f tWelv e m o nths a f ter initia l p rep a ra tio n) . Af ter
p rep a ra tio n, s ta nda rd s o lu tio ns m u s t b e a liq u o ted into v ia ls
to redu c e the a m o u nt o f s ta nda rd ex p o s ed to ro o m tem p era
tu re. Prio r to u s e in a n HPLC s a m p le a s s a y , the req u ired
US 7, 344, 736 B2
2 1
nu m b er o f s ta nda rd v ia ls a re rem o v ed a nd a llo w ed to
eq u ilib ra te to ro o m tem p era tu re.
S a m p le Prep a ra tio n:
In a ll p rep a ra tio ns , a lterna tiv e Weig hts a nd v o lu m es m a y
b e u s ed to g iv e the s a m e ?na l dilu tio ns .
Bo ta nic a l Ra W Ma teria l
Ac c u ra tely Weig h a p p ro x im a tely 1 00 m g o f c ho p p ed
dried ho m o g eneo u s m a teria l into a 1 0 m l v o lu m etric
?a s k .
Dis p ers e m a teria l in m etha no l: c hlo ro f o rm ( 9 : 1 v / v ) a nd
m a k e to v o lu m e in the s a m e s o lv ent.
Ex tra c t s a m p le in a n u ltra s o nic b a th f o r 1 5 m inu tes .
Centrif u g e a n a liq u o t a t 300 rp m f o r a b o u t 2 m inu tes .
Dilu te 1 00 p l o f the s u p erna ta nt to 1 m l With m etha no l in
a s u ita b le HPLC s a m p le v ia l. ( Fu rther dilu tio n m a y b e
req u ired if the p rinc ip a l c a nna b ino id c o nc entra tio n is
o u ts ide the linea r Wo rk ing ra ng e) .
Dec a rb o x y la ted Bo ta nic a l Ra W Ma teria l:
As f o r Bo ta nic a l Ra W Ma teria l.
Bo ta nic a l Dru g S u b s ta nc e:
Ac c u ra tely Weig h a p p ro x im a tely 8 0 m g o f BDS into a 5 0
m l v o lu m etric ?a s k .
Dis s o lv e BDS a nd m a k e u p to v o lu m e With m etha no l.
Dilu te 1 00 p l o f the p rep a red s u p erna ta nt to 1 m l With
m etha no l in a s u ita b le HPLC a u to s a m p ler v ia l.
Chro m a to g ra p hy Pro c edu re:
S a m p les a re p la c ed in the a u to s a m p ler ra c k in the o rder
entered into the s eq u enc e lis t o n the Ag ilent c hem s ta tio n.
S ta nda rd s o lu tio ns a re u s ed to p ro v ide q u a ntita tiv e a nd
retentio n tim e da ta . Thes e m a y b e ty p ic a lly inj ec ted in
du p lic a te o r trip lic a te p rio r to the inj ec tio n o f a ny s a m p le
s o lu tio ns a nd then s ing u la rly a t s u ita b le interv a ls du ring the
ru n, With a m a x im u m o f 1 0 tes t s a m p les in b etWeen s ta n
da rds .
Chro m a to g ra p hy Ac c ep ta nc e Criteria :
TABLE 1 0
Retentio n tim e Windo w s a nd Rela tiv e Retentio n Tim e ( RRT) to AQ THC
f o r ea c h a na lv te:
Retentio n Tim e
c a nna b ino id ( Minu tes ) RRT( THC)
CBD 5 . 1 5 . 8 0. 5 8
CBN 7. 44; . 3 0. 8 3
A9 THC 9 . 0i1 0. 0 1 . 00
CBDA 5 . 5 4. 2 0. 61 5
A9 THCV 5 . 9 4. 6 0. 645
CBC 1 1 642 . 8 1 . 30
A9 THCA 1 4. 6* 1 6. 0 1 . 605
S a m p le THC c o nc entra tio n:
2 0
2 5
30
35
40
45
5 0
5 5
TABLE 1 1
Pea k S ha p e ( S y m m etry Fa c to r a c c o rding to Britis h Pha rm a c o p o eia
m etho d) :
c a nna b ino id S y m m etry Fa c to r
CBD <l. 30
CBN <l. 2 5
A9 THC <1 . 35
Ca lc u la tio n:
Bo ta nic a l Ra W Ma teria l:
The f o llo Wing eq u a tio n is u s ed to o b ta in a res u lt f o r the
p u rity o f the p rinc ip a l c a nna b ino id a s a % o f the c u rrently
a s s a y a b le c a nna b ino ids ( CBD, CBDA, CBN , A9 THC & A9
THCA) in the b a tc h:
Fo r hig h A9 THC m a teria l:
p ea k a rea s u m o f THC & THCA
7 THC : i
0 p ea k a rea s u m o f a s s a y a b le c a nna b ino ids
X 1 00
Fo r hig h CBD m a teria l, CBD & CBDA rep la c e THC &
THCA in the to p line o f the eq u a tio n.
Dec a rb o x y la ted Bo ta nic a l Ra W Ma teria l:
The f o llo Wing eq u a tio n is u s ed to c a lc u la te the ef ?c ienc y
o f the dec a rb o x y la tio n p ro c es s :
Fo r hig h A9 THC m a teria l:
Pea k a rea o f THC
Pea k a rea s u m o f THC & THCA X1 00
% dec a rb o x y la tio n ef f ic ienc y :
Fo r hig h CBD m a teria l, CBD & CBDA rep la c e THC &
THCA in the eq u a tio n.
Bo ta nic a l Dru g S u b s ta nc e:
The f o llo Wing eq u a tio ns a re u s ed to c a lc u la te the c o n
c entra tio n o f dru g s u b s ta nc e s a m p le, the indiv idu a l s a m p le
c a nna b ino id c o nc entra tio n, the % c o ntent o f the a s s a y a b le
c a nna b ino ids in the dru g s u b s ta nc e, the q u a ntity o f p rinc ip a l
c a nna b ino id a s a % o f c u rrently a s s a y a b le c a nna b ino ids a nd
the a m o u nt o f p rinc ip a l c a nna b ino id in the Who le Weig ht o f
ex tra c ted dru g s u b s ta nc e.
Fo r hig h A9 THC m a teria l:
Weig ht o f s a m p le
Co nc entra tio n o f dru s u b s ta nc e s a rn le= . .
g p D1 lu t1 o n f a c to r
Where dilu tio n f a c to 1 : 5 0><l0: 5 00
THC s ta nda rd c 0nc ><m ea n THC s a rnp le a rea
m ea n THC s ta nda rd a rea
THC s a rnp le c o nc entra tio n
% w / w THC c o ntent o f dru g s u b s ta nc e : i X1 00
dru g s u b s ta nc e s a rnp le c o nc entra tio n
US 7, 344, 736 B2
2 3
CBD a nd CBN c a n b e s u b s titu ted into a ll o f thes e
eq u a tio ns ins tea d o f A9 THC to o b ta in q u a ntita tiv e res u lts
f o r b o th. A9 THCA a nd CBDA a re a ls o c a lc u la ted u s ing the
s ta nda rd c o nc entra tio ns f o r A9 THC o r CBD in the a b s enc e
o f s p ec i?c ref erenc e s ta nda rds o f their o Wn.
Rela ted S u b s ta nc es a re de?ned a s the s u m o f the m ea n %
W/ W v a lu es f o r CBN , A9 THCA a nd CBDA.
THC a s % o f to ta l % w / w THC c o ntent
: ><1 00
a s s a y a b le c a nna b ino ids s u m o f % w / w o f a ll a s s a y a b le c a nna b ino ids
The to ta l a m o u nt o f A9 THC p res ent in the Who le dru g
s u b s ta nc e ex tra c t is o b ta ined.
Ex a m p le 2
Inv es tig a tio n o f the S ta b ilis a tio n o f Bo ta nic a l Dru g
S u b s ta nc e ( BDS ) b y Pa rtia l Pu ri?c a tio n Us ing
Ac tiv a ted Cha rc o a l
Res u lts f ro m s ta b ility s tu dies o n THC f o rm u la tio ns indi
c a te tha t THC in the f o rm o f BDS is u ns ta b le ev en a t s to ra g e
tem p era tu res a s lo W a s 5 C. This c o ntra s ts With the b eha v
io u r o f the p u ri?ed THC ( Dro na b ino l US P) in Ma rino l s o f t
g el c a p s u les , f o r Whic h a s helf lif e o f 2 y ea rs a t c o o l a m b ient
tem p era tu re is a c c ep ted. It s ho u ld a ls o b e no ted tha t the s helf
lif e o f THC s ta nda rd s o lu tio ns in m etha no l s u p p lied b y
S ig m a - Aldric h is c la im ed to b e 4 y ea rs When s to red ref rig
era ted a nd p ro tec ted f ro m lig ht.
This a p p a rent dis c rep a nc y b etWeen the s ta b ility o f BDS
( THC) a nd p u ri?ed THC p ro m p ted s p ec u la tio n tha t s o m e
c o m p o nent o f BDS Wa s des ta b ilis ing the p rinc ip a l c a nna b
ino id.
A s o lu tio n to this p ro b lem Wo u ld b e to p u rif y the BDS
( THC) to y ield hig h p u rity , p ref era b ly c ry s ta lline c a nna b
ino id. Ho Wev er, the a dditio na l p ro c es s ing c o s ts inc u rred o n
tra ns f o rm ing BDS to p u re c a nna b ino id Wo u ld s u b s ta ntia lly
inc rea s e the c o s t o f ?nis hed p ha rm a c eu tic a l p ro du c ts inc o r
p o ra ting the c a nna b ino id.
Henc e, the a p p lic a nt s o u g ht to dev elo p a s im p le p u ri?
c a tio n s tep Whic h Wo u ld p ro du c e BDS With enha nc ed s ta
b ility b u t Whic h did no t inc rea s e p ro c es s ing c o s ts to a
p ro hib itiv e ex tent.
The a p p lic a nt ha s determ ined tha t a c ha rc o a l c lea n- u p
s tep m a y b e c o nv eniently c a rried o u t in c lo s e c o nj u nc tio n
With the Winteris a tio n p ro c es s b y p a s s ing the etha no lic
Winteris a tio n s o lu tio n thro u g h a ?lter b ed to rem o v e p re
c ip ita ted Wa x es a nd then direc tly thro u g h a c ha rc o a l c o lu m n
in a s ing le s tep a nd tha t the u s e o f a c tiv a ted c ha rc o a l
s ig ni?c a ntly im p ro v es s helf lif e.
Ex p erim enta l Deta il.
S o lu tio ns o f either BDS ( THC) o r BDS ( CBD) a t a
c o nc entra tio n o f 1 00 m g / m l in a b s o lu te etha no l BP Were
p a s s ed thro u g h a c o lu m n p a c k ed With a c tiv a ted c ha rc o a l a nd
the elu a te c o llec ted. Thes e Were then dilu ted With f u rther
a b s o lu te etha no l to a c hiev e a c o nc entra tio n o f c a . 2 5 m g / m l
c a nna b ino id. The s o lu tio n Wa s then tra ns f erred into a 1 0 m l
ty p e AXl ( i. e. a m b er g la s s ) v ia l a nd c rim p s ea led. Thes e
s a m p les Were des ig na ted c ha rc o a l p u ri?ed BDS .
S a m p les o f the BDS ( THC) a nd BDS ( CBD) s o lu tio ns
Whic h ha d no t b een p a s s ed thro u g h the c ha rc o a l c o lu m n
Were s im ila rly dilu ted to g iv e a c a nna b ino id c o nc entra tio n o f
2 5 m g / m l a nd Were then s ea led in a n a m b er g la s s v ia l o f the
2 5
30
35
40
45
5 0
5 5
60
65
2 4
s a m e ty p e. Thes e s a m p les Were des ig na ted s ta nda rd BDS
a nd s erv ed a s a c o ntro l f o r the s ta b ility s tu dy .
The v ia ls c o nta ining s ta nda rd BDS ( s td BDS ) a nd
c ha rc o a l p u ri?ed BDS o f ea c h ty p e Were s to red in a s ta b ility
inc u b a to r a t 400 C. a nd s a m p les then p erio dic a lly Withdra Wn
o v er the p erio d 1 41 2 m o nths f o r HPLC a na ly s is o f c a nna b
ino id c o ntent a nd TLC p ro ?ling .
N o rm a l p ha s e TLC a na ly s is em p lo y ed the f o llo Wing c o ndi
tio ns :
S ta tio na ry Pha s e: S ilic a G el G
Mo b ile Pha s e: 8 0: 2 0 hex a ne/ a c eto ne
Dev elo p m ent: 2 x 8 c m i. e. do u b le dev elo p m ent
Vis u a lis a tio n: Dip in 0. 1 % W/ v Fa s t Blu e B ( a q )
Rev ers e p ha s e TLC a na ly s is em p lo y ed the f o llo Wing c o n
ditio ns :
S ta tio na ry Pha s e: C1 8 c o a ted S ilic a G el
Mo b ile Pha s e: 6: 7: 1 6 0. 2 5 % v / v a c etic a c id ( a q ) / m etha no l/ a c eto nitrile
Dev elo p m ent: 2 x 8 c m i. e. do u b le dev elo p m ent
Vis u a lis a tio n: Dip in 0. 1 % W/ v Fa s t Blu e B ( a q )
Fo r ea c h s a m p le a v o lu m e o f s o lu tio n c o nta ining a p p ro x i
m a tely 5 p g to ta l c a nna b ino id Wa s a p p lied to the TLC p la te.
Res u lts a nd Dis c u s s io n
The etha no lic s o lu tio ns o f s td BDS ( THC) a nd s td BDS
( CBD) a re a f a irly intens e y ello W. Pa s s a g e o f the BDS
s o lu tio ns thro u g h the a c tiv a ted c ha rc o a l ef f ec tiv ely dec o
lo u ris ed the s o lu tio ns , p res u m a b ly b y the a ds o rp tio n o f p la nt
p ig m ents c o - ex tra c ted With the c a nna b ino ids du ring the
p rep a ra tio n o f BDS f ro m c a nna b is herb b y liq u id CO2
ex tra c tio n.
The HPLC a na ly s is res u lts f o r the dif f erent BDS s o lu tio ns
a re ta b u la ted b elo W a s Ta b le 1 2 a nd a re a ls o p res ented in
g ra p hic a l f o rm ( FIG S . 1 43) . All da ta is rep o rted a s % o f the
t0 a s s a y . CBN v a lu es a re inc lu ded f o r the BDS ( THC)
s o lu tio ns a s this c o m p o u nd ha s b een identi?ed a s a m a rk er
o f therm a l deg ra da tio n o f THC in p rev io u s s ta b ility s tu dies .
TABLE 1 2
Ca nna b ino id As s a y Va lu es f o r S td a nd Pu ri?ed BDS S o lu tio ns o v er the
Perio d 1 41 2 Mo nth a t 400 C.
Mo nths
S o lu tio n Ca nna b ino id 1 4 6 1 2
S td BDS THC 9 7. 3% 9 2 . 4% 8 5 . 3% 74. 0%
( THC) CBN 1 04% 1 1 9 % 1 33% 1 5 4%
Pu ri?ed THC 1 02 . 9 % 1 07. 4% 9 6. 0% 8 8 . 6%
BDS ( THC) CBN 9 4% 1 1 1 % 1 1 1 % 1 2 0%
S td BDS CBD 1 00. 3% 1 03. 6% 9 3. 3% 9 1 . 0%
( CBD)
Pu ri?ed CBD 1 01 . 0% 1 00. 7% 9 7. 2 % 9 6. 9 %
BDS ( CBD)
US 7, 344, 736 B2
2 5
Fro m the a b o v e da ta it is q u ite c lea r tha t f o r b o th BDS
( THC) a nd BDS ( CBD) there is s o m e c o m p o nent o f the
b a lla s t, Whic h c a n b e rem o v ed b y c ha rc o a l, Whic h is des ta
b ilis ing the c a nna b ino ids .
Co m p a ris o n o f the lev els o f deg ra da tio n rea c hed a f ter 1 2
m o nths a t 400 C. f o r the s td BDS a nd the c o rres p o nding
c ha rc o a l p u ri?ed BDS indic a te tha t f o r b o th the THC a nd the
CBD ex tra c ts the c ha rc o a l p u ri?c a tio n inc rea s es the res is
ta nc e to therm a l deg ra da tio n b y o v er 5 0% .
Fo r BDS ( THC) , the lev el o f CBN is s een to inc rea s e a s
a f u nc tio n o f the p rinc ip a l c a nna b ino id lo s t ( FIG . 3) . As
o b s erv ed f o r o ther f o rm u la tio ns c o nta ining THC, the lev el o f
CBN is a g a in c o n?rm ed to b e a m a rk er o f therm a l deg ra
da tio n.
Co m p a ris o n b etWeen c a nna b ino id reg io ns o f HPLC c hro
m a to g ra m s o f s ta nda rd BDS ( CBD) a nd p u ri?ed BDS
( CBD) s a m p les a f ter 1 2 m o nths a t 400 C. ( da ta no t s ho Wn)
rev ea led no s ig ni?c a nt inf o rm a tio n. Ho Wev er, s im ila r c o m
p a ris o n o f HPLC c hro m a to g ra m s o f the s ta nda rd a nd p u ri
?ed BDS ( THC) a f ter deg ra da tio n Wa s inf o rm a tiv e.
The CBN Wa s a t a hig her lev el in the m o re hig hly
deg ra ded u np u ri?ed s ta nda rd BDS , b u t a s ec o nd s ig ni?c a nt
deg ra da tio n p ro du c t Wa s a ls o o b s erv ed, Whic h is a g a in
p res ent in b o th s a m p les b u t Whic h is m o re a b u nda nt in the
m o re deg ra ded s a m p le. The s p ec tru m o f this deg ra da tio n
p ro du c t Wa s a g a in es s entia lly identic a l to tha t o f CBN a nd
o n the b a s is o f this a nd the retentio n tim e a p p ea red to b e o ne
o f the CBN a na lo g u es .
Co nc lu s io n
S ig ni?c a nt im p ro v em ent in res is ta nc e to therm a l deg ra
da tio n is a c hiev ed b y a s im p le c ha rc o a l trea tm ent.
Ex a m p le 3
Ef f ec t o f Additio n o f Org a nic Mo di?er o n CO2
Ex tra c tio n o f Ca nna b is Pla nt Ma teria l
The f o llo Wing ex a m p le des c rib es a n inv es tig a tio n into the
ef f ec t o f the a dditio n o f a p o la r c o - s o lv ent o n the c ha ra c
teris tic s o f a n ex tra c t p ro du c ed f ro m c a nna b is p la nt m a teria l
( G 5 c hem o v a r) u s ing liq u id CO2 ex tra c tio n, a nd illu s tra tes
the dif f erenc e in s elec tiv ity o b ta ined u s ing s u b - c ritic a l v s
s u p er- c ritic a l CO2 ex tra c tio n.
Ex p erim enta l Deta il.
Ex tra c tio n ex p erim ents Were c a rried o u t u s ing a 1 liter
c a p a c ity CO2 ex tra c tio n a p p a ra tu s . Fo o d g ra de CO2 a nd BP
g ra de a b s o lu te etha no l Were em p lo y ed a s s o lv ents .
A b a tc h o f G 5 c a nna b is ( a hig h CBD c hem o v a r) Wa s
u s ed. The CBD c o ntent Wa s 7. 3% W/ W a f ter dec a rb o x y la
tio n. Ana ly s is o f the c a nna b ino id c o ntent o f the ex tra c ts Wa s
c a rried o u t b y HPLC.
Res u lts a nd Dis c u s s io n.
The da ta rela ting to the c o m p o s itio n o f the ?na l ex tra c t
o b ta ined a f ter a 4 ho u r ex tra c tio n tim e u nder the s p ec i?ed
c o nditio ns is p res ented b elo W in Ta b le 1 3:
TABLE 1 3
Co m p o s itio n a nd Yield Da ta f o r Ex tra c ts Pro du c ed u nder Dif f erent
Ex tra c tio n Co nditio n .
Ex tra c tio n % W/ W % CBD % Rec o v ery
S a m p le Co nditio ns Ex tra c t ( W/ W) o f CBD
AC470 1 0 C. / 60 BAR 8 . 4% 63. 6% 72 . 9 %
AC471 40 C. / 1 00 BAR 1 0. 7% 5 4. 4% 79 . 5 %
2 0
2 5
30
35
40
45
5 0
5 5
60
65
2 6
TABLE 1 3 - c o ntinu ed
Co m p o s itio n a nd Yield Da ta f o r Ex tra c ts Pro du c ed u nder Dif f erent
Ex tra c tio n Co nditio ns .
Ex tra c tio n % W/ W % CBD % Rec o v ery
S a m p le Co nditio ns Ex tra c t ( W/ W) o f CBD
AC472 40 C. / 1 00 BAR + 1 0. 3% 64. 6% 9 1 . 0%
2 % ETHAN OL
The rec o v ery ef ?c ienc y is b a s ed o n the CBD a v a ila b le in
dec a rb o x y la ted p la nt m a teria l c ha rg ed to the v es s el f o r ea c h
ex tra c tio n.
The res u lts illu s tra te tha t c ha ng ing the ex tra c tio n c o ndi
tio ns f ro m s u b - c ritic a l to s u p er- c ritic a l inc rea s es the s o lv a t
ing p o Wer o f the CO2 a nd res u lts in a hig her rec o v ery o f the
a v a ila b le CBD. Ho Wev er, the s u p erc ritic a l CO2 c a n no W
s o lu b ilis e a Wider ra ng e o f c o m p o u nds a nd the ex tra c tio n o f
thes e a dditio na l c o m p o u nd ha s the ef f ec t o f dilu ting the
c o nc entra tio n o f CBD in the ex tra c t to s u c h a n ex tent tha t it
is no W lo Wer tha n tha t o b ta ined f o r the s u b - c ritic a l ex tra c
tio n. Co ns eq u ently , the m a rg ina l a dditio na l rec o v ery o f
a v a ila b le CBD f ro m the ra W m a teria l Wo u ld no t o u tWeig h
this dis a dv a nta g e a nd dem o ns tra tes the u s e o f s u p erc ritic a l
c o nditio ns is no t des ira b le.
The a dditio n o f 2 % W/ W a b s o lu te etha no l to s u p erc ritic a l
CO2 a s a m o di?er inc rea s es the rec o v ery o f the a v a ila b le
CBD to >9 0% . Pres u m a b ly the rela tiv ely p o la r c a nna b ino id
is m o re s o lu b le in the ex tra c t o f inc rea s ed p o la rity .
Interes ting ly , the c o nc entra tio n o f CBD in the ex tra c t is
inc rea s ed s lig htly b y the a dditio n o f p o la r m o di?er. This
Wo u ld s eem to indic a te tha t the c o - ex tra c ta b le no n- c a nna b
ino id m a teria l p res ent in the p la nt m a teria l is les s p o la r tha n
the ta rg et c a nna b ino id a nd henc e the ex tra c tio n o f this
m a teria l ( the b a lla s t ) is des elec ted When p o la rity is
inc rea s ed.
Thu s , ex tra c tio n o f c a nna b is p la nt m a teria l With s u p erc riti
c a l CO2 +2 % W/ W etha no l p ro v ides a n inc rea s e in rec o v ery
o f the ta rg et a c tiv e With no a ttenda nt p ena lty o f lo s s o f
s elec tiv ity .
In S u m m a ry :
1 . A s Witc h f ro m s u b - c ritic a l to s u p er- c ritic a l c o nditio ns
p ro du c es little a dv a nta g e in term s o f o v era ll rec o v ery o f
c a nna b ino id f ro m the ra W m a teria l b u t do es res u lt in the
dis a dv a nta g e o f redu c ing the a c tiv e c o ntent o f the ex tra c t.
2 . The a dditio n o f 2 % a b s o lu te etha no l m o di?er to s u p er
c ritic a l CO2 res u lts in a s ig ni?c a nt im p ro v em ent in the
rec o v ery o f c a nna b ino id f ro m the ra W m a teria l With no
p ena lty o f dilu tio n o f a c tiv e c o ntent b y c o - ex tra c ted m a te
ria l.
Tho s e s k illed in the a rt Will rec o g niZ e, o r b e a b le to
a s c erta in u s ing no m o re tha n ro u tine ex p erim enta tio n, m a ny
eq u iv a lents to the s p ec i?c em b o dim ents o f the inv entio n
des c rib ed herein. S u c h eq u iv a lents a re intended to b e
enc o m p a s s ed b y the f o llo Wing c la im s .
All ref erenc es dis c lo s ed herein a re inc o rp o ra ted b y ref
erenc e in their entirety .
The inv entio n c la im ed is :
1 . A m etho d o f ex tra c ting c a nna b ino ids f ro m p la nt m a te
ria l c o m p ris ing a dec a rb o x y la tio n s tep , a n ex tra c tio n With
liq u id c a rb o n dio x ide ( CO2 ) , a nd a s tep to redu c e the
p ro p o rtio n o f no n- ta rg et m a teria ls in the ex tra c t, c ha ra c ter
is ed in tha t the ex tra c tio n With liq u id CO2 is c o ndu c ted u nder
US 7, 344, 736 B2
2 7
s u b - c ritic a l c o nditio ns a t a tem p era tu re in the ra ng e o f f ro m
5 to 1 5 C. a nd a p res s u re in the ra ng e o f f ro m 5 0 to 70 b a r.
2 . A m etho d a s c la im ed in c la im 1 Wherein the dec a r
b o x y la tio n s tep is c a rried o u t a f ter ex tra c tio n With liq u id
C02 .
3. A m etho d a s c la im ed in c la im 1 Wherein the dec a r
b o x y la tio n s tep is c a rried o u t p rio r to ex tra c tio n With liq u id
C02 .
4. A m etho d a s c la im ed in c la im 1 Wherein the tem p era
tu re is in the ra ng e o f f ro m 8 to 1 2 C.
5 . A m etho d a s c la im ed in c la im 1 Wherein the p res s u re is
in the ra ng e o f f ro m 5 5 to 65 b a r.
6. A m etho d a s c la im ed in c la im 1 Wherein the CO2 ha s a
m a s s Ho w in the ra ng e o f 1 00041 5 00 Kg / h.
7. A m etho d a s c la im ed in c la im 1 Wherein the ex tra c tio n
is ru n f o r u p to 1 0 ho u rs .
8 . A m etho d a s c la im ed in a ny o f c la im s 1 44, 5 , 6 a nd 7
Wherein the CO2 is rem o v ed b y dep res s u ris a tio n a nd the
rec o v ered ex tra c t held a t a tem p era tu re in the ra ng e f ro m
1 5 C. to 2 0 C.
9 . A m etho d a s c la im ed in c la im 1 Wherein the s tep to
redu c e the p ro p o rtio n o f no n- ta rg et m a teria ls in the ex tra c t
is a p rec ip ita tio n With a C1 4C5 a lc o ho l, Wherein the m a teria l
to b e trea ted is Wa rm ed to a b o v e ro o m tem p era tu re b ef o re
the C1 4C5 a lc o ho l is a dded.
1 0. A m etho d a s c la im ed in c la im 9 Wherein the C1 4C5
a lc o ho l is etha no l.
1 1 . A m etho d a s c la im ed in c la im 9 Wherein the ex tra c t is
Wa rm ed to a tem p era tu re in the ra ng e f ro m 36 C. to 44 C.
1 2 . A m etho d a s c la im ed in c la im 1 0 Wherein the C1 4C5
a lc o ho l is a dded in a n a m o u nt o f f ro m 3: 1 to 1 : 1 C1 4C5
a lc o ho l v o lu m e to Weig ht o f the m a teria l to b e trea ted.
1 3. A m etho d a s c la im ed in c la im 9 Wherein the s o lu tio n
res u lting f ro m a dditio n o f C1 4C5 a lc o ho l to the m a teria l to
b e trea ted is c hilled a nd ins o lu b le m a teria ls a llo Wed to
p rec ip ita te o u t.
1 4. A m etho d a s c la im ed in c la im 1 3 Wherein the s o lu tio n
res u lting f ro m a dditio n o f C1 4C5 a lc o ho l to the m a teria l to
b e trea ted is c hilled to a tem p era tu re in the ra ng e f ro m 1 5
C. to 2 5 C.
5
1 5
2 0
2 5
30
35
2 8
1 5 . A m etho d a s c la im ed in c la im 1 3 Wherein the p re
c ip ita te o f ins o lu b le m a teria ls is rem o v ed b y ?ltra tio n.
1 6. A m etho d a s c la im ed in c la im 9 f u rther c o m p ris ing a
m u lti- s tep ev a p o ra tio n u nder redu c ed p res s u re.
1 7. A m etho d a s c la im ed in c la im 1 6 Wherein the ?rs t
C1 5 a lc o ho l is rem o v ed a nd then Wa ter is rem o v ed.
1 8 . A m etho d a s c la im ed in c la im 3 Wherein the dec a r
b o x y la tio n s tep is c a rried o u t p rio r to ex tra c tio n With liq u id
CO2 a nd is c o ndu c ted b y hea ting the p la nt m a teria l to
tem p era tu res a nd f o r tim es Whic h ens u re a t lea s t 9 5 %
c o nv ers io n o f the a c id c a nna b ino ids to their neu tra l f o rm
Whils t ens u ring therm a l deg ra da tio n o f THC to CBN is les s
tha n 1 0% .
1 9 . Am etho d a s c la im ed in c la im 1 8 in Whic h a m u lti s tep
hea ting p ro c es s is c o ndu c ted in Whic h the p la nt m a teria l is :
i) hea ted to a ?rs t tem p era tu re f o r a ?rs t tim e p erio d to
ev a p o ra te o if reta ined Wa ter a nd a llo W f o r u nif o rm
hea ting o f the p la nt m a teria l; a nd
ii) the tem p era tu re is inc rea s ed to a s ec o nd tem p era tu re
f o r a s ec o nd tim e p erio d u ntil a t lea s t 9 5 % c o nv ers io n
o f the a c id c a nna b ino ids to their neu tra l f o rm ha s
o c c u rred.
2 0. A m etho d a s c la im ed in c la im 1 f u rther c o m p ris ing the
s tep o f trea ting a n ex tra c t deriv ed f ro m the p la nt m a teria l
With a c tiv a ted c ha rc o a l.
2 1 . A m etho d o f m a k ing a p ha rm a c eu tic a l c o m p o s itio n
c o m p ris ing , a s a n a c tiv e a g ent, a b o ta nic a l dru g s u b s ta nc e
Whic h is a n ex tra c t f ro m a t lea s t o ne c a nna b is p la nt, Whic h
m etho d c o m p ris es p rep a ring a b o ta nic a l dru g s u b s ta nc e
c o nta ining c a nna b ino ids f ro m a t lea s t o ne c a nna b is p la nt
u s ing a n ex tra c tio n m etho d a c c o rding to c la im 1 , a nd
f o rm u la ting the b o ta nic a l dru g s u b s ta nc e With o ne o r m o re
p ha rm a c eu tic a lly a c c ep ta b le dilu ents , c a rriers o r ex c ip ients
to p ro du c e a p ha rm a c eu tic a l c o m p o s itio n.